Australian Handbook for the Identification of Fruit Flies

Transcription

THE AUSTRALIAN HANDBOOK
FOR THE IDENTIFICATION OF
FRUIT FLIES
Version 2.1
Species: Bactrocera bryoniae
Species: Bactrocera frauenfeldi
Species: Bactrocera kandiensis
Species: Bactrocera tau
Species: Bactrocera trilineola
Species: Bactrocera umbrosa
Species: Bactrocera xanthodes
Species: Bactrocera newmani
Department of Agriculture
and Water Resources
For more information on Plant Health Australia
Phone:
+61 2 6215 7700
Fax:
+61 2 6260 4321
Email:
Visit our website
http://www.planthealthaustralia.com.au/
An electronic copy of this plan is available from the website listed above.
© Plant Health Australia 2016
This work is copyright except where attachments are provided by other contributors and referenced,
in which case copyright belongs to the relevant contributor as indicated throughout this document.
Apart from any use as permitted under the Copyright Act 1968, no part may be reproduced by any
process without prior permission from Plant Health Australia.
Requests and enquiries concerning reproduction and rights should be addressed
to: Communications Manager
Plant Health Australia
1/1 Phipps Close
DEAKIN ACT 2600
ISBN 978-0-9872309-0-4
In referencing this document, the preferred citation is:
Plant Health Australia (2016). The Australian Handbook for the Identification of Fruit Flies. Version
2.1. Plant Health Australia. Canberra, ACT.
Disclaimer:
The material contained in this publication is produced for general information only. It is not intended
as professional advice on any particular matter. No person should act or fail to act on the basis of
any material contained in this publication without first obtaining specific, independent professional
advice.
Plant Health Australia and all persons acting for Plant Health Australia in preparing this publication,
expressly disclaim all and any liability to any persons in respect of anything done by any such person
in reliance, whether in whole or in part, on this publication. The views expressed in this publication
are not necessarily those of Plant Health Australia.
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1
Contributors
This document has been made possible through consultation with and input from the following fruit fly
entomologists, scientists, academics and diagnosticians:
Organisation
Contributor
Bio-Protection Research Centre, Lincoln University
Karen Armstrong
Department of Agriculture and Fisheries, Queensland
Jane Royer
Department of Agriculture and Food, Western Australia
Andras Szito, Bill Woods
Department of Agriculture and Water Resources
Carol Quashie-Williams, Craig Hull
Department of Economic Development, Jobs,
Transport and Resources, Victoria
Mali Malipatil, Linda Semeraro, Mark Blacket
Department of Resources, Northern Territory
Austin McLennan, Mary Finlay-Doney
Griffith University
Dick Drew, Meredith Romig
Ministry for Primary Industries NZ
Dongmei Li
New South Wales DPI
Chris Bloomfield, Peter Gillespie, Bernie Dominiak
Northern Australia Quarantine Strategy
James Walker, Sally Cowan
Queensland University of Technology
Matt Krosch, Mark Schutze
South Australian Museum
Mark Adams
South Australian Research and Development Institute
(SARDI)
Peter Crisp
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Foreword
The accurate identification of fruit flies is a key component of Australia’s
biosecurity system that underpins the domestic movement of fruit and
vegetables, maintains international market access for Australian producers
and protects Australia’s borders from exotic pest incursion.
In Australia’s tropics, routine surveillance of coastal and island communities
results in a requirement to process and identify thousands of adult flies per
hour. In some parts of southern Australia fruit fly sampling numbers are
smaller, however diagnosticians still have to be skilled and equipped to
identify a single fly of economic importance amongst a large range of native
fruit flies that have no impact on commercial fruits and vegetables.
For this second edition we have completely reviewed and updated all the diagnostic techniques
currently used in Australia for the identification of fruit flies. A new set of descriptions and photographs
have been prepared to assist the identification of flies by adult morphology. In addition, updated and
new protocols used for the identification of fruit flies using molecular biology techniques are presented.
A new section, Diagnostician’s Notes, is included in this edition which provides informal tips on adult
identification.
This document has been written by Australia’s fruit fly diagnosticians for diagnosticians and it is my
hope that the dialogue, sharing of information and experience, and constructive discourse that brings
together this publication will continue to grow. Together the combined expertise and knowledge of
Australia’s fruit fly researchers, academics, surveillance officers, diagnosticians and laboratory scientists
make up a formidable national resource, which when networked and coupled with extensive fruit fly
reference collections, provides a world-class national capability.
This valuable document provides a useful benchmark against which future updates, protocols and
revisions can be developed and training programs can be delivered.
I would like to thank all the entomologists and scientists who have brought this document together and
have the greatest pleasure in endorsing its adoption and use by practitioners and jurisdictions in
Australia.
Professor Dick Drew
International Centre for Management of Pest Fruit Flies
Griffith University
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Preface
The Australian Handbook for the Identification of Fruit Flies (v2.1)

Was written by diagnosticians for diagnosticians;

Collates current and existing practices and knowledge into a single document;

Pools experience from Australia’s network of fruit fly experts;

Establishes a resource that can support and develop the confidence and expertise of all users;

Provides a mechanism to possibly identify future information and research needs; and,

Considers the potential of both morphological and molecular techniques.
The Handbook has been an important part of building a network of fruit fly diagnosticians across
Australia and it is hoped that both the network and this document continue to grow and develop in the
future. We also welcome feedback from fruit fly experts around the world.
The current Handbook is a compilation of diagnostic techniques for 59 fruit fly species, most of which
are exotic to Australia, as well as the non-tephritid fruit infesting Drosophila suzukii. The Handbook is
intended to facilitate rapid diagnosis of fruit fly species and be a comprehensive guide for Australian
diagnosticians and field officers.
A copy of the Handbook can be downloaded by following the link below.
http://www.planthealthaustralia.com.au/national-programs/fruit-fly/handbook-for-theidentification-of-fruit-fly/
This is the second version of The Australian Handbook for the Identification of Fruit Flies. It is provided
freely as a reference resource with an expectation that it is appropriately acknowledged when it is
used. As a living document it is designed to be continuously updated as more information becomes
available through Australia’s skilled network of fruit fly diagnosticians. For further information, please
contact Plant Health Australia, 1 Phipps Close Deakin, ACT, 2600 Australia, [email protected]
Funding for this important initiative was provided by the Department of Agriculture and Water Resources
and the Plant Biosecurity Cooperative Research Centre (PBCRC). The PBCRC and Department of
Agriculture and Water Resources would like to recognise the significant in-kind contribution made by
researchers, academics, surveillance officers, diagnosticians and laboratory scientists who have
collectively updated this valuable document in 2015. Project facilitation and coordination was provided
by Plant Health Australia.
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Contents
1
Contributors ................................................................................................................................... 2
2
Foreword ........................................................................................................................................ 3
3
Preface............................................................................................................................................ 4
Diagnostician’s notes .......................................................................................................................... 13
Grouping of fruit fly species by morphological similarity ...................................................................... 13
TEPHRITIDAE: DACINAE ................................................................................................................... 13
Distinctive feature: Black scutum & T/ wrap T on abdomen ........................................................... 13
Distinctive feature: Black scutum & incomplete costal band ........................................................... 17
Distinctive feature: Black scutum & broad costal band ................................................................... 19
Distinctive feature: Black scutum & abdomen, no vittae ................................................................. 21
Distinctive feature: Red- brown to golden flies ................................................................................ 23
Distinctive feature: Glossy black scutum & band on wing ............................................................... 25
Distinctive feature: Glossy or matte black scutum & faint band on wing ........................................ 27
Distinctive feature: Black scutum, heavy abdominal markings, no band on wing ........................... 28
Distinctive feature: Medial vitta........................................................................................................ 29
Distinctive feature: Other wing pattern ............................................................................................ 33
Dacus spp........................................................................................................................................ 35
Ceratitis spp..................................................................................................................................... 36
TEPHRITIDAE: TRYPETINAE ............................................................................................................ 37
Rhagoletis spp. ................................................................................................................................ 39
Anastrepha spp. .............................................................................................................................. 41
DROSOPHILIDAE ............................................................................................................................... 43
4
Introduction.................................................................................................................................. 44
Background ............................................................................................................................. 44
Coverage of this diagnostic handbook ................................................................................... 45
5
Detection ...................................................................................................................................... 48
Plant products affected ........................................................................................................... 48
Signs and symptoms .............................................................................................................. 48
Development stages ............................................................................................................... 48
Methods for detection ............................................................................................................. 49
5.4.1
Trap types ......................................................................................................................... 49
5.4.2
Attractants ......................................................................................................................... 52
5
6
Inspection of material, sample preparation and storage ........................................................ 54
Infested fruit ............................................................................................................................ 54
Sample collection and handling .............................................................................................. 54
Sample transportation............................................................................................................. 54
Sample processing, identification and preservation ............................................................... 54
6.4.1
Molecular Specimens ........................................................................................................ 55
7
Identification ................................................................................................................................ 57
Overview ................................................................................................................................. 57
Morphological identification .................................................................................................... 63
7.2.2
Identification ...................................................................................................................... 64
Molecular identification ........................................................................................................... 68
7.3.1
DNA extraction for adults and immature stages ............................................................... 69
7.3.2
DNA Barcoding ................................................................................................................. 71
7.3.3
PCR-RFLP ........................................................................................................................ 80
7.3.4
Real-time PCR detection of Bactrocera tryoni complex ................................................... 92
7.3.5
Allozyme electrophoresis .................................................................................................. 97
8
Diagnostic Information ............................................................................................................. 100
Key to family Tephritidae ...................................................................................................... 101
Simplified key to major pest fruit fly genera (after White and Elson-Harris 1992): Adults .... 101
Species diagnosis: Anastrepha (Tephritidae: Trypetinae: Toxotrypanini) ............................ 102
8.3.1
Anastrepha distincta (Greene) ........................................................................................ 102
8.3.2
Anastrepha fraterculus (Wiedemann) ............................................................................. 106
8.3.3
Anastrepha grandis (Macquart) ...................................................................................... 109
8.3.4
Anastrepha ludens (Loew) .............................................................................................. 112
8.3.5
Anastrepha obliqua (Macquart)....................................................................................... 115
8.3.6
Anastrepha serpentina (Wiedemann) ............................................................................. 118
8.3.7
Anastrepha striata (Schiner) ........................................................................................... 121
8.3.8
Anastrepha suspensa (Loew) ......................................................................................... 123
Species diagnosis: Bactrocera & Dacus (Tephritidae: Dacinae: Dacini) .............................. 126
8.4.1
Bactrocera (Bactrocera) albistrigata (de Meijere) ......................................................... 126
8.4.2
Bactrocera (Bactrocera) aquilonis (May) ........................................................................ 131
8.4.3
Bactrocera (Paratridacus) atrisetosa (Perkins) ............................................................... 135
8.4.4
Bactrocera (Bactrocera) bryoniae (Tryon) ...................................................................... 138
8.4.5
Bactrocera (Bactrocera) carambolae (Drew and Hancock) ............................................ 141
8.4.6
Bactrocera (Bactrocera) caryeae (Kapoor) ..................................................................... 144
8.4.7
Bactrocera (Bactrocera) correcta (Bezzi) ....................................................................... 147
8.4.8
Bactrocera (Austrodacus) cucumis (French) .................................................................. 150
8.4.9
Bactrocera (Zeugodacus) cucurbitae (Coquillett) ........................................................... 153
8.4.10 Bactrocera (Bactrocera) curvipennis (Froggatt).............................................................. 157
8.4.11 Bactrocera (Paradacus) decipiens (Drew) ...................................................................... 160
8.4.12 Bactrocera (Zeugodacus) depressa (Shiraki) ................................................................. 163
8.4.13 Bactrocera (Bactrocera) dorsalis (Hendel) ..................................................................... 165
8.4.14 Bactrocera (Bactrocera) facialis (Coquillett) ................................................................... 169
8.4.15 Bactrocera (Bactrocera) frauenfeldi (Schiner) ................................................................ 172
8.4.16 Bactrocera (Afrodacus) jarvisi (Tryon) ............................................................................ 175
8.4.17 Bactrocera (Bactrocera) kandiensis (Drew and Hancock) .............................................. 178
8.4.18 Bactrocera (Bactrocera) kirki (Froggatt) ......................................................................... 181
8.4.19 Bactrocera (Bactrocera) kraussi (Hardy) ........................................................................ 184
8.4.20 Bactrocera (Bactrocera) latifrons (Hendel) ..................................................................... 187
6
8.4.21 Bactrocera (Bactrocera) melanotus (Coquillett) .............................................................. 190
8.4.22 Bactrocera (Tetradacus) minax (Enderlein) .................................................................... 192
8.4.23 Bactrocera (Bactrocera) musae (Tryon) ......................................................................... 194
8.4.24 Bactrocera (Bactrocera) neohumeralis (Hardy) .............................................................. 198
8.4.25 Bactrocera (Bactrocera) obliqua (Malloch) ..................................................................... 201
8.4.26 Bactrocera (Bactrocera) occipitalis (Bezzi) ..................................................................... 203
8.4.27 Bactrocera (Daculus) oleae (Rossi) ................................................................................ 206
8.4.28 Bactrocera (Bactrocera) passiflorae (Froggatt) .............................................................. 208
8.4.29 Bactrocera (Bactrocera) psidii (Froggatt) ........................................................................ 211
8.4.30 Bactrocera (Bactrocera) pyrifoliae (Drew & Hancock) .................................................... 214
8.4.31 Bactrocera (Zeugodacus) tau (Walker)........................................................................... 216
8.4.32 Bactrocera (Bactrocera) trilineola (Drew) ....................................................................... 220
8.4.33 Bactrocera (Bactrocera) trivialis (Drew) .......................................................................... 223
8.4.34 Bactrocera (Bactrocera) tryoni (Froggatt) ....................................................................... 226
8.4.35 Bactrocera (Tetradacus) tsuneonis (Miyake) .................................................................. 231
8.4.36 Bactrocera (Bactrocera) tuberculata (Bezzi) ................................................................... 233
8.4.37 Bactrocera (Bactrocera) umbrosa (Fabricius) ................................................................. 235
8.4.38 Bactrocera (Notodacus) xanthodes (Broun) ................................................................... 238
8.4.39 Bactrocera (Bactrocera) zonata (Saunders) ................................................................... 241
Species diagnosis: Ceratitis (Tephritidae: Dacinae: Ceratitidini).......................................... 244
8.5.1
Ceratitis capitata (Wiedemann)....................................................................................... 244
8.5.2
Ceratitis (Pterandrus) rosa (Karsch) ............................................................................... 247
Species diagnosis: Dacus (Tephritidae) ............................................................................... 250
8.6.1
Dacus (Callantra) longicornis (Wiedemann) ................................................................... 250
8.6.2
Dacus (Callantra) solomonensis (Malloch) ..................................................................... 252
Species diagnosis: Dirioxa (Tephritidae: Phytalmiinae) ....................................................... 254
8.7.1
Dirioxa pornia (Walker) ................................................................................................... 254
Species diagnosis: Rhagoletis (Tephritidae: Trypetinae: Trypetini) ..................................... 256
8.8.1
Rhagoletis cerasi (Linnaeus) .......................................................................................... 256
8.8.2
Rhagoletis cingulata (Leow)............................................................................................ 258
8.8.3
Rhagoletis completa (Cresson)....................................................................................... 260
8.8.4
Rhagoletis fausta (Osten-Sacken) .................................................................................. 262
8.8.5
Rhagoletis indifferens (Curran) ....................................................................................... 264
8.8.6
Rhagoletis mendax (Curran) ........................................................................................... 266
8.8.7
Rhagoletis pomonella (Walsh) ........................................................................................ 268
Species diagnosis: (Diptera: Drosophilidae)......................................................................... 270
8.9.1
Drosophila (Sophophora) suzukii (Matsumura 1931) ..................................................... 270
9
Diagnostic resources ................................................................................................................ 273
Key contacts and facilities .................................................................................................... 273
Reference collections ........................................................................................................... 277
Printed and electronic resources .......................................................................................... 278
9.3.1
Morphological keys ......................................................................................................... 278
9.3.2
Electronic resources ....................................................................................................... 279
Supplier details ..................................................................................................................... 280
10
References ................................................................................................................................. 281
11
a.
b.
Appendices ................................................................................................................................ 288
Specificity test for Tephritidae specimens in the real-time PCR assay .................................... 312
Real-time PCR assay for B. tryoni and B. curvipennis interceptions ........................................ 313
7
Tables
Table 1. Fruit flies covered in this diagnostic handbook .......................................................................... 46
Table 2. Commonly employed sample preservation techniques for molecular identification. Methods that
are suitable for a particular lifestage are indicated by “+”, those that are less suitable are indicated by a
“–“. ............................................................................................................................................................ 56
Table 3. Diagnostic methods used to identify fruit fly species ................................................................. 60
Table 4. Approximate size of the ITS1 amplicon for each species. Potential diagnostic size ranges are
contained in a block ................................................................................................................................. 85
Table 5. Analysis of RFLP products from ITS1 fragments from fruit flies ................................................ 88
Table 6. TaqMan real-time PCR for B. tryoni complex protocol using SsoAdvanced Universal Probe
Supermix (BioRad) ................................................................................................................................... 94
Table 7. Enzymes most commonly used for fruit-fly genetic identifications using allozyme
electrophoresis. ........................................................................................................................................ 99
Table 8. Bactrocera frauenfeldi complex and similar species (best diagnostic characters in bold). ..... 128
Figures
Figure 1. Types of fruit fly traps ............................................................................................................... 51
Figure 2. Chemical structure of cue lure and methyl eugenol. ................................................................ 53
Figure 3. Overview of fruit fly diagnostic procedures (adult specimens). ................................................ 58
Figure 4. Overview of fruit fly diagnostic procedures (larval specimens). ............................................... 59
Figure 5. Adult morphology; head (top) and wing (bottom) (White and Elson-Harris 1992). ................... 65
Figure 6. Adult morphology, Thorax; Dorsal features (White and Elson-Harris 1992). ........................... 66
Figure 7. Adult morphology, thorax; lateral features (White and Elson-Harris 1992). ............................. 67
Figure 8. Adult morphology, abdomen; male with features of typical dacini (left), Female, with extended
ovipositor (right) (White and Elson-Harris 1992). ..................................................................................... 67
Figure 9. BOLD generated identification results for the closest matches for an Island Fly (Dirioxa pornia)
COI sequence (accessed June 2015)...................................................................................................... 76
Figure 10. Specimen confidently assigned to species (Lamprolonchaea brouniana), (BOLD tree June
2015). ....................................................................................................................................................... 77
Figure 11. Specimen only confidently assigned to species group (Bactrocera tryoni complex), due to
three closely related species (B. tryoni, B. aquilonis, B. neohumeralis) being “mixed together” (i.e. nonmonophyletic) on the phylogenetic tree (BOLD tree June 2015). ............................................................ 78
Figure 12. Closely related species (B. curvipennis) genetically distinct from the species complex (B.
tryoni) but within the <2% assignment limit set by BOLD (BOLD July 2015). ......................................... 79
Figure 13. Part of the ribosomal RNA operon with the location of primer positions for Tests 1 and 2 .... 80
Figure 14. Workflow of PCR-RFLP procedures for fruit fly identification ................................................. 81
Figure 15. Amplification curves of real-time PCR assay for B. tryoni complex. The sample tested
positive for B. tryoni complex, DNA sequences has further confirmed the results. ................................. 96
8
Figure 16. Anastrepha distincta ............................................................................................................. 103
Figure 21. Anastrepha fraterculus .......................................................................................................... 108
Figure 22. Anastrepha fraterculus .......................................................................................................... 108
Figure 23. Anastrepha grandis female ................................................................................................... 110
Figure 24. Anastrepha grandis wing ...................................................................................................... 110
Figure 25. Anastrepha grandis ............................................................................................................... 111
Figure 26. Anastrepha grandis ............................................................................................................... 111
Figure 27. Anastrepha ludens ................................................................................................................ 114
Figure 28. Anastrepha ludens ................................................................................................................ 114
Figure 29. Anastrepha obliqua ............................................................................................................... 117
Figure 30. Anastrepha obliqua ............................................................................................................... 117
Figure 31. Anastrepha serpentina .......................................................................................................... 120
Figure 32. Anastrepha serpentina .......................................................................................................... 120
Figure 33. Anastrepha striata ................................................................................................................. 122
Figure 34. Anastrepha suspensa ........................................................................................................... 124
Figure 35. Anastrepha suspensa ........................................................................................................... 125
Figure 36. Bactrocera albistrigata .......................................................................................................... 129
Figure 37. Bactrocera albistrigata .......................................................................................................... 130
Figure 38. Bactrocera aquilonis ............................................................................................................. 133
Figure 39. Bactrocera aquilonis ............................................................................................................. 134
Figure 40. Bactrocera atrisetosa ............................................................................................................ 136
Figure 41. Bactrocera atrisetosa ............................................................................................................ 137
Figure 42. Bactrocera bryoniae .............................................................................................................. 139
Figure 43. Bactrocera bryoniae .............................................................................................................. 140
Figure 44. Bactrocera carambolae ......................................................................................................... 143
Figure 45. Bactrocera carambolae ......................................................................................................... 143
Figure 46. Bactrocera caryeae ............................................................................................................... 146
Figure 47. Bactrocera correcta .............................................................................................................. 149
Figure 48. Bactrocera cucumis .............................................................................................................. 152
Figure 49. Bactrocera cucumis .............................................................................................................. 152
Figure 50. Bactrocera cucurbitae ........................................................................................................... 155
9
Figure 53. Bactrocera curvipennis ......................................................................................................... 158
Figure 54. Bactrocera curvipennis ......................................................................................................... 159
Figure 55. Bactrocera decipiens ............................................................................................................ 161
Figure 56. Bactrocera decipiens ............................................................................................................ 162
Figure 57. Bactrocera depressa ............................................................................................................. 164
Figure 58. Bactrocera dorsalis ............................................................................................................... 167
Figure 59. Bactrocera dorsalis ............................................................................................................... 168
Figure 60. Bactrocera facialis ................................................................................................................ 171
Figure 61. Bactrocera facialis ................................................................................................................ 171
Figure 62. Bactrocera frauenfeldi ........................................................................................................... 174
Figure 63. Bactrocera frauenfeldi ........................................................................................................... 174
Figure 64. Bactrocera jarvisi .................................................................................................................. 177
Figure 65. Bactrocera jarvisi .................................................................................................................. 177
Figure 66. Bactrocera kandiensis .......................................................................................................... 180
Figure 67. Bactrocera kirki ..................................................................................................................... 183
Figure 68. Bactrocera kirki ..................................................................................................................... 183
Figure 69. Bactrocera kraussi ................................................................................................................ 186
Figure 70. Bactrocera latifrons ............................................................................................................... 189
Figure 71. Bactrocera melanotus ........................................................................................................... 191
Figure 72. Bactrocera minax .................................................................................................................. 193
Figure 73. Bactrocera musae ................................................................................................................. 196
Figure 76. Bactrocera neohumeralis ...................................................................................................... 200
Figure 77. Bactrocera neohumeralis ...................................................................................................... 200
Figure 78. Bactrocera obliqua ................................................................................................................ 202
Figure 79. Bactrocera occipitalis ............................................................................................................ 205
Figure 80. Bactrocera occipitalis ............................................................................................................ 205
Figure 81. Bactrocera oleae ................................................................................................................... 207
Figure 82. Bactrocera oleae ................................................................................................................... 207
Figure 83. Bactrocera passiflorae .......................................................................................................... 210
Figure 84. Bactrocera passiflorae .......................................................................................................... 210
Figure 85. Bactrocera psidii ................................................................................................................... 213
Figure 86. Bactrocera psidii ................................................................................................................... 213
Figure 87. Bactrocera pyrifoliae ............................................................................................................. 215
Figure 88. Bactrocera tau....................................................................................................................... 218
10
Figure 91. Bactrocera trilineola .............................................................................................................. 221
Figure 92. Bactrocera trilineola .............................................................................................................. 222
Figure 93. Bactrocera trivialis ................................................................................................................ 224
Figure 94. Bactrocera trivialis ................................................................................................................ 225
Figure 95. Bactrocera tryoni ................................................................................................................... 229
Figure 99. Bactrocera tsuneonis ............................................................................................................ 232
Figure 100. Bactrocera tuberculate ........................................................................................................ 234
Figure 101. Bactrocera umbrosa ........................................................................................................... 236
Figure 102. Bactrocera umbrosa ........................................................................................................... 237
Figure 103. Bactrocera xanthodes ......................................................................................................... 240
Figure 104. Bactrocera xanthodes ......................................................................................................... 240
Figure 105. Bactrocera zonata ............................................................................................................... 243
Figure 106. Ceratitis capitata ................................................................................................................. 246
Figure 107. Ceratitis capitata ................................................................................................................. 246
Figure 108. Ceratitis rosa ....................................................................................................................... 249
Figure 109. Ceratitis rosa ....................................................................................................................... 249
Figure 110. Dacus longicornis ............................................................................................................... 251
Figure 111. Dacus solomonensis ........................................................................................................... 253
Figure 112. Dirioxa pornia ...................................................................................................................... 255
Figure 113. Dirioxa pornia ...................................................................................................................... 255
Figure 114. Rhagoletis cerasi ................................................................................................................ 257
Figure 115. Rhagoletis cingulata ........................................................................................................... 259
Figure 116. Rhagoletis completa ........................................................................................................... 261
Figure 117. Rhagoletis fausta ................................................................................................................ 263
Figure 118. Rhagoletis indifferens ......................................................................................................... 265
Figure 119. Rhagoletis mendax ............................................................................................................. 267
Figure 120. Rhagoletis pomonella ......................................................................................................... 269
Figure 121. Drosophila suzukii A. Female (left), Male (right). B. Female ovipositor. C. Male basal
tarsal segments. ..................................................................................................................................... 272
11
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12
Diagnostician’s notes
Grouping of fruit fly species by morphological similarity
TEPHRITIDAE: DACINAE
Distinctive feature: Black scutum & T/ wrap T on abdomen
1
1
6
2
5
4
3
2
Bactrocera dorsalis complex
(see page 165)
1 – fore tibia usu. dark
2 – narrow costal band dips in at end of R2+3
3 - very narrow anal streak
4 – T on abdomen, can be very reduced
5 - black scutum, can have patches of brown to
being mostly brown
6 - broader parallel sided vittae enclosing ia bristle
Bactrocera carambolae
(see page 141)
1 – spot on fore femora
2 – flared costal band
- similar to B. dorsalis (p. 165) except has flared costal
band and spot on fore femora
- similar to B. endiandrae (Qld rainforests, ME) which
can be separated by having tapered shorter vittae,
wide anal streak and diffuse wraparound T
- similar to B. musae (Nth Qld, ME, p. 193) which
can be separated by having a broad costal band
overlapping R2+3, narrow but wider anal streak,
slightly tapered vittae (See p. 197)
13
1
5
2
4
3
Bactrocera occipitalis
(see page 203)
3
Bactrocera caryeae
(see page144)
1 – apical spots on femora
2 – narrow costal band
3 – very narrow anal streak
4 – wide basal band
5 – very narrow vittae
-may appear superficially similar to B. endiandrae (E.
Aus. rainforests, ME) in having a black scutum and
diffuse wrap T but differs in the characters on the
diagram
1
2
Bactrocera kandiensis
(see page 178)
1 – spots on femora
2 – narrow vittae
3 – anteromedial corners of ppn lobes redbrown
Bactrocera pyrifoliae
(see page 214)
14
1
2
3
Bactrocera trivialis
(see page 223)
1 - black mesonotum
2 - colourless costal cells
3 - black across T3
15
16
Distinctive feature: Black scutum & incomplete costal band
Bactrocera oleae
(see page 206)
Bactrocera tuberculata
(see page 233)
1
2
Bactrocera correcta
(see page 147)
1 - scutum black or red-brown with black band or
lanceolate
2 - spot on end of wing
- squat fly with wide abdomen
- similar to B. dorsalis (Exotic, ME, p. 165) but
costal band reduced to apical spot and transverse
facial spots.
- similar to B. zonata (Exotic, ME, p. 241) but
scutum black or with black band.
17
18
Distinctive feature: Black scutum & broad costal band
1
1
3
2
2
Bactrocera bryoniae
(see page 138)
1 - wide costal band to R4+5
2 - distinct T
3 - black scutum
- usually larger fly, may appear superficially similar
to the exotic B. curvipennis (Cue, p. 157) except has
T shape on abdomen and no infuscation on r-m
vein.
Bactrocera curvipennis
(see page 157)
1 - Infuscation r-m vein
2 - broad costal band to R4+5
- superficially similar to B. bryoniae (widespread Aus.,
Cue, p. 138) except no T on abdomen, has infuscation
on r-m vein and tapered vittae.
19
1
1
3
3
2
2
Bactrocera latifrons
(see page 187)
1 - flared costal band, may appear to be spot at
R4+5
2 - broad abdomen, usu. no markings
3 - narrower parallel sided vittae enclosing ia
- may appear similar to B. bancroftii (E. Qld, weakly
attracted to ME) in having a black scutum and broad
abdomen without patterning, but differs in having a
flared costal band, narrow anal streak and no apical
band on the scutellum.
- may appear similar to B. musae (Nth Qld, ME, p.
193) but has a uniformly orange abdomen that is
quite wide and flared costal band.
- also separated from the above species by host
range (Solanaceae, instead of Musaceae for B.
musae and Maclura cochinchinensis for B.
bancroftii) and no response to ME.
Bactrocera musae
(see page 194)
1 - narrow costal band overlaps
2 - narrow anal streak
3 - blunt ended vittae
note: variation in abdomen
20
Distinctive feature: Black scutum & abdomen, no vittae
1
3
2
Bactrocera melanotus
(see page 190)
Bactrocera passiflorae
(see page 208)
1 - Humeral calli entirely black
2 - Mesonotum entirely black
3 - Lacks lateral and medial vittae
21
22
Distinctive feature: Red- brown to golden flies
4
2
1
1
3
5
2
3
2
4
Bactrocera neohumeralis
(see page 198)
1 – costal cells dark, microtrichia in both cells
2 – pointed vittae
3 – mesonotum red-brown
4 – dark humeral calli
-similar to B. tryoni (widely distributed in E Aus., Cue,
p. 226) except has dark humeral calli (= postprotonotal
lobes).
Bactrocera kraussi
(see page 184)
1 – yellow tinted costal cells
2 – faint smoky tint to wing
3 – dark ‘knees’
4 – lateral dark spots usu. most defined on T5
5 – broad basal band
6 – narrow longer vittae to ia, slightly taper
- similar to B. tryoni (Cue, p. 226) except doesn’t have
microtrichia in both costal cells (though often tinted),
has longer less tapered vittae, lateral spots on
abdomen (without medial line), narrow mesopleural
stripe, broad basal band on scutellum and dark apices
of the femora.
23
1
4
2
1
3
2
Bactrocera aquilonis
(see page 131)
Bactrocera tryoni
(see page 226)
1 – microtrichia in both costal cells
2 – faint wrap T or no patterning
- overall golden-orange
- similar to B. tryoni (widespread E. Aus., Cue, p. 226)
but paler and less patterning on abdomen
1 – short pointed vittae
2 – microtrichia in both costal cells
3 – mesonotum red-brown
4 – humeral calli entirely yellow
1
2
1
2
3
3
Bactrocera jarvisi
(see page 175)
1 – joining band between npl calli and ppn lobes
2 – flared costal band
3 – distinct T
Bactrocera zonata
(see page 241)
1 – costal cells colourless and lacking microtrichia
2 – narrow costal band ending at apex of R2+3
3 – small spot at apex of wing
24
Distinctive feature: Glossy black scutum & band on wing
2
2
1
1
5
5
3
3
4
4
Bactrocera albistrigata
(see page 126)
1 - pale ppn lobes
2 - paler legs than B. frauenfeldi
3 - band on wing
4 - usu. narrower medial line & less lateral colouring
than B. frauenfeldi
5 - long vittae
-similar to B. frauenfeldi (Nth Qld, Cue, p. 171) except
has pale ppn lobes, paler legs and less heavy
patterning on abdomen.
Note: pic 20B from Drew & Romig 2013 shows
variation in abdomen
Bactrocera frauenfeldi
(see page 172)
1 – vittae short to long
2 – dark ppn lobes, can be partly paler
3 – legs with red-brown to black markings
4 – broad medial line
5 – black triangle shallow or to end of scutellum
- may appear similar to the exotic B. albistrigata (Cue,
p. 126) except generally dark ppn lobes (or if lighter
not as pale as vittae), darker patterning on legs, thicker
medial line and more distinct lateral bands on T3-T5
- may appear similar to the exotic B. trilineola (Cue, p.
220) except vittae present and facial spots present
(instead of mask)
25
2
1
Bactrocera trilineola
(see page 220)
1 – vittae absent
2 – dark humeral calli
-glossy black face
NB: Bactrocera trilineola is similar to B. frauenfeldi
(Nth Qld, Cue, p.146) but differs in having a glossy
black face and no lateral vittae
26
Distinctive feature: Glossy or matte black scutum & faint band on wing
2
3
1
5
4
Bactrocera kirki
(see page 181)
1 – vittae absent
2 – pale ppn lobes
3 – slight infuscation on r-m & dm-cu veins
4 – very wide medial line
5 – very wide band on scutellum
- may appear superficially similar to B. frauenfeldi (Nth
Qld, Cue, p. 171) but no distinct band on wing (though
can have faint infuscation on r-m and dm-cu cross
veins), lacks vittae, has yellow ppn lobes, very wide
band on the scutellum and very thick medial line on
abdomen.
Bactrocera obliqua
(see page 201)
Bactrocera psidii
(see page 211)
27
Distinctive feature: Black scutum, heavy abdominal markings, no band on
wing
1
2
4
3
Bactrocera facialis
(see page 169)
1 – facial spots absent
2 – pale ppn lobes
3 – narrow costal band, no transverse band on wing
4 – vittae small or absent
- may appear superficially similar to B. frauenfeldi (N. Qld,
Cue, p.171) except has no band on wing, legs pale, ppn
lobes pale and distinct costal band.
28
Distinctive feature: Medial vitta
1
1
4
4
3
3
2
2
Bactrocera atrisetosa
(see page 135)
1 – medial vitta
2 – markings can be reduced to black spot on T5 or no
marking
3 – 4 sc bristles
4 – sa & prsc bristles
- similar to B. cucumis (Qld, NT, NSW, p. 150) but sa
and prsc bristles present
Bactrocera cucumis
(see page 150)
1 – medial vitta
2 – black spot
3 – 4 sc bristles
4 – sa & prsc bristles absent
- similar to the exotic B. atrisetosa (Exotic, p. 135) but
no sa or prsc bristles
29
2
1
5
3
4
Bactrocera cucurbitae
(see page 153)
Bactrocera depressa
(see page 163)
1 – medial vitta
2 – Semi-circle on end of wing
3 – Infuscation dm-cu vein
4 – T shape
5 – 2 sc bristles
- similar to B. chorista (E. Qld, Cue) except more
golden instead of orange, only 2 sc bristles instead of
4, more distinct spot on end of wing, narrower vittae,
less black markings on scutum and less heavy wrap T
on abdomen.
30
1
2
3
Bactrocera tau
(see page 216)
Bactrocera minax
(see page 192)
1 – lateral and medial vitta present
2 – apical spot at end of wing
3 – yellow scutellum with 4 scutellar bristles
NB. This species is similar to Bactrocera cucurbitae
(Exotic, Cue, p. 126) but lacks the pale infuscation
along r-m crossvein and has four scutellar bristles
instead of two
2
1
3
Bactrocera tsuneonis
(see page 231)
Bactrocera decipiens
(see page 160)
1 – medial vitta
2 – large S shape marking on wing
3 – scutum red-brown to black
- usually larger fly with distinctive wing pattern
31
Bactrocera xanthodes
(see page 238)
32
Distinctive feature: Other wing pattern
1
2
Bactrocera umbrosa
(see page 235)
1 - black mesonotum
2 - three obvious bands across wing
Note: abdomen can vary on this species, often with
longer medial line and dark markings on tergite 3 at
edges.
33
34
Dacus spp.
Dacus longicornis
(see page 250)
Dacus solomonensis
(see page 252)
35
Ceratitis spp.
Ceratitis capitata
(see page 244)
Ceratitis rosa
(see page 247)
36
TEPHRITIDAE: TRYPETINAE
Dirioxa pornia
(see page 254)
37
38
Rhagoletis spp.
Rhagoletis cingulata
(see page 258)
Rhagoletis completa
(see page 260)
Rhagoletis fausta
(see page 262)
Rhagoletis indifferens
(see page 264)
39
Rhagoletis mendax
(see page 266)
Rhagoletis cerasi
(see page 256)
Rhagoletis pomonella
(see page 268)
40
Anastrepha spp.
Anastrepha fraterculus
Anastrepha distincta
Anastrepha grandis
(see page 106)
(see page 102)
(see page 109)
Anastrepha ludens
Anastrepha obliqua
Anastrepha serpentina
(see page 112)
(see page 115)
(see page 118)
Anastrepha striata
Anastrepha suspensa
(see page121)
(see page 123)
41
42
DROSOPHILIDAE
Drosophila suzukii
(see page 270)
43
4
Introduction
Fruit flies are one of the world’s most destructive horticultural pests and pose risks to most commercial
fruit and vegetable crops. This has major implications for the sustainable production and market access
of Australia’s $4.8 billion horticultural industry. Worldwide there are some 4,000 species of fruit flies in
the family Tephritidae of which around 350 species are of economic importance.
More than 280 species of fruit fly occur in Australia although only a few of these have been found to
have any degree of economic impact, with Queensland fruit fly and Mediterranean fruit fly being the
species of economic concern. It is therefore important to be able to distinguish between those endemic
species that pose a threat to production and domestic market access from those that do not.
Furthermore, Australia is free from many species that impact production elsewhere. Neighbouring
countries in Southeast Asia and the South Pacific are home to numerous species of fruit fly that pose an
immediate incursion risk to Australian quarantine. Rapid diagnosis of these flies, should they arrive in
Australia, is therefore a critical prerequisite to containing and eradicating the populations before they
establish.
Although a range of diagnostic methods are available that can be undertaken by a number of
laboratories in Australia, there has not been an established agreement on (a) the number and type of
tests that should be conducted to establish a positive identification, (b) the exact protocols that should
be followed for specific diagnostic tests, and (c) agreement on the number and type of protocols that
should be retained and maintained to facilitate a diagnosis at short notice.
This project was therefore undertaken to establish an agreed national standard that is able to facilitate
rapid diagnosis and streamline a national response when suspected incursions occur, and to include
taxonomic identifications using morphological and molecular approaches.
Plant Health Australia (PHA) would like to acknowledge the support, encouragement and professional
advice contributed by all participants to this process.
Background
Australia has a strong, internationally recognised capacity to diagnose fruit fly species and maintains a
wide network of fruit fly traps as part of a national surveillance system. From the Northern Territory and
the Torres Strait Islands to Tasmania, and from Perth to Melbourne, significant expertise is maintained
in state and federal government departments, universities and in the private sector to support the
identification of fruit fly species.
Supported by an extensive world-class fruit fly collection (albeit split across various interstate locations),
Australia is fortunate to have a group of entomologists and other scientists with extensive experience
and knowledge of fruit fly diagnostics.
Not surprisingly, given the range of endemic and exotic fruit flies that can be encountered in different
climatic zones, many jurisdictions have developed specialist expertise to identify species pertinent to
regional production and quarantine requirements.
Against this background, this project was undertaken to establish a diagnostic procedure that has a
national focus and can assist all stakeholders to maintain the strongest capability to identify fruit flies.
44
This project also provides an opportunity to:

collate current (existing) practices and knowledge base into a single document

pool experience from all of Australia’s experts in a collegiate manner

facilitate and improve the constructive exchange of ideas and material across jurisdictions and
entities

establish a resource that can support and develop the confidence and expertise of all users

provide a mechanism to possibly identify future information and research needs, and

consider the potential of both morphological and molecular techniques as they are developed
and become available
Coverage of this diagnostic handbook
To develop this document, a review was firstly conducted to establish those fruit fly species being
targeted by jurisdictions in their current surveillance programs. These species were also reviewed
against diagnostic tools (e.g. electronic and internet keys) already available and in use to support
routine diagnosis. This review, supported by a diagnosticics workshop in 2015, enabled the
development of a revised species list to be covered by this publication (Table 1).
45
Table 1. Fruit flies covered in this diagnostic handbook
Scientific name1
Common name
Exotic
Anastrepha
fraterculus2
South American fruit fly
Exotic
Exotic
Anastrepha grandis
Anastrepha ludens
Mexican fruit fly
Exotic
Anastrepha obliqua
West Indian fruit fly
Exotic
Sapote fruit fly
Exotic
Anastrepha striata
Guava fruit fly
Exotic
Anastrepha suspensa
Caribbean fruit fly
Exotic
Exotic
Bactrocera
aquilonis3
Northern Territory fruit fly
Present in Australia
Exotic
Bactrocera bryoniae
Carambola fruit fly
Exotic
Exotic
Bactrocera caryeae
Bactrocera correcta
Guava fruit fly
Exotic
Bactrocera cucumis
Cucumber fruit fly
Melon fly
Exotic
Bactrocera
cucurbitae4
Exotic
Pumpkin fruit fly
Exotic
Bactrocera depressa4
Pumpkin fruit Fly
Exotic
Oriental fruit fly
Exotic
Bactrocera dorsalis
s.s.5
Exotic
Bactrocera facialis
Bactrocera
frauenfeldi6
Bactrocera jarvisi
Mango fruit fly
Jarvis's fruit fly
Exotic
Bactrocera kirki
Exotic
Bactrocera kraussi
Krauss’s fruit fly
Solanum fruit fly
Exotic
Exotic
Bactrocera minax
Exotic
Note: others that belong to species complexes, but which there is only one representative for in this handbook are:
B. bryoniae (7 species)
B trivialis (8 species)
B. musae (17 species)
B. tau (21 species)
C. rosa (3 species)
1
Note: This is now accepted as a complex of species that are only recognised as morphotypes and some at least as distinct
biological entities, but none described per se with names. Morphological and molecular identifications can only be down to the
level of the complex.
2
3
Belongs to a species complex of which two others are listed in this table: tryoni and neohumeralis.
Zeugodacus has been elevated from sub-generic rank (under genus Bactrocera) to full generic status based on molecular
genetic data (Virgilio et al. 2015). This is yet to be widely accepted by the scientific community.
4
Includes B. papayae, philippinensis and invadens since their synonymisation with dorsalis sensu stricto as per Schutze et al.
(2015) in Systematic Entomology. This is a taxonomic update of the previous list as opposed to an omission of these species.
5
6
Bactrocera frauenfeldi belongs to a complex of five species, including B. albistrigata and B. trilineola.
46
Scientific name1
Common name
Bactrocera musae
Banana fruit fly
Lesser Queensland fruit fly
Bactrocera obliqua
Exotic
Exotic
Bactrocera oleae
Olive fruit fly
Exotic
Fijian fruit fly
Exotic
Bactrocera psidii
South sea guava fruit fly
Exotic
Exotic
Bactrocera
Exotic
tau4
Exotic
Exotic
Bactrocera tryoni7
Queensland fruit fly
Exotic
Exotic
Bactrocera umbrosa
Breadfruit fruit fly
Exotic
Pacific fruit fly
Exotic
Bactrocera zonata
Peach fruit fly
Exotic
Ceratitis capitata
Mediterranean fruit fly
Ceratitis rosa
Natal fruit fly
Exotic
Dacus longicornis
Exotic
Dacus solomonensis
Exotic
Dirioxa pornia
Drosophila
suzukii8
Island fly
Spotted-wing drosophila
Exotic
Exotic
Rhagoletis cerasi
Rhagoletis
Exotic
cingulata9
Rhagoletis completa
Walnut husk fly
Exotic
Rhagoletis fausta
Black cherry fruit fly
Exotic
Western cherry fruit fly
Exotic
Exotic
Rhagoletis mendax
Rhagoletis
7
pomonella10
Apple maggot
Exotic
Belongs to a species complex of which two others are listed in this table: aquilonis and neohumeralis
A vinegar fly that belongs to the Drosophilidae family. Drosophila flies are sometimes called fruit flies, however, true fruit flies
belong to the family Tephritidae
8
9
Belongs to a species complex which also includes R. indifferens
10
Belongs to a species complex which also includes R. mendax
47
5
Detection
Plant products affected
Fruit flies can infest a wide range of commercial and native fruits and vegetables. Lists of hosts are
provided in the data sheets contained in Section 8.
Fruit is increasingly likely to be attacked as it becomes more mature and as fruit fly populations
increase during summer and autumn. A wide range of fruits are potentially vulnerable to fruit fly attack.
In urban home gardens, and in orchards close to urban areas, fruit fly populations are often much
higher than in outlying orchards.
Plant parts liable to carry the pest in trade or transport include fruiting bodies, in which eggs or larvae
can be borne internally. The illegal movement or smuggling of non-commercially produced fruit is the
major risk pathway for exotic fruit fly incursions (CABI 2007).
Signs and symptoms
The oviposition-site punctures in the fruit are commonly referred to as ‘stings’. Stings are usually
identified by making a shallow cut through the skin of the fruit and looking for the egg cavity containing
eggs, larvae or the remains of hatched eggs. In fruits such as peaches, the stings are not very
noticeable, while in pale, smooth-skinned fruits, the sting mark may be easily detected and can
disfigure the fruit when marked by ‘gum bleed’. Some fruits, such as avocado and passionfruit,
develop hard, thickened areas where they are stung. In mature citrus, the sting mark may be a small
brown depressed spot, or have an indistinct, bruised appearance, while on green citrus fruit the skin
colours prematurely around the sting mark. In humid conditions, the fungi responsible for green mould
in citrus and brown rot in stone fruit will readily infect stung fruit.
Fruit will fall from the tree as a result of larval infestation. The extent of the damage caused by larvae
tunnelling through fruit varies with the type and maturity of the fruit, the number of larvae in it, and the
prevailing weather conditions. Larvae burrow towards the centre in most fruits, with internal decay
usually developing quickly in soft fruits. In hard fruit a network of channelling is usually seen, followed
by internal decay. Larval development can be very slow in hard fruits such as Granny Smith apples.
Development stages
The following life history, from McKenzie et al. (2004), is based on the much-studied Queensland fruit
fly. This is also relevant to most other fruit flies, although differences may occur with regard to host
preference and the relationship between developmental rate and temperature.
Typically, fruit flies lay their eggs in semi-mature and ripe fruit. The female fruit fly has a retractable,
sharp egg-laying appendage (the ovipositor) at the tip of the abdomen that is used to insert up to six
eggs into a small chamber about 3 mm under the fruit skin.
Tephritid fruit fly eggs are white, banana shaped and nearly 1 mm long. Infested fruit may show ‘sting’
marks on the skin and may be stung more than once by several females. In two or three days larvae
(maggots) hatch from the eggs and burrow through the fruit. To the naked eye, the larvae resemble
blowfly maggots. They are creamy white, legless, blunt-ended at the rear and tapered towards the
front where black mouth hooks (cephalopharyngeal skeleton) are often visible. Female flies may have
an association with bacteria resident in their gut in some regions of Australia, which they regurgitate
onto the fruit before ovipositing (see Appendix 5). Most of the damage sustained by the fruit is caused
by the bacteria and the maggots simply lap up the juice.
48
A pair of mouth hooks allows the larvae to readily tear the fruit flesh. The larvae develop through three
larval stages to become about 9 mm long and pale yellow when fully grown. Several larvae can
develop in each fruit, and when fully developed they leave the fruit, falling to the soil beneath the tree
and burrowing down about 5 cm to form a hard, brown, barrel-like pupal case from its own skin where
it completes its development. Many flies leave the fruit while it is on the ground. Most insects cannot
pupate successfully in the presence of excess moisture and fruit flies have a prepupal stage when
they can 'flick' themselves over some distance, presumably to distance themselves from the host fruit
or predators.
The duration of pupal developmental is dependent on temperature with each stage taking from nine
days to several weeks to complete. Adult flies emerge from their pupal cases in the soil and burrow
towards the surface where they inflate their wings and fly away. Adults are able to mate within a week
of emerging, living for many weeks with females continuing to lay eggs throughout their lifecycle. Adult
fruit flies feed on carbohydrates from sources such as damaged fruit and honeydew, the sweet
secretion from aphids and scale insects, as well as natural protein sources, including bird droppings
and bacteria.
Fruit fly larvae can be attacked by parasitoids although they appear to have little impact on
populations of most fruit flies, with 0-30% levels of parasitism typical (CABI 2015). However, mortality
due to vertebrate consumption of infested fruit can be very high, as can pupal mortality in the soil,
either due to predation or environmental factors.
Methods for detection
Monitoring is carried out largely by setting traps in areas of interest. However, there is evidence that
some fruit flies have different host preferences in different parts of their range (CABI 2015). As such,
host fruit surveys may be required in the event of an exotic incursion. Where known, specific lures are
provided for each species in the data sheets contained in Section 8. The following information and
images are taken from Lawson et al. (2003).
5.4.1
Trap types
All traps are designed to lure flies into a container, kill them and retain the dead flies in some manner.
Traps should not touch foliage as ants may remove dead flies if an easy pathway is available. Foliage
and twigs in the immediate area of the trap should be checked and pruned regularly. Spiders may also
set up residence in or on traps and remove dead flies. It may be necessary to apply sticky coating
such as Tanglefoot to the wire or string on which the trap is suspended to minimise fly predation or
removal. All traps are hung by wire or string in shady host trees.
DRY TRAPS
Lynfield, Steiner and Paton traps are all ‘dry traps’ used with male attractants (cue-lure, methyl
eugenol (ME) or capilure). They have a cotton dental wick impregnated with a lure and insecticide mix
hung inside the trap body. The cotton dental wicks are inexpensive, provide absorption of the
attractant and insecticide mix, yet still allow evaporation of the lure over relatively long periods. These
traps have small holes in the bottom for drainage. Cone traps are new dry traps that normally are used
with a male lure sachet. Usually information labels are placed on the outside of the trap body (Poison,
lure type and contact info).
5.4.1.1
Lynfield Trap
The Lynfield trap (see Figure 1a) is a pot type trap that has been in use since about 1990. It consists
of a modified clear plastic container, e.g. a 1 litre container with a 100 mm base, a 90 mm diameter
49
top and depth 115 mm. There are four entry holes 25 mm in diameter evenly spaced 15 mm below the
lip of the trap.
Cowley et al. (1990) claimed the Lynfield trap was cheap and extremely easy to operate and inspect,
compared to the Jackson sticky trap and that surveillance costs could be halved using the same
density of Lynfield versus Jackson traps. The Lynfield trap is the standard trap for south eastern
Australia and is the only trap type mentioned in the Code of Practice for the Management of
Queensland fruit fly (Anon. 1996).
5.4.1.2
Steiner trap
The Steiner trap (see Figure 1b) is an open horizontal plastic cylinder. It has openings at each end of
the trap for entry of flies and to allow free movement of the attractant vapour from the cotton wick. The
modified Steiner trap has ingress tubes to prevent rain getting in and flies escaping, with mesh
halfway across the entry to prevent dead flies falling out.
Steiner traps are used in areas of high rainfall such as far north Queensland. This design may be less
effective than Jackson sticky traps in some circumstances in southern Australia (O’Loughlin et al.
1983).
5.4.1.3
Paton trap
The Paton trap (see Figure 1c) is used in areas of high rainfall or wind or where traps may be set for
longer periods (e.g. a month) between collections. Generally, they are used on Cape York Peninsula
and the Torres Strait Islands in Queensland. They are very rain resistant, prevent flies falling out in
windy situations and are able to hold about 10 000 flies (where the Steiner trap can only hold about 6
000). They often are used with cardboard spacers to maintain the samples in good condition.
5.4.1.4
Cone trap
The cone trap (Figure 1e) was recently developed in Spain and has several design advantages. The
ingress holes are short tunnels (similar to Steiners and Patons) rather than plain holes as found in
Lynfield traps. The base of the cone trap is yellow which may be attractive to some species. Cone
traps have a trap door in the bottom of the trap to facilitate removal of dead flies. Initial Australian
research has demonstrated that cone traps are as effective, if not more so, at trapping fruit flies
compared with Lynfield traps (pers. comm. Dominiak 2015 and Royer 2015) and work is underway
comparing Steiner traps with cone traps (pers. comm. Royer 2015). This research will be published in
the immediate future.
WET TRAPS
5.4.1.5
MCPhail trap
The McPhail trap (see Figure 1d) is a plastic flask-shaped container with an invaginated entrance at
the base. It is generally used with liquid lures such as orange ammonia, proteinaceous solutions and
fruit juices, which attract and kill both males and females by drowning them. They can also be used
with solid lures suspended from the lid (e.g. cucumber volatile lure) and a propylene glycol solution for
drowning flies (Royer et al. 2014). McPhails attract flies in far fewer numbers than male lure traps due
to the short range of lure attraction (Dominiak and Nicol 2010). However, they are useful in trapping
species that do not respond to male lures. They need to be cleared regularly to avoid deterioration of
the specimens and attractant liquids need to be replaced at least weekly making these traps time
consuming and messy to service.
These traps may also be used with suspended male lures (e.g. cue-lure or ME) and propylene glycol
when fresh specimens are required for DNA sequencing. Specimens are then transferred to absolute
ethanol.
50
STICKY TRAPS
5.4.1.6
Jackson Trap
The Jackson trap is a delta trap with a sticky base insert (Fig. 1f). Usually these traps are made from
stiffened cardboard or plastic. A robust version was made from galvanised metal with a metal sliding
base for easy removal. Sticky bases are sent to identification centres and flies identified either on the
base or removed with a glue dissolver (e.g. Orange Power). Sticky bases can cause problems in
transport and when trying to remove flies from the glue for identification and are therefore not ideal
when good specimens are required. The Jackson trap was demonstrated to be more effective than
other trap designs in the 1980s (O’Loughlin et al. 1983) and was a standard trap until the design of the
Lynfield trap (Cowley 1990). They appear to have been withdrawn from use in the early 1990s as it
took about four times as long to service compared with dry traps.
Figure 1. Types of fruit fly traps
(a) Lynfield trap (Image courtesy of NSW
I&I)
(b) Steiner trap
(c) Paton trap (Image courtesy of AQIS)
(d) McPhail trap
51
(e) Cone trap
5.4.2
(f) Jackson trap (Image courtesy of
Western Australian Agriculture Authority)
Attractants
Attractants or lures are commonly used to trap fruit flies as they provide an easy way to collect large
numbers of flies in a short period of time.
Food-based attractants, such as those used in McPhail traps, were widely used in the past. These are
still in use today as they offer the advantage of attracting both sexes of many species, including those
not attracted to male lures.
Males of many species respond to male attractants, which attract flies from large distances. Many
species of Dacini are attracted to either cue-lure or methyl eugenol, however, many species have no
known male lure attraction. There has been recent work on male lures expanding the range of
attractants available for trapping.
The different male lures are:

Methyl eugenol (ME) (Figure 2b) - widely used in collecting Bactrocera and Dacus spp. fruit
flies

The ‘cue-group’ (Figure 2a) - cue-lure, raspberry ketone and raspberry ketone formate all are
similar in structure and range of species attracted and are referred to as the ‘cue-group’.
These male attractants are widely used in collecting Bactrocera and Dacus spp. fruit flies with
cue-lure being the most widely used.
o
Raspberry ketone (or Willison’s lure) - one of the breakdown products of cue-lure and
is itself an attractant (Metcalf et al. 1983) attracting the same range of species as cuelure but not as strongly.
o
Raspberry ketone formate or Melolure attracts cue-responsive species and is a
stronger attractant to some species than cue-lure e.g. B. cucurbitae, B. tryoni
([Casana-Giner et al. 2003, Royer 2015], Dominiak et al. 2015a) but less attractive to
others (e.g. B. neohumeralis [Royer 2015]).

Zingerone - a relatively new male attractant which was found to be highly attractive to the
Australian pest species B. jarvisi (Tan & Nishida 2000, Fay 2012) and attracts several species
that do not respond to cue-lure or ME in addition to some cue and ME-responsive species
(Fay 2012, Royer 2015, Dominiak et al. 2015b).

Capilure – widely used for trapping Ceratitis spp.
52

A newer lure Isoeugenol has been found to be more attractive to the cue-responsive pest B.
kraussi than cue-lure, as well as attracting several species thought to be non-responsive to
male lures such as B. halfordiae (Royer 2015).
Cue-lure, ME and capilure are used in Lynfield and Steiner traps, while only cue-lure and ME are used
in Paton traps in Australia (because these traps are used in the tropics and Ceratitis spp. has a lower
likelihood of establishing there).
Figure 2. Chemical structure of cue lure and methyl eugenol.
(a) Cue lure
(b) Methyl eugenol
Male attractants are generally highly volatile chemicals and need only to be used in small amounts to
be effective. Methyl eugenol needs to be replaced every 3-4 months while cue lure can remain
effective in traps for six months. Generally, a wick is impregnated with a mixture of 4 mL attractant and
1 mL or less of 50% w/v concentrate of malathion or dichlorvos and is then suspended within the trap.
It is very important that lure or wick contamination does not occur during the setup process, from when
the wicks are prepared through to when the traps are emptied. If this occurs, then some flies that are
attracted to one lure may also end up in traps containing flies attracted to another. This can lead to
confusion if the lure compound to which they are attracted is also used as part of the diagnostic
identification process. Some flies attracted to methyl eugenol are repelled by cue lure: contamination
of these lures may lead to a failure to detect methyl eugenol attracted flies.
53
6
Inspection of material, sample preparation and
storage
This section was modified/extended by Jane Royer, Mark Blacket, Matt Krosch and Karen Armstrong.
Infested fruit
Fruits (locally grown or samples of fruit imports) should be inspected for puncture marks and any
associated necrosis. Suspect fruits should be cut open and checked for larvae. If immature stages are
being raised to adults, infested fruit should be held in a container which has a gauze cover to allow
aeration (See Appendix 1. Rearing fruit fly larvae to adults for the purpose of identification). The
detection of small larvae can be difficult using the slicing technique.
Sample collection and handling
Fruit fly adults, larvae and eggs should never be handled live if there is any chance of the sample
being involved with a quarantine breach. For the purposes of this protocol, all fruit fly samples, where
the fruit fly adult, larva or egg has been removed from its substrate, should be placed in a sealed vial
or container and be suitably preserved (see below). The sample vial should have labels stating the
collection details including (at a minimum) the collector, collection date, host (if known), place of
collection and where possible accession number(s).
Fruit flies for molecular analysis need to be ‘repacked’ for that purpose, e.g. either as legs removed or
whole specimens dry (with silica pouches or frozen) or in preservative.
Sample transportation
Samples should be collected and despatched in a manner compliant with PLANTPLAN (with particular
reference to sampling procedures and protocols for confirmation). If any live material is to be
transported, appropriate control measures must be employed – including double sealing samples to
ensure quarantine-control is maintained – before placing in (the third level of containment) the postal
packaging prior to dispatch (Hall 2011). Postal transport of more than 30 ml of ethanol may require
special conditions (Hall 2011). Appropriately packed volumes of ethanol less than 30 ml are permitted
to be posted within Australia under “Special Provision A180”,
(http://www.casa.gov.au/scripts/nc.dll?WCMS:STANDARD:1001:pc=PC_100324).
Sample processing, identification and preservation
When samples are delivered to the reception area of a laboratory, the sample and relevant information
should be recorded in a log to ensure they are not lost is subsequent processes. If they become lost,
the loss will be detected and investigated. Upon delivery to the identification area, sample numbers
and their contents should be recorded to ensure the samples sent from the field match the samples
received. This information will be important for audits of laboratory processes and procedures.
Identifications should be conducted using standard identification keys and texts. Part of the recording
should be the identifier’s name and keys used to conduct the identification. These fruit fly
identifications need to be matched with the data sent from the field by monitoring staff. Misidentification or attributing identifications to the wrong location or time could have market access
implications. If the organisation has a paper based system, this information needs to be sent by email
54
or fax to the surveillance staff on the same day as identifications are completed. If the organisation
has an electronic system, identification staff need to find the appropriate electronic surveillance entries
and match the date and location with field records sent with the samples and update the electronic
records. Electronic systems frequently have a reporting protocol, often the following day. This
reporting step completes the full circle for the sample being physically detected in the field to the paper
or electronic report on species identification returned to field staff and managers.
All steps should be able to be audited and audits should be conducted at least annually. Where
inconsistencies or errors are detected, a corrective action report should be issued and processes
improved. If samples are to be retained in the laboratory, samples should be assigned an accession
number and stored appropriately.
Laboratory reared adult specimens are usually preserved as dried pinned reference specimens (e.g.
Mantel & Upton 2010), while bulk field-caught (trap sample) adults are usually initially preserved using
the other options listed in Table 2.
If an adult fly sample is to be preserved for morphological identification, it should be kept dry in a
suitably labelled clean container (as above). Adult specimens are commonly stored for pinning by
freezing which doesn’t adversely affect the morphology of the specimen (e.g. Mantel & Upton 2010),
or greatly affect the DNA yield (e.g. Dean and Ballard 2001).
Do not store specimens for morphological identification in ethanol/alcohol/propylene glycol. They leach
diagnostic colours and patterns necessary for identification.
The key to maintaining good DNA for molecular analyses is to protect the tissues from moisture. Once
removed from the substrate, samples from separate traps or pieces of fruit should be placed in
separate sealed vials and preserved dehydrated by air drying (adults only, not recommended for
humid climates) or freezing, or with the preservatives ethanol (95-100%) or propylene glycol.
Note:

If adults are to be used for morphological identification as well as molecular identification, they
should not be placed in preservative but maintained dry to retain colour (see Section 0)

Live larvae need to be placed directly into boiling water for fixing prior to preservation.
6.4.1
Molecular Specimens
If a sample is being preserved for molecular identification it is important that suitable preservation
methods are applied as soon as possible (Table 2). Moisture is known to damage DNA – a high
concentration of alcohol, propylene glycol or freezing are suitable ways of preventing DNA
degradation. Generally DNA yields are reduced as specimens age (e.g. Dean and Ballard 2001).
However, all of the preservatives outlined below appear more effective when combined with low
temperature (i.e. refrigeration or freezing), increasing the preservative effect (e.g. Moreau et al. 2013).
Cross-contamination could be a potential issue when storing multi-species specimens in bulk, care
must be taken with some types of samples (e.g. dry-frozen adult flies) that the sample used for
molecular testing (e.g. leg) is from that actual specimen.
Freezing: Adult specimens are commonly frozen (-20oC or -80oC) to preserve them. This is a good
way to maintain DNA quality and retain a useful morphological specimen (see alcohol note below).
Freezing however can be of limited use when collecting in the field.
Alcohol: Highly concentrated ethanol (95-100%) is commonly employed to preserve specimens for
molecular testing (all lifestages). However, please note that adult flies preserved in ethanol lose their
colours over time (i.e. losing morphologically diagnostic information). Isopropanol is also sometimes
55
employed as a preservative (e.g. for midges); while other forms of alcohol, such as methylated spirits
are not suitable for DNA preservation. Larval specimens are commonly blanched in hot water prior to
preservation (to aid morphological identification), this practice does not seem to adversely affect DNA
quality. DNA appears to degrade over time when specimens are stored in a lower concentration of
ethanol (<80%), commonly employed for morphological preservation.
Propylene Glycol: Adult material collected from traps is commonly collected into propylene glycol
(e.g. Vink et al. 2005, Moreau et al. 2013), especially in the tropics. Propylene glycol does not
evaporate (unlike ethanol) and can thus be used both for wet trapping and for subsequent
preservation of specimens (e.g. Moreau et al. 2013). An additional advantage of propylene glycol is
that it is non-flammable making transportation of samples easier (e.g. Moreau et al. 2013).
RNAlater: This solution can be used to preserve not only the DNA but also the RNA (e.g. Vink et al.
2005). Overall, it appears that DNA yield might be reduced in RNAlater preserved material compared
with ethanol or propylene glycol preservation (Moreau et al. 2013), although for some specimens
RNAlater appears equally effective (e.g. Vink et al. 2005). Preservation of RNA is critical for
transcriptome-based applications such as differential expression analysis and candidate gene
discovery. RNAlater is not an economical option for large bulk samples and is best used to store small
numbers of conspecific specimens. Critically, RNA degrades much more rapidly than DNA, thus any
trapping design intended to capture RNA profiles from specimens needs to minimise the time between
a specimen entering a trap and preservation.
FTA cards: These can be used for preserving DNA from larval samples, and these have been shown
to be suitable for use under tropical conditions (e.g. Blacket et al. 2015 a). Please note that material
preserved this way does not retain any morphological characters.
Table 2. Commonly employed sample preservation techniques for molecular identification. Methods that are
suitable for a particular lifestage are indicated by “+”, those that are less suitable are indicated by a “–“.
Dry-ambient
Dry-freezing
>95%
Ethanol
Propylene
Glycol
RNA later
Adults
-*
+
+
+
+
Larvae
-
-
+
+
Pupae
-
+
+
+
Eggs
-
-
+
+
FTA card
+
* Storage of dry adults (e.g. trap samples) short-term at ambient temperatures is often employed until
one of the more effective preservation methods is used; wet adults stored at ambient temperature
rapidly degrade (both morphologically and molecularly).
56
7
Identification
Overview
The National Handbook for the Identification of Fruit Flies in Australia (overview presented in Figure 3
and Figure 4) proposes that primary identification is undertaken using conventional taxonomy with the
support of molecular genetic techniques for some species or immature stages. The diagnostic
methods available for each species are presented in Table 3 and covered in greater detail in sections
7.2 (Morphological), 7.3 (Molecular) and Section 8 (data sheets with the specific morphological and
molecular diagnostic information for each species). These techniques are currently in use in Australia
and form the basis of this national protocol.
Tephritid fruit fly adult specimens are primarily identified through an examination of diagnostic
morphological characters. Other life stages are more problematic, with only third instar larvae (and
sometimes pupae) of some species usually identified through visual examination (e.g. White & ElsonHarris 1992). Identification of earlier life stages (early instars, eggs), and adults of morphologically
ambiguous species, generally requires the use of molecular techniques.
Molecular techniques are best used to support or augment morphological identification. They can be
used to identify early larval stages (which are hard to identify reliably on morphological features) and
eggs. They also can be used for incomplete adults that may be missing specific anatomical features
required for morphological keys, or specimens that have not fully developed their features (especially
colour patterns). It should be recognised, however, that the success of a molecular diagnosis can be
impacted by factors such as life stage, specimen quality or any delays in processing. As a result, the
suitability of each method has been identified (See Section 7).
The molecular protocols require a laboratory to be set up for molecular diagnostics, but can be
conducted by almost any laboratory so equipped. Access to on-line published sequences is required
for the DNA barcoding protocol.
Most molecular techniques presented in this standard involve the amplification of particular region(s)
of the fly genome using a polymerase chain reaction (PCR), while the other technique covered
(allozymes) examines protein variation. The PCR target is either the mitochondrial gene for
cytochrome oxidase subunit I (COI), known as the DNA barcode region, or a region of the ribosomal
RNA operon, either just the first internal transcribed spacer (ITS1) or part of the 18S subunit plus the
ITS1.
DNA barcoding is now available in this manual as an alternative to the original ITS-based techniques.
This technology, in contrast, incorporates population variation and has an international, independently
growing reference dataset. However, difficulties can still arise among closely related species within
species complexes.
For the ITS1, the size of the PCR amplicon is useful for identification of a few species. However,
restriction digestion of the ITS1 PCR amplicon, which denotes the actual sequence in defined regions
of the amplicon, is recommended for all analyses as a more robust method of identification. This is
referred to as restriction fragment length polymorphisms (RFLP) analysis. Reference data has been
developed for the economically important species. However, the RFLP does not necessarily eliminate
non-economic fruit flies for which reference data have not been developed.
This national protocol is presented on the premise that most species can generally be resolved using
traditional taxonomy without ambiguity. The molecular methods described here are recommended for
use alongside morphological methods where there is any ambiguity; although they may be equally
ambiguous in the case of species complexes. They can be used on their own for morphologically
cryptic immature life stages, or when only part of a specimen is available, or when access to the
appropriate taxonomic expertise is lacking.
57
Figure 3. Overview of fruit fly diagnostic procedures (adult specimens).
58
Figure 4. Overview of fruit fly diagnostic procedures (larval specimens).
59
Table 3. Diagnostic methods used to identify fruit fly species
Scientific name
Morphological
description
Anastrepha fraterculus

PCRRFLP1
PCRRFLP 2
PCR- DNA
Barcoding4

49/141e*
0/27e*

3/35

b
68/174e*



52/133e*



8/52*
Anastrepha striata


18/59
Anastrepha suspensa

b
5/76e*


Bactrocera aquilonis

a

Bactrocera bryoniae




Bactrocera caryeae

1/1g
Bactrocera correcta

54/54
Bactrocera cucumis



2/11



244/289*



2/2

Bactrocera depressa

Bactrocera dorsalis s.s. 12



>1000/1000g
Bactrocera facialis



1/1
Bactrocera frauenfeldi


d
2/16
Bactrocera jarvisi



3/7

Bactrocera kirki13

Anastrepha grandis

Anastrepha ludens

Anastrepha obliqua
Bactrocera kraussi
11
Allozyme
qPCR11
electrophoresis
24/24
c
3/38f

1/11

110/111

17/17g

d,3
5/5
3/3
Real-time PCR
Includes B. papayae, philippinensis and invadens since their synonymisation with dorsalis sensu stricto as per Schutze et al.
(2015). This is a taxonomic update of the previous list as opposed to an omission of these species.
12
13
B. kirki can be distinguished from B. frauenfeldi and B. trilineola using an additional enzyme Acc I
60
Scientific name
Morphological
description
PCRRFLP1
PCRRFLP 2
PCR- DNA
Barcoding4



82/86


3/3
Allozyme
qPCR11
electrophoresis
58/58
Bactrocera minax
Bactrocera musae14



21/255

a
c
2/4f

Bactrocera obliqua

Bactrocera oleae


Bactrocera psidii

31/34g

109/114


0/1


2/2
85/100*
Bactrocera tau


d
21/21


3/3
Bactrocera tryoni

c
48/55f
a
21/21
5/5
Bactrocera umbrosa



39/39



6/7
Bactrocera zonata



36/61
Ceratitis capitata



208/292
Ceratitis rosa



46/50*


2/2
Dacus longicornis
Dacus solomonensis
Dirioxa pornia





3/3

The 17 species within the B. musae complex (Drew et al. 2011) have not been tested by the RFLP method, but DNA
barcodes will distinguish B. musae s.s. from at least the non-pest species B. conterminal, B. prolixa, B. rufivitta, and B.
tinomiscii; adults of all species can be distinguished morphologically, see section 7.2.
14
61
Scientific name
Morphological
description
PCRRFLP1
PCRRFLP 2
PCR- DNA
Barcoding4
24/29
Rhagoletis completa

Rhagoletis fausta



21/21*
1/1
Rhagoletis mendax
2/2h
Rhagoletis cerasi
17/29
Rhagoletis pomonella
Allozyme
qPCR11
electrophoresis

23/24h
1
Species with the same superscript letter cannot be distinguished from each other with this test.
2
This test is best used when the unknown samples may be suspected as one for which the PCR-RFLP test 1 has not been
developed; all RFLP patterns available in Armstrong & Cameron (1998) are listed in Appendices 3 and 4, and includes other
species not listed in this table, A. bistrigata, A. sorocula, A. zenildae and B. quadriestosa.
3
Numbers in brackets (x/y) refer to the x number of individuals of that species with publically accessible DNA (COI) barcodes
on the Barcode of Life (BOLD) website and y number of total sequences available if using the BOLD identification engine; the
difference represents private barcodes unavailable for download
(www.boldsystems.org/views/taxbrowser.php?taxid=439) as of 18th September 2015.
Where there are no specific DNA barcodes available, others for other species within the genus could be used to at least
determine genus-level identification; as of 18th September 2015, there are 203 species of Bactrocera, 66 of Ceratitis, 82 of
Dacus, 28 species of Rhagoletis and 1 of Toxotrypana that have barcodes available.
Species with the same superscript letter cannot be distinguished from each other with DNA barcodes, or can only be
identified to the level of a species complex.
Species with * are not well resolved amongst a number of other species in BOLD, although low divergence clades may be
apparent. Used in conjunction with other information about the specimen this may still be sufficient to make a confident
identification. Complimentary identification by ITS1 PCR-RFLP could also be considered.
62
Morphological identification
Approximately 90% of the dacine pest species can be identified accurately, and quickly, by
microscopic examination of the adult. For these species there is no need for supporting evidence. The
remaining 10% (mainly some dorsalis complex species) can be identified with this same method but
require expert examination and may require additional supporting evidence such as the molecular
diagnosis or host association records. Methods here are updated according to Drew & Romig (2013).
Only morphological diagnostic procedures and information for adult fruit flies are contained in this
document. Aside from molecular techniques, larval diagnosis has been excluded from this protocol.
For routine morphological identification:
 Collect flies in dry traps.
 When clearing traps collect samples into a tissue. Put tissue in a small box with collection
details on the outside. Samples in tissue can also be collected into a vial though this is less
preferred particularly in the tropics as samples can sweat causing specimens to deteriorate. If
collecting into vial a pencil or permanent pen label (not biro as it runs) should be put inside as
writing on the outside can rub off.
 Store samples in freezer until ready for identification.
 Sort dry specimens in a petri dish under a binocular microscope.
 If keeping specimens after identification store in freezer to prevent deterioration.
 Do not store specimens in ethanol/alcohol/propylene glycol unless being kept for DNA
analysis. They leach diagnostic colours and patterns necessary for morphological
identification.
For suspect specimens requiring further identification
 Store the specimen in a small vial with tissue to protect it until ready to pin or ship. Add a
pencil or permanent ink label (not biro as it runs) detailing collection location, collection date,
collection method, collector, tentative identification and identifier.
 As manipulating loose specimens with forceps tends to damage them, suspect specimens
should ideally be pinned to keep them as intact as possible. If the specimen is a suspect
exotic and needs to be shipped to a specialist ASAP they can be sent unpinned in a tube as
above.
For pinning:
 Put specimen in relaxing chamber with thymol (to prevent mould growth) for 6-12 hours.
 Using a micropin, pin through RHS of scutum. Mount micropinned specimen on a pith stage
on a pin.
 Add label to pin.
 Store pinned specimens in reference collection conditions i.e. 21 OC and 50% RH.
SAMPLE PREPARATION
7.2.1.1
Procedure
The following apparatus and procedures should be used to prepare the specimen for identification
(adapted from QDPIF 2002):
Apparatus:

Stereoscopic microscope or Stereomicroscope with magnification range of 7X to 35X.

Light source

90mm diameter petri dishes
63
Forceps (Inox #4)

Preparation procedure:
1)
Ensure the workstation is clean and clear of all flies before commencing.
2)
Adjust chair height and microscope, and turn on the light source (refer to specific operating
procedures for the microscope in use).
3)
If applicable, record the lure and trap type or host material in which the specimen was found.
4)
Carefully place the fruit fly into a plastic petri dish. If examining more than one fly at once
ensure there is a single layer of flies only, with room to move flies from one side of the dish
to the other.
5)
While looking through the microscope check each fly individually. Manipulate them with the
forceps so that diagnostic features are visible.
7.2.2
Identification
Key features (Figure 5, Figure 6, Figure 7 and Figure 8) used for the morphological diagnosis of adult
fruit flies include:

Overall colour and colour patterning

Wing morphology and infuscation

Presence, shape and colour of thoracic vittae. A vitta is a band or stripe of colour.

Presence or absence of various setae, and relative setal size (for Trypetinae). (Note:
chaetotaxy, the practice of setal taxonomy, is not as important for Dacinae, i.e Bactrocera and
Dacus)
Use the morphological diagnostic key and descriptions contained in Section 8 to identify the species of
fruit fly under microscopic examination.
If identification cannot be made using this diagnostic procedure and/or the specimen is suspected to
be of quarantine concern, it should be referred to either a State or National authority (see section 9.1
Key contacts and facilities). If the specimen is identified as an exotic fruit fly, it should be referred to a
National Authority within 24 hours and the appropriate National Authority notified as required in
PLANTPLAN.
64
Figure 5. Adult morphology; head (top) and wing (bottom) (White and Elson-Harris 1992).
ar – arista
comp eye – compound eye
fc – face
flgm 1 – 1st flagellomere
fr – frons
fr s – frontal setae
gn – gena (plural: genae)
gn grv – genal groove
g ns – genal seta
i vt s – inner vertical seta
lun – lunule
oc – ocellus
oc s – ocellar seta
o vt s – outer vertical seta
orb s – orbital setae
pafc – arafacial area
ped – pedicel
poc s – postocellar seta
pocl s – postcular setae
ptil fis – ptilinal fissure
scp – scape
vrt – vertex
65
Figure 6. Adult morphology, Thorax; Dorsal features (White and Elson-Harris 1992).
a npl s – anterior notopleural seta
hlt – halter or haltere
prepst – propisternum
a sctl s – apical scutellar seta
ial s – intra-alar seta
presut area – presutural area
a spal s – anterior supra-alar seta
kepst – katepisternum
a spr – anterior spiracle
kepst s – katepisternal seta
presut spal s – preutural supraalar seta
anatg – anatergite
ktg – katatergite
anepm – anepimeron
npl – notopleuron
anepst – anepisternum
p npl s – posterior notopleural
seta
anepst s – upper anepisternal
seta
b sctl s – basal scutellar seta
cx – coax
dc s – dorsocentral seta
psctl acr s – prescutellar
acrostichal seta
psut sct – postcutural scutum
sbsctl – subscutellum
p spal s – posterior supra-alar
seta
scape – scapula setae
p spr – posterior spiracle
trn sut – transverse scuture
sctl – scutellum
pprn lb – postpronotal lobe
pprn s – postporontal seta
66
Figure 7. Adult morphology, thorax; lateral features (White and Elson-Harris 1992).
See Figure 5 for abbreviations.
Figure 8. Adult morphology, abdomen; male with features of typical dacini (left), Female, with extended ovipositor
(right) (White and Elson-Harris 1992).
acul – aculeus
ovsc – oviscape
ev ovp sh – eversible ovipositer
sheath
st – sternites numbered 1-5 in
the male and 1-6 in the female
tg – tergites where 1+2 are fused to
form syntergosternite 1+2, followed
by tergites 3-5 in the male and 3-6 in
the female
67
Molecular identification
Three methods are presented here, all of which can use a common sample storage and handling
technique (Section 6), DNA extraction method (Section 7.3.1), and are based on PCR (Polymerase
Chain Reaction) analysis.
Choice of method: The DNA barcoding method (Section 7.3.2) is generally recommended over the PCRRFLP (restriction fragment length polymorphism) methods (Section 7.3.3) because:
(a) DNA barcoding can produce better resolution between species as it utilises variation in the
complete sequence amplified. PCR-RFLP is limited to variation at just a few or several 4-6 bp
restriction sites within the amplicon, the suite of which are dependent on the nature of the
restriction enzymes used
(b) DNA barcoding uses a very large reference sequence database. This is international, publicly
accessible and constantly being added-to by unrelated institutions and projects. Consequently
there is greater inclusion of taxonomically comparative species and population data to improve
confidence in identification. PCR-RFLP generally relies on in-house developed reference
restriction patterns; therefore comparative species and population data are incorporated at a
significantly reduced rate without contributions from independent and international laboratories
(c) DNA barcoding is quantifiable and accessible to bioinformatics analyses. PCR-RFLP is
essentially qualitative, relying on visual inspection against control samples and molecular weight
markers.
If access to a laboratory with DNA sequencing equipment is difficult, PCR-RFLP is a useful alternative
for the majority of species. There are also some species for which identification is ambiguous with DNA
barcoding but not for PCR-RFLP (Table 3); however, while this may be a function of the different gene
regions used, it may also be a result of the many more species and populations included in the DNA
barcode database covering more of the variation.
Gene regions used: DNA barcoding utilises a mitochondrial locus within the cytochrome oxidase I (COI)
gene, while the RFLP methods both utilise a ribosomal DNA (rDNA) gene region that includes the first
internal transcribed spacer (ITS1). Both gene regions have been chosen for the suitability of their
sequences to be distinct between species (Jinbo et al 2011; Wang 2015).
Preparation of general reagents is provided in Appendix 2.
68
7.3.1
DNA extraction for adults and immature stages
The preferred method presented here uses the DNeasy® Blood and Tissue kit (Qiagen). This produces
DNA of very high quality suitable for archiving, should any future work be needed. Other cheaper and
more rapid methods, such as prepGEM (ZyGEM, AKL, NZ) and ZR Tissue & Insect DNA MicroPrep
(Zymo Research, CA, USA) kits, are also highly suitable for diagnostic purposes, but the DNA is of
poorer quality and not so well suited to long-term archiving.
DNA should ideally be obtained using relatively non-destructive techniques, to ensure that a voucher
specimen is available for future morphological examination (Floyd et al. 2010). For fruit fly adults a leg
or the head of a specimen can be used, therefore retaining many other valuable morphological features
of the specimen (such as wings, thorax and abdomen). For larvae, anterior and posterior sections can
be sectioned off and retained, preserving the morphologically valuable mouthparts and spiracles.
Alternative even less destructive methods, involving Proteinase K digestion of internal tissues, are also
very effective (e.g. Gilbert et al. 2007).
DNA should be extracted in a room or a bench separated from PCR set up, and from post-PCR product
analysis, i.e. electrophoresis, sequencing or RFLP tests (Sections 7.3.4 and 7.3.3).
Suitable DNA can be obtained from dry, frozen or ethanol / propylene glycol preserved specimens,
(see Section 6.4.1).
Equipment and/or material needed
•
Blotting paper or Kimwipe tissues
•
Scalpel blades (if sub-sampling each sample)
•
Benchtop Microcentrifuge
•
Microcentrifuge tubes (1.5 mL), (certified DNA free, or autoclaved)
•
QIAGEN extraction kit (DNeasy® Blood and Tissue kit)
•
Heating block or water bath to 56°C
•
Forceps
•
Sterile micro pestle
•
Vortexer
•
Bead-Mill with 3 mm solid glass beads, or plastic micropestle and matching microcentrifuge tube,
(certified DNA free, or autoclaved)
•
QIAGEN DNeasy® Blood and Tissue Kit Handbook, July 2006 (for reference if required)
METHOD
The DNA is extracted by following the manufacturer’s instruction in the DNeasy Blood and Tissue kit
handbook (July 2006) for animal tissue (spin column). Slight modifications are highlighted here, with
additional notes, for DNA used in the real-time or quantitative PCR (qPCR) assay.
Prior to starting the extraction:
 Check if the ATL and AL buffers have precipitated during storage and, if so, warm them until
the precipitate has fully dissolved.
 Ensure ethanol has been added to the AW1 and AW2 buffers as specified on the reagent
bottles.
 Heat the heating block or water bath to 56°C
 Pipette 100 µl x (n+1) of AE buffer into a clean 1.5 mL Eppendorf tube, and place the tube in
the 56°C heating block. (n=the number of samples to be extracted). Eluting the DNA with pre69
warmed AE buffer can increase DNA yield and this is recommended for use with the real-time
PCR protocol.
Starting the extraction:
1)
Allow several hours for processing (Proteinase K digest time dependent).
2)
If using a “Bead-Mill” to homogenise the tissue add two 3 mm solid glass beads (acid-washed in
10 % HCl prior to use) to a clean microcentrifuge tube and add 20 µL of proteinase K. If using a
micro pestle to homogenise the tissue, add 20 µL of Proteinase K directly to the paired microfuge
tube.
3)
Remove samples (e.g. larva, adult head or leg) from ethanol and dry on blotting “tissue” until
ethanol evaporates (approximately 1 min). If samples were “dry” frozen or air dry omit this step.
Eggs can be used, but usually require multiple eggs (>5) to be processed together to obtain enough
DNA (K. Armstrong pers. comm.). Note: 1-2 eggs are sufficient for use in the real-time PCR assay.
4)
Add sample to the Proteinase K tube, cleaning forceps (ethanol wipe) in between samples to
prevent cross-contamination. Homogenise the specimen, either in a Bead-Mill (1 min @ 30 MHz)
or with plastic micro pestle in a microfuge tube.
5)
Quick-spin in centrifuge (up to 10,000 x g / rpm).
6)
If processing large numbers of specimens, rotate previously “outer” samples to “inner” position of
the Bead-Mill (i.e. swap the inner microcentrifuge tube insert around). Repeat Bead-Mill shaking (1
min @ 30 MHz).
7)
Repeat Bead-Mill and centrifuge steps until samples contain no large visible fragments. If using a
plastic pestle, initially grind until this point is reached.
8)
Add 180 µL of Buffer ATL and vortex. Note: the tissue can alternatively be homogenised in 180 µl
of ATL buffer plus the 20 µl of proteinase K together.
9)
Incubate for 0.5-1 ½ h at 56oC, or possibly overnight for complete digestion. For empty pupal cases,
or aged and dried samples, an overnight incubation is recommended.
10) Vortex, quick-spin to remove liquid from inner lid of microcentrifuge tubes.
11) Add 200 µL of Buffer AL and 200 µL of ethanol (Buffer AL and ethanol can be premixed in a large
tube for multiple samples, and then dispensed [400 µL] to each sample). Vortex.
12) Pipette mixture to QIAGEN kit column to bind DNA and centrifuge at ~6000 x g for 1 min. Discard
lower collection tube.
13) Place column into a new collection tube. Add 500 µL of Buffer AW1 to wash. Centrifuge at ~6,000
x g for 1 min. Discard lower collection tube.
14) Place column into a new collection tube. Add 500 µL of Buffer AW2 to wash. Centrifuge at ~20,000
x g (17,000 x g is acceptable) for 3 min. Discard lower collection tube (making sure no AW2 Buffer
splashes onto the base of column).
15) Label the top and side of clean 1.5 mL microcentrifuge tube with the sample’s identification /
database number.
16) Place column into the new microcentrifuge tube. Add 100 µL of AE buffer (this buffer must come
into direct contact with the column filter). Incubate at room temperature for 1 min, then centrifuge
for 1 min at 6000 x g to elute the DNA from the column. Note: 50 µl of pre-warmed AE buffer should
be used for DNA to be used in the real-time PCR protocol.
17) Repeat elution a second time.
18) Discard column and retain microcentrifuge tube containing ~200 µL (or ~100 µL) of DNA in AE
Buffer.
20) Store DNA in -20°C or -80oC freezer. Ideally, DNA can be aliquoted for long-term storage.
70
7.3.2
DNA Barcoding
This test was developed by Mark Blacket, Linda Semeraro and Mali Malipatil, Victorian Department of
Economic Development, Jobs, Transport and Resources (Blacket et al. 2012). This section (7.3.2) was
written by Mark Blacket and Karen Armstrong.
INTRODUCTION
Tephritid fruit fly adult specimens are primarily identified through an examination of diagnostic
morphological characters (Table 2). Other life stages are more problematic, with only third instar
larvae (and sometimes pupae) of some species usually identified through visual examination (e.g.
White & Elson-Harris 1992). Identification of earlier life stages (early instars, eggs), and adults of
morphologically ambiguous species, generally requires the use of molecular techniques.
DNA barcoding is a diagnostic molecular method that is routinely applied at the DEDJTR (Vic.) and
Ministry for Primary Industries (MPI) laboratories (NZ) to identify morphologically problematic
specimens and confirm new fruit fly incursions. The Australian Department of Agriculture and Water
Resources (DAWR) also regularly uses it to screen immature stages (e.g. eggs) collected from fruit to
exclude the non-tephritid exotic pest Drosophila suzukii (A. Broadley pers. comm.). DNA barcoding of
an unknown insect specimen involves obtaining a DNA sequence of a specific region of the
mitochondrial Cytochrome Oxidase I (COI) gene, and then comparing it with a database of sequences
from positively identified reference specimens.
There are currently many reference DNA barcoding sequences available. The Barcode of Life Data
Systems (BOLD) (http://www.boldsystems.org, accessed June 2015) holds 833 x Tephritidae,
including 198 x Bactrocera “species with barcodes”. This also includes data from NCBI GenBank and
of regional studies covering a broad range of endemic or intercepted tephritid species – e.g. 60
species from Oceania (Armstrong and Ball 2005), 153 species from Africa (Virgilio et al. 2012), 135
species from Europe (Smit et al. 2013). However, there is no single peer-reviewed published DNA
barcoding laboratory protocol that covers all of the targeted tephritid species listed in Table 3.
DNA barcoding should ideally obtain DNA using relatively non-destructive techniques, to ensure that a
voucher specimen is available for future morphological examination (Floyd et al. 2010). For fruit fly
adults a leg or the head of a specimen can be used, therefore retaining many other valuable
morphological features of the specimen (such as wings, thorax and abdomen). For larvae, anterior
and posterior sections can be sectioned off and retained, preserving the morphologically valuable
mouthparts and spiracles. Alternative even less destructive methods, involving Proteinase K digestion
of internal tissues, are also very effective (e.g. Gilbert et al. 2007).
There are a number of potential issues that should be taken into consideration when applying a DNA
barcoding approach to fruit fly species identification:
1) Many studies have not used the same gene region rendering the data unusable (Boykin et al.
2012);
2) Some commonly applied combinations of standard polymerase chain reaction (PCR) primers
(e.g. LCO/HCO) may amplify a nuclear copy (a numt pseudogene) of the COI barcode region
in some fruit flies (Blacket et al. 2012);
3) The presence of fruit fly species complexes can limit precise species identification (e.g.
Blacket et al. 2012, Jiang et al. 2014), enabling identification only to the complex level (e.g. B.
tryoni complex);
4) The confidence of assigning an unknown sequence to a reference species can be problematic
if all relevant potential taxa are not able to be included in the reference dataset (e.g. Virgilio et
al. 2012). The degree of identification uncertainty is dependent upon a number of factors,
71
including the accuracy of the taxonomic reconstruction (Lou & Goulding 2010), how recently
the sister species diverged (e.g. van Velzen et al. 2012), and the geographic scale of
reference samples (Bergsten et al. 2012).
Other applications of DNA barcoding data in addition to species identification are beginning to be
explored. For example, in a biosecurity context the DNA sequence data generated can also be used to
trace lineages (Blacket 2011) and possible source populations (Barr et al. 2014).
AIM
This test aims to identify fruit fly species, from any life stage, through DNA sequencing and
comparison with reference sequences of the DNA barcoding region using the publicly available
reference information in BOLD.
TARGETS
Almost all of the relevant species of fruit flies from the Australasian region are represented in BOLD
(Table 3). The small number of species that have not been sequenced to date belong to genera where
many other species have been examined; this allows a DNA barcoding approach to at least place
these species to the appropriate genus. Note: for some species complexes COI or ITS DNA
sequencing is unable to differentiate individual species (see Table 3).
PROCEDURE
This document provides supporting information for a three-step DNA barcoding process involving:
1) DNA extraction for fruit flies (adults and immature stages)
2) PCR of the mitochondrial COI barcode gene region from the fruit fly DNA.
3) Data analysis of the COI barcode gene region for species assignment using BOLD.
7.3.2.1
PCR OF THE MITOCHONDRIAL COI BARCODE GENE REGION FROM FRUIT FLY
DNA
The DNA barcode region of the COI gene is PCR amplified using a fruit fly-specific priming system,
producing a ~550 bp amplicon for sequencing. The PCR Master Mix reagents and set up must be in a
separate storage and at a separate bench (respectively) to the DNA extraction and post-PCR product
analysis (electrophoresis or RFLP, Sections 7.3.3 and 7.3.4).
Equipment and/or material needed

Primers:
o Forward: FruitFlyCOI-F (FFCOI-F) 5’-GGAGCATTAATYGGRGAYG-3’ (Blacket et al.
201215)
o Reverse: HCO 5’- TAAACTTCAGGGTGACCAAAAATCA-3’ (Folmer et al. 1994)
This primer is a fly-specific primer that was initially successfully tested on Bactrocera, Ceratitis (Tephritidae) and Calliphora
(Calliphoridae) species.
15
72

PCR Master Mix:
o BSA [1X] (diluted with distilled H2O from 100X BSA stock)
o NEB 10X Buffer (Cat# M0267S)
o dNTP’s [2.5 mM]
o MgCl2 [25 mM]
o Primers [10 µM] (working primer concentration is 10 µM, store stocks at 100 µm, -20oC)
o NEB Taq DNA Polymerase (Cat# M0267S)
o QIAGEN QIAquick Spin Handbook, March 2008 (for reference if required)

DNA template of unknown specimen(s) (see Section 7.3.1)

Fruit fly DNA to use as a positive control (to confirm that the PCR amplification is working)

AE Buffer (or TE) from the DNA extraction kit can be used to replace DNA as a negative
control (to confirm that no PCR reagents are contaminated).

Thermal cycler, e.g. “T800”, Eppendorf (epgradient S) Thermocycler

Additional equipment / materials: Submerged gel electrophoresis power pack, gel tank, TBE
buffer, agarose etc.
Methods
1) Extract fruit fly DNA for use as template (see Section 7.3.1).
2) Set up Master mix (see below), keeping all reagents on ice during setup and adding the DNA
template last.
3) The Master Mix consists of 23 µL reaction volume for each sample (plus a negative and
positive control). Note, if dealing with large numbers of samples (e.g. >10) an extra reaction
may be required to account for retention of liquid in pipette tips:
MasterMix
Reagent
x 1 reaction (µL)
x 3 reactions (µL)
1X BSA
15.3
45.9
10X Buffer
2.5
7.5
dNTP’s
2
6
MgCl2
0.5
1.5
FFCOI-F
1.25
3.75
HCO
1.25
3.75
NEB Taq
0.2
0.6
DNA Template
2
-
TOTAL:
25 µL
69 µl (x 23 µL per reaction)
73
4) PCR Conditions (“T800”, Eppendorf (epgradient S) Thermocycler):
1 x cycle
94 °C, 2 min
40 x cycles
94 °C, 30 s
52 °C, 30 s
72 °C, 30 s
1 x cycles
72 °C, 2 min
1 x hold
15 °C, indefinitely
5) After PCR is complete, load 5 µL of the PCR product (plus 2 µL loading dye) onto a 2%
agarose checking gel (use 5 µL of SYBR Safe [Cat# S33102] per 50 mL liquid gel mix, before
casting gel). Mix PCR product and dye together in plastic “gel loading” plate, using a new
pipette tip for each sample. Run agarose gel at 100 V, for 30 min. Visualise and photograph
gel on a UV light box. Estimate PCR product concentration for DNA sequencing from the
agarose gel photo (weak PCR ~20 ng µL-1, strong PCR >100 ng µL-1).
6) If necessary, clean successful PCR products using QIAquick PCR Purification Kit (QIAGEN,
Cat# 28104), elute final volume in 30 µL of EB Buffer (some external sequencing companies,
e.g. Macrogen do this step).
7) Send to external facility (e.g. Micromon, Monash University or Macrogen, Korea) for DNA
sequencing in both directions using the PCR primers above (Note, depending on each
laboratories procedures, and quality of the sequence obtained therein, sequencing may be
conducted in one direction only).
8) After high quality DNA sequences have been obtained (preferably with a QV or Phred score of
greater than 20), and with PCR primer sequence removed and the forward and reverse
consensus sequence determined (if sequencing both directions), they can be compared with a
public database (i.e. BOLD, Barcode of Life website16) to identify species as outlined in
Section 7.3.2.2.
16
http://www.boldsystems.org
74
7.3.2.2
DATA ANALYSIS – DNA BARCODING SPECIES IDENTIFICATION
The DNA sequence data for each specimen is compared with other publicly available reference
sequences for species identification. Some species, such as Bactrocera tryoni, are members of a
species complex, where the very closely related species within the complex cannot be distinguished
by their DNA barcode sequence; these specimens are thus reported as being identified to a species
group (e.g. B. tryoni complex, rather than B. tryoni).
Method
1) Go to the Barcode of Life website (http://www.boldsystems.org/).
2) Click the “Identification” tab (http://www.boldsystems.org/index.php/IDS_OpenIdEngine).
3) Paste the DNA sequence (use only the high quality section of the DNA sequence) into the
“Enter sequences in fasta format:” box (the sequence can just be entered as simple text and
does not need to be in FASTA format).
4) Select the “Species Level Barcode Records” (default); this selects reference sequences that
are >500 bp from named (identified) specimens, but some species may be represented by
only one or two specimens or be included with interim taxonomy.
5) Click the “Submit” button.
6) The top 20 matches are displayed (default), 99 best matches are available, together with the
“Specimen Similarity” score (as a percentage).
7) The matches with the highest percentage similarity (listed from highest to lowest) are the
reference sequences that best match the unknown specimen being identified. The best
matches are listed under “Search Results” and in a “Identification Summary” table together
with the percentage “Probability of Placement” (Figure 9).
8) The matches should also be viewed as a phylogenetic tree using the “Tree based
Identification” button.
9) Click “View Tree” to view a PDF of the phylogenetic tree.
10) On the tree the specimen being identified is referred to as the “Unknown Specimen” (written in
red) on the tree (indicated with arrows in Figure 10 and Figure 11), and is shown closest to the
reference specimens that it best matches.
11) The specimen can now be assigned to the species that it is most similar to, with three caveats:
a. Some specimens can only be assigned to a species group (i.e. a closely related
complex of species, Figure 11), where the species within it are unable / too similar to
be distinguished using DNA barcoding. The indications for this are when different
specimens of the same species are mixed among other species and there is no clear
single-species clade (Figure 11); this is known to occur for the complexes of B. tryoni,
B. dorsalis and Anastrepha fraterculus.
b. The assignments made by BOLD are based on a generic <2% within-species
divergence (i.e BINs, Ratnasingham & Hebert 2013). This may not be appropriate for
all of the species being analysed. For example Figure 12 shows divergences within
the B. tryoni complex are very similar (<2%) to another closely related distinct species,
B. curvipennis.
c.
The closest match is not always with the correct species. BOLD recommends that an
“identification is solid unless there is a very closely allied congeneric species that has
not yet been analysed”. While most pest fruit fly species of interest are represented in
BOLD (e.g. Table 2), closely related congeners may not. Indications for this are
difficult to observe and this caveat is dependent on knowledge of the local taxonomy.
75
Figure 9. BOLD generated identification results for the closest matches for an Island Fly (Dirioxa pornia) COI
sequence (accessed June 2015).
76
Figure 10. Specimen confidently assigned to species (Lamprolonchaea brouniana), (BOLD tree June 2015).
77
Figure 11. Specimen only confidently assigned to species group (Bactrocera tryoni complex), due to three closely
related species (B. tryoni, B. aquilonis, B. neohumeralis) being “mixed together” (i.e. non-monophyletic) on the
phylogenetic tree (BOLD tree June 2015).
78
Figure 12. Closely related species (B. curvipennis) genetically distinct from the species complex (B. tryoni) but
within the <2% assignment limit set by BOLD (BOLD July 2015).
79
7.3.3
PCR-RFLP
OVERVIEW
Two tests are described. Both are based on fruit fly-specific PCR of the rDNA ITS region (Figure 13).
Differences in method details (sections 7.3.3.1 and 7.3.3.2) reflect subtle differences in the standard
operating procedures of the original laboratories.
Test 1, developed by McKenzie et al. (2004), utilises a 0.6 to 1.2 kb PCR fragment of the ITS1 only
(Figure 13). As the ITS1 can be variable in length, for some species the size of the PCR fragment can
be an additional diagnostic character. Separate aliquots of the PCR amplicon are digested with each
of up to six different restriction enzymes to produce a species-specific RFLP profile. This number may
be reduced if the potential species could be narrowed down by knowledge of likely origin and/or host,
and by reference to Table 4 which illustrates which enzymes distinguish which species. This test can
be used for identification of 26 fruit fly species (Table 3).
Test 2, originally developed by Armstrong and Cameron (1998), is similar to Test 1 but amplifies a
larger 1.5-1.8 kb DNA fragment, encompassing the 18S and the ITS1 gene regions (Figure 13).
Because of this, the variability in PCR fragment length is not so discernible and is therefore not useful
as an additional character. Various combinations of three or four out of 10 restriction enzymes are
recommended to produce a species-specific RFLP profile. The specific suite will depend on the likely
species identity if it can be narrowed down by likely country of origin or the fruit. This test can be used
for identification of 33 fruit fly species (Table 3); details are available in Armstrong & Cameron (1998).
In the specific circumstance to distinguish just B. tryoni complex from C. capitata the enzymes AluI,
DdeI, RsaI and SspI are diagnostic.
Figure 13. Part of the ribosomal RNA operon with the location of primer positions for Tests 1 and 2
ITS1
18S
baITS1f
5.8S
baITS1r
ITS2
28S
Test 1
Test 2
NS15
ITS6
The general workflow is presented in Figure 14.
80
Figure 14. Workflow of PCR-RFLP procedures for fruit fly identification
1. Choose standards to
run in conjunction with
unknown and up to four
restriction enzymes for
diagnosis
Start
10. Set up fresh reactions;
incubate for longer
No
2. DNA extraction from
unknown sample
Section 7.3.3
8. Test 1 & 2
Restriction analysis.
Section 7.3.3.1 or
7.3.3.2
9. Have restriction
reactions gone to
completion?
Yes
3. PCR amplification
Section 7.3.3.1 or 7.3.3.2
5. Electrophoresis test for
amplified DNA
11. Compare fragment
number and length with
reference data
13. Consider additional
enzymes, or conclude it
is a different species to
any of the reference
species
Test 1 Table 5
Test 2 Appendix 4
12. Document fruit fly
identification
No
6. Has DNA been
amplified?
Yes
No
7. Test 1. Record if ITS1
fragment length is
diagnostic (Table 4).
9. Is identity consistent
across enzymes; does
identity make
sense re
Yes
other information on
fruit host or origin?
Yes
End
81
SPECIES TARGETS
Test 1 can distinguish 25 of 27 target species within the current list of 59 species (Table 1); three species
within the Bactrocera tryoni group (including B. tryoni, B. neohumeralis and B. aquilonis) cannot be
distinguished from each other.
Test 2 can distinguish 24 of 33 target species within the current list of 59 species (Table 1); nine species
form four species groups.
Test 2a distinguishes B. tryoni complex from C. capitata only.
All tests should be considered in terms of the host fruit (for immature life stage samples) and likely
country/place of origin to (a) narrow the possible species at the outset for choice of restriction enzymes
and (b) to check the results for reliability. Host records (Section 7) for the target taxa may assist in the
elimination of possible non-target species.
CHOICE OF RFLP TEST
Test 1 is more rapid, the RFLP patterns less complex to interpret and the variation in size of the ITS1
region easier to detect than Test 2. Therefore this test is recommended when the potential species is
within that listed for this test (Table 3).
Test 2 takes slightly longer, the RFLP patterns are sometimes more complex to interpret and the
variation in size of the ITS1 region not as easy to detect is in Test 1. However, a slightly modified version
(Semeraro & Malipatil 2005) provided below, is recommended instead of Test 1 specifically for the
diagnosis of the high-risk pest species B. tryoni (although indistinguishable from B. neohumeralis and
B. aquilonis within the species complex) and Ceratitis capitata. The original method provided in
Armstrong and Cameron (1998) (not included in this Handbook) is also recommended for a further 11
species not included in Test 1 but within the now extended species list in this version of the Handbook
(Table 3); the diagnostic data for Test 2 is however provided in Appendices 3 and 4.
Potential for misidentification:
 False positives are possible if other less economically important species, not included during
development of the tests, have the same RFLP pattern. This scenario is reduced by increasing
the number of restriction sites screened by using several restriction enzymes for an
identification. It is also unlikely for exotic fruit fly interceptions given the low potential for nonpest species to reach Australia. For Australian methyl eugenol or cue lure trapped specimens,
the range of taxonomically close local non-target species has not been included in development
of either test. Therefore care is needed in interpretation of the results in light of other information
about the sample, e.g. data on host or geographic origin (Section 8).
 False negatives could arise if a non-conforming RFLP pattern is produced by a population of
a species that has an aberrant polymorphism at a diagnostic restriction site. This is unlikely
given the range of populations included during the development of these tests. Also, the ITS1
is generally not useful for detecting population-level variation (cf COI barcodes, Section 7.3.3).
False negatives could also arise if the restriction fails or fails to go to completion. DNA from a
positive control (known species) should therefore be included to detect this.
CHOICE OF STANDARDS
All analyses should incorporate at least one standard, i.e. DNA from a known species. This can include
one of the anticipated species for identity of the unknown specimen, which is useful as a positive control
to help with sizing the restriction fragments of the unknown. Alternatively, it can be from a completely
unrelated species, which is useful to minimise any question of contamination or mix up. Standards are
essential to provide confidence that the PCR and restriction steps are operating correctly.
82
7.3.3.1
PCR-RFLP TEST 1
A) PCR
Equipment
 Pipettors and tips
 Sterile disposable microcentrifuge tubes
 Microcentrifuge
 Gel tank and power pack
 Latex or Nitrile gloves
 Microwave
 UV transilluminator with camera
 Thermocycler
 Personal protective equipment including lab coat, eye protection, gloves
Reagents (see APPENDIX 2 for reagent compositions)
 Manufacturer’s polymerase enzyme buffer 10X
 Taq polymerase enzyme (5U μL-1)
 Primer (McKenzie et al. 1999) are:
o baITS1f 5’ GGA AGG ATC ATT ATT GTG TTC C 3’, 10 μM
o baITS1r 5’ ATG AGC CGA GTG ATC CAC C 3’, 10 μM
 dNTP’s (2 mM)
 MgCl2 (50 mM)
 Sterile water
 1X TBE buffer
 1% (w/v) agarose gel: 1 g DNA grade agarose per 100 mL 1X TBE
 6X Loading dye
 DNA molecular weight marker (aka 100 bp ladder)
 Gel staining solution, e.g. Ethidium bromide (final concentration 800 ng/μL), or non-toxic options
such as Syber Safe, or Red Safe according to manufacturer’s instructions
Method
In a pre-PCR cabinet:
1) label sterile 0.2 ml PCR tubes
2) make a Reagent Master Mix for the total number of samples to be analysed according to the
table below, all reagents except for the polymerase (note, recommend adding an extra volume
for more than 10 samples to allow for any loss of solution via adherence to the outside of the
pipette tip):
Reagent
Final concentration
Vol per reaction (μl)
Manufacturer’s reaction buffer (10X)
1X
5
MgCl2 (50 mM)
1.5 mM
1.5
dNTP’s (2 mM)
200 μM
5
Forward primer (10 μM)
1 μM
5
Reverse primer (10 μM)
1 μM
5
H2O
20.25
Taq polymerase enzyme (5U μL-1)
0.25
Total volume
42
83
3) Store Master Mix on ice in sterile 1.5 mL centrifuge tube.
4) Add 8 μL of sterile distilled H2O or DNA buffer to the first negative control tube.
In a laminar flow hood or PCR cabinet:
5) Add the Taq polymerase to the Master Mix
6) Aliquot 42 μL Taq Master Mix to each PCR tube
7) Add 8 μL of DNA extract/control to each sample tube as appropriate
8) PCR amplify the DNA in a thermal cycler using the following program:

Cycle 1
- denature 94°C 2 min

Cycles 2 to 35 - denature 94°C 1 min, anneal 60°C 1 min, extend 72°C 1 min

Cycle 36
- extend 72°C 5 min
9)
10)
11)
12)
13)
Place reaction products on ice or freeze until ready to analyse.
Mix 3 μL of each PCR sample with 2 μL loading dye.
Load samples and 100 bp DNA ladder onto separate wells of 1% (w/v) agarose gel in 1X TBE.
Electrophorese in 1X TBE buffer at 100 V for around 40 min.
Stain the gel in ethidium bromide or other stain, according to local Standard Operation
Procedure.
14) Visualise bands and capture image using the Gel Documentation System.
Diagnostic use of ITS1 amplicon size
The expected size of the amplified product is between 500 and 1000 bp, depending on the species
(Table 4). For some species the size can be an additional diagnostic character. However, relying on
the resolution available on a gel to distinguish the majority of these amplicon sizes is not advisable as
the only diagnostic character, but is useful in conjunction with restriction analysis as described below.
Flies producing fragments of less than 700 bp or greater than 900 bp are segregated and then
restriction enzymes are used in series to differentiate the species. Sizes are given as a range to reflect
that sizing is approximate when using low-resolution gel electrophoresis systems as here.
84
Table 4. Approximate size of the ITS1 amplicon for each species. Potential diagnostic size ranges are contained
in a block
Species Fragment range
ITS1 amplicon ~size range (bp)
Potential species
500-520
D. pornia
590-610
B. cucurbitae
640-680
A. ludens
650-690
A. obliqua
670-700
B. xanthodes
740-760
A. serpentina
740-780
R. pomonella
750-780
B. umbrosa, B. facialis
760-770
B. cucumis
760-780
B. latifrons
770-790
B. musae
770-800
B. endiandrae
780-800
B. psidii
790-840
B. bryoniae, B. tryoni sp. complex
800-820
B. moluccensis
800-840
B. dorsalis sp., B. jarvisi
810-840
B. passiflorae
820-850
B. zonata
830-860
B. carambolae, B. curvipennis, B. frauenfeldi
840-860
B. albistrigata, B. kirki
890-900
C. capitata
1000-1040
C. rosa
85
B) RESTRICTION DIGESTION OF PCR PRODUCT
The use of a combination of enzymes, in series, allows definitive identification of the majority of the
species. This also eliminates the reliance on discrete restriction sites and minimises the likelihood of
false negatives that may arise through a rare recombination event.
Restriction endonucleases used are VspI, HhaI, SspI, HinfI, BsrI, SnaBI and/or Sau3aI. During the
development of this test standard enzymes purchased from New England Biolabs were used but other
brands would work equally well. Enzymes were also selected based on the requirement for differences
in fragment sizes to be easily detected by visual examination of an agarose gel.
If the likely species can be narrowed down, e.g. by fruit or geographic region, then a reduced number
of enzymes could be used. However, for robust diagnoses, it is recommended that at least four enzymes
are used, even if there is a diagnostic pattern for one enzyme that distinguishes a species from all
others.
Equipment
 Pipettors and tips
 Sterile disposable microcentrifuge tubes
 Microcentrifuge
 Dry heating block, waterbath or similar
 Gel tank and power pack
 Latex or nitrile gloves
 Microwave
 UV transilluminator with camera and image capture and analysis software
 Personal protective equipment including lab coat, eye protection, gloves
Reagents
 Sterile distilled water
 Bovine serum albumin (BSA, 10 μg/μL) (comes supplied with NEB enzymes)
 Restriction enzymes VspI, HhaI, SspI, HinfI, BsrI, SnaBI, and Sau3aI
 Restriction buffer supplied with enzyme
 Ethidium bromide solution, 800 ng/μL final concentration, or non-toxic options such as Syber
Safe, or Red Safe according to manufacturer’s instructions
Method
1) Label microcentrifuge tubes, including one for the positive control.
2) To each centrifuge tube add:
Water
2.3 μL
10X buffer
2 μL
BSA (10 ug/μL)
0.2 μL
PCR product
5 μL
Restriction enzyme
0.5 μL
3) Mix reagents and place tubes in a water bath preheated to 37oC for 2 h.
4) Store tubes on ice or at -20oC until ready to load on agarose gel.
5) Add 3 μL of 6X loading buffer to each tube.
6) Load the entire volume of each sample (23 μL) into a lane of a 2% (w/v) high resolution agarose gel.
7) Load 100 bp DNA molecular weight marker into one or two wells of the gel.
8) Analyse products by electrophoresis at 100 V for 50 min.
86
9) Stain the gel with ethidium bromide, or alternative non-toxic stain.
10) Visualise fragments using a UV transilluminator.
11) Capture gel image using gel documentation system.
Analysis of RFLP products
For diagnostic purposes, RFLP bands under 100 bp and over 1500 bp in size are disregarded due to
difficulty in accurate sizing. The molecular weights of the restriction fragments are estimated with
reference to the DNA molecular weight standard loaded on the same gel.
Tables 4 and 5 summarise the expected fragment lengths for the six restriction enzymes used in this
method. All species listed can be differentiated from each other, with the exception of those within the
B. tryoni complex. Care should be taken with fragments produced for B. albistrigata and B. kirki,
which are very similar for all six restriction enzymes and potentially difficult to confidently distinguish on
an electrophoresis gel.
Species
87
Table 5. Analysis of RFLP products from ITS1 fragments from fruit flies
ITS1*
Species
˂700
700-900
HinfI
˃900
DNC
VspI
Cuts
DNC
Hhal
Cuts
DNC
A. ludens
X
550
550
X
A. obliqua
X
450, 270
550
X
420, 250
X
A. serpentina
X
X
B. albistrigata
X
X
B. aquilonis
X
B. bryoniae
X
B. carambolae
X
X
B. cucumis
X
X
B. cucurbitae
X
Cuts
SspI
DNC
BsrI
Cuts
DNC
X
550, 150
X
Cuts
SnaBI
DNC
Sau3aI
Cuts
DNC
X
X
X
450, 200
X
X
530, 200
X
X
450, 400
X
X
670, 180
620, 180
770
X
640, 190
570, 180
600, 200
X
415
760
X
620, 200
560, 180
600, 230
X
400
680, 200
X
X
550, 180
X
X
X
X
X
X
400, 180
X
X
X
480, 350
X
X
X
620, 170
X
650, 190
X
650, 260
600, 180
X
600, 200
B. curvipennis
B. dorsalis sp.
complex
X
B. facialis
X
X
X
B. frauenfeldi
X
X
X
B. jarvisi
X
B. kirki
X
X
770
770
650, 250
550, 200
620, 180
570, 250
X
600, 250
530, 350
450, 400
X
420
540, 320
X
X
390
X
450, 400
X
420
X
450, 400
X
640, 180
700
X
X
680, 190
620, 180
B. latifrons
X
X
X
600, 190
X
600, 200
B. musae
X
X
X
635, 220
X
600, 250
B. neohumeralis
X
770
X
640, 190
570, 180
600, 200
X
B. passiflorae
X
770
X
650, 190
750
650, 270
X
X
B. psidii
X
X
640, 190
570, 250
X
X
X
Cuts
X
X
X
X
X
520, 320
X
420
88
Species
ITS1*
˂700
700-900
HinfI
DNC
770
X
640, 190
570, 180
B. umbrosa
X
730
X
600, 190
680
B. xanthodes
X
680
X
670, 200
380, 250
B. zonata
X
X
680, 190
750
X
X
X
C. rosa
X
800, 200
X
X
DNC
Cuts
DNC
BsrI
Cuts
X
Cuts
SspI
X
C. capitata
DNC
Hhal
B. tryoni
D. pornia
˃900
VspI
Cuts
DNC
SnaBI
Cuts
600, 200
DNC
Cuts
Sau3aI
DNC
Cuts
X
420
X
X
380
X
X
600, 200
X
535, 330
X
650, 200
X
520, 160
X
X
X
600, 300
X
570, 480
X
X
X
X
300, 220
X
X
X
The length of the ITS1 fragment and the response of each to seven restriction enzymes (HinfI, VspI, HhaI, SspI, BsrI, SnaBI, Sau3aI) are indicated for each of the target
species. ITS1 fragment length is scored as one of three classes (approximate length in bp). Enzyme responses are measured in two classes - either does not cut (DNC) or
cuts.
(Cuts – this column shows the length of each fragment in bp). Highlighted boxes denote diagnostic RFLP patterns, which are more than 20 bp different to other fragment produced
by that restriction enzyme.
89
7.3.3.2
PCR-RFLP TEST 2
A) PCR
Materials and equipment
As for PCR-RFLP Test 1 Section 7.3.3.1
Reagents
As for PCR-RFLP Test 1 Section 7.3.3.1 with the exception of primers, which are here:
 NS15 5’ CAATTGGGTGTAGCTACTAC 3’
 ITS6 5’ AGCCGAGTGATCCACCGCT 3’
Method
In a PCR (laminar flow or clean bench) cabinet:
1) label sterile 0.2 ml PCR tubes
2) make a Reagent Master Mix for the total number of samples to be analysed according to the
table below, adding the polymerase last (note, recommend adding an extra volume for more
than 10 samples to allow for any loss of solution via adherence to the outside of the pipette tip):
Reagent
Final concentration
Double-distilled H2O
30.6
Expand High Fidelity polymerase buffer
1X
5
dNTPs (2.5 μM)
200 μM
4
Forward primer 10 μM (NS16)
0.5 μM
2.5
Reverse primer 10 μM (ITS6)
0.5 μM
2.5
Expand Hi Fidelity Taq Polymerase
2 Units
0.4
Total
45
3) Store Master Mix on ice.
4) Vortex DNA extractions for 5 s.
5) Aliquot 5 μL of each unknown DNA template to labelled 0.2 mL tubes, plus at least one positive
control DNA and one negative control (water or the buffer in which the DNA is suspended).
6) Aliquot 45 μL of master mix to each 0.2 mL tubes (containing 5 μL of template DNA).
7) Mix product and reagents well (or vortex briefly) and centrifuge for 3-5 s.
8) Place samples in PCR machine and program the following temperature profile (based on
Armstrong and Cameron 1998):
•
Cycle 1
- denature 94°C 2 min
•
Cycles 2 to 35 - denature 94°C 15 s, anneal 60°C 30 s, extend 68°C 2 min
•
Cycle
- extend 72°C 5 min
9) Add 1 μL of loading dye to 5 μL of PCR product
10) Load onto a 1.5% agarose gel, together with a 100 bp molecular ladder at either side and
electrophorese according to Appendix 2. If product visible at 1.5-1.8 kb then proceed to Section
B) – restriction digestion.
90
B) RESTRICTION DIGESTION OF PCR PRODUCT
Equipment
See 7.3.3.1 B
Reagents
 Restriction enzymes, on ice.
o NB. Choose at least four of the enzymes listed in Appendix 3 according to which combination
will provide the best discrimination for the potential species; the letters represent RFLP
patterns in Appendix 4
o Note: In the specific circumstance of distinguishing only B. tryoni complex and C. capitata the
enzyme combination of AluI, DdeI and RsaI (10 U/μl) and SspI (5U/μl) is diagnostic.
 Restriction enzyme buffers, 10X
 Sterile nuclease free H2O
Method
1) Prepare master mix (following recipe below) for each enzyme; multiplying volumes plus one
for the number of reactions required.
Adapted from Armstrong & Cameron (1998)
Final concentration
Double-distilled H2O
5.6*
10X Buffer
1X
1.0
Enzyme†
4U
0.4
Total
7.0
* volume of water can be varied to accommodate any change to DNA volumes added, see below
† Standard stock concentrations are usually 10 U/µl; if not, this volume will change accordingly
2) Aliquot 7 µL of each master mix into labelled tubes
3) Add 2-3 μL (100-200 ng) PCR product into each tube depending on how strong or weak the
PCR products are; adjust water in reaction mix accordingly
4) Flick to mix reagents and PCR product, centrifuge briefly for 3-5 s.
5) Place samples in incubator at 37°C 2-3 h unless otherwise recommended by the enzyme
manufacturer
6) Prepare a 2-3% agarose gel according to Appendix 2, at least 10 cm in length, to visualise
fragment pattern and use a 100 bp ladder for determining fragment sizes
7) Compare results with positive controls and fragment patterns in Appendix 4
8) Species diagnosis is considered positive if all four enzyme patterns agree.
91
7.3.4
Real-time PCR detection of Bactrocera tryoni complex
This test was developed and validated by Dhami et al., (2015), further validation was conducted by
David Waite and Dongmei Li during 2014-2015 at the Plant Health and Environment Laboratory
(PHEL), Ministry for Primary Industries (MPI), New Zealand. This section (7.3.4) was written by
Dongmei Li, David Waite and Disna Gunawardana.
INTRODUCTION
Genetically similar but morphologically different species have been reported in several fruit fly species
(Clarke et al., 2005; Krosch et al., 2012). Such species are considered as belonging to “species
complexes” (Drew, 2004). The commonly called ‘Queensland fruit fly’ (Q-fly) is one such species
complex, consisting of four species; B. tryoni, B. aquilonis, B. neohumeralis and B. melas (Drew, 1989).
However, while B. melas exists in historic documents, researchers now consider it a cryptic form of B.
tryoni (Clarke et al., 2011).
Analysis of microsatellite markers has shown that there is only intraspecies level of differentiation
present between B. tryoni and B. aquilonis (Wang et al., 2003). Those analyses also reflect that B.
neohumeralis and B. tryoni represent separate species (Clarke et al., 2005; Wang et al., 2003). The
closely related species B. curvipennis, however, is difficult to distinguish from B. tryoni complex based
upon DNA sequencing of the cytochrome c oxidase subunit 1 (COI) gene (Armstrong & Ball, 2005),
although it is morphologically distinct. More comprehensive molecular analysis has similarly indicated
that B. curvipennis has a paraphyletic relationship with the B. tryoni complex and is currently considered
very close to, but not within, the B. tryoni complex sensu Drew 1989 (Blacket et al., 2012; Jiang et al.,
2014).
DNA barcoding relies on PCR of predetermined marker genes, DNA sequencing and comparison of
those sequences to a database of reference sequences (Armstrong & Ball, 2005). Currently, DNA
barcoding of the COI gene is the most commonly used technique for the molecular identification of fruit
flies (Armstrong & Ball, 2005; Blacket et al., 2012). This process is still relatively time consuming, with
the quickest possible identification requiring a full day. In contrast, real-time PCR, or quantitative PCR
(qPCR), drastically reduces the end-to-end time of analysis. This technique is based on the amplification
of DNA monitored in real time and therefore without the need of post-PCR processing (e.g. gel
electrophoresis), thus it enables identification within a few hours. The premise for this technology is that
highly species-specific PCR primers or probes are designed to give positive reactions only with DNA of
the target species. A positive identification is determined by the measurably lower number of PCR cycles
taken for the amplicon to reach a given concentration, and for fluorescence to be detected (the
quantitation cycle, Cq), than would occur for a negative or suboptimal amplification, as would be
expected from DNA of the wrong species. The TaqMan chemistry has the potential for greater specificity
through the incorporation of a third oligonucleotide in the reaction. The method has previously been
applied to other fruit fly species on the list in this manual (Burgher-MacLellan et al., 2009; Yu et al.,
2005; Yu et al., 2004).
Here, a real-time PCR assay for the B. tryoni complex has been developed, validated and applied by
PHEL in the routine diagnostics of intercepted fruit fly material at borders, post borders and in recent
New Zealand Q-fly responses.
AIM
This assay aims to provide a rapid method for the identification of the B. tryoni complex, from any life
stage, using the real-time TaqMan technique.
TARGET
This assay can be used identify B. tryoni complex, (B. tryoni, B. aquilonis, and B. neohumeralis), and
the closely related species B. curvipennis.
92
PROCEDURE
This document provides all the supporting information for conducting the real-time PCR assay and
analysis of the results, including:
1. DNA extraction and subsequent PCR from a range of life stages of the fruit flies including eggs,
part of an adult (one leg is commonly used in diagnostics), part of a larva, part of a pupa, empty
pupal case
2. PCR competency test
3. Real-time PCR assay for the B. tryoni complex (FAM probe)
4. Results - analysis of Real-time PCR assays.
7.3.4.1
DNA Extraction protocols
See section 7.3.1.
7.3.4.2
Real-time PCR assay for B. tryoni complex
Set up the PCR assay in a PCR workstation. The assay is described as run on the CFX1000 TM realtime PCR system (BioRad), with the data analysed in the CFX manager 3.0 analysis software
(BioRad).
Equipment and material
 Primers and Probe (Dhami et al., 2015)
o Forward primer: Btry2F, 5’-AATTGTAACAGCCCATGC-3’
o Reverse primer: Btry1R, 5’- GTGGGAATGCTATATCGG-3’
o Probe: Btry2PL: 6FAM-AG[+C]CA[+G]TTTCC[+G]AA[+A]CC-BBQ (or BHQ1)
 Real-time PCR mastermix
o SsoAdvanced Universal Probe Supermix (Cat#1725280, Biorad), 2x qPCR mix,
containing dNTPs, Sso7d fusion polymerase, MgCl2, stabilizers, ROX normalization
dyes.
o Primers/probe, working concentration of 5 µM (primer stock concentration 100 µM,
stored at -20°C in dark)
o BSA (Bovine serum albumin, working concentration of 10ng/µl) (Sigma Cat# A78850g)
 DNA samples of unknown Tephritidae species extracted as in section 7.3.1.
 Internal positive control to test unknown DNA samples for PCR competency to avoid false
negative results
o Conventional PCR with Folmer primers LCO1490 and HCO2198 (Folmer et al., 1994)
o TaqMan ribosomal RNA control reagents (VIC probe) (Applied Biosystems, cat #
4308329), see Appendix 1 for details.
 Controls for real-time PCR setup:
o Positive control to monitor the performance of the real-time PCR
 DNA samples of B. tryoni complex or
 Plasmid DNA of COI insert of B. tryoni complex (available from MPI PHEL,
NZ on request)
o Non-template control
 Sterile water or PCR-clean water
 PCR work station or DNA/RNA-free area for PCR setup
 Thermal cycler: CFX96 or CFX1000 Touch Real-time PCR (BioRad). Other brands of realtime thermal cycler can be used.
 Additional equipment: centrifuge, pipettes, plugged PCR tips, PCR tube, PCR plates, plate
seals.
Method
1. PCR competency test: It is recommended prior to any qPCR analyses to test all the DNA
extractions for PCR competency, using either conventional PCR or 18S internal control
TaqMan assay (see Appendix 1 for details)
93
2. Set up the real-time PCR Mastermix in a PCR work station or clean area, see below for the
compositions (Table 6). Include enough volume to run the samples in duplicate or triplicate
plus positive and non-template controls. It can also be run at 20 µl or 10 µl volume.
a. For 20 µl volume, aliquot 18 µl of master mix to each PCR tube
b. For 10 µl volume, aliquot 9 µl of master mix to each PCR tube
Table 6. TaqMan real-time PCR for B. tryoni complex protocol using SsoAdvanced Universal Probe Supermix
(BioRad)
Reagents
1 x reaction (µl)a
10x reactions (µl) b
Sterile distilled H2O
2.2
22
2 x Probe Supermix
10.0
100
5 µM Btry2F (400 nM)
1.6
16
5 µM Btry1R (400 nM)
1.6
16
5 µM Btry2PL(250 nM)
1.0
10
BSA (10mg/ml)
1.6
16
DNA template
2.0
Note: aThe compositions for 20 µl are listed in this table, halve the volumes for each reagent if using in 10 µl volume. bUsing 10x
reaction as an example, calculate the volumes of each reagents using the number of reactions you are going to test when
conducting the assay; note, one or two extra reaction volumes should be included to account for loss due to retention of liquid in
pipette tips.
3. Set up the Mastermix, mix well and spin briefly, aliquot 18 µl of the Mastermix to each labelled
PCR tube or well of a plate.
4. Add the appropriate DNA template or control solution to the tubes or the wells of a plate with
the aliquoted Mastermix, close the tubes or seal the plate, mix well and spin the tubes or plate
briefly
5. Put the tubes/plate into real-time PCR machine and run the program below:

PCR cycling parameter (CFX 1000TM Thermal cycler)
1 x cycle
95 °C, 2 min
40 x cycles
95 °C, 15 sec
65 °C, 60 sec
Plate read after each cycle
6. Open CFX manager 3.0 program, set up the protocol and run the real-time PCR assay.
7. Name the assay and save the data in a folder.
8. Once the run is finished, the data can be opened in the CFX manager 3.0 and analysed.
94
7.3.4.3
Analysis of Real-time PCR results
The real-time PCR results are analysed here with the CFX Manager 3.0 (BioRad). If different real-time
PCR systems are used, analysis of the amplification curves will need to be carried out according to
that manufacturers’ manual.
1. Open BioRad CFX Manager 3.0.
2. Click file, open the data file, and choose the real-time PCR data you have saved for the assay.
3. Click on quantification tab, the amplification curve will appear on the screen, see Figure 15 for
an example.
4. Set up the baseline for 50.
5. Report: Click Tools tab and choose reports, an analysis report for the real-time PCR including
amplification curves and Cq values will be generated. It can be saved as pdf format or print it
out.
6. Interpretation of the results - 18S internal control:
a. If Cq values ≤30 cycles, the DNA extraction is PCR competent and the samples can
be used for real-time PCR assay against B. tryoni complex.
b. If Cq values >30 cycles, this indicates that inhibitors are in the DNA sample or there is
insufficient DNA. Re-extraction of the samples will be needed.
7. Interpretation of the results - B. tryoni complex assay (use the PCR competent DNA samples
only):
a. For Cq values ≤30 cycles, the sample is considered as positive for B. tryoni complex,
and possible for closely related B. curvipennis (Cq values between 25-30 cycles)
i. DNA barcoding needs to be conducted to confirm the species if the Cq values
>25 cycles.
ii. The origin of the specimen might assist in distinguishing B. tryoni and B.
curvipennis (e.g. B. curvipennis is endemic to New Caledonia and not present
in Australia).
b. Samples with Cq values between 30-35 cycles could be interpreted as either a
different species with suboptimal match to the primers or that there is a very low copy
number of the target species DNA. These should be considered as questionable and
require further investigation by either DNA barcoding or re-extraction of DNA.
c. The negative threshold for the assay is Cq values >35, at which point the samples
should be considered as negative for B. tryoni complex, and identification by another
method is necessary.
8. Validation of the real-time PCR assay for B. tryoni complex used DNA extracted from fresh
and aged (up to 2 year olds) samples. This included B. tryoni from NSW, VIC, QLD (n>80)
and B. neohumeralis from QLD (n>20). The following results were observed;
a. Cq values ≤25 cycles were obtained with the DNA extracted from the B. tryoni
complex except those with empty pupal cases.
b. Cq values between 25-35 cycles were observed for the DNA extracted from B.
curvipennis samples. DNA barcode sequencing confirmed the identification.
c. Negative results (Cq >35) were obtained for all other Tephritidae species tested (see
Appendix 2).
95
Sample tested
B. tryoni
Positive control
Water control
Figure 15. Amplification curves of real-time PCR assay for B. tryoni complex. The sample tested positive for B.
tryoni complex, DNA sequences has further confirmed the results.
96
7.3.5
Allozyme electrophoresis
This section was written by Mark Adams.
AIM
Allozyme electrophoresis provides a method for the rapid molecular identification of various species of
fruit fly.
TARGETS
Routinely targets Bactrocera tryoni, Ceratitis capitata, Dirioxia pornia, Bactrocera dorsalis s.s. and
Bactrocera jarvisi (Table 3). Additional species can be incorporated where suitable reference
material is provided as freshly-frozen specimens (either as larvae or adults).
SUITABILITY
Suitable for the comparison of genetic profiles, as expressed in a range of soluble proteins, from
live, recently-dead, or freshly-frozen larvae or adults. The service is currently routinely provided
by the South Australia Museum's Evolutionary Biology Unit laboratory. The procedures take 2-3
hours to complete for a single screen of up to 20 specimens for 10 different genes.
Given the comparative nature of the technique and its continued reliance on reference samples, it is
important to note that additional species can only be identified as "new" (i.e. not one of the five
reference species) unless suitable, known-identity samples can also be provided for a putative
match. Moreover, the incorporation of additional species into the routine screening procedure may
also require a re-evaluation of which enzyme markers are diagnostic for these additional species, in
order to satisfy the minimum requirement of three diagnostic genetic differences between every pair
of species.
PROCEDURE OVERVIEW
Crude extracts of soluble protein from live, recently-dead, or freshly-frozen larvae or adults
are compared electrophoretically against known-identity extracts representing these five
species.
Test samples are readily identifiable by their comparative allozyme profile (i.e. relative band mobility)
at a suite of at least six enzyme markers, encoded by a minimum of 10 independent genes, and
together able to unambiguously diagnose the five reference species from one another at a minimum
of three genes.
Neither B. aquilonis nor B. neohumeralis are listed in the five target species, so this method is
not designed to differentiate between B. tryoni and these other two, genetically very similar
species.
7.3.5.1
Specimen preparation
Test specimens

Need to be supplied either (a) alive, (b) freshly dead and kept cool and moist, or (c) frozen
when alive and not allowed to thaw until tested (dry ice required for transport; ice is not
suitable)

Can represent any life history stage
97
Reference specimens

Frozen specimens representing the species requiring discrimination must be available. When
kept at -70oC, these remain suitable for use as controls for at least 10 years.

A single homogenized larva provides enough homogenate to act as a reference specimen on
up to eight separate occasions. Once prepared, these reference homogenates can be stored
at -20 oC as separate ~5 µL aliquots inside glass capillary tubes. Thus reference specimens
for any test run are usually available as pre-prepared homogenates straight from the freezer.
Specimen Preparation (ideally in cold room at 4oC)

Specimens are hand-homogenized in an equal volume of a simple homogenizing solution
(0.02 M Tris-HCl pH 7.4 containing 2 g PVP-40, 0.5 mL 2-mercaptoethanol and 20 mg NADP
per 100 mL).

~0.5 µL of homogenate loaded directly onto each gel.

The remaining homogenate is transferred as a series of ~5 µL aliquots into individual glass
capillary tubes and stored at -20oC. These samples can either be subjected to further
allozyme analysis if doubt remains as to species identity, or used as fresh reference material
for the species thus identified (activity declines over a 12 month period at -20oC).
7.3.5.2
Electrophoresis (ideally in cold room at 4oC)
Allozyme analyses are conducted on cellulose acetate gels (CellogelTM) according to the principles
and procedures of Richardson et al. (1986). Table 7 indicates the suite of enzymes most commonly
used for fruit-fly genetic identifications and details the electrophoretic conditions employed for each.
7.3.5.3
Gel interpretation
The interpretation of allozyme gels requires some expertise; the intensity of allozyme bands changes
over time after a gel is stained, plus banding patterns can be affected by the “freshness” of the
specimen and by what type of gut contents are present (some plants contain compounds which affect
fruit fly enzymes once the sample is homogenised). Richardson et al. (1986) devote an entire section
to the interpretation of allozyme gels, but there is no substitute for experience.
7.3.5.4
Recording of results
All gels are routinely scanned several times over the time course of stain incubation and the resultant
JPG files archived as a permanent record.
98
Table 7. Enzymes most commonly used for fruit-fly genetic identifications using allozyme electrophoresis.
Enzyme
Abbr
E.C.
No.
No. of
genes
Buffer1 Run time
Stain
Aconitase
hydratase
ACON
4.2.1.3
2
I17
Richardson
et al. (1986)
C. capitata vs B. tryoni vs
B. jarvisi vs B. dorsalis vs
D. pornia
Aminoacylase
ACYC
Manchenko
(1994)
C. capitata vs B. tryoni
vs B. jarvisi / B. dorsalis
vs D. pornia
Richardson
et al. (1986)
C. capitata vs B. tryoni vs
B. jarvisi vs D. pornia
method 3;
Manchenko
(1994)
C. capitata vs B. tryoni /
B. dorsalis vs B. jarvisi
vs D. pornia
Richardson
et al. (1986)
B. jarvisi / B. dorsalis vs
D. pornia
Richardson
et al. (1986)
B. dorsalis vs D. pornia
Richardson
et al. (1986)
B. jarvisi vs B. papaya vs
D. pornia
1.2 h
@ 250 V
3.5.1.14 1
C
1.5 h
@ 250 V
Alcohol
dehydrogenase
ADH
Aspartate
aminotransferase
GOT
Glycerol-3phosphate
dehyrogenase
GPD
Glucose-6phosphate
isomerase
GPI
3Hydroxybutyrate
dehydrogenase
HBDH
Isocitrate
dehydrogenase
IDH
Malate
dehydrogenase
MDH
Dipeptidase
PEPA
1.1.1.1
2
C
1.5 h
@ 250 V
2.6.1.1
2
B
1.5 h
@ 250 V
1.1.1.8
1
C
1.5 h
@ 250 V
5.3.1.9
1
B
1.5 h
@ 250 V
1.1.1.30 1
B
1.5 h
@ 250 V
1.1.1.42 2
B
1.5 h
@ 250 V
1.1.1.37 2
C
1.5 h
@ 250 V
3.4.13
2
B
1.3 h
@ 250 V
1Code
17
Species delineated
Richardson
et al. (1986)
B. tryoni vs B. dorsalis
Richardson
et al. (1986)
B. dorsalis / B. jarvisi vs
D. pornia
Richardson
et al. (1986)
B. jarvisi / B. dorsalis vs
D. pornia
for buffers follows Richardson et al. (1986).
Sample origin is placed in the centre of the gel
99
8
Diagnostic Information
The data sheets for all diagnostic methods are presented here. They are compiled for each species,
although not all methods are available for all species (Table 3).
Morphological data follow that of White & Elson Harris (1992) and Drew & Romig (2013).
Molecular data for the DNA barcoding method is not provided here and the user is guided to submit
their COI barcode sequence to the BOLD identification engine at
http://www.barcodinglife.com/index.php/IDS_OpenIdEngine.
Alternatively, an in-house reference dataset can be composed from the public data in BOLD (see
Table 3). Note: any qualifying statements made under the individual species sections to indicate
issues with diagnosis were accurate as of October 2015, but may have changed subsequently if
additional data has been submitted to BOLD.
Molecular data for the PCR-RFLP Test 1 provided under the individual species sections refers to
information in Table 5, and given as fragment lengths in bp or DNC (does not cut). Molecular data for
the PCR-RFLP Test 2 is provided in Appendices 3 and 4. PCR-RFLP patterns for unknown samples
should be compared within those reference datasets to that of any other species likely to have been
trapped (if known) or to any closely related species for which data may be available.
100
Key to family Tephritidae
The family Tephritidae can be separated from all other Diptera by the shape of the subcostal vein,
which bends abruptly through a right-angle and fades to a fold before reaching the wing edge,
combined with the presence of setulae (small setae) along the dorsal side of vein R1.
* Zeugodacus has been elevated from sub-generic rank (under genus Bactrocera) to full generic
status based on molecular genetic data (Virgilio et al. 2015). This is yet to be widely accepted by the
scientific community.
Simplified key to major pest fruit fly genera (after White and
Elson-Harris 1992): Adults
1
Vein Sc abruptly bent forward at nearly 90°, weakened beyond the bend and ending at
subcostal break; dorsal side of vein R1, with setulae. Wing usually patterned by coloured bands.
Wing cell cup with an acute extension…………………......................................TEPHRITIDAE .2
-
Vein Sc not abruptly bent forward, except in the Psilidae, which lack both dorsal setulae on vein
R1, and frontal setae. Species associated with fruit very rarely have any wing patterning. Wing
cell cup usually without an acute extension (exceptions include some Otitidae and
Pyrgotidae)…………………..……………………………………..Families other than Tephritidae
2
Cell cup very narrow and extension of cell cup very long. 1st flagellomere (3rd segment of
antenna) at least three times as long as broad. Wing pattern usually confined to a costal band
and an anal streak. (Tropical and warm temperate Old World; adventive species in Hawaii and
northern South America)……………….……………………BACTROCERA* and DACUS (p. 102)
-
Cell cup broader and the extension shorter. 1st flagellomere shorter. Wing pattern usually
includes some coloured crossbands……………………………………….…………………………..3
3
The wing vein that terminates just behind the wing apex (vein M) is curved forwards before
merging into the wing edge. Wing pattern usually similar to Figure 22. (South America, West
Indies and southern USA)…………………………..………….… ………….ANASTREPHA (p.
102)
-
The wing vein that terminates just behind the wing apex (vein M) meets the wing edge at
approximately a right angle. Wing pattern usually similar to Figure 107………………….............4
4
Cell cup, including its extension, shaped as Figure 107. Basal cells of wing usually with spotand fleck-shaped marks, giving a reticulate appearance. Scutellum convex and shiny. (Ceratitis
capitata is found in most tropical and warm temperate areas; other spp. are
African)…………….……………………………………..……..………………….CERATITIS (p. 244)
-
Cell cup, including its extension, shaped as Figure 32-Figure 116 Basal area of wing not
reticulate. Scutellum fairly flat and not shiny. (Larvae develop in the fruits of Berberidaceae,
Caprifoliaceae, Cornaceae, Cupressaceae, Elaeagnaceae, North temperate regions and South
America).……………………………………………………………………….RHAGOLETIS (p. 266)
101
Species diagnosis: Anastrepha (Tephritidae: Trypetinae:
Toxotrypanini)
8.3.1 Anastrepha distincta (Greene)
TAXONOMIC INFORMATION
Common Name: Inga fruit fly
Previous scientific name:
Anastrepha silvai Lima
DIAGNOSIS
8.3.1.1
Morphological – Adult
Anastrepha distincta is characterized by the presence of a dark brown spot medially over the
scutoscutellar suture (sometimes absent); the mediotergite yellow to orange-brown medially and dark
brown laterally; scutum entirely covered with microtrichia; wing with cell bc with microtrichia on anterior
margin; male aedeagus length 2.7-4.3 mm; female aculeus length 0.34-0.43 mm at tip; dorsobasal
scales on eversible membrane; large, hook-like and in a triangular pattern; wing pattern typical but
similar to that in Anastrepha edentata Stone.
8.3.1.2
Morphological – Larvae
See Pest Fruit Flies of the World – Larvae
Via delta-intkey.com/ffa Website
8.3.1.3
Molecular
DNA barcoding: BOLD reference data available, but cannot be distinguished from A. fraterculus, A.
ludens, A. obliqua, or A. suspensa.
PCR-RFLP Test 1: data not available
Allozyme eletrophoresis: data not available
HOST RANGE
Anastrepha distincta utilizes Inga species as its preferred host. Also recorded from species of
Chrysophyllum, Eugenia and Mangifera.
DISTRIBUTION
Distributed from Southern Mexico to Peru and Brazil
REMARKS
While Anastrepha distincta is a distinct species it can be confused with A. ludens and A. edentata.
PEST STATUS
 Exotic
 A serious pest of Inga species
102
ATTRACTANT
No known record
FIGURES
Figure 16. Anastrepha distincta
Image courtesy of the United States Department of Agriculture
103
Image courtesy of Carroll et al., Pest fruit flies of the world, http://delta-intkey.com/ffa (as of 22 November
2015)
104
105
8.3.2 Anastrepha fraterculus (Wiedemann)
Common name: South American fruit fly
Previous scientific names:
Dacus fraterculus
Trypeta fraterculus
Acrotoxa fraterculus
Trypeta (Acrotoxa) fraterculus
DIAGNOSIS
8.3.2.1
Morphological - Adult
Among all Anastrepha species found in the Americas, A. fraterculus, A. obliqua and A. suspensa
present the most difficult identification problems in the genus; these three species are likely to be
confused because of the similarity of their external features. Critical differences between A. fraterculus
and A. obliqua are:
A. A. frateculus:
a. Aculeus usually longer than the distance on vein M from the junction of MP and M to
vein r-m.
b. Subscutellum darkened laterally
B. A. obliqua:
a. Aculeus always shorter than the distance on vein M from the junction of MP and M to
vein r-m.
b. Subscutellum not darkened laterally.
The apical arm of the S band of A. fraterculus is narrow compared with that of A. suspensa. There is
frequently a distinct scutoscutellar black spot, but it is usually smaller than in A. suspensa. One of the
most important distinguishing features is the nature of the aculeus tip, which has serrations only on its
apical third in contrast to that of A. obliqua (Foote, Blanc and Norrbom 1993).
8.3.2.2
Morphological - Larvae
- Not available/included in this edition 8.3.2.3
Molecular
DNA barcoding: BOLD reference data available, but cannot be distinguished from A. distincta, A.
ludens, A. obliqua, or A. suspensa.
PCR-RFLP Test 2: No individual restriction enzymes are diagnostic, but a selection will differentiate A.
fraterculus from other Anastrepha and from species within other genera
106
HOST RANGE
Anastrepha fraterculus has been recorded on hosts from a wide range of families. These include:
Actinidiaceae, Anacardiaceae, Annonaceae, Combretaceae, Ebenaceae, Fabaceae, Juglandaceae,
Lauraceae, Lythraceae, Malvaceae, Moraceae, Myrtaceae, Oleaceae, Oxalidaceae, Rosaceae,
Rubiaceae, Rutaceae, Sapotaceae and Vitaceae (for a full list of recorded hosts see CABI 2007).
Major commercial hosts (UF & FDACS 2009; CABI 2007):
Scientific name
Common name
Scientific name
Common name
Annona cherimola
cherimoya
Prunus persica
peach
Citrus spp.
citrus
Psidium guajava
guava
Eugenia uniflora
Surinam cherry
Syzygium jambos
rose apple
DISTRIBUTION
Northern Mexico south to northern Argentina, Trinidad; introduced to Galapagos Is.; occasionally
trapped in USA (southern Texas), but not currently established (Carroll et al. 2002).
REMARKS
Anastrepha fraterculus is believed to belong to a group of closely related sibling species which, to
date, have not been identified and described. In addition, it is very close to A. obliqua and A.
suspensa. Consequently, A. fraterculus is difficult to diagnose and its exact area of distribution
uncertain. It is regarded as a species of major economic importance (pers. comm. Drew 2010).
PEST STATUS

Exotic

High priority pest identified in the Citrus IBP

Anastrepha fraterculus is an important pest of guavas and mangoes, and also to some extent
of Citrus spp. and Prunus spp.
ATTRACTANT
No known record, but can be captured in traps emitting ammonia.
107
FIGURES
Figure 21. Anastrepha fraterculus
Image courtesy of Carroll et al., Pest fruit flies of the world, http://delta-intkey.com/ffa (as of 22 August 2011)
Figure 22. Anastrepha fraterculus
108
8.3.3 Anastrepha grandis (Macquart)
Common name: South American cucurbit fruit fly
Tephritis grandis
Anastrepha latifasciata
Anastrepha schineri
DIAGNOSIS
8.3.3.1
Wing entirely infuscated from anterior margin to vein R4+5, a transverse band across r-m and curving
back to anal streak, infuscation around dm-cu; ovipositor eversible membrane with large hook-like
scales, apex of aculeus with 2 ridges, one on dorsal surface and the other on ventral surface.
8.3.3.2
See Pest Fruit Flies of the World - Larvae via delta-intkey.com/ffa Website
8.3.3.3
Molecular
DNA barcoding: diagnostic BOLD reference data available
PCR-RFLP Test 2: DdeI and Sau3A are fully diagnostic
DdeI:850, 270, 220,120; Sau3A: 770, 530, 110, 80
HOST RANGE
Attacks fruit of Cucurbitaceae and Myrtaceae
DISTRIBUTION
Venezuela to Brazil, Argentina
REMARKS
A distinct species based on wing patterns, not likely to pose a threat to Australia
PEST STATUS
 Exotic
 A pest of cucurbitaceae but cucurbit pest species of Tephritidae have a poor record for being
distributed round the world
ATTRACTANT
No known record
109
FIGURES
Figure 23. Anastrepha grandis female
Image courtesy of Carroll et al., Pest fruit flies of the world, http://delta-intkey.com/ffa (as of 14 October 2015)
Figure 24. Anastrepha grandis wing
110
Figure 25. Anastrepha grandis
Figure 26. Anastrepha grandis
111
8.3.4 Anastrepha ludens (Loew)
Common name: Mexican fruit fly
Trypeta ludens
Acrotoxa ludens
Trypeta (Acrotoxa) ludens
DIAGNOSIS
8.3.4.1
Anastrepha ludens is characterized by a relatively long aculeus and oviscape, the former 3.4 - 4.7mm
long and the latter correspondingly long and tapering in its apical third. This external character alone
will alert the identifier to the possibility of A. ludens. The apical third of the aculeus tip is slightly
expanded in the area of the lateral serrations, which are relatively few and not prominent. Anastrepha
suspensa and A. fraterculus differ in having a much shorter aculeus and aculeus tip with more
prominent lateral serrations and by other characters as well. Anastrepha ludens usually has a pair of
lateral dark spots on the subcutellum which typically extend ventrally onto the mediotergite. The V
band is usually not connected to the S band and is faint anteriorly in most specimens (Foote, Blanc
and Norrbom 1993).
8.3.4.2
Molecular
distincta, A. obliqua or A. suspensa.
PCR-RFLP Test 1: approximate ITS1 fragment length - gel: 650 bp
BsrI:
DNC
SnaBI: DNC
HhaI:
DNC
SspI:
DNC
HinfI:
550
Vspl:
550
Sau3AI: DNC
PCR-RFLP Test 2: There are no fully diagnostic restriction enzymes for A. ludens. HinfI will
distinguish the group A. ludens and A. suspensa, species which cannot be distinguished from each
other
Allozyme electrophoresis: data not available
HOST RANGE
Anastrepha ludens has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Caricaceae, Clusiaceae, Ebenaceae, Lauraceae, Lythraceae,
Myrtaceae, Passifloraceae, Rosaceae, Rubiaceae, Rutaceae and Sapotaceae (for a full list of
recorded hosts see CABI 2007).
112
Scientific name
Common name
Scientific name
Common name
Annona cherimola
cherimoya
Mangifera indica
mango
Citrus spp.
citrus
Prunus persica
peach
DISTRIBUTION
Texas, United States, south through Mexico to Costa Rica (Foote, Blanc and Norrbom 1993).
REMARKS
Anastrepha ludens is a well-defined and clearly distinct species, although there is a possibility of a
separate but nearly indistinguishable form in the extreme southern part of its distribution in Costa Rica
(UF & FDACS 2009).
PEST STATUS

Exotic

High priority pest identified in Citrus IBP

Anastrepha ludens is serious pest of Citrus spp. and mangoes
ATTRACTANT
113
FIGURES
Figure 27. Anastrepha ludens
Figure 28. Anastrepha ludens
114
8.3.5 Anastrepha obliqua (Macquart)
Common name: West Indian fruit fly
Tephritis obliqua
Anastrepha obliqua
DIAGNOSIS
8.3.5.1
Externally, Anastrepha obliqua quite closely resembles A. fraterculus and A. suspensa, thereby
presenting problems in their separation. However, a number of characters exist that appear to be
critical in separating A. obliqua from A. fraterculus. The aculeus is subtly different from those of A.
fraterculus and A. suspensa, having lateral serrations on more than two-thirds of the tip in contrast to
those of the other species, where they are limited to the apical two-fifths to three-fifths of the tip. In A.
obliqua, the tip also is relatively wider at the base of the serrations compared with the width at the
genital opening. The white medial vitta on the scutum is wider in A. obliqua than in A. suspensa and A.
fraterculus, and no scutoscutellar black spot or lateral dark marks on the subscutellum are present,
although the mediotergite usually has a lateral dark stripe (Foote, Blanc and Norrbom 1993).
8.3.5.2
- Not available / included in this edition 8.3.5.3
Molecular
distincta, A. ludens, or A. suspensa.
BsrI:
DNC
SnaBI: DNC
HhaI:
DNC
SspI:
150, 550
HinfI:
270, 450
Vspl:
550
Sau3AI: 200, 450
PCR-RFLP Test 2: HinfI, Sau3A and SspI are diagnostic
HinfI: 610, 270, 240,150,110, 80, 70; Sau3A: 770, 450, 200-170, 110; SspI: 1400,110
HOST RANGE
Anastrepha obliqua has been recorded on hosts from a range of families. These include:
Anacardiaceae, Ebenaceae, Malpighiaceae, Moraceae, Myrtaceae, Oxalidaceae, Passifloraceae,
Rosaceae, Rubiaceae, Rutaceae and Sapotaceae (for a full list of recorded hosts see CABI 2007).
115
Major commercial hosts (CABI 2007):
Scientific name
Common name
Scientific name
Common name
Mangifera indica
mango
Spondias purpurea
purple mombin
DISTRIBUTION
Throughout the greater and lesser Antilles, Jamaica, Trinidad, the Rio Grande Valley of Texas, Mexico
to Panama, Venezuela, Ecuador, and the vicinity of Rio de Janeiro, Brazil (UF & FDACS 2009).
REMARKS
Anastrepha obliqua, along with A. fraterculus and A. suspensa, is best recognised by the wing colour
pattern (Figure 30). It is one of the most widely distributed Anastrepha species, having been recorded
from Florida (USA), Southern and Central America and the West Indian islands. It is an important pest
of mangoes, guava, rose apple and Spondias (pers. comm. Drew 2010).
PEST STATUS

Exotic

Anastrepha obliqua is one of the most important fruit fly pests of mango in Central and
Southern America
ATTRACTANT
116
FIGURES
Figure 29. Anastrepha obliqua
Figure 30. Anastrepha obliqua
117
8.3.6 Anastrepha serpentina (Wiedemann)
Common name: Sapote fruit fly
Dacus serpentinus
Acrotoxa serpentinus
DIAGNOSIS
8.3.6.1
As in Anastrepha ocresia and a few non-U.S. Anastrepha species, the very dark wing markings of A.
serpentina contrast strongly with the light hyaline areas of the wing. The S band is quite slender and is
not connected to the proximal area of the V band, the apical arm of which is absent in all specimens.
Anastrepha serpentina and A. ocresia are the only species of Anastrepha occurring in the United
States that have a distinct pale yellow to hyaline area in cell r 1 immediately posterior to the
pterostigma, but the former may be distinguished from the latter by the complete absence of the distal
arm of the V band and the difference in abdominal markings. The scutum of the species is
characterised by contrasting light and dark markings; the subscutellum and mediotergite are very dark,
with a lighter brownish or yellowish spot or stripe dorsally (Foote et al. 1993).
8.3.6.2
Molecular
DNA barcoding: BOLD reference data available, but not well resolved amongst a number of other
species in BOLD. Low divergence clades may be apparent and used in conjunction with other
information about the specimen may still be sufficient to make a confident identification.
BsrI:
DNC
SnaBI: DNC
HhaI:
DNC
SspI:
DNC
HinfI:
DNC
Vspl:
250, 420
Sau3AI: 200, 530
PCR-RFLP Test 2: AluI and DdeI are fully diagnostic
AluI: 800-790, 340-330, 240, 120; DdeI: 830-800, 270, 220, 210
HOST RANGE
Anastrepha serpentina has been recorded on hosts from eight families. These include:
Anacardiaceae, Annonaceae, Clusiaceae, Lauraceae, Myrtaceae, Rosaceae, Rutaceae and
Sapotaceae (for a full list of recorded hosts see CABI 2007).
118
Scientific name
Common name
Scientific name
Common name
Chrysophyllum cainito
cainito
Manilkara zapota
sapodilla
Citrus spp.
citrus
DISTRIBUTION
Northern Mexico south to Peru and northern Argentina. Also known from Trinidad & Tobago and
Curaçao (Norrbom 2003).
REMARKS
Anastrepha serpentina is distinguished by its very dark wing patterns (Figure 32). It is most prevalent
in Mexico, Southern and Central America, as far south as Brazil. It has a wide host range but is not
considered to be of significant economic importance (pers. comm. Drew 2010).
PEST STATUS

Exotic

Not considered to be of significant economic importance
ATTRACTANT
119
FIGURES
Figure 31. Anastrepha serpentina
Figure 32. Anastrepha serpentina
120
8.3.7 Anastrepha striata (Schiner)
Common name: Guava fruit fly
DIAGNOSIS
8.3.7.1
A small to medium-sized species with a “normal” Anastrepha wing pattern, A. striata is one of the few
species occurring in the United States that has distinct dark scutal markings in addition to darkening
along the scutoscutellar suture. On the sublateral dark scutal areas, a pair of dense patches of short,
brownish black setae is present, as well as some hoary pile visible only when viewed from in front, but
the lateral half of the scutal brown stripe is denuded. Anastrepha striata is the only U.S. species
having such scutal characters. The aculeus tip is distinctly broad and wedge-shaped with a very blunt
apex and extremely fine lateral serrations. The size of the hyaline mark at the apex of vein R1 varies
considerably (Foote et al. 1993).
8.3.7.2
Molecular
PCR-RFLP Test 2: RsaI is diagnostic, but additional restriction enzymes are needed for a robust
identification
RsaI: 480-450, (370-360) x2, 290
HOST RANGE
Anastrepha striata has been recorded on hosts from a range of families. These include:
Anacardiaceae, Annonaceae, Combretaceae, Ebenaceae, Euphorbiaceae, Lauraceae, Myrtaceae,
Rosaceae, Rutaceae and Sapotaceae (for a full list of recorded hosts see CABI 2007).
Scientific name
Common name
Psidium guajava
guava
DISTRIBUTION
Southern Texas, Mexico, Central America, south to Peru, Bolivia and Brazil. Also found in Trinidad,
West Indies (UF & FDACS 2009).
REMARKS
Anastrepha striata is a smaller species of Anastrepha and best diagnosed by the distinct dark colour
markings on the scutum, composed of a U-shaped black pattern. It is present in Mexico, Central
America and most of Southern America. It is primarily a pest of guava (pers. comm. Drew 2010).
121
PEST STATUS

Exotic
ATTRACTANT
FIGURES
Figure 33. Anastrepha striata
Carroll et al., Pest fruit flies of the world, http://delta-intkey.com/ffa (as of 22 August 2011)
122
8.3.8 Anastrepha suspensa (Loew)
Common name: Caribbean fruit fly
Trypeta suspensa
Acrotoxa suspensa
Trypeta (Acrotoxa) suspensa
DIAGNOSIS
8.3.8.1
Anastrepha suspensa possesses external characters that closely resemble those of A. fraterculus and
A. obliqua; therefore, the separation of these three species is sometimes difficult. One of the most
obvious distinguishing marks in A. suspensa is the presence (except in some specimens from
Jamaica) of a dark spot at the junction of the scutum and scutellum. This spot is sometimes present in
A. fraterculus but is usually smaller, and it is absent in A. obliqua. The apical part of the S band in A.
suspensa is relatively wide compared with that in the other two species, and its inner margin is less
concave. It covers the apex of vein M or ends immediately anterior to it, whereas in the other two
species it normally ends well anterior to the apex of vein M, or its inner margin is strongly concave. As
in the identification of other species of Anastrepha, the shape of the aculeus tip is important. In A.
suspensa, as in A. fracterculus, the serrations occupy no more than three-fifths of the tip, whereas
those in A. obliqua occupy at least two-thirds; this character is variable and should be used with care
(Foote et al. 1993).
8.3.8.2
Molecular
DNA barcoding: BOLD reference data available, but cannot be distinguished from A. distincta, A.
fraterculus, A. ludens, or A. obliqua
PCR-RFLP Test 2: There are no fully diagnostic restriction enzymes for A. ludens. HinfI will
distinguish the group A. ludens and A. suspensa, species which cannot be distinguished from each
other
HOST RANGE
Anastrepha suspensa has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Arecaceae, Canellaceae, Caricaceae, Chrysobalanaceae, Clusiaceae,
Combretaceae, Curcurbitaceae, Ebenaceae, Lauraceae, Lythraceae, Malpighiaceae, Moraceae,
Myrtaceae, Oxalidaceae, Polygonaceae, Rosaceae, Rutaceae, Salicaceae, Sapindaceae, Sapotaceae
and Solanaceae (for a full list of recorded hosts see CABI 2007).
123
Major commercial hosts (CABI 2007; UF & FDACS 2009):
Scientific name
Common name
Scientific name
Common name
Annona reticulata
bullock's heart
Prunus persica
peach
Eugenia uniflora
Surinam cherry
Psidium guajava
guava
Fortunella margarita
oval kumquat
Syzygium jambos
rose apple
Manilkara zapota
sapodilla
Terminalia catappa
Singapore almond
DISTRIBUTION
Cuba, Jamaica, Hispaniola, Puerto Rico, southern Florida, (United States) (UF & FDACS 2009).
REMARKS
Along with Anastrepha obliqua and A. fraterculus, A. suspensa is very difficult to identify. These
species all have similar wing colour patterns and A. fraterculus is suspected of belonging to a complex
of closely related species. Generally, A. suspensa possesses a dark spot on the posterocentral area
of the scutum where it joins the scutellum. A. suspensa is distributed in Florida (USA), the Bahamas
and the West Indies, has a wide host range and is considered to be of major economic importance
(pers. comm. Drew 2010).
PEST STATUS

Exotic

Major economic importance
ATTRACTANT
FIGURES
Figure 34. Anastrepha suspensa
124
Figure 35. Anastrepha suspensa
125
Species diagnosis: Bactrocera & Dacus (Tephritidae: Dacinae:
Dacini)
8.4.1 Bactrocera (Bactrocera) albistrigata
(de Meijere)
Common name:
Dacus albistrigatus
Dacus (Bactrocera) albistrigata
DIAGNOSIS
8.4.1.1
A medium sized species; face fulvous with a pair of circular to oval black spots; postpronotal lobes
yellow (anteromedial corners black); notopleura yellow; scutum mostly black; lateral postsutural vittae
present; medial postsutural vitta absent; mesopleural stripe reaching to anterior npl. seta dorsally;
scutellum yellow with a broad black basal band; wing with a narrow fuscous costal band which is
extremely pale beyond extremity of cell sc to apex of wing, a narrow dark fuscous transverse band
across wing enclosing r-m and dm-cu crossveins, a broad fuscous to dark fuscous anal streak; cells bc
and c pale fuscous; microtrichia in outer corner of cell c only; abdominal terga III-V orange-brown with
a narrow to medium width medial longitudinal dark fuscous to black band over all three terga and
lateral dark markings which vary from narrow anterolateral dark fuscous to black corners on all three
terga to broad lateral longitudinal dark fuscous to black bands over all three terga; posterior lobe of
male surstylus short; female with aculeus tip needle shaped (Drew and Romig 2013). For potentially
morphologically confounding species see REMARKS or Table 8.
8.4.1.2
See White and Elson-Harris 1992 p. 176.
8.4.1.3
Molecular
PCR–RFLP Test 1: approximate ITS1 fragment length: 850 bp
Bsr1:
DNC
SnaB1: DNC
HhaI:
670, 180
SspI:
180, 620
HinfI:
DNC
Vspl:
DNC
Sau3a1: 400, 450
126
HOST RANGE
Bactrocera albistrigata has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Apocynaceae, Combretaceae, Moraceae, Myrtaceae and Verbenaceae (for a full list
of recorded hosts see Allwood et al. 1999).
Major commercial hosts (Allwood et al. 1999; Drew and Romig 2013):
Scientific name
Common name
Scientific name
Common name
Mangifera indica
mango
Syzygium malaccense
malay-apple
Syzygium aqueum
watery rose-apple
Syzygium samarangense
water apple
DISTRIBUTION
Andaman Islands, central to southern Thailand, Peninsular Malaysia, East Malaysia and Kalimantan
(Borneo), Singapore, Indonesia east to Sulawesi, Christmas Island (Drew and Romig 2013).
REMARKS
Bactrocera albistrigata belongs to the frauenfeldi complex described by Drew (1989). The other
species in the complex, B. caledoniensis, B. frauenfeldi, B. parafrauenfeldi and B. trilineola all possess
the same basic body and wing colour patterns. See Table 8 for characters separating these species.
127
Table 8. Bactrocera frauenfeldi complex and similar species (best diagnostic characters in bold).
B. frauenfeldi
B. albistrigata
B. trilineola
B.
parafrauenfeldi
B.
caledoniensis
B. kirki *
B. psidii*
exotic/
Australian/
pest
pest, present
QLD
pest, exotic (SE
Asia)
pest, exotic
(Vanuatu)
host not known,
present NT
no economic
hosts, exotic
(New
Caledonia)
pest, exotic
(Samoa
Tonga, Niue,
Tahiti)
pest, exotic
(New
Caledonia)
face
facial spots
facial spots
black mask
facial spots
no markings
facial spots
facial spots
lateral
vittae
very short
tapered or
medium
tapered or
moderately
broad & long
moderately
broad & long
absent
none or present
as short narrow
tawny coloured
spots
moderately
broad & long
absent
medium
length
tapered
vittae
ppn lobes
black or
fuscous or
partly to mostly
orangey yellow
mostly yellow
fuscous to
black
black
mostly yellow
mostly
yellow
mostly
yellow
scutellum
broad black
basal band or
triangle or
broad band to
apex
broad black
basal band
broad black
band to
apex
broad black band
to apex
broad black
basal band
about 1/3 of
scutellum
broad black
band to
apex
large black
triangle
legs
fuscous to
black hind
tibiae, mid &
hind femora
dark apically
hind tibiae
fuscous,
femora
generally
fulvous
tending redbrown apically
hind tibiae
fuscous,
hind femora
fuscous
apically
all tibiae fuscous,
mid and hind
femora black
apically
hind tibiae
fuscous
fuscous
tibiae,
femora
fulvous
all
segments
pale
abdomen
broad medial
line and 2
broad lateral
longitudinal
bands
narrow or
medium medial
line , darkened
lateral margins
varying from
narrow
elongate
antero-lateral
corners on T3T5 to 2 broad
lateral
longitudinal
bands
very broad
medial line
and 2 broad
lateral
longitudinal
bands
broad medial line
and 2 broad
lateral
longitudinal
bands
all terga
black except
for narrow
fuscous band
on T5
broad
medial line
and 2 broad
lateral
longitudinal
bands
all terga
black
except for
narrow
fuscous
band on T5
wing
transverse
band
transverse band
transverse
band
transverse band
transverse
band
tinge
around r-m
and dm-cu
cross veins
that may
appear to
be faint
band
tinge
around r-m
and dm-cu
cross veins
that may
appear to
be faint
band
*not frauenfeldi complex
128
The B. frauenfeldi complex can be separated from other members of the subgenus (Bactrocera
(Bactrocera)) by the presence of a dark crossband from the pterostigma (cell sc), which also
includes both the r-m and dm-cu crossvein. This runs roughly parallel to the anal stripe (diagonal
mark across wing base). However, the costal band is very pale and often not visible at all beyond
apex of R2 + 3.
PEST STATUS

Exotic

Medium level pest species
ATTRACTANT
Cue lure.
FIGURES
Figure 36. Bactrocera albistrigata
Images courtesy of Ken Walker, Museum Victoria, www.padil.gov.au (as of 22 August 2011)
129
Figure 37. Bactrocera albistrigata
Image courtesy of S. Phillips and the International Centre for the Management of Pest Fruit Flies, Griffith
University
130
8.4.2 Bactrocera (Bactrocera) aquilonis (May)
Common name: Northern Territory fruit fly
Strumeta aquilonis
Dacus (Bactrocera) aquilonis
DIAGNOSIS
8.4.2.1
Medium sized species; large black facial spots present; postpronotal lobes and notopleura yellow;
scutum pale red-brown with fuscous markings, mesopleural stripe reaching almost to anterior npl.
seta, lateral postsutural vittae present, medial postsutural vitta absent, scutellum yellow; wing with a
narrow fuscous costal band and broad fuscous anal streak, cells bc and c fuscous, microtrichia
covering cell c and most of cell bc; abdominal terga III-V pale orange-brown with pale fuscous along
anterior margin of tergum III and widening over lateral margins of that tergum, a medial longitudinal
pale fuscous band on terga III to V; posterior lobe of male surstylus short; female with aculeus tip
needle shaped (Drew 1989; pers. comm. Drew 2010).
8.4.2.2
8.4.2.3
Molecular
DNA barcoding: BOLD reference data available, but cannot be distinguished from B. neohumeralis or
B. tryoni
BsrI:
600, 200
SnaBI: DNC
HhaI:
650, 200
SspI:
570, 180
HinfI:
770
Vspl:
DNC
Sau3a1: 420
PCR-RFLP Test 2: SspI produces a B. tryoni species complex diagnostic restriction pattern; fragment
sizes (bp) are 1000, 550 and 100. This species cannot be distinguished from B. neohumeralis or B.
tryoni. Other restriction patterns useful for diagnosis of the complex are listed in Appendix 3, with
choice based on relative distinctiveness from other species potentially trapped; descriptions of the
patterns are given in Appendix 4. Cameron et al. (2010) found that there is no genetic evidence
supporting the separation of B. aquilonis and B. tryoni as distinct species.
131
HOST RANGE
Bactrocera aquilonis has been recorded on hosts from a wide range of families. These include:
Annonaceae, Arecaceae, Chrysobalanaceae, Combretaceae, Curcurbitaceae, Ebenaceae,
Elaeocarpaceae, Euphorbiaceae, Lauraceae, Meliaceae, Myrtaceae, Rosaceae, Rubiaceae,
Rutaceae, Santalaceae and Sapotaceae (for a full list of recorded hosts see Smith et al. 1988;
Hancock et al. 2000).
Major commercial hosts (CABI 2015; Hancock et al. 2000; pers. comm. Drew 2010):
Scientific name
Common name
Scientific name
Common name
Anacardium occidentale
cashew nut
Flacourtia rukam
rukam
Annona muricata
soursop
Fortunella x crassifolia
meiwa kumquat
Annona reticulata
bullock's heart
Lycopersicon
esculentum
tomato
Annona squamosa
sugarapple
Malpighia glabra
acerola
Averrhoa carambola
carambola
Malus domestica
apple
Blighia sapida
akee apple
Mangifera indica
mango
Capsicum annuum
bell pepper
Manilkara zapota
sapodilla
caimito
Prunus persica
peach
Citrus limon
lemon
Psidium guajava
guava
Citrus maxima
pummelo
Psidium littorale var.
longipes
strawberry guava
Citrus reticulata
mandarin
Spondias dulcis
otaheite apple
Citrus x paradisi
grapefruit
Syzygium aqueum
watery rose-apple
Eriobotrya japonica
loquat
Syzygium jambos
rose apple
Eugenia uniflora
Surinam cherry
Syzygium malaccense
malay-apple
Flacourtia jangomas
Indian plum
Ziziphus mauritiana
jujube
DISTRIBUTION
Northern areas of Western Australia and the Northern Territory (Hancock et al. 2000).
REMARKS
In the Northern Territory this species dramatically increased its host range during 1985. Since
B. aquilonis and B. tryoni will produce viable offspring when crossed in the laboratory (Drew and
Lambert 1986), hybridisation with B. tryoni was strongly suspected and might explain this increase
(Smith and Chin 1987; Smith et al. 1988). By 1997, most but not all commercial production areas and
larger towns supported populations of this fly, which attacks a wide range of cultivated hosts.
Therefore, many of the Northern Territory host records for B. aquilonis since March 1985 are attributed
to the suspected hybrid B. aquilonis x B. tryoni and are recorded under B. tryoni and not under B.
aquilonis in the above table (Hancock et al. 2000).
Bactrocera aquilonis and B. tryoni are very similar in general body and wing colour patterns.
Bactrocera aquilonis differs in being an overall paler colour with the scutum pale red-brown and the
abdominal terga generally fulvous without distinct fuscous markings. However, these differences are
not easily observed. Also these species can be separated on the differences on the ovipositors: apex
of aculeus rounded and spicules with 7-10 uniform dentations in B. tryoni compared with the more
pointed aculeus and uneven dentations in B. aquilonis (Drew 1989).
132
A recent molecular genetic study of northwestern Australian fruit fly populations (Cameron et al. 2010)
concluded that there is no genetic evidence supporting B. aquilonis as a distinct species from B.
tryoni. They conclude that the recent increase in host range of fruit flies in northwestern Australia is
due to local populations of B. tryoni (= B. aquilonis) utilising additional food resources from increased
agricultural production in this region.
Currently, B. aquilonis is regarded as belonging to a species complex including B. tryoni and
neohumeralis. Further research is currently being undertaken.
PEST STATUS

Endemic

Minor pest species
ATTRACTANT
Cue lure.
FIGURES
Figure 38. Bactrocera aquilonis
Image courtesy of the International Centre for the Management of Pest Fruit Flies, Griffith University
133
Figure 39. Bactrocera aquilonis
Image courtesy of M. Romig, International Centre for the Management of Pest Fruit Flies, Griffith University
134
8.4.3 Bactrocera (Paratridacus) atrisetosa
(Perkins)
Common name:
Zeugodacus atrisetosus
Melanodacus atrisetosus
DIAGNOSIS
8.4.3.1
Medium sized species; small fuscous facial spots present; postpronotal lobes and notopleura yellow;
scutum red-brown with irregularly shaped fuscous markings, mesopleural stripe reaching midway
between anterior margin of notopleuron and anterior npl. seta, lateral postsutural vittae beginning
anterior to mesonotal suture, medial postsutural vitta present, scutellum yellow; wing with a narrow
fuscous costal band and anal streak, cells bc and c pale fulvous with microtrichia in outer ½ of cell c
only; abdominal terga III-V orange-brown occasionally with fuscous on lateral margins of tergum III
and generally with narrow medial fuscous band on tergum V; posterior lobe of male surstylus long;
female with aculeus tip blunt trilobed (Drew 1989; pers. comm. Drew 2010).
8.4.3.2
Molecular
DNA barcoding: BOLD reference data not available
HOST RANGE
This species has been reared from eight host species in seven genera and three families, and is
mainly associated with cucurbits: watermelons, honeydew and rock melons, cucumbers, pumpkins,
zucchini, luffa and tomatoes (Leblanc et al. 2012).
Major commercial hosts (Drew 1989):
Scientific name
Common name
Scientific name
Common name
Citrullus lanatus
watermelon
Cucurbita pepo
ornamental gourd
Cucumis sativus
cucumber
Lycopersicon esculentum
tomato
DISTRIBUTION
Known only from mainland Papua New Guinea (LeBlanc et al. 2001)
135
REMARKS
Bactrocera atrisetosa is distinguished in having the costal band narrow (just overlapping R 2+3), scutum
red-brown, wings colourless, cells bc and c pale fulvous, abdominal terga III-V orange-brown except
for a narrow medial longitudinal fuscous band on tergum V and lateral margins of tergum III fuscous
(Drew 1989).
Bactrocera atrisetosa is very similar in appearance to the endemic B. cucumis but differs in having
prescutellar and supra-alar setae present. In common with B. cucumis it also lacks pecten.
PEST STATUS

Exotic

Medium level pest species
ATTRACTANT
No known record.
FIGURES
Figure 40. Bactrocera atrisetosa
Image courtesy of Mr. S. Wilson and the International Centre for the Management of Pest Fruit Flies, Griffith
University
136
Figure 41. Bactrocera atrisetosa
137
8.4.4 Bactrocera (Bactrocera) bryoniae (Tryon)
Common name:
Chaetodacus bryoniae
Strumeta bryoniae
Dacus (Strumeta) bryoniae
DIAGNOSIS
8.4.4.1
Large species; irregularly circular black facial spots present; postpronotal lobes and notopleura yellow;
scutum dull black, mesopleural stripe slightly wider than notopleuron, lateral postsutural vittae present,
medial postsutural vitta absent, scutellum yellow; wing with a broad fuscous costal band and anal
streak, cells bc and c fulvous, microtrichia covering outer ½ of cell c only; abdominal terga III-V
orange-brown with a medial and two lateral longitudinal dark bands joined along anterior margin of
tergum III; posterior lobe of male surstylus short; female with aculeus tip needle shaped (Drew 1989;
pers. comm. Drew 2010).
8.4.4.2
Molecular
BsrI:
600, 230
SnaBI: DNC
HhaI:
620, 200
SspI:
560, 180
HinfI:
760
Vspl:
DNC
Sau3A1: 400
HOST RANGE
Bactrocera bryoniae has been recorded on hosts from five families. These include: Curcurbitaceae,
Loganiaceae, Musaceae, Passifloraceae and Solanaceae (for a full list of recorded hosts see Hancock
et al. 2000).
Major commercial host (Drew 1989; Hancock et al. 2000):
Scientific name
Common name
Capsicum annuum
chilli
Infests wild species of Cucurbitaceae, Loganiaceae and Passifloraceae (Hancock et al. 2000).
Records from banana and capsicum (Drew 1989) were from Papua New Guinea, not Queensland.
138
DISTRIBUTION
Widespread and common all over Indonesia (Papua, formerly part of Irian Jaya), Papua New Guinea
(every province except Bougainville and Manus), and Australia (Northern Western Australia, Northern
Territory, east coast south to Sydney, New South Wales, and the Torres Strait Islands) (Drew and
Romig 2013).
REMARKS
There are a number of species in Southeast Asia and the South Pacific with broad costal bands.
However, Bactrocera bryoniae differs from these species in having costal band confluent with R4+5,
lateral postsutural vittae ending at upper pa. seta, abdominal terga III-V red-brown with a broad, dark
fuscous band along anterior margin of tergum III and covering lateral margins, anterolateral corners of
terga IV and V fuscous and a medial longitudinal dark fuscous band over all 3 terga (Drew 1989).
PEST STATUS

Endemic

Low level pest species in Queensland but not in Western Australia or the Northern Territory
ATTRACTANT
Cue group. In north Qld this species was significantly more attracted to raspberry ketone formate
(Melolure) than cue-lure (Royer 2015).
FIGURES
Figure 42. Bactrocera bryoniae
Image courtesy of S. Wilson, the International Centre for the Management of Pest Fruit Flies, Griffith University
and the Queensland Museum
139
Figure 43. Bactrocera bryoniae
140
8.4.5 Bactrocera (Bactrocera) carambolae
(Drew and Hancock)
Common name: Carambola fly
DIAGNOSIS
8.4.5.1
Face fulvous with a pair of medium sized oval black spots; scutum dull black with brown behind lateral
postsutural vittae, around mesonotal suture and inside postpronotal lobes; postpronotal lobes and
notopleura yellow; mesopleural stripe reaching midway between anterior margin of notopleuron and
anterior npl. seta dorsally; two broad parallel sided lateral postsutural vittae ending at or behind ia.
seta; medial postsutural vitta absent; scutellum yellow; legs with femora fulvous with a large elongate
oval dark fuscous to black preapical spot on outer surfaces of fore femora in some specimens, tibiae
dark fuscous (except mid tibiae paler apically); wings with cells bc and c colourless, microtrichia in
outer corner of cell c only, a narrow fuscous costal band slightly overlapping R2+3 and expanding
slightly beyond apex of R2+3 across apex of R4+5, a narrow fuscous anal streak; supernumerary lobe of
medium development; abdominal terga III-V orange-brown with a ‘T’ pattern consisting of a narrow
transverse black band across anterior margin of tergum III and widening to cover lateral margins, a
medium width medial longitudinal black band over all three terga, anterolateral corners of terga IV dark
fuscous to black and rectangular in shape and anterolateral corners of tergum V dark fuscous, a pair
of oval orange-brown shining spots on tergum V; abdominal sterna dark coloured; posterial lobe of
male surstylus short; female with aculeus tip needle shaped (Drew and Romig 2013).
8.4.5.2
8.4.5.3
Molecular
BsrI:
650, 250
SnaBI: 350, 530
HhaI:
680, 200
SspI:
DNC
HinfI:
DNC
Vspl:
355, 485
Sau3A1: 400, 450
PCR-RFLP Test 2: All restriction enzymes listed in Appendix 3, except SspI, distinguish B.
carambolae from others in the B. dorsalis species complex. Choice of enzyme will be based on
relative distinctiveness from other species potentially trapped; descriptions of the patterns are given in
Appendix 4.
141
HOST RANGE
Bactrocera carambolae has been recorded on hosts from a wide range of families. These include:
Alangiaceae, Anacardiaceae, Annonaceae, Apocynaceae, Arecaceae, Clusiaceae, Combretaceae,
Euphorbiaceae, Lauraceae, Loganiaceae, Meliaceae, Moraceae, Myristicaceae, Myrtaceae,
Oleaceae, Oxalidaceae, Polygalaceae, Punicaceae, Rhamnaceae, Rhizophoraceae, Rutaceae,
Sapindaceae, Sapotaceae, Simaroubaceae, Solanaceae and Symplocaceae (for a full list of recorded
species see Allwood et al. 1999).
Major commercial hosts (Allwood et al. 1999):
Scientific name
Common name
Scientific name
Common name
Averrhoa carambola
carambola
Syzygium jambos
rose apple
Manilkara zapota
sapodilla
Syzygium malaccense
malacca apple
Psidium guajava
guava
wax apple
Syzygium aqueum
watery rose-apple
DISTRIBUTION
Southern Thailand, Southern Vietnam, Peninsular Malaysia, East Malaysia, Kalimantan (Borneo),
Singapore, Indonesian islands east to Sumbawa, Andaman Islands, Surinam, French Guiana,
Guyana, Brazil (Drew and Romig 2013).
REMARKS
Bactrocera carambolae is similar to B. propinqua and some specimens of B. dorsalis in possessing
broad parallel sided or subparallel lateral postsutural vittae, costal band slightly overlapping R 2+3,
abdominal terga III-V with narrow to medium width dark lateral margins, shining spots on abdominal
tergum V pale (orange-brown to fuscous), femora entirely fulvous or with, at most, subapical dark
spots on fore femora only, in addition to the general characteristics of the dorsalis complex.
It differs from B. dorsalis in having a broad medial longitudinal black band on abdominal terga III-V, a
broader costal band apically, and shorter male aculeus and female ovipositor and from B. propinqua in
having a narrower medial longitudinal black band on abdominal terga III-V (in B. propinqua this band is
very broad) and apex of the aculeus needle shaped (in B. propinqua the apex of the aculeus is
trilobed).
PEST STATUS

Exotic

High priority pest identified in the Tropical Fruit Industry Biosecurity Plan (IBP; Plant Health
Australia)

This species is a major economic pest throughout the region where it occurs
ATTRACTANT
Methyl eugenol.
142
FIGURES
Figure 44. Bactrocera carambolae
Figure 45. Bactrocera carambolae
Image courtesy of M. Romig, the International Centre for the Management of Pest Fruit Flies,
Griffith University
143
8.4.6 Bactrocera (Bactrocera) caryeae (Kapoor)
Common name:
Dacus (Strumeta) caryeae
Dacus (Bactrocera) caryeae
DIAGNOSIS
8.4.6.1
Face fulvous with a pair of large elongate oval black spots; scutum black with a small area of dark
brown posterolateral to lateral postsutural vittae; postpronotal lobes yellow (except anterodorsal
corners fuscous); notopleura yellow; mesopleural stripe reaching midway between anterior margin of
notopleuron and anterior npl. seta dorsally; two narrow lateral postsutural vittae which are either
parallel sided or narrowing slightly posteriorly to end at or just before ia. seta; medial postsutural vitta
absent; scutellum yellow with a broad black basal band; legs with femora fulvous with large dark
fuscous to black preapical spots on outer surfaces of fore femora and inner surfaces of mid and hind
femora, fore tibiae fuscous, mid tibiae fulvous, hind tibiae dark fuscous; wings with cells bc and c
colourless, sparse microtrichia in outer corner of cell c only, a very narrow fuscous costal band
confluent with R2+3 and remaining very narrow around apex of wing, a narrow fuscous anal streak
contained within cell cup; supernumerary lobe of medium development; abdominal terga III-V orangebrown with dark fuscous to black across anterior 1/3 to 1/2 of tergum III, two broad lateral longitudinal
dark fuscous to black bands and a narrow medial longitudinal black band over all three terga, a pair of
oval orange-brown shining spots on tergum V; abdominal sterna dark coloured; posterial lobe of male
surstylus short; female with aculeus tip needle shaped (Drew and Romig 2013).
8.4.6.2
Molecular
DNA barcoding: BOLD reference data available. Diagnostic to the level of the B. dorsalis species
complex, but difficult to resolve amongst several other species within the complex. See Table 3.
144
HOST RANGE
Bactrocera caryeae has been recorded on hosts from six families. These include: Anacardiaceae,
Lecythidaceae, Malpighiaceae, Myrtaceae, Rutaceae and Sapotaceae (for a full list of recorded
species see Allwood et al. 1999).
Scientific name
Common name
Scientific name
Common name
Citrus maxima
pummelo
Mangifera indica
mango
Citrus reticulata
mandarin
Psidium guajava
guava
DISTRIBUTION
Southern India (Drew and Romig 2013).
REMARKS
Bactrocera caryeae is similar to B. kandiensis and B. arecae in possessing narrow parallel sided
lateral postsutural vittae, preapical dark markings on at least one pair of femora in addition to the
general characteristics of the dorsalis complex. It differs from B. arecae in possessing preapical dark
markings on all femora (in B. arecae the preapical dark markings are on fore femora only) and from B.
kandiensis in possessing a broad medial longitudinal dark band and broad lateral longitudinal dark
bands over abdominal terga III-V (Drew and Romig 2013).
It may appear similar to the Australian ME-responsive species B. endiandrae (see diagnostician’s
notes).
PEST STATUS

Exotic

Bactrocera caryeae occurs in very large populations in many fruit growing areas of southern
India and is probably responsible for much of the damage generally attributed to Bactrocera
dorsalis
ATTRACTANT
Methyl eugenol.
145
FIGURES
Figure 46. Bactrocera caryeae
146
8.4.7 Bactrocera (Bactrocera) correcta (Bezzi)
Common name: Guava fruit fly
Chaetodacus correctus
Dacus (Strumeta) correctus
DIAGNOSIS
8.4.7.1
Face fulvous with a pair of transverse elongate black spots almost meeting in centre; scutum black
with dark red-brown along lateral and posterior margins; postpronotal lobes and notopleura yellow;
mesopleural stripe reaching almost to anterior npl. seta dorsally; broad parallel sided lateral
postsutural vittae ending behind ia. seta; medial postsutural vitta absent; scutellum yellow with narrow
black basal band; legs with all segments entirely fulvous except hind tibiae pale fuscous; wings with
cells bc and c colourless, both cells entirely devoid of microtrichia, a narrow pale fuscous costal band
confluent with R2+3 and ending at apex of this vein, a small oval fuscous spot across apex of R4+5, anal
streak absent but with a pale fuscous tint within cell cup; supernumerary lobe of medium development;
abdominal terga III-V red-brown with a ‘T’ pattern consisting of a narrow transverse black band across
anterior margin of tergum III and a narrow medial longitudinal black band over all three terga, narrow
black anterolateral corners on terga IV and V, a pair of oval red-brown shining spots on tergum V;
posterior lobe of male surstylus short; female with aculeus tip needle shaped (Drew and Romig 2013).
8.4.7.2
Molecular
147
HOST RANGE
Bactrocera correcta has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Apocynaceae, Arecaceae, Cactaceae, Capparaceae, Caricaceae,
Combretaceae, Curcurbitaceae, Dipterocarpaceae, Elaeocarpaceae, Euphorbiaceae, Flacourtiaceae,
Lecythidaceae, Malpighiaceae, Meliaceae, Moraceae, Musaceae, Myristicaceae, Myrtaceae,
Olacaceae, Oxalidaceae, Rhamnaceae, Rosaceae, Rutaceae, Sapindaceae, Sapotaceae and
Simaroubaceae (for a full list of recorded hosts see Allwood et al. 1999).
Major commercial hosts (Allwood et al. 1999; Drew and Romig 2013):
Scientific name
Common name
Scientific name
Common name
cashew nut
Psidium guajava
guava
Mangifera indica
mango
water apple
Manilkara zapota
sapodilla
Terminalia catappa
Singapore almond
Mimusops elengi
Spanish cherry
Ziziphus jujuba
common jujube
Muntingia calabura
Jamaican cherry
DISTRIBUTION
Sri Lanka, India, Nepal, Pakistan, Myanmar, northern Thailand, southern China, Bhutan, Peninsular
Malaysia, Vietnam (Drew and Romig 2013).
REMARKS
Bactrocera correcta is similar to B. dorsalis in the general colour patterns of the body, wings and legs
but differs from B. dorsalis in possessing transverse facial spots and an incomplete costal band. It is
also similar to B. penecorrecta in the general colour patterns of the body and wings but differs from
this species in having abdominal terga III-V mostly pale coloured (not mostly black as in B.
penecorrecta) and the scutellum with a narrow black basal band (Drew and Romig 2013).
PEST STATUS

Exotic

Major pest species, particularly in Vietnam
ATTRACTANT
Methyl eugenol.
148
FIGURES
Figure 47. Bactrocera correcta
University
149
8.4.8 Bactrocera (Austrodacus) cucumis (French)
Common name: Cucumber fruit fly
Dacus tryoni var. cucumis
Dacus cucumis
Austrodacus cucumis
Dacus (Austrodacus) cucumis
DIAGNOSIS
8.4.8.1
Medium sized species; small fuscous to black facial spots present; postpronotal lobes and notopleura
yellow; scutum orange-brown without dark markings, mesopleural stripe reaching almost to anterior
npl. seta, lateral postsutural vittae beginning anterior to mesonotal suture, broad medial postsutural
vitta present, scutellum yellow; wing with a narrow fuscous costal band and anal streak, cells bc and c
pale fulvous (cell c slightly paler than cell bc), microtrichia in outer corner of cell c only; abdominal
terga I and II orange-brown, terga III-V fulvous except for two broad lateral longitudinal orange-brown
bands over all three terga and a narrow medial longitudinal band which is orange-brown on tergum III
and orange-brown to dark fuscous on tergum IV and V (this band is broader on tergum V); posterior
lobe of male surstylus long; female with aculeus tip blunt trilobed (Drew 1989; pers. comm. Drew
2010).
8.4.8.2
8.4.8.3
Molecular
BsrI:
DNC
SnaBI: DNC
HhaI:
550, 180
SspI:
DNC
HinfI:
DNC
Vspl:
DNC
Sau3AI DNC
PCR-RFLP Test 2: DraI, RsaI and SspI produce diagnostic restriction patterns (Appendix 3 and 4);
DraI: 1250, 200, 120; RsaI: 570, 410, 300, 290; SspI: 880, 650
HOST RANGE
While cucurbits are the major hosts of this species, it has been reared at moderate to high levels from
several other species in different plant families, including pawpaw and tomato (Hancock et al. 2000).
The rare or incidental hosts (usually a single rearing) include mango, avocado, guava, carambola,
apricot, some species of citrus, and capsicum. B. cucumis attacks wild cucurbits such as Diplocyclos
palmatus and these may be reservoir hosts (CABI 2007).
150
Bactrocera cucumis has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Caricaceae, Combretaceae, Curcurbitaceae, Ebenaceae, Euphorbiaceae, Lauraceae,
Myrtaceae, Oxalidaceae, Passifloraceae, Rosaceae, Rubiaceae, Rutaceae, Solanaceae and Vitaceae
(for a full list of recorded hosts see Hancock et al. 2000).
Major commercial hosts (Drew, 1989; Hancock et al. 2000):
Scientific name
Common name
Scientific name
Common name
Carica papaya
papaw
Passiflora edulis
passionfruit
Cucumis sativus
cucumber
Solanum lycopersicum
tomato
Cucurbita moschata
pumpkin
Trichosanthes anguinea
guada bean
Cucurbita pepo
squash and zucchini
DISTRIBUTION
Eastern Queensland appears to be the home range with Rockhampton as the centre of the range. It
has been trapped very occasionally in New south Wales however these are likely to be transient
populations or incursions that do not establish (Dominiak and Senior, in prep.). Hancock et al. (2000)
list it as present in the Northern Territory and Torres Strait Islands but its presence there cannot be
proven (pers. comm. Drew 2011). Although morphologically indistinguishable from Queensland
specimens (Drew pers. comm. 2011), the Northern Territory strain does not infest commercial crops
and in laboratory culture, failed to develop on undamaged cucurbit, solanaceous or other commercial
hosts, but could be reared on sliced cucumber (Smith and Chin 1987). Vargas et al. 2015 place the
species in Northern Territory and Queensland but the population is not continuous along the Gulf of
Carpentaria.
REMARKS
Bactrocera cucumis is a pale orange-brown species with medial and lateral postsutural vittae present,
a yellow scutellum, prsc. and sa. setae absent, 4 sc. seta present and a small elongate-oval black spot
centrally on tergum V (Drew 1989). It appears similar to the exotic B. atrisetosa from PNG but lacks
supra-alar and pre-scutellar bristles.
Other remarks:
In common with most species in, or close to, subgenus Zeugodacus, the scutum has three yellow
vittae (lateral and medial stripes), four setae on the margin of the scutellum, and the males lack a
deep V-shaped notch in posterior margin of 5th sternite. This species is unusual in that it also lacks
both anterior supra-alar setae and prescutellar acrostichal setae, and the males lack a pecten (comb
of setae on each postero-lateral corner of tergite 3) (CABI 2007).
PEST STATUS

Endemic

Major pest species in Queensland. Regarded as a potential pest of fruit in the National
Tropical Fruit IBP (page 26)
ATTRACTANT
No known attraction to male lures, although occasionally trapped at cue lure. It can be trapped using
cucumber volatile blend or orange-ammonia in McPhail traps (Royer et al. 2014).
151
FIGURES
Figure 48. Bactrocera cucumis
Image courtesy of Mr. S. Wilson, the International Centre for the Management of Pest Fruit Flies, Griffith
University and Queensland Museum
Figure 49. Bactrocera cucumis
152
8.4.9 Bactrocera (Zeugodacus) cucurbitae
(Coquillett)
Common name: Melon fly
Dacus cucurbitae
Chaetodacus cucurbitae
Strumeta cucurbitae
Dacus (Strumeta) cucurbitae
Dacus (Zeugodacus) cucurbitae
DIAGNOSIS
8.4.9.1
Medium sized species; large black facial spots present; postpronotal lobes and notopleura yellow;
scutum red-brown with or without fuscous markings, mesopleural stripe reaching midway between
anterior margin of notopleuron and anterior npl. seta, lateral postsutural vittae beginning anterior to
mesonotal suture, narrow medial postsutural vitta present, scuttelum yellow; wing with a broad fuscous
costal band expanding into a fuscous spot at wing apex, a broad fuscous anal streak, dark fuscous
along dm-cu crossvein, pale infuscation along r-m crossvein, cells bc and c colourless, microtrichia in
outer corner of cell c only; abdominal terga III-V orange-brown except for a narrow transverse black
band across anterior margin of tergum III which expands over anterolateral corners, a narrow medial
longitudinal dark fuscous to black band over all three terga and anterolateral corners of terga IV and V
fuscous; posterior lobe of male surstylus long; female with aculeus needle shaped (Drew 1989; Drew
and Romig 2013).
8.4.9.2
8.4.9.3
Molecular
BsrI:
DNC
SnaBI: DNC
HhaI:
400, 180
SspI:
DNC
HinfI:
DNC
Vspl:
DNC
Sau3AI: DNC
PCR-RFLP Test 2: RsaI produces a highly diagnostic restriction pattern; other useful enzymes are
AluI, CfoI, DraI, HinfI, Sau3A, SspI (Appendix 3 and 4)
RsaI: 410, 400, 310, 290
153
HOST RANGE
Bactrocera cucurbitae is primarily a pest of Cucurbitaceae, however it has also been recorded from
eleven other families. These include: Agavaceae, Capparaceae, Fabaceae, Malvaceae, Moraceae,
Myrtaceae, Rhamnaceae, Rutaceae, Sapotaceae, Solanaceae and Vitaceae (for a full list of recorded
hosts see Allwood et al. 1999; Leblanc et al. 2012).
Major commercial hosts (Drew 1989, Allwood et al. 1999):
Scientific name
Common name
Scientific name
Common name
Coccinia grandis
ivy gourd
Cucurbita pepo
ornamental gourd
Cucumis melo
melon
Momordica charantia
Trichosanthes cucumerina var.
anguinea
Cucumis satinus
Cucurbita maxima
snakegourd
giant pumpkin
DISTRIBUTION
Widely distributed over Southeast Asia, the Indian subcontinent, southern China, northern Africa and
Papua New Guinea. Introduced into the Mariana Islands, the Hawaiian Islands and from Papua New
Guinea to the Solomon Islands. Present in Indonesia and East Timor.
REMARKS
Bactrocera cucurbitae is similar to B. emittens in possessing only a slight widening of the costal band
in wing apex, a narrow infuscation along dm-cu crossvein and abdominal terga with ground colour
fulvous but differs in having the spot on apex of costal band not reaching M, cells bc and c colourless,
abdominal tergum III with a narrow transverse black band across base and tip of piercer of ovipositor
needle shaped. The most distinctive characteristic of the adult is the wing pattern (Drew 1989).
B. cucurbitae can appear similar to the endemic B. chorista widespread in eastern Qld. (see
diagnostician’s notes)
PEST STATUS

Exotic

High priority pest identified in the Tropical fruit and Vegetables IBPs

Bactrocera cucurbitae is a very serious pest of cucurbit crops
ATTRACTANT
Cue-group. Melolure (raspberry ketone formate) was found to be a stronger attractant than cue-lure
(Casana-Giner et al. 2003, Jang et al. 2007).
154
FIGURES
Figure 50. Bactrocera cucurbitae
Image courtesy of Ken Walker, Museum Victoria, www.padil.gov.au (as of 22 August 2011)
155
156
8.4.10 Bactrocera (Bactrocera) curvipennis
(Froggatt)
Common name:
Dacus curvipennis
Strumeta curvipennis
Dacus (Strumeta) curvipennis
DIAGNOSIS
8.4.10.1
Small species; very small pale fuscous facial spots present; postpronotal lobes and notoluera yellow;
scutum black, mesopleural stripe reaching midway between anterior margin of notoplueron and
anterior npl. seta, lateral postsutural vittae present, medial postsutural vitta absent, scutellum yellow;
wing with a broad fuscous costal band and anal streak, a broad fuscous band along r-m crossvein,
cells bc and c pale fuscous, microtrichia covering cell c and outer corner of cell bc; abdominal terga IIIV orange-brown with a narrow transverse fuscous band along anterior margin of tergum III merging
into broad lateral black margins and with anterolateral corners of terga IV and V fuscous; posterior
lobe of male surstylus short; female with aculeus tip needle shaped (Drew 1989; pers. comm. Drew
2010).
8.4.10.2
Molecular
BsrI:
570, 250
SnaBI: DNC
HhaI:
620, 170
SspI:
550, 200
HinfI:
DNC
Vspl:
DNC
Sau3AI: 420
PCR-RFLP Test 2: RsaI and SspI produce diagnostic restriction patterns (Appendix 3 and 4);
RsaI: 490, 460, 410, 290; SspI: 1020, 580, 100
157
HOST RANGE
Bactrocera curvipennis has been recorded on hosts from 20 families (for a full list of recorded hosts
see Leblanc et al. 2012).
Major commercial hosts (Drew 1989; Leblanc et al. 2012):
Scientific name
Common name
Scientific name
Common name
Citrus reticulata
mandarin
Mangifera indica
mango
DISTRIBUTION
New Caledonia and one remote island in Vanuatu (Drew 1989).
REMARKS
Bactrocera curvipennis is distinct in having the mesopleural stripe not extending to the postpronotal
lobes, microtrichia covering cell c and outer corner of cell bc, and abdominal terga III-V orange-brown
with a very narrow transverse fuscous band across anterior margin of tergum III which merges into
broad lateral black margins and the anterolateral corners of terga IV and V fuscous (Drew 1989).
PEST STATUS

Exotic
ATTRACTANT
Cue lure, Willison's lure.
FIGURES
Figure 53. Bactrocera curvipennis
Image courtesy of Mr. S. Wilson and the International Centre for the Management of Pest Fruit Flies, Griffith
University
158
Figure 54. Bactrocera curvipennis
159
8.4.11 Bactrocera (Paradacus) decipiens (Drew)
Common name: Pumpkin fruit fly
Dacus (Paradacus) decipiens
DIAGNOSIS
8.4.11.1
Large species; medium sized fuscous to black facial spots present; postpronotal lobes and notopluera
yellow; scutum red-brown with two broad lateral longitudinal fuscous bands, mesoplural stripe
reaching almost to anterior npl. seta, lateral postsutural vittae beginning anterior to mesonotal suture,
broad medial postsutural vitta present, scutellum yellow: wing with a broad fucsous costal band and
anal streak, an irregular recurved pale fuscous marking across wing, cells bc and c extremely pale
fuscous (cell c paler in centre), microtrichia in outer 1/3 of cell c only; abdominal terga I-V fulvous
except for broad lateral fuscous margins on tergum I and a narrow medial longitudinal fuscous band
on tergum V; posterior lobe of male surstylus long; female with aculeus tip trilobed (Drew 1989; pers.
comm. Drew 2010).
8.4.11.2
- Not available / included in this edition 8.4.11.3
Molecular
Allozyme: data not available
HOST RANGE
Pumpkin (Cucurbita pepo) is the only recorded host.
Scientific name
Common name
Cucurbita pepo
pumpkin
DISTRIBUTION
Papua New Guinea (New Britain) (Drew 1989).
REMARKS
Bactrocera decipiens is similar to B. perplexa in possessing infuscation on wings in addition to costal
band and anal streak but differs in having an irregular S-shaped pale fuscous marking across wing,
microtrichia in outer 1/3 of cell c only, mesoplueral stripe not extending to postpronotal lobes,
abdominal terga mostly fulvous with broad lateral fuscous margins on tergum I, a narrow medial
longitudinal fuscous band on tergum V and apex of piercer of ovipositor with one pair of subapical
lobes (Drew 1989).
160
PEST STATUS

Exotic

A major pest of pumpkins in New Britain
ATTRACTANT
No known record.
FIGURES
Figure 55. Bactrocera decipiens
161
Figure 56. Bactrocera decipiens
162
8.4.12 Bactrocera (Zeugodacus) depressa (Shiraki)
Common Name:
Zeugodacus depressus Shiraki
Paradacus depressus (Shiraki)
Bactrocera (Paradacus)depressa (Shiraki)
DIAGNOSIS
8.4.12.1
A large species; face fulvous with a pair of small to medium ill-defined fuscous to dark fuscous spots;
postpronotal lobes and notopleura yellow; scutum basically red-brown with dark markings; lateral and
medial postsutural yellow vittae present; a yellow spot anterior to notopleural suture in front of lateral
postsutural yellow vittae; mesopleural stripe reaching midway between anterior margin of notopleuron
and anterior npl. seta dorsally; scutellum yellow; wing with cells bc and c with a pale fuscous tint,
microtrichia in outer corner of cell c only; a narrow fuscous costal band slightly overlapping R2+3; a
broad fuscous anal streak; a pale tint around medial posterior area of wing margin; abdominal terga
III-V fulvous with a black ‘T’ pattern consisting of a narrow transverse black band across anterior
margin of tergum III and a medium-width medial longitudinal black band over all three terga that is
broken on posterior margin of tergum III, anterolateral corners of terga IV and V dark fuscous.
8.4.12.2
No information available
8.4.12.3
Molecular
HOST RANGE
Family Cucurbitaceae (Cucurbita moschata, Cucurbita pepo, Trichosanthes quadricirrha)
DISTRIBUTION
Taiwan, Japan (all major islands), Korea
REMARKS
Bactrocera depressa is similar to Bactrocera (Zeugodacus) okunii (Shiraki) and Bactrocera
(Zeugodacus) uncinata Drew & Romig in possessing a basic red-brown scutum, wings colourless
except for costal band and anal streak and with the costal band expanded into an apical spot, lateral
postsutural vittae ending before ia. seta. It differs from B. uncinata in lacking a recurved black barb on
tergum IV and from B. okunii in having cells bc and c with a pale tint and a smaller apical spot on apex
of costal band.
163
PEST STATUS
 Exotic
 A pest of cucurbit crops
ATTRACTANT
No known record
FIGURE
Figure 57. Bactrocera depressa
Image courtesy of A.Carmichael, International Centre for the Management of Pest Fruit Flies, Griffith University
164
8.4.13 Bactrocera (Bactrocera) dorsalis (Hendel)
Common name: Oriental fruit fly
Dacus dorsalis
Dacus (Strumeta) dorsalis
Strumeta dorsalis
Dacus (Bactrocera) dorsalis
DIAGNOSIS
8.4.13.1
Face fulvous with a pair of medium sized circular black spots; scutum colour variable red-brown to
black, with lanceolate dark patterning in intermediate forms; postpronotal lobes and notopleura yellow;
mesopleural stripe reaching midway between anterior margin of notopleuron and anterior npl. seta
dorsally; broad parallel sided lateral postsutural vittae ending behind ia. seta; medial postsutural vitta
absent; scutellum yellow; legs with femora entirely fulvous, fore tibiae pale fuscous and hind tibiae
fuscous; wings with cells bc and c colourless, microtrichia in outer corner of cell c only, a narrow
fuscous costal band confluent with R2+3 and remaining very narrow around apex of wing (occasionally
there can be a very slight swelling around apex of R4+5), a narrow pale fuscous anal streak;
supernumerary lobe of medium development; abdominal terga III-V exhibits a range of colour patterns
(see Drew and Hancock 1994) but possesses the basic pattern of a black ‘T’ consisting of a narrow
transverse black band across anterior margin of tergum III, a narrow medial longitudinal black band
over all three terga, narrow anterolateral fuscous to dark fuscous corners on terga IV and V; a pair of
oval orange-brown to pale fuscous shining spots on tergum V; abdominal sterna dark coloured;
posterior lobe of male surstylus short; female with aculeus tip needle shaped (Schutze et al. 2015).
8.4.13.2
8.4.13.3
Molecular
BsrI:
650, 260
SnaBI: 326, 540
HhaI:
656, 192
SspI:
DNC
HinfI:
770
Vspl:
DNC
Sau3AI: DNC
PCR-RFLP Test 2: No distinctive restriction enzymes but a combination of several can be chosen to
distinguish the complex from other Bactrocera and species in other genera. Choice of enzyme will be
based on relative distinctiveness from other species potentially trapped. See Appendices 3 and 4
165
HOST RANGE
Bactrocera dorsalis has been recorded on hosts from a wide range of families. These include:
Alangiaceae, Anacardiaceae, Annonaceae, Apocynaceae, Arecaceae, Burseraceae, Capparaceae,
Caprifoliaceae, Caricaceae, Celastraceae, Chrysobalanaceae, Clusiaceae, Combretaceae,
Convolvulaceae, Curcurbitaceae, Ebenaceae, Elaeocarpaceae, Euphorbiaceae, Fabaceae,
Flacourtiaceae, Lauraceae, Lecythidaceae, Malpighiaceae, Meliaceae, Moraceae, Musaceae,
Myrtaceae, Olacaceae, Oleaceae, Oxalidaceae, Polygalaceae, Rhamnaceae, Rosaceae, Rubiaceae,
Rutaceae, Sapindaceae, Sapotaceae, Simaroubaceae and Solanaceae (for a full list of recorded hosts
see Allwood et al. 1999; Leblanc et al. 2012).
Scientific name
Common name
Scientific name
Common name
cashew nut
Mimusops elengi
spanish cherry
Annona reticulata
bullock's heart
Muntingia calabura
Jamaican cherry
Annona squamosa
sugarapple
Musa
banana
Averrhoa carambola
carambola
Prunus armeniaca
apricot
Capsicum annuum
bell pepper
Prunus avium
sweet cherry
Carica papaya
papaw
Prunus cerasus
sour cherry
caimito
Prunus domestica
plum
Citrus reticulata
mandarin
Prunus persica
peach
Coffea arabica
arabica coffee
Psidium guajava
guava
Dimocarpus longan
longan tree
Pyrus communis
European pear
Diospyros kaki
persimmon
Syzygium aqueum
watery rose-apple
Malpighia glabra
acerola
Syzygium cumini
black plum
Malus domestica
apple
Syzygium jambos
rose apple
Mangifera foetida
bachang
Syzygium malaccense
malay-apple
Mangifera indica
mango
water apple
Manilkara zapota
sapodilla
DISTRIBUTION
Africa, India, Sri Lanka, Nepal, Bhutan, Myanmar, southern China, Taiwan, Hong Kong, northern
Thailand, Vietnam, Cambodia, Laos, Hawaii, Mariana Islands, Tahiti (pers. comm. Drew 2010).
REMARKS
Bactrocera dorsalis is similar to B. carambolae and B. verbascifoliae in possessing broad parallel
sided lateral postsutural vittae, costal band confluent with or very slightly overlapping R 2+3 and to B.
verbascifoliae in having the costal band remaining very narrow beyond apex of R 2+3, femora entirely
fulvous and abdominal terga III-V with a narrow medial longitudinal dark band.
It differs from B. carambolae in possessing a very narrow apical section of the costal band, narrow
medial longitudinal dark band on abdominal terga III-V and triangular shaped anterolateral dark
corners on abdominal terga IV and V (these markings are rectangular in B. carambolae).
It differs from B. verbascifoliae in possessing narrow lateral dark margins on abdominal terga IV and
V.
166
Other dorsalis complex species that are similar to B. dorsalis are B. hantanae, B. irvingiae, B. raiensis
and B. syzygii; however, all of these species possess a broad medial longitudinal dark band on
abdominal terga III-V and have not been recorded as having males responding to methyl eugenol. See
Drew and Hancock (1994) for a full discussion of type specimens, relationships and synonymies.
Bactrocera dorsalis can appear similar to the Australian species B. endiandrae (eastern Australia to
northern NSW) and B. musae (north Qld to Townsville) (see diagnostician’s notes).
Following the publication on the dorsalis complex by Drew and Hancock (1994), there has been
considerable research to investigate the integrity of many of the morphologically close species in the
dorsalis complex. The review of Clarke et al. (2005) summarised the bulk of this research and has
demonstrated that most taxa within the complex can be satisfactorily resolved and that the complex is
undergoing rapid morphological change. Drew & Romig (2013) synonymised Bactrocera philippinensis
with B. papayae; Schutze et al. (2015) synonymised B. papayae and B. invadens with B. dorsalis.
PEST STATUS

Exotic

High priority pest identified in the Apple and Pear, Avocado, Banana, Citrus, Summerfruit,
Tropical fruit and Vegetable IBPs

Bactrocera dorsalis is a major economic pest and utilises a wide range of commercial, edible
and rainforest fruits
ATTRACTANT
Methyl eugenol.
FIGURES
Figure 58. Bactrocera dorsalis
Image courtesy of the University of Florida and the Florida Department of Agriculture and Consumer Services
http://entomology.ifas.ufl.edu/creatures/index.htm (as of 22 August 2011)
167
Figure 59. Bactrocera dorsalis
168
8.4.14 Bactrocera (Bactrocera) facialis (Coquillett)
Common name:
Dacus facialis
Chaetodacus facialis
Strumeta facialis
Dacus (Strumeta) facialis
DIAGNOSIS
8.4.14.1
Small species; facial spots absent; postpronotal lobes and notopleura yellow; scutum dark fuscous to
black, mesopleural stripe reaching almost to postpronotal lobes, narrow short lateral postsutural vittae
present, medial postsutural vitta absent, scutellum yellow; wing with a narrow fuscous costal band and
narrow pale fuscous anal streak, cells bc and c colourless with microtrichia in outer corner of cell c
only; abdominal terga III-V orange-brown with a moderately broad medial longitudinal fuscous to black
band over all three terga, broad lateral fuscous to black margins on tergum III and anterolateral
corners of terga IV and V; posterior lobe of male surstylus short; female with aculeus tip needle
shaped (Drew 1989; pers. comm. Drew 2010).
8.4.14.2
Molecular
BsrI:
580, 250
SnaBI: DNC
HhaI:
600, 180
SspI:
DNC
HinfI:
DNC
Vspl:
DNC
Sau3AI: 400
PCR-RFLP Test 2: AluI, DraI and RsaI produce diagnostic restriction patterns for B. facialis and B.
passiflorae which cannot be distinguished (Appendix 3 and 4);
AluI: 770-760, 270, 170, 130, 120, 110; DraI: 1200, 290, 110; RsaI: 500, 420, 410, 290
Allozyme electrophoresis: Data not available
169
HOST RANGE
Bactrocera facialis has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Combretaceae, Fabaceae, Lauraceae, Moraceae, Myrtaceae, Passifloraceae,
Rosaceae, Rutaceae, Sapindaceae and Solanaceae (for a full list of recorded hosts see Leblanc et al.
2012).
Scientific name
Common name
Scientific name
Common name
Capsicum annuum
bell pepper
Psidium guajava
guava
Mangifera indica
mango
Syzygium spp.
Carica papaya
papaya
Citrus spp.
Persea americana
avocado
DISTRIBUTION
Known from the Tongatapu I. and the Ha’apai Group, Tonga (Drew 1989).
REMARKS
Bactrocera facialis is distinct in having broad black lateral margins on abdominal tergum III and
anterolaterally on terga IV and V, a moderately broad medial longitudinal black band on terga III-V and
lateral postsutural vittae very short and narrow ending at level of sa. setae (Drew 1989).
It can appear superficially similar to B. frauenfeldi which is established in north Queensland (see
diagnostician’s notes).
PEST STATUS

Exotic

Bactrocera facialis is a major pest, which causes up to 100% fruit loss in Capsicum species in
Tonga
ATTRACTANT
Cue lure.
170
FIGURES
Figure 60. Bactrocera facialis
Figure 61. Bactrocera facialis
171
8.4.15 Bactrocera (Bactrocera) frauenfeldi
(Schiner)
Common name: Mango fruit fly
Dacus frauenfeldi
Strumeta frauenfeldi
Dacus (Strumeta) frauenfeldi
DIAGNOSIS
8.4.15.1
Medium sized species; large black facial spots present; postpronotal lobes black; notopleura yellow;
scutum glossy black, mesopleural stripe reaching midway between anterior margin of notopleuron and
anterior npl. seta, lateral postsutural vittae present, medial postsutural vitta absent, scutellum yellow
with a black triangle on dorsal surface; wing with a narrow extremely pale fuscous costal band and
broad fuscous anal streak, a narrow fuscous transverse band across wing, cells bc and c pale
fuscous, microtrichia covering most of cell c; abdominal terga III-V orange-brown with a broad medial
and 2 broad lateral longitudinal black bands; posterior lobe of male surstylus short; female with
aculeus tip needle shaped (Drew 1989; Drew and Romig 2013). For potentially morphologically
confounding species see REMARKS and Table 8.
8.4.15.2
8.4.15.3
Molecular
BsrI:
DNC
SnaBI: DNC
HhaI:
600, 200
SspI:
180, 620
HinfI:
DNC
Vspl:
DNC
Sau3AI: 400, 450
PCR-RFLP Test 2: B. frauenfeldi cannot be distinguished from B. kirki or B. trilineola. No distinctive
restriction enzymes, but a combination of several can be chosen to distinguish this group from other
Bactrocera and species in other genera. See Appendices 3 and 4.
172
HOST RANGE
Bactrocera frauenfeldi has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Caricaceae, Clusiaceae, Combretaceae, Ebenaceae, Euphorbiaceae,
Lecythidaceae, Loganiaceae, Malpighiaceae, Meliaceae, Moraceae, Musaceae, Myrtaceae,
Olacaceae, Oxalidaceae, Passifloraceae, Rubiaceae, Rutaceae, Sapotaceae and Solanaceae (for a
full list of recorded hosts see Hancock et al. 2000; Leblanc et al. 2004; Leblanc et al. 2012).
Scientific name
Common name
Scientific name
Common name
Mangifera indica
mango
Psidium guajava
guava
Syzygium malaccense
malay-apple
Manilkara kauki
Citrus spp.
DISTRIBUTION
Widely distributed in Papua New Guinea and across the Bismark Archipelago to the Solomon Islands,
and established in the Torres Strait and northern Queensland as far south as Townsville (Royer et al.
2015; Drew and Romig 2013). This is an exotic pest species that was detected at the tip of
Queensland in the 1970s. It appears to have reached the limit of its spread south to Townsville without
any controls due to climatic limitations (Royer et al. 2015).
REMARKS
Bactrocera frauenfeldi is similar to B. albistrigata (see diagnostician’s notes) but differs in having dark
postpronotal lobes (though some specimens can have paler ppn lobes) darker leg patterning and
wider medial line on the abdomen with broad lateral bands (though B. albistrigata can also have
similar abdominal patterning). Bactrocera albistrigata, regarded as a synonym of B. frauenfeldi by
Hardy and Adachi (1954), is a distinct species. It possesses yellow postpronotal lobes and is confined
to South-east Asia (Drew 1989) and can be separated by DNA barcoding (see Armstrong pers. comm.
in Royer et al. 2015). See Table 8 under B. albistrigata entry for differences with other frauenfeldi
complex species.
PEST STATUS

Established

A major pest fruit fly species in Papua New Guinea, attacking most locally grown tropical fruits
and nuts (with the exception of banana which is a rare host)
ATTRACANT
Cue-group. Raspberry ketone formate (Melolure) was found to be a stronger attractant for this species
in north Queensland (Royer 2015).
173
FIGURES
Figure 62. Bactrocera frauenfeldi
Figure 63. Bactrocera frauenfeldi
174
8.4.16 Bactrocera (Afrodacus) jarvisi (Tryon)
Common name: Jarvis’ fruit fly
Chaetodacus jarvisi
Chaetodacus jarvisi var. careya
Dacus (Afrodacus) jarvisi
Afrodacus jarvisi
DIAGNOSIS
8.4.16.1
Medium sized species; medium sized irregularly oval black facial spots present; postpronotal and
notopluera yellow and connected by a broad yellow band; scutum red-brown, mesopleural stripe
reaching almost to anterior npl. seta, lateral postsutural vittae present, medial postsutural vittae
absent, wing with a narrow fuscous costal band and broad fuscous anal streak, cells bc and c
colourless with microtrichia in outer corner of cell c only; abdominal terga III-V orange-brown except
for a fuscous to black transverse band across anterior margin of tergum III and fuscous to black
medial longitudinal band generally over all three terga but often variable; posterior lobe of male
surstylus long; female with aculeus tip needle shaped (Drew 1989; pers. comm. Drew 2010).
8.4.16.2
8.4.16.3
Molecular
BsrI:
600, 250
SnaBI: DNC
HhaI:
650, 180
SspI:
700
HinfI:
770
Vspl:
DNC
Sau3AI: 420
PCR-RFLP Test 2: SspI produces a diagnostic pattern. Additional restriction enzymes can be chosen
based on relative distinctiveness from other species potentially trapped; see Appendices 3 and 4.
SspI: 880, 640, 100
Allozyme electrophoresis: (Section 7.3.5).
175
HOST RANGE
Bactrocera jarvisi has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Arecaceae, Cactaceae, Caricaceae, Celastraceae, Chrysobalanaceae,
Clusiaceae, Combretaceae, Curcurbitaceae, Ebenaceae, Elaeocarpaceae, Lauraceae,
Lecythidaceae, Malpighiaceae, Meliaceae, Moraceae, Musaceae, Myrtaceae, Oleaceae, Oxalidaceae,
Passifloraceae, Punicaceae, Rosaceae, Rubiaceae, Rutaceae, Sapindaceae, Sapotaceae and
Solanaceae (for a full list of recorded hosts see Hancock et al. 2000).
Scientific name
Common name
Scientific name
Common name
Mangifera indica
mango
Prunus persica
peach
Psidium guajava
guava
Musa sp.
banana
DISTRIBUTION
Northern Australia from Broome, Western Australia to eastern Arnhem Land, Northern Territory and
northwest Queensland, Torres Strait islands and eastern Australia from Cape York to the Sydney
district, New South Wales (Hancock et al. 2000). In recent tests using zingerone, B. jarvisi was not
detected in Sydney and the southern boundary is considered to be somewhere north of Sydney
(Dominiak et al. 2015b). Has been recorded from Indonesia (Irian Jaya) by White and Elson-Harris on
one occasion but is not established there and should not be regarded as a permanent record (pers.
comm. Drew 2010).
REMARKS
Bactrocera jarvisi is not morphologically similar to any Australian native species that respond to
zingerone or cue-lure, or that would be reared from commercial hosts. It is distinctive by being a
golden-orange colour with a joining band between the notopleural calli and postpronotal lobes, T on
the abdomen and flared costal band.
PEST STATUS

Endemic

A major pest in Queensland and the Northern Territory where it attacks a large number of fruit
and vegetable crops
ATTRACTANT
Attracted to zingerone (Fay 2012) and also known to be attracted to cue-lure (Drew 1989, Royer &
Hancock 2012, IAEA 2003).
176
FIGURES
Figure 64. Bactrocera jarvisi
Figure 65. Bactrocera jarvisi
177
8.4.17 Bactrocera (Bactrocera) kandiensis
(Drew and Hancock)
Common name:
DIAGNOSIS
8.4.17.1
Face fulvous with a pair of large oval black spots; scutum black except brown below and behind lateral
postsutural vittae, around mesonotal suture, inside postpronotal lobes, around prsc. setae and on
anterocentral margin; postpronotal lobes yellow (anteromedial corners red-brown); notopleura yellow;
mesopleural stripe slightly wider than notopleuron dorsally; narrow parallel sided lateral postsutural
vittae ending at ia. seta; medial postsutural vitta absent; scutellum yellow with a moderately broad
black basal band; legs with femora fulvous with dark fuscous on outer apical 2/3 of fore femora, inner
apical 1/2 of mid and inner apical 1/3 of hind femora, fore tibiae fuscous, mid tibiae fulvous and hind
tibiae dark fuscous; wings with cells bc and c colourless, microtrichia in outer corner of cell c only, a
narrow fuscous costal band confluent with R2+3 and remaining narrow around margin of wing to end
between extremities of R4+5 and M, a narrow fuscous cubital streak; supernumerary lobe of medium
development; abdominal terga III-V orange-brown with a narrow transverse black band across anterior
margin of tergum III but not covering lateral margins, a very narrow medial longitudinal fuscous to dark
fuscous band over all three terga (occasionally interrupted at intersegmental lines) and very narrow
fuscous to dark fuscous anterolateral corners on terga IV and V, a pair of oval orange-brown shining
spots on tergum V; abdominal sterna dark coloured; posterior lobe of male surstylus short; female with
aculeus tip needle shaped (Drew and Romig 2013).
8.4.17.2
Molecular
178
HOST RANGE
Bactrocera kandiensis has been recorded on hosts from six families, Anacardiaceae and Clusiaceae
(for a full list of recorded hosts see Allwood et al. 1999).
Major commercial hosts (Allwood et al. 1999; Tsuruta et al.1997):
Scientific name
Common name
Scientific name
Common name
cashew nut
Psidium guajava
guava
Annona glabra
pond apple
Spondias cytherea
jew plum
Citrus maxima
pummelo
Syzygium aromaticum
clove
Mangifera indica
mango
Syzygium jambos
rose apple
Averrhoa carambola
carambola
Carica papaya
papaya
DISTRIBUTION
Bactrocera kandiensis is confined to Sri Lanka (Drew and Hancock 1994).
REMARKS
Bactrocera kandiensis is similar to B. caryeae and B. neoarecae in possessing narrow parallel sided
lateral postsutural vittae and dark patterns on the apices of all femora or, at least, on fore and mid
femora.
It differs from B. neoarecae in possessing a single ‘T’ pattern over abdominal terga III-V (not on each
of the three separate terga), a narrow black basal band on the scutellum and dark markings on the
apices of all femora and from B. caryeae in possessing a very narrow medial longitudinal dark band on
abdominal terga III-V and narrow dark anterolateral corners on terga IV and V (pers. comm. Drew
2010).
It is also similar to B. dorsalis but has narrow vittae, spots on the apices of all femora and very narrow
costal band.
PEST STATUS

Exotic

Bactrocera kandiensis is a major pest of mangoes and is probably responsible for much of the
damage generally attributed to B. dorsalis in Sri Lanka
ATTRACTANT
Methyl eugenol.
179
FIGURES
Figure 66. Bactrocera kandiensis
180
8.4.18 Bactrocera (Bactrocera) kirki (Froggatt)
Common name:
Dacus kirki
Strumeta kirki
Dacus (Strumeta) kirki
DIAGNOSIS
8.4.18.1
Medium sized species; large black facial spots present; postpronotal lobes yellow (anterodorsal
margins black); notopluera yellow; scutum glossy black, mesoplueral stripe slightly wider than
notopleuron, lateral and medial postsutural vittae absent, scutellum glossy black with extreme lateral
margins yellow; wing with a narrow pale fuscous costal band and narrow fuscous anal streak, a
narrow pale fuscous tinge around r-m and dm-cu crossveins, cells bc and c with extremely pale
fuscous tinge and microtrichia in outer ½ of cell c only; abdominal terga glossy black except for two
longitudinal orange-brown bands over terga II-V either side of a broad medial longitudinal glossy black
band; posterior lobe of male surstylus short; female with aculeus tip needle shaped (Drew 1989; pers.
comm. Drew 2010). For potentially morphologically confounding species see REMARKS or Table 8.
8.4.18.2
Molecular
BsrI:
DNC
SnaBI: DNC
HhaI:
680, 190
SspI:
180, 620
HinfI:
DNC
Vspl:
DNC
Sau3AI: 400, 450
PCR-RFLP Test 2: B. kirki cannot be distinguished from B. frauenfeldi or B. trilineola. No distinctive
Bactrocera and species in other genera. See Appendices 3 and 4
181
HOST RANGE
Bactrocera kirki has been recorded on hosts from a range of families. These include: Anacardiaceae,
Bromeliaceae, Combretaceae, Fabaceae, Myrtaceae, Oxalidaceae, Passifloraceae, Rosaceae,
Rutaceae, Solanaceae (for a full list of recorded hosts see Leblanc et al. 2012).
Scientific name
Common name
Scientific name
Common name
Mangifera indica
mango
Persea americana
avocado
Psidium guajava
guava
Syzygium spp.
Citrus spp.
DISTRIBUTION
Widespread in the South Pacific islands: Western Samoa, American Samoa, Tonga, Niue and Tahiti
(Drew 1989).
REMARKS
Bactrocera kirki is similar to members of the B. frauenfeldi complex but differs in not having a distinct
transverse band on the wing, although it does have infuscation on the r-m and dm-cu cross veins. It
may appear similar to B. frauenfeldi (see diagnostician’s notes and Table 8 on page 128 for other
separating characters).
PEST STATUS

Exotic

High priority pest identified in the Avocado IBP

Bactrocera kirki is considered a major pest, and perhaps the most significant in the South
Pacific region
ATTRACTANT
Cue lure, Willison's lure (raspberry ketone).
182
FIGURES
Figure 67. Bactrocera kirki
Figure 68. Bactrocera kirki
183
8.4.19 Bactrocera (Bactrocera) kraussi (Hardy)
Common name:
Dacus (Strumeta) kraussi
Strumeta kraussi
DIAGNOSIS
8.4.19.1
Medium sized species; medium sized oval facial spots present; postpronotal lobes and notopleura
yellow; scutum red-brown with irregularly shaped lateral longitudinal pale fuscous to fuscous bands,
mesopleural stripe reaching midway between anterior margin of notopleron and anterior npl. seta,
lateral postsutural vittae present, medial postsutural vitta absent, scutellum yellow with a broad redbrown to fuscous basal band; wing colourless or with a pale fulvous tint and a narrow fuscous costal
band and broad fuscous anal streak, cells bc and c pale fulvous to fulvous with microtrichia in outer
corner of cell c only, abdominal terga III and IV fuscous and tergum V fulvous except for broad lateral
dark fuscous margins on terga III and IV and broad fuscous lateral margins on tergum V; posterior
lobe of male surstylus short; female with aculeus tip needle shaped (Drew 1989; pers. comm. Drew
2010).
8.4.19.2
Molecular
PCR-RFLP Test 1: Data not available
HOST RANGE
Bactrocera kraussi has been recorded on hosts from a wide range of families. These include:
Agavaceae, Anacardiaceae, Annonaceae, Apocynaceae, Clusiaceae, Combretaceae, Cunoniaceae,
Davidsoniaceae, Elaeocarpaceae, Euphorbiaceae, Flacourtiaceae, Icacinaceae, Lauraceae,
Lecythidaceae, Loganiaceae, Malpighiaceae, Meliaceae, Menispermaceae, Moraceae, Musaceae,
Myrtaceae, Oleaceae, Oxalidaceae, Passifloraceae, Rosaceae, Rubiaceae, Rutaceae, Sapindaceae,
Sapotaceae, Solanaceae and Thymeliaceae (for a full list of recorded hosts see Hancock et al., 2000).
Major commercial hosts (Drew 1989, Hancock et al. 2000):
Scientific name
Citrus sp.
Mangifera indica
Common name
Grapefruit, mandarin,
orange
mango
Scientific name
Common name
Musa sp.
banana
Psidium guajava
guava
It should be noted that fruit flies are not known to attack hard green bananas (Hancock et al., 2000).
184
DISTRIBUTION
Torres Strait Islands and northeast Queensland, as far south as Townsville (Hancock et al. 2000,
Royer & Hancock 2012).
REMARKS
Bactrocera kraussi is similar to all other species in the fagraea complex being a general red-brown fly,
scutellum with a broad dark basal band and cells bc and c not covered in dense microtichia. It differs
from B. rufescens in lacking a medial dark band on abdomen, from B. fagraea and B. russeola in
having lateral fuscous markings on abdominal terga III and IV and from B. halfordiae in having a redbrown scutum with or without fuscous markings, abdomen usually fuscous over terga III and IV and
laterally on tergum V, mesopleural stripe 1 ½ times the width of notopleuron and lateral postsutural
vittae parallel sided (Drew 1989).
In cue traps it is most likely to be confused with B. tryoni but differs in having tint only in both costal
cells (instead of microtrichia in both), longer less tapered vittae, lateral markings only on the abdomen
(instead of a wraparound T), narrow mesopleural stripe, broad basal band on the scutellum and dark
apices of the femora.
PEST STATUS

Endemic

A major pest species in North Queensland
ATTRACTANT
Cue lure, but is more strongly attracted to isoeugenol (Royer 2015).
185
FIGURES
Figure 69. Bactrocera kraussi
Image courtesy S. Sands, International Centre for the Management of Pest Fruit Flies, Griffith University
186
8.4.20 Bactrocera (Bactrocera) latifrons (Hendel)
Common name: Solanum fruit fly
Chaetodacus latifrons
DIAGNOSIS
8.4.20.1
A medium sized species; face fulvous with a pair of large oval black spots; postpronotal lobes and
notopleura yellow; scutum dull black; lateral postsutural vittae present; medial postsutural vitta absent;
mesopleural stripe extending to anterior npl. seta dorsally; scutellum yellow; wing with a narrow
fuscous costal band overlapping R2+3 and expanding into a small spot around apex of R4+5, a medium
width fuscous anal streak; cells bc and c colourless; microtrichia in outer corner of cell c only; all
abdominal terga entirely dark orange-brown, posterior lobe of male surstylus short; female with apex
of aculeus trilobed (Drew and Romig 2013).
8.4.20.2
8.4.20.3
Molecular
Bsr1:
600, 200
SnaB1: DNC
HhaI:
600, 190
SspI:
DNC
HinfI:
DNC
Vspl:
DNC
Sau3a1: DNC
PCR-RFLP Test 2: Sau3A produces a diagnostic pattern. Other useful enzymes DraI, NcoI and RsaI
Sau3A: 760, 730,110
HOST RANGE
Bactrocera latifrons has been recorded on hosts from a wide range of families. These include:
Lythraceae, Myrtaceae, Oleaceae, Passifloraceae, Punicaceae, Rhamnaceae, Rutaceae,
Sapindaceae, Solanaceae and Verbenaceae (for a full list of recorded hosts see Allwood et al. 1999).
Scientific name
Common name
Scientific name
Common name
Capsicum sp.
peppers
tomato
Capsicum annuum
bell pepper
Solanum melongena
eggplant
187
DISTRIBUTION
Sri Lanka, India, Pakistan through to Southern China, Japan, Taiwan, Thailand, Laos, Vietnam,
Peninsular Malaysia, Indonesia, Hawaii, Tanzania (Drew and Romig 2013).
REMARKS
Bactrocera latifrons can be confused with species in the B. musae complex and the B. dorsalis
complex in possessing similar body colour patterns. However, it is distinct in having a trilobed apex on
the aculeus and uniformly dark orange-brown abdominal terga. Bactrocera musae can also have a
uniformly orange abdomen but it also differs from this species in having a flared costal band. It is
similar to B. citima in possessing a generally black scutum, costal band overlapping R 2+3, cells bc and
c colourless and parallel sided lateral postsutural vittae but differs from this species in having redbrown around the lateral and posterior margins of the scutum, femora entirely fulvous and abdominal
terga III-V entirely red-brown (Drew and Romig 2013).
PEST STATUS

Exotic

This is a pest of solanaceous crops throughout its range
ATTRACTANT
Alpha-ionol and cade oil is a moderate attractant for this species. Although not as attractive as cuelure and ME are to other species it is significantly more attractive than protein bait (Flath et al. 1994,
McQuate & Peck 2001, McQuate et al. 2013).
188
FIGURES
Figure 70. Bactrocera latifrons
Image courtesy of Y. Martin and International Centre for the Management of Pest Fruit Flies, Griffith University
189
8.4.21 Bactrocera (Bactrocera) melanotus
(Coquillett)
Common name:
Dacus melanotus
Chaetodacus melanotus
Strumeta melanotus
Dacus (Strumeta) melanotus
DIAGNOSIS
8.4.21.1
Medium sized species; facial spots absent or small and pale; postpronotal lobes yellow (anterolateral
corners black); notopleura glossy black; scutum glossy black, mesopleural stripe reaching to
postpronotal lobe, lateral and medial postsutural vittae absent, scutellum glossy black; wing with a
narrow pale fuscous costal band and narrow fuscous tint in anal cell, narrow pale fuscous markings
along r-m and dm-cu crossveins, cells bc and c colourless or with a very pale fuscous tint, microtrichia
in outer corner of cell c only; all abdominal terga entirely glossy black; posterior lobe of male surstylus
short; female with aculeus tip needle shaped (Drew 1989; pers. comm. Drew 2010).
8.4.21.2
8.4.21.3
Molecular
PCR-RFLP Test 2: No fully diagnostic restriction enzymes, but a combination of several can be
chosen to distinguish B. melanotus from other Bactrocera and species in other genera. See
Appendices 3 and 4.
HOST RANGE
Bactrocera melanotus has been recorded on hosts from seven families. These include:
Anacardiaceae, Caricaceae, Combretaceae, Fabaceae, Myrtaceae, Rutaceae and Sapindaceae (for a
full list of recorded hosts see Leblanc et al. 2012).
Scientific name
Common name
Citrus sp.
Mangifera indica
mango
Scientific name
Common name
Psidium guajava
guava
Carica papaya
papaya
Syzygium spp.
190
DISTRIBUTION
Restricted to Cook Is (Drew 1989).
REMARKS
Bactrocera melanotus is similar to B. atra and B. perfuscain possessing an entirely black scutellum,
scutum black with medial and lateral postsutural vittae absent, abdominal terga black but differs from
these species in having infuscation around r-m and dm-cu crossveins. In addition, it can be separated
from B. atra in having postpronotal lobes mostly yellow, yellow mesopleural stripe and black femora
(Drew 1989). Bactrocera melanotus is unusual in that its scutum, scutellum and abdomen are entirely
dark coloured (black or very dark brown) (CABI 2007).
PEST STATUS

Exotic

High priority pest identified in the Avocado IBP

Bactrocera melanotus is considered a major pest of papaw and citrus crops
ATTRACTANT
Cue lure
FIGURES
Figure 71. Bactrocera melanotus
191
8.4.22 Bactrocera (Tetradacus) minax (Enderlein)
Common Name: Chinese Citrus Fruit Fly
Polistomimetes minax Enderlein
Mellesis citri Chen
DIAGNOSIS
8.4.22.1
A large Callantra-like species; face fulvous with a pair of large elongate black spots filling most of
antennal furrow; postpronotal lobes and notopleura yellow; a lateral yellow band running from
posterior margin of postpronotal lobe towards notopleuron but not reaching notopleuron; scutum redbrown with a mottled appearance from dorsoventral flight muscles and occasional irregular dark
fuscous markings; lateral and medial postsutural yellow vittae present; no yellow marking anterior to
notopleural suture; mesopleural stripe reaching anterior npl. seta dorsally; scutellum yellow; wing with
cells bc and c fuscous, microtrichia covering outer l/2 of cell c only; a broad fuscous costal band
overlapping R4+5, and with a dark fuscous spot within apex of this band; anal streak absent but with
pale fuscous within cell cup; abdominal terga III-V fulvous to red-brown and generally with a dark
fuscous to black ‘T’ pattern and dark fuscous to black anterolateral corners on terga IV and V.
8.4.22.2
8.4.22.3
Molecular
HOST RANGE
Family Rutaceae (Citrus spp. and Fortunella spp.)
DISTRIBUTION
Bhutan, Nepal, northern India, southern China
REMARKS
Bactrocera (Tetradacus) minax (Enderlein) is similar to Bactrocera (Tetradacus) tsuneonis (Miyake) in
the general shape of the thorax and abdomen and in the basic body colour patterns to a point where
these species could be confused in diagnosis. However, B minax differs in lacking a.sa. setae, an
incomplete lateral yellow band between the postpronotal lobe and notopleuron, a broader costal band
overlapping R4+5 with a distinct dark fuscous spot within its apex and a needle-shaped apex on the
piercer (not trilobed).
192
PEST STATUS
 Exotic
 Major pest of citrus in its native geographic range
ATTRACTANT
Weak response to methyl eugenol (Drew & Romig 2007)
FIGURES
Figure 72. Bactrocera minax
193
8.4.23 Bactrocera (Bactrocera) musae (Tryon)
Common name: Banana fruit fly
Chaetodacus musae
Chaetodacus tryoni var. musa
Chaetodacus musae var. dorsopicta
Dacus (Strumeta) musae
Strumeta musae
DIAGNOSIS
8.4.23.1
Medium sized species; medium sized black facial spots present; postpronotal lobes and notopleura
yellow; scutum dull black (although some Cape York Peninsula specimens have a lot of brown on the
scutum), mesopleural stripe reaching midway between anterior margin of notopleuron and anterior npl.
seta, lateral postsutural vittae present; medial postsutural vitta absent, scutellum yellow; wing with a
narrow fuscous costal band and anal streak, cells bc and c colourless with microtrichia in outer corner
of cell c only; abdominal terga III-V may vary from uniformly orange-brown to orange-brown with a
fuscous to black medial longitudinal band and fuscous to black anterolateral corners on tergum III;
posterior lobe of male surstylus short; female with aculeus tip needle shaped (Drew 1989; pers. comm.
Drew 2010).
8.4.23.2
8.4.23.3
Molecular
DNA barcoding: BOLD reference data available to distinguish B. musae s.s. from at least the nonpest species in the B. musae complex, B. conterminal, B. prolixa, B. rufivitta, and B. tinomiscii (Drew et
al. 2011); however other species within the complex that are not represented in BOLD could confound
diagnosis
BsrI:
600, 250
SnaBI: 320, 520
HhaI:
630, 220
SspI:
DNC
HinfI:
DNC
Vspl:
DNC
Sau3AI: DNC
PCR-RFLP Test 2: CfoI and NcoI are diagnostic amongst other Bactrocera species and genera (see
Appendices 3 and 4), but species within the B. musae complex have not been tested
194
HOST RANGE
Bactrocera musae has been recorded on hosts from nine families. These include: Capparaceae,
Caricaceae, Musaceae, Myrtaceae, Olacaceae, Passifloraceae, Rubiaceae, Rutaceae and
Solanaceae (for a full list of recorded hosts see Hancock et al. 2000).
Major commercial hosts (Drew, 1989; Hancock et al., 2000):
Scientific name
Common name
Musa sp.
banana
DISTRIBUTION
Torres Strait Islands and northeast Queensland, as far south as Townsville (Hancock et al. 2000),
Papua New Guinea and associated islands, Bismark Archipelago, the Solomon Islands (Drew 1989)
and Indonesia (West Papua, formerly part of Irian Jaya) (Drew and Romig 2013).
REMARKS
There are a large number of species similar to Bactrocera musae, all placed in the musae complex. It
is similar to B. bancroftii an endemic ME-responsive species in Queensland in the musae complex, but
differs in not having an apical band on the scutellum, having a wider costal band and narrower anal
streak.
B. musae has a considerable intraspecific variation and can appear similar to the ME-responsive B.
endiandrae (a Queensland rainforest species) and B. dorsalis. It differs from B. endiandrae in having a
wider costal band, narrower anal streak, longer less tapered vittae and less of a distinct T on the
abdomen. It differs from B. dorsalis in having a wider costal band that doesn’t dip in at the end of
R2+3, wider anal streak and slightly tapering, slightly shorter vittae that usually end at the ia bristle,
and usually less patterning on the abdomen (although some B. dorsalis specimens can have reduced
abdominal markings similar to B. musae).
PEST STATUS

Endemic

Major pest of commercial bananas.
ATTRACTANT
Methyl eugenol.
195
FIGURES
Figure 73. Bactrocera musae
Image courtesy of Ken Walker, Museum Victoria, www.padil.gov.au (as of 22 August 2011))
196
197
8.4.24 Bactrocera (Bactrocera) neohumeralis
(Hardy)
Common name: Lesser Queensland fruit fly
Chaetodacus humeralis
Strumeta humeralis
Dacus (Strumeta) tryoni var. neohumeralis
Dacus (Strumeta) neohumeralis
Dacus (Bactrocera) neohumeralis
DIAGNOSIS
8.4.24.1
Medium sized species; medium sized black facial spots present; postpronotal lobes dark brown to
fuscous; notopleura yellow; scutum dark red-brown with dark fuscous to black markings, mesopleural
stripe reaching midway between anterior margin of notopleuron and anterior npl. seta, lateral
postsutural vittae present, medial postsutural vitta absent, scutellum yellow; wing with a narrow
fuscous costal band and broad fuscous anal streak, cells bc and c fuscous, microtrichia covering cell c
and outer ½ of cell bc; abdominal terga III-V generally dark fuscous to dull black and tending redbrown medially; posterior lobe of male surstylus short; female with aculeus tip needle shaped (Drew
1989; pers. comm. Drew 2010).
8.4.24.2
8.4.24.3
Molecular
DNA barcoding: BOLD reference data available, but cannot be distinguished from B. aquilonis or B.
tryoni
BsrI:
200, 600
SnaBI: DNC
HhaI:
640, 190
SspI:
180, 570
HinfI:
770
Vspl:
DNC
Sau3AI: 420
sizes (bp) are 1000, 550 and 100. This species cannot be distinguished from B. aquilonis or B. tryoni.
Other restriction patterns useful for diagnosis of the complex are listed in Appendix 3, with choice
based on relative distinctiveness from other species potentially trapped; descriptions of the patterns
are given in Appendix 4.
198
HOST RANGE
Bactrocera neohumeralis has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Apocynaceae, Arecaceae, Basellaceae, Cactaceae, Capparaceae,
Caricaceae, Celastraceae, Chrysobalanaceae, Clusiaceae, Combretaceae, Davidsoniaceae,
Ebenaceae, Elaeocarpaceae, Euphorbiaceae, Flacourtiaceae, Hippocraterceae, Lauraceae,
Leeaceae, Lecythidaceae, Malpighiaceae, Melastomataceae, Meliaceae, Moraceae, Musaceae,
Myrtaceae, Olacaceae, Oleaceae, Oxalidaceae, Passifloraceae, Piperaceae, Rhamnaceae,
Rhizophoraceae, Rosaceae, Rubiaceae, Rutaceae, Santalaceae, Sapindaceae, Sapotaceae,
Smilacaceae, Solanaceae, Verbenaceae and Vitaceae (for a full list of recorded hosts see Hancock
et al. 2000).
Major commercial hosts:
A large number of important commercial/edible host fruits and vegetables (see Drew 1989; Hancock
et al. 2000).
DISTRIBUTION
Common pest in Eastern Australia, south to Coffs Harbour, Torres Strait Islands and mainland Papua
New Guinea (Drew 1989). It is not found in central and southern NSW (Osborne et al. 1997, Gillespie
2003, Dominiak et al. 2003) or in Sydney (Dominiak et al. 2011, 2015a, 2015b). Vargas et al. 2015
place this species along the Queensland coast and perhaps the far north coast of New South Wales.
REMARKS
B. neohumeralis differs from B. tryoni in having dark postprotonotal lobes (this is a distinct character)
in addition to being generally darker. Although these two species are very similar morphologically,
their different daily mating periods (B. tryoni at dusk and B. neohumeralis during the middle of the day)
are good reason to keep them separate (Drew 1989).
PEST STATUS

Endemic

Bactrocera neohumeralis is a major pest of commercial fruit crops in Queensland, Australia,
and in some crops it occurs in equal abundance to B. tryoni
ATTRACTANT
Cue group. More attracted to cue-lure than raspberry ketone formate (Melolure) in north Queensland
(Royer 2015).
199
FIGURES
Figure 76. Bactrocera neohumeralis
Figure 77. Bactrocera neohumeralis
200
8.4.25 Bactrocera (Bactrocera) obliqua (Malloch)
Common Name:
Dacus obliquus Malloch
DIAGNOSIS
8.4.25.1
A dark coloured species; face black; postpronotal lobes and notopleura yellow; scutum black; lateral
potsutural vittae short and tapering posteriorly; medial postsutural vitta absent; scutellum yellow with a
broad medial longitudinal black band; wing with a narrow fuscous costal band and anal streak, r-m
crossvein oblique and enclosed by a pale infuscation, cells bc and c colourless; all abdominal terga
entirely black.
8.4.25.2
8.4.25.3
Molecular
HOST RANGE
Recorded from Myrtaceae (Guava and Syzygium species)
DISTRIBUTION
Bismarck Archipelago; Bougainville Island
REMARKS
B. obliqua is similar to B. psidii (Froggatt) in possessing a broad black triangular marking on dorsal
surface of scutellum, infuscation on crossveins and mesonotum black with medial postsutural vitta
absent but differs in having a black face, r-m crossvein longer than dm-cu crossvein, lateral
postsutural vittae very short and narrow ending just behind sa. setae, legs with all tibiae and apices of
femora and apical 4 segments of tarsi fuscous.
PEST STATUS
 Exotic
 A minor pest of Myrtaceae (Guava, Syzygium species)
ATTRACTANT
No known record
201
FIGURES
Figure 78. Bactrocera obliqua
202
8.4.26 Bactrocera (Bactrocera) occipitalis (Bezzi)
Common name:
Chaetodacus ferrugineus var. occipitalis
Dacus (Strumeta) dorsalis var. occipitalis
Dacus (Strumeta) occipitalis
Dacus (Bactrocera) occipitalis
DIAGNOSIS
8.4.26.1
Face fulvous with a pair of large oval black spots; scutum black except dark red-brown along posterior
margin and enclosing prsc. setae, below and behind lateral postsutural vittae, around mesonotal
suture, around anterior margin of notopleura and inside postpronotal lobes; postpronotal lobes and
notopleura yellow; mesopleural stripe reaching midway between anterior margin of notopleuron and
anterior npl. seta dorsally; broad parallel sided or subparallel lateral postsutural vittae ending at ia.
seta (in some specimens the vittae end behind the ia. seta); medial postsutural vitta absent; scutellum
yellow; legs with femora entirely fulvous, fore tibiae pale fuscous to fuscous, mid tibiae pale fuscous to
fuscous basally tending paler apically, hind tibiae fuscous; wings with cells bc and c colourless,
microtrichia in outer corner of cell c only, a narrow fuscous costal band distinctly overlapping R 2+3 and
widening markedly across apex of wing, a narrow fuscous anal streak; supernumerary lobe of medium
development; abdominal terga III-V with a narrow transverse black band across anterior margin of
tergum III and expanding to cover lateral margins, dark fuscous to black rectangular markings
anterolaterally on tergum IV which sometimes continue to cover posterolateral margins of this tergum,
dark fuscous to black anterolateral corners on tergum V, a very broad medial longitudinal black band
over all three terga, a pair of oval orange-brown shining spots on tergum V; abdominal sterna dark
coloured; posterior lobe of male surstylus short; female with apex of aculeus needle shaped (Drew
and Romig 2013).
8.4.26.2
Molecular
203
HOST RANGE
Allwood et al. (1999) host records are incomplete due to a lack of field host survey work through the
area of distribution of the species (pers. comm. Drew 2010). Bactrocera occipitalis has been recorded
on hosts from three families, Anacardiaceae, Myrtaceae and Rutaceae (Allwood et al. 1999).
Scientific name
Common name
Scientific name
Common name
Citrus microcarpa
musk lime
Psidium guajava
guava
Mangifera indica
mango
DISTRIBUTION
Philippines, East Malaysia (Sabah), Brunei, Indonesia (Kalimantan) (Drew and Romig 2013).
REMARKS
Bactrocera occipitalis is similar to some specimens of B. fuscitibia in possessing broad parallel sided
lateral postsutural vittae, costal band overlapping R2+3, narrow to medium width dark patterns on
lateral margins of abdominal terga III-V, shining spots on abdominal tergum V pale coloured, femora
entirely fulvous or with a dark spot on outer apical surfaces of fore femora only and a broad medial
longitudinal dark band on abdominal terga III-V.
It differs from B. fuscitibia in having the anterolateral bare area on the scutum broad and lateral dark
markings on abdominal terga IV and V of medium width (not narrow). Some populations of fruit flies
throughout South-East Asia have been misidentified as B. occipitalis in previous literature. See Drew
& Hancock (1994) for a complete discussion on this species and previous misidentifications (Drew and
Romig 2013).
PEST STATUS

Exotic

High priority pest identified in the Tropical fruit IBP

Bactrocera occipitalis is a major pest species within the dorsalis complex of South-east Asia
ATTRACTANT
Methyl eugenol.
204
FIGURES
Figure 79. Bactrocera occipitalis
Figure 80. Bactrocera occipitalis
205
8.4.27 Bactrocera (Daculus) oleae (Rossi)
Common Name: Olive fruit fly
Musca oleae Rossi
Musca oleae Gmelin
DIAGNOSIS
8.4.27.1
Face fulvous with a pair of black spots; scutum black; postpronotal lobes and notopleura yellow;
mesopleural stripe reaching midway between anterior margin of notopleuron and anterior npl. seta;
lateral and medial postsutural yellow vittae absent, scutellum yellow; wing with cells bc and c
colourless, cell sc. fuscous, a small fuscous spot around apex of vein R4+5 (costal bland absent), anal
streak absent; abdominal terga III – V red-brown with dark fuscous to black anterolateral corners on
terga III and IV; posterior lobe of male surstylus short; female with aculeus tip needle shaped.
8.4.27.2
8.4.27.3
Molecular
PCR-RFLP Test 2: A combination of AluI, DraI, RsaI and SspI produces a diagnostic pattern.
AluI: 680, 240, 170, 130, 120, 110; DraI: 1120,170, 130, 100; RsaI: 430, 400, 370, 290; SspI: 1150,
350
HOST RANGE
Recorded from family Oleaceae (olives)
DISTRIBUTION
Palearctic, Afrotropical and Oriental Regions
REMARKS
Bactrocera oleae is distinguished by the black scutum lacking lateral and medial postsutural yellow
vittae, yellow postpronotal lobes and notopleura, yellow scutellum, wing with fuscous cell sc and a
small spot around apex of R4+5 (costal band and anal streak lacking), abdominal terga III – V redbrown with dark fuscous to black on antero-lateral corners of terga III and IV.
PEST STATUS
 Exotic
 High priority
 A major pest of olives in the Mediterranean Region
206
ATTRACTANT
No known record
FIGURES
Figure 81. Bactrocera oleae
Figure 82. Bactrocera oleae
207
8.4.28 Bactrocera (Bactrocera) passiflorae
(Froggatt)
Common name: Fijian fruit fly
Dacus passiflorae
Chaetodacus passiflorae
Strumeta passiflorae
Dacus (Strumeta) passiflorae
DIAGNOSIS
8.4.28.1
Small species; facial spots absent; postpronotal lobes glossy black; notopleura yellow; scutum glossy
black, mesopleural stripe reaching to or beyond anterior npl. seta, lateral and medial postsutural vittae
absent, scutellum yellow; wing with a narrow fuscous costal band and narrow pale fuscous anal
streak, cells bc and c colourless with microtrichia in outer corner of cell c only; abdominal terga I-IV
glossy black and tergum V either glossy black with posterior margin dark fuscous or fuscous with a
medial longitudinal black band; posterior lobe of male surstylus short; female with aculeus tip needle
shaped (Drew 1989; pers. comm. Drew 2010).
It may appear similar to B. melanotus from the Cook Islands in being all black but differs in having pale
legs and scutellum and black postpronotal lobes.
8.4.28.2
8.4.28.3
Molecular
BsrI:
650, 270
SnaBI: DNC
HhaI:
650, 190
SspI:
750
HinfI:
770
Vspl:
DNC
Sau3AI: DNC
PCR-RFLP Test 2: AluI, DraI and RsaI produce diagnostic restriction patterns for B. facialis and B.
passiflorae which cannot be distinguished (Appendix 3 and 4);
AluI: 770-760, 270, 170, 130, 120, 110; DraI: 1200, 290, 110; RsaI: 500, 420, 410, 290
208
HOST RANGE
Reared from host fruits in 28 plant families (see Leblanc et al. 2012).
Major commercial hosts (Leblanc et al. 2012):
Scientific name
Common name
Scientific name
Common name
cashew nut
Passiflora quadrangularis
giant granadilla
Carica papaya
papaw
Persea americana
avocado
Citrus aurantiifolia
lime
Psidium guajava
guava
Citrus reticulata
mandarin
Solanum melongena
eggplant
Mangifera indica
mango
Theobroma cacao
cocoa
Passiflora edulis
passionfruit
DISTRIBUTION
Fiji Islands, Niue, Wallis and Futuna. There is also a separate form of B. passiflorae with paler
abdomen. This is probably an undescribed new species which occurs in Fiji, Tuvalu, Tokelau and
possibly the Niuas group in Tonga. Its host range and potential pest status have not yet been well
studied (SPC 2006).
REMARKS
Bactrocera passiflorae is similar to B. thistleoni in possessing black postpronotal lobes, scutellum
entirely yellow, scutum black with lateral and medial postsutural vittae absent but differs in having
facial spots absent and legs entirely fulvous (Drew 1989).
PEST STATUS

Exotic

High priority pest identified in the Avocado and Tropical Fruit IBPs
ATTRACTANT
Cue lure.
209
FIGURES
Figure 83. Bactrocera passiflorae
Image courtesy of the Secretariat of the Pacific Community Pacific Fruit Fly Web, www.spc.int/pacifly (as of 22
August 2011)
Figure 84. Bactrocera passiflorae
210
8.4.29 Bactrocera (Bactrocera) psidii (Froggatt)
Common name: South sea guava fruit fly
Tephritis psidii
Dacus psidii
Strumeta psidii
Dacus (Strumeta) psidii
DIAGNOSIS
8.4.29.1
Medium sized species; generally small fuscous to dark fuscous facial spots present; postpronotal
lobes yellow except anterodorsal corner black; notopleura yellow; scutum glossy black, mesopleural
stripe equal in width to notopleuron, short lateral postsutural vittae present, medial postsutural vitta
absent, scutellum yellow with a broad triangular black marking on dorsal surface; wing with a narrow
tint of extremely pale fuscous colouration around costal margin and a narrow fulvous anal streak, a
narrow tint of fuscous colouration around r-m and dm-cu crossveins, cells bc and c colourless to
extremely pale fulvous with microtrichia in outer corner of cell c only; abdominal terga entirely glossy
black; posterior lobe of male surstylus short; female with aculeus tip needle shaped (Drew 1989; pers.
comm. Drew 2010). For potentially morphologically confounding species see Table 8.
8.4.29.2
Molecular
Bsr1:
DNC
SnaBI: DNC
HhaI:
640, 190
SspI:
200, 550
HinfI:
DNC
Vspl:
DNC
Sau3AI: DNC
chosen to distinguish B. psidii from other Bactrocera and species in other genera. See Appendices 3
and 4
211
HOST RANGE
Bactrocera psidii has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Apocynaceae, Caricaceae, Combretaceae, Ebenaceae,
Euphorbiaceae, Malpighiaceae, Moraceae, Myrtaceae, Oxalidaceae, Passifloraceae, Punicaceae,
Rosaceae, Rutaceae and Vitaceae (Leblanc et al. 2013).
Scientific name
Common name
Citrus sp.
Mangifera indica
Scientific name
Common name
Psidium guajava
guava
mango
DISTRIBUTION
Restricted to New Caledonia (Drew 1989).
REMARKS
Bactrocera psidii is similar to B. obliqua in possessing infuscation on crossveins and the scutelum
yellow with a broad black triangular marking on dorsal surface. It differs from this species in having the
face fulvous with small pale spots in 75% of specimens, costal band narrow and not overlapping R 2+3,
r-m crossvein shorter than dm-cu crossvein, infuscation around crossveins very narrow and pale, legs
entirely fulvous, lateral postsutural vittae elongated and ending before ia. setae; posterior lobe of male
surstylus short; female with apex of aculeus needle shaped. This species is unusual in having wing
patterning very pale (including a mark along the r-m crossvein), scutellum marked with a large black
triangle and the abdomen entirely dark (black or dark orange-brown) (Drew 1989; pers. comm. Drew
2010).
It may appear superficially similar to the cue-responsive B. frauenfeldi in north Queensland but differs
in not having a distinct band on the wing, not having distinct medial line and lateral bands on the
abdomen, having pale legs and pale ppn lobes. See Table 8 on page 128.
PEST STATUS

Exotic

Bactrocera psidii is a major pest
ATTRACTANT
212
FIGURES
Figure 85. Bactrocera psidii
Figure 86. Bactrocera psidii
213
8.4.30 Bactrocera (Bactrocera) pyrifoliae
(Drew & Hancock)
Common Name:
DIAGNOSIS
8.4.30.1
Face fulvous with a pair of medium-sized circular black spots; postpronotal lobes and notopleura
yellow; scutum black except dark brown posterolateral to lateral postsutural vittae and dark red-brown
anterior to notopleural suture and inside postpronotal lobes, narrow lateral postsutural yellow vittae
tapering posteriorly to end before ia. seta; medial postsutural yellow vitta absent; mesopleural stripe
equal in width to notopleuron dorsally, scutellum yellow; legs with femora fulvous except for a small
subapical oval black spot on outer surfaces of fore femora and dark fuscous around apices of mid and
hind femora; fore and mid tibiae dark fuscous, hind tibiae black; wings with cells bc and c colourless,
microtrichia in outer corner of cell c only; a narrow fuscous costal band confluent with R 2+3 and with a
sight swelling around apex of R4+5; a narrow fuscous anal streak; supernumerary lobe weak in
females, of medium development in males; abdominal terga III and IV orange-brown and generally
each with its own ‘T’ pattern consisting of a narrow- to medium-width transverse dark fuscous to black
band across anterior margin which expands to cover outer 1/3 of lateral margins; a narrow- to
medium-width medial longitudinal dark fuscous to black band, tergum V with a narrow- to mediumwidth medial longitudinal dark fuscous to black band and dark fuscous to black anterolateral corners
which may also meet along anterior margin, a pair of oval dark fuscous shining spots on tergum V;
abdominal sterna dark coloured.
8.4.30.2
8.4.30.3
Molecular
HOST RANGE
Recorded from five plant families, Araliaceae, Euphorbiaceae, Myrtaceae, Polygalaceae, Rosaceae.
More common in the Rosaceae (Peach, Pear and Himalayan Cherry).
DISTRIBUTION
Northern Thailand and Northern Vietnam
REMARKS
Bactrocera pyrifoliae is a pest species within the dorsalis complex and is morphologically unique in
possessing a dark ‘T’ shaped pattern on each tergum III, IV and V.
214
PEST STATUS
 Exotic
 A minor pest of stone and pome fruits
ATTRACTANT
Possible weak response to cue lure (Drew & Romig 2007)
FIGURES
Figure 87. Bactrocera pyrifoliae
215
8.4.31 Bactrocera (Zeugodacus) tau (Walker)
Common name:
Dasyneura tau
Dacus (Zeugodacus) tau
DIAGNOSIS
8.4.31.1
A medium sized species; face fulvous with a pair of medium sized circular to oval black spots;
postpronotal lobes and notopleura yellow; scutum black with large areas of red-brown centrally and
anterocentrally; lateral and medial postsutural vittae present; yellow spot anterior to mesonotal suture
in front of lateral postsutural vittae; mesopleural stripe reaching midway between anterior margin of
notopleuron and anterior npl. seta; scutellum entirely yellow; wing with a narrow dark fuscous costal
band overlapping R2+3 and expanding into a distinct apical spot and broad dark fuscous anal streak;
cells bc and c colourless; microtrichia in outer corner of cell c only; abdominal terga III-V fulvous with a
black ‘T’ pattern and anterolateral corners of terga IV and V with broad black markings; posterior lobe
of male surstylus short; female with apex of aculeus trilobed (Drew and Romig 2013).
8.4.31.2
8.4.31.3
Molecular
species in BOLD. Low divergence clades are apparent and when used in conjunction with other
information about the specimen this may be sufficient to make a confident identification.
HOST RANGE
Bactrocera tau has been recorded on hosts from nine families. These include: Arecaceae,
Curcurbitaceae, Fabaceae, Loganiaceae, Moraceae, Myrtaceae, Oleaceae, Sapotaceae and Vitaceae
(for a full list of recorded hosts see Allwood et al. 1999).
Scientific name
Common name
Scientific name
Common name
Cucumis melo
melon
Manilkara zapota
sapodilla
Cucumis sativus
cucumber
Momordica charantia
bitter gourd
Cucurbita maxima
giant pumpkin
Psidium guajava
guava
Luffa acutangula
angled luffa
216
DISTRIBUTION
India, Sri Lanka, Bhutan, Vietnam, Southern China, Taiwan, Thailand, Peninsular Malaysia,
Singapore, East Malaysia, Brunei and Indonesian provinces (Drew and Romig 2013).
REMARKS
Bactrocera tau is a very common species throughout southeast Asia. It is an economic pest species,
mainly in cucurbit crops, but can be misidentified as it belongs to a complex of closely related species.
The tau-complex includes Zeugodacus species with a black scutum, wings colourless except for a
costal band and cubital streak, cells bc and c colourless or with an extremely pale tint, costal band
overlapping R2+3 and expanding into a distinct spot at apex. Bactrocera tau is distinct in having an
entirely yellow scutellum, abdominal terga III-V with a distinct dark ‘T’ pattern and all femora with dark
preapical spots (Drew and Romig 2013).
PEST STATUS

Exotic

A major pest of cucurbit crops
ATTRACTANT
Cue lure.
217
FIGURES
Figure 88. Bactrocera tau
218
University
219
8.4.32 Bactrocera (Bactrocera) trilineola (Drew)
Common name:
Dacus (Strumeta) triseriatus
DIAGNOSIS
8.4.32.1
Medium sized species; face entirely glossy black; postpronotal lobes fuscous to black; notopleura
yellow; scutum glossy black; mesopleural stripe reaching midway between anterior margin of
notopleuron and anterior npl. seta, lateral and medial postsutural vittae absent, scutellum glossy black
with lateral margins yellow; wing with a narrow extremely pale fuscous costal vein and broad fuscous
anal streak, a narrow fuscous transverse band across wing, cells bc and c extremely pale fuscous,
microtrichia covering outer ½ of cell c only; abdominal terga mostly glossy black except for two broad
longitudinal fulvous bands on terga II-V either side of a broad medial longitudinal glossy black band;
posterior lobe of male surstylus short; female with aculeus tip needle shaped (Drew 1989; pers. comm.
Drew 2010). For potentially morphologically confounding species see REMARKS or Table 8.
8.4.32.2
Molecular
PCR-RFLP Test 2: B. trilineola cannot be distinguished from B. kirki or B. frauenfeldi. No distinctive
Bactrocera and species in other genera. See Appendices 3 and 4.
HOST RANGE
Bactrocera trilineola has been recorded on hosts from 17 plant families. These include:
Anacardiaceae, Annonaceae, Caricaceae, Caesalpinaceae, Combretaceae, Lauraceae, Moraceae,
Musaceae, Myrtaceae, Oxalidaceae, Rutaceae and Sapindaceae (for a full list of recorded hosts see
Leblanc et al. 2012).
Major commercial hosts:
Scientific name
Common name
Mangifera indica
mango
DISTRIBUTION
Restricted to Vanuatu where it is common over nearly every island (Drew 1989). The record from New
Caledonia (Leblanc et al. 2012) is doubtful.
220
REMARKS
Bactrocera trilineola belongs to the frauenfeldi complex. It differs from B. caledoniesis and B.
frauenfeldi in possessing a glossy black face and in lacking lateral postsutural vittae and from B.
parafrauenfeldi in having a glossy black face, cells bc and c extremely pale fuscous, microtrichia in
outer ½ of cell c only, costal band present but very pale beyond subcostal cell and legs fulvous except
apical 1/3 of hind femora and hind tibiae fuscous. The apex of piercer and the spicules on the middle
segment of the ovipositor are similar in B. frauenfeldi and B. trilineola, however the apex of the
aculeus is slightly more pointed in B. trilineola. See Table 8 on page 128 for comparisons with other
frauenfeldi complex species.
PEST STATUS

Exotic
ATTRACTANT
FIGURES
Figure 91. Bactrocera trilineola
221
Figure 92. Bactrocera trilineola
222
8.4.33 Bactrocera (Bactrocera) trivialis (Drew)
Common name:
Dacus (Strumeta) trivialis
DIAGNOSIS
8.4.33.1
Medium sized species; medium sized pear shaped facial spots present; postpronotal lobes and
notopleura yellow; scutum black, mesopleural stripe ending midway between anterior margin of
notopleuron and anterior npl. seta, lateral postsutural vittae present, medial postsutural vitta absent,
scutellum yellow; wing with a narrow fuscous costal band and anal streak, cells bc and c colourless,
microtrichia in outer corner of cell c only; males with all leg segments fulvous except hind tibiae
fuscous, females with dark colour patterns on femora and tibiae; abdominal terga III-V generally black
with a medial longitudinal fulvous area from posterior margin of tergum III to tergum V; posterior lobe
of male surstylus short; female with apex of aculeus needle shaped (Drew 1989; pers. comm. Drew
2010).
8.4.33.2
Molecular
chosen to distinguish B. trivialis from other Bactrocera and species in other genera. See Appendices 3
and 4
HOST RANGE
Bactrocera trivialis has been recorded on hosts from ten families. These include: Anacardiaceae,
Combretaceae, Euphorbiaceae, Myrtaceae, Rosaceae, Rutaceae, Santalaceae and Solanaceae (for a
full list of recorded hosts see Leblanc et al. 2012).
Scientific name
Common name
Scientific name
Common name
Capsicum frutescens
chilli
Prunus persica
peach
Citrus x paradisi
grapefruit
Psidium guajava
guava
Magnifera indica
mango
223
DISTRIBUTION
Mainland Papua New Guinea (less common in the Highlands than at low elevations), Indonesia (Irian
Jaya) (Drew 1989). The record from Sulawesi (White and Elson-Harris 1992) is doubtful.
Although Bactrocera trivialis is not established in the Torres Strait Islands, occasional incursions do
occur. They are promptly eradicated under the Exotic Fruit Fly in Torres Strait Response Plan jointly
run by the Federal Department of Agriculture and Queensland Department of Agriculture and
Fisheries.
REMARKS
A large collection of specimens reared from grapefruit at Mt. Hagen, 1980, 1981, show sexual
dimorphism in leg colour patterns: females possess fore, mid and apical 1/3 of hind femora dark
fuscous, fore tibiae and apical four segments of fore tarsi fuscous, hind tibiae dark fuscous; males
have all segments fulvous except hind tibiae fuscous.
B. trivialis can appear similar to B. rufofuscula, a cue-responsive north Queensland rainforest species.
However, B. trivialis has a black scutum whereas B. rufofuscula has a dark brown scutum.
PEST STATUS

ATTRACTANT
Cue lure.
FIGURES
Figure 93. Bactrocera trivialis
224
Figure 94. Bactrocera trivialis
225
8.4.34 Bactrocera (Bactrocera) tryoni (Froggatt)
Common name: Queensland fruit fly
Tephritis tryoni
Dacus tryoni
Chaetodacus tryoni
Chaetodacus tryoni var. juglandis
Chaetodacus tryoni var. sarcocephali
Dacus (Strumeta) tryoni
Strumeta tryoni
Dacus (Bactrocera) tryoni
DIAGNOSIS
8.4.34.1
yellow; scutum red-brown with fuscous markings, mesopleural stripe reaching midway between
anterior margin of notopleuron and anterior npl. seta, lateral postsutural vittae present, medial
postsutural vitta absent, scutellum yellow; wing with a narrow fuscous costal band and broad fuscous
anal streak, cells bc and c fuscous, microtrichia covering cell c and outer ½ of cell bc; abdominal terga
III-V generally red-brown with a medial and two broad lateral longitudinal fuscous bands over all three
terga and joined along anterior margin of tergum III; paler forms of the abdomen are often present;
posterior lobe of male surstylus short; female with apex of aculeus needle shaped (Drew 1989; pers.
comm. Drew 2010).
8.4.34.2
8.4.34.3
Molecular
DNA barcoding: BOLD reference data available, but cannot be distinguished from B. neohumeralis or
B. aquilonis
BsrI:
200, 600
SnaBI: DNC
HhaI:
640, 190
SspI:
180, 570
HinfI:
770
Vspl:
DNC
Sau3AI: 420
sizes (bp) are 1000, 550 and 100. This species cannot be distinguished from B. aquilonis or B.
neohumeralis. Other restriction patterns useful for diagnosis of the complex are listed in Appendix 3,
with choice based on relative distinctiveness from other species potentially trapped; descriptions of the
patterns are given in Appendix 4.
226
Allozyme electrophoresis: Section 7.3.5.
HOST RANGE
Bactrocera tryoni has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Apocynaceae, Arecaceae, Cactaceae, Capparaceae, Caricaceae,
Celastraceae, Clusiaceae, Combretaceae, Curcurbitaceae, Cunoniaceae, Davidsoniaceae,
Ebenaceae, Elaeocarpaceae, Ericaceae, Euphorbiaceae, Fabaceae, Flacourtiaceae, Goodeniaceae,
Hippocraterceae, Juglandaceae, Lauraceae, Lecythidaceae, Loganiaceae, Malpighiaceae,
Melastomataceae, Meliaceae, Moraceae, Musaceae, Myrtaceae, Oleaceae, Oxalidaceae,
Passifloraceae, Punicaceae, Rhamnaceae, Rosaceae, Rubiaceae, Rutaceae, Santalaceae,
Sapindaceae, Sapotaceae, Smilacaceae, Solanaceae, Thymeliaceae, Tiliaceae, Verbenaceae,
Vitaceae (for a full list of recorded hosts see Hancock et al. 2000; Leblanc et al. 2012).
Major hosts (Hancock et al. 2000):
Scientific name
Common name
Scientific name
Common name
cashew
Mangifera indica
mango
Annona atemoya
atemoya
Manikara zapota
sapodilla
Annona glabra
pond apple
Morus nigra
mulberry
Annona muricata
soursop
Passiflora edulis
passionfruit
Annona reticula
bullock’s heart
Passiflora suberosa
corky passionfruit
Averrhoa carambola
carambola
Prunus persica
peach
Capsicum annuum
capsicum
Prunus persica var.
nucipersia
nectarine
Capsicum annuum
chilli
Psidium cattleianum
(=littorale)
cherry guava
Carica papaya
papaya
Psidium guajava
guava
Casimiroa edulis
white sapote
tomato
Chryosphyllum cainito
star apple
Syzgium aqueum
water apple
Coffea arabica
coffee
Syzygium forte ssp. forte
white apple
Eugenia uniflora
Brazilian cherry
Syzygium jambos
wax jambu
Eriobotrya japonica
loquat
Syzygium malacense
Malay apple
Fortunella japonica
kumquat
Syzygium suborbiculare
red bush apple
Malus sylvestris
apple
Syzygium tierneyanum
river cherry
DISTRIBUTION
Occurs in large populations throughout eastern Australia from Northern Territory, Cape York
(Queensland) to East Gippsland (Victoria) (Hancock et al. 2000; Dominiak and Daniels 2011, Royer &
Hancock 2012). The distribution was artificially altered by the pest free region known as the Fruit Fly
Exclusion Zone (FFEZ) however following the closure of the FFEZ, this species is deemed endemic in
Northern Territory, Queensland, Victoria and New South Wales (Dominiak and Mapson in prep.). It is
also established in New Caledonia, Austral Islands, many islands of the society group, and has been
eradicated from Easter Island (Drew et al. 1982). Despite three specimens being recorded from Papua
New Guinea, it is most doubtful that this species is established there (Drew 1989).
227
REMARKS
Bactrocera tryoni is similar to B. aquilonis (tryoni complex) in the general patterns of the wing, thorax
and abdomen but Bactrocera tryoni differs in having dark fuscous patterns on the scutum and the
abdomen. In B. aquilonis the scutum and abdomen are generally pale red-brown (Drew 1989). These
species can also be separated on the differences on the ovipositors: apex of aculeus rounded and
spicules with 7-10 uniform dentations in B. tryoni compared with the more pointed aculeus and uneven
dentations in B. aquilonis (Drew 1989). However, these differences are not easily observed (Cameron
et al. 2010).
In north Queensland cue traps B. tryoni can also appear similar to B. breviaculeus and B. kraussi. It
differs from both species in having tapered slightly shorter vittae and microtrichia in both costal cells.
Bactrocera kraussi also has a broad basal band on the scutellum, narrow mesopleural stripe and dark
apices of the femora.
PEST STATUS

Endemic

Bactrocera tryoni is the major fruit fly pest species in eastern Australia and is the target of
major control and quarantine programmes
ATTRACTANT
Cue group, Raspberry ketone formate (Melolure) was found to be a stronger attractant than cue-lure in
north Queensland (Royer 2015) but less attractive than cue-lure in Sydney (Dominiak et al. 2015a).
Combinations of cue lure and methyl eugenol are more attractive in the drier inland but less so in
Sydney (Dominiak et al. 2011). Zingerone is equally attractive as cue lure in Sydney (Dominiak et al.
2015b). Wet food lures such as citrus juice or protein attracted more males than females but remain
the only lures attractive to female B. tryoni (Dominiak and Nicol 2010). Male lures are much more
effective lures compared with wet food lures (Dominiak and Nicol 2010).
228
FIGURES
Figure 95. Bactrocera tryoni
Image courtesy of the Department of Economic Development, Jobs, Transport and Resources
229
230
8.4.35 Bactrocera (Tetradacus) tsuneonis (Miyake)
Common Name: Citrus fruit fly
Dacus tsuneonis Miyake
Dacus cheni Chao
DIAGNOSIS
8.4.35.1
A large species; face fulvous with a pair of small to medium-sized oval black spots; postpronotal lobes
and notopleura yellow; a lateral yellow band connecting postpronotal lobe and notopleuron; scutum
red-brown; lateral and medial postsutural yellow vittae present; no yellow spot anterior to notopleural
suture; mesopleural stripe reaching anterior npl. seta dorsally; scutellum yellow; wing with cells bc and
c with a strong pale fuscous tint, microtrichia in outer corner of cell c only; a broad fuscous to pale
fuscous costal band confluent with R4+5 at base and overlapping this vein towards apex of wing; anal
streak reduced to pale fuscous within cell cup; abdominal terga III-V orange-brown to fulvous with a
dark ’T’ pattern consisting of a narrow transverse dark fuscous band across anterior margin of tergum
III and a narrow medial longitudinal black band over all three terga and in some specimens fuscous to
dark fuscous in anterolateral corners of tergum IV.
8.4.35.2
8.4.35.3
Molecular
HOST RANGE
Recorded from family Rutaceae (Citrus and Fortunella species)
DISTRIBUTION
Japan (Ryukyu Is.); Southern China
REMARKS
Bactrocera tsuneonis is similar to B. minax in general body shape and colour patterns and these two
species may be confused. B. tsuneonis can be distinguished in possessing a yellow band running
between the postpronotal lobe and notopleuron, a.sa. setae and the costal band lacking a dark
fuscous spot in the apex.
PEST STATUS
 Exotic
 A minor pest of citrus
231
ATTRACTANT
No known record
FIGURES
Figure 99. Bactrocera tsuneonis
Image courtesy of M. Baker, International Centre for the Management of Pest Fruit Flies, Griffith University
232
8.4.36 Bactrocera (Bactrocera) tuberculata (Bezzi)
Common Name:
Chaetodacus tuberculatus Bezzi
DIAGNOSIS
8.4.36.1
A medium-sized species; face fulvous with a pair of large circular black spots; postpronotal lobes and
notopleura yellow, scutum mostly black; lateral postsutural yellow vittae present; medial postsutural
yellow vitta absent; no yellow spot anterior to notopleural suture; mesopleural stripe reaching midway
between anterior margin of notopleuron and anterior npl. seta dorsally; scutellum yellow; wing with
cells bc and c colourless, microtrichia in extreme outer corner of cell c only; a narrow pale fuscous to
fuscous costal band confluent with R2+3 and broken beyond apex of this vein and with a small fuscous
to dark fuscous spot around apex of R4+5; a narrow pale fuscous anal streak confined within cell cup;
abdominal terga III-V entirely black.
8.4.36.2
8.4.36.3
Molecular
HOST RANGE
Recorded from 8 plant families, some of which include commercial/edible fruit crops
DISTRIBUTION
Bhutan, Myanmar, Southern China, Thailand, Vietnam
REMARKS
Bactrocera tuberculata is similar to B. correcta and B. zonata in possessing an incomplete costal band
that ends at the apex of cell sc. and a fuscous spot around apex of vein R4+5. It differs from these
species in having abdominal terga III-V mostly black.
PEST STATUS
 Exotic
 A minor pest of mango, papaya and some Syzygium species
ATTRACTANT
Methyl eugenol
233
FIGURES
Figure 100. Bactrocera tuberculate
Image courtesy of Y. Martin, International Centre for the Management of Pest Fruit Flies, Griffith University
234
8.4.37 Bactrocera (Bactrocera) umbrosa
(Fabricius)
Common name: Breadfruit fruit fly
Dacus umbrosus
Strumeta umbrosa
Dacus (Strumeta) umbrosus
Dacus (Bactrocera) umbrosus
DIAGNOSIS
8.4.37.1
yellow; scutum black, mesopleural stripe reaching to postpronotal lobe, lateral postsutural vittae
present, medial postsutural vitta absent, scutellum yellow; wing with a broad fuscous costal band and
anal streak, three transverse reddish-fuscous bands across wing with the basal one joining with the
anal streak, cells bc and c fulvous with microtrichia in outer ½ of cell c only; abdominal terga varying
from orange-brown with a medial longitudinal black stripe on terga IV and V to orange-brown with a
broad medial and two broad longitudinal black bands over terga III-V; posterior lobe of male surstylus
short; female with apex of aculeus needle shaped (Drew 1989; Drew and Romig 2013).
8.4.37.2
8.4.37.3
Molecular
BsrI:
DNC
SnaBI: DNC
HhaI:
600, 190
SspI:
680
HinfI:
730
Vspl:
DNC
Sau3AI: 380
PCR-RFLP Test 2: AluI, DraI and RsaI produce diagnostic patterns (Appendix 3 and 4)
HOST RANGE
Bactrocera umbrosa has been recorded on hosts from only the family Moraceae (for a full list of
recorded hosts see Allwood et al. 1999; Leblanc et al. 2012).
Scientific name
Common name
Scientific name
Common name
Artocarpus altilis
breadfruit
Artocarpus heterophyllus
jackfruit
235
DISTRIBUTION
Widespread and very common in Malaysia, southern Thailand, Philippines, Indonesia, Palau, Papua
New Guinea (much less common in the Highlands), Solomon Islands, Vanuatu and New Caledonia
(Drew and Romig 2013).
REMARKS
Bactrocera umbrosa bears no close resemblance to other species. It is easily recognised by the three
broad transverse bands across the wings which are red-brown, not the usual fuscous colour (Drew
1989; Drew and Romig 2013). However, there are a few other PNG ME-responsive non-pest species
that have bands on the wings and similar pattern on the abdomen that may superficially appear similar
e.g. B. seguyi and B. confluens.
PEST STATUS

Exotic

Major pest of Artocarpus species
ATTRACTANT
Methyl eugenol.
FIGURES
Figure 101. Bactrocera umbrosa
236
Figure 102. Bactrocera umbrosa
Image courtesy of S. Sands and the International Centre for the Management of Pest Fruit Flies, Griffith
University
237
8.4.38 Bactrocera (Notodacus) xanthodes (Broun)
Common name: Pacific fruit fly
Tephrites (Dacus) xanthodes
Dacus (Tephrites) xanthodes
Chaetodacus xanthodes
Dacus xanthodes
Notodacus xanthodes
Dacus (Notodacus) xanthodes
DIAGNOSIS
8.4.38.1
Medium sized species; small black facial spots present; postpronotal lobes fulvous except for a broad
yellow band on posterior 2/3; notopleura orange-brown; scutum transparent with a shining orangebrown colouration and with irregular dark markings, broad lateral yellow band running from
postpronotal lobe to end just before anterior end of lateral postsutural vitta, large yellow spot on
pleural region in place of the normal mesopleural stripe, lateral postsutural vittae present and
beginning anterior to mesonotal suture, medial postsutural vitta present, scutellum orange-brown with
lateral yellow margins, wing with a narrow fuscous costal band and a broad fulvous anal streak, cells
bc and c extremely pale fulvous with microtrichia in outer corner of cell c only, abdominal terga
transparent and shining orange-brown with no dark markings; posterior lobe of male surstylus short;
female with apex of aculeus needle shaped (Drew 1989; pers. comm. Drew 2010).
8.4.38.2
8.4.38.3
Molecular
BsrI:
DNC
SnaBI: DNC
HhaI:
670, 200
SspI: 380, 250
HinfI:
680
Vspl:
DNC
Sau3AI: DNC
PCR-RFLP Test 2: AluI, DraI, RsaI and SspI produce diagnostic patterns.
AluI: 700-660, 200,170,130,120,110; DraI: 1150-1100, 250-240. <100, RsaI: 750,370,360; SspI:
1200, 210, 80
238
HOST RANGE
Bactrocera xanthodes has been recorded on hosts from twenty plant families. These include:
Anacardiaceae, Annonaceae, Apocynaceae, Caricaceae, Combretaceae, Euphorbiaceae, Lauraceae,
Lecythidaceae, Moraceae, Passifloraceae, Rutaceae and Sapotaceae (for a full list of recorded hosts
see Leblanc et al. 2012).
Scientific name
Common name
Scientific name
Common name
Artocarpus altilis
breadfruit
Carica papaya
pawpaw
DISTRIBUTION
Fiji Islands, Tonga, Niue, Samoa, American Samoa, Southern group of Cook Islands, Wallis and
Futuna. Introduced on Nauru (first detected in 1992) but subsequently eradicated by male annihilation.
Detected in April 1998 on Raivavae (French Polynesia) but subsequently eradicated by male
annihilation (Drew 1989; Leblanc et al. 2012).
REMARKS
Bactrocera xanthodes is a unique species having a pair of well-developed postpronotal lobe setae, the
transparent integument on the head, thorax and abdomen, a soft integument particularly noticeable on
the abdomen where the terga fold ventrally in dead specimens (Drew 1989).
Other remarks:
Bactrocera xanthodes belongs to subgenus Notodacus, an unusual feature of which is the presence of
a seta on each postpronotal lobe (i.e. shoulder). It has a very distinct V-shaped notch in the apex of its
scutellum. Bactrocera paraxanthodes has this to a lesser extent. Another unusual feature of B.
xanthodes is that the lateral stripes (vittae) on the scutum extend forward to the postpronotal lobes
and back down the sides of the scutellum. There is also a medial yellow stripe that extends to the
posterior edge of the scutum (immediately before the scutellum); this stripe is shorter in B.
paraxanthodes. The most obvious difference between the closely related B. paraxanthodes and B.
xanthodes is that B. xanthodes has yellow lateral margins to the scutellum while B. paraxanthodes has
dark margins (CABI 2007).
PEST STATUS

Exotic

High priority pest identified in the Avocado and Tropical fruit IBPs
ATTRACTANT
Methyl eugenol.
239
FIGURES
Figure 103. Bactrocera xanthodes
Figure 104. Bactrocera xanthodes
240
8.4.39 Bactrocera (Bactrocera) zonata (Saunders)
Common name: Peach fruit fly
Dasyneura zonatus
Dacus (Strumeta) zonatus
DIAGNOSIS
8.4.39.1
Face fulvous with a pair of medium sized oval black spots; scutum red-brown with pale fuscous
patterning posteriorly; postpronotal lobes and notopleura yellow; mesopleural stripe reaching to or
almost to anterior npl. seta dorsally; medium width parallel sided lateral postsutural vittae ending at or
just behind ia. seta; medial postsutural vitta absent; scutellum yellow; legs with all segments entirely
fulvous except apices of femora red-brown and hind tibiae pale fuscous to fuscous; wings with cells bc
and c colourless and entirely devoid of microtrichia, a narrow fuscous costal band confluent with R 2+3
and ending at apex of this vein, a small oval fuscous spot across apex of R 4+5, anal streak reduced to
a pale tint within cell cup; supernumerary lobe of medium development; abdominal terga III-V redbrown with a ‘T’ pattern consisting of a narrow transverse black band across anterior margin of tergum
III (this band is often broken in the central region) and a narrow medial longitudinal black band over all
three terga (this band is often reduced to a stripe over parts of terga IV and V), narrow anterolateral
fuscous corners on terga IV and V, a pair of oval red-brown shining spots on tergum V; posterior lobe
of male surstylus short; female with apex of aculeus needle shaped (Drew and Romig 2013).
8.4.39.2
Molecular
BsrI:
600, 200
SnaBI: 535, 330
HhaI:
680, 190
SspI: 750, 120
HinfI:
DNC
Vspl:
DNC
Sau3AI: DNC
chosen to distinguish B. zonata from other Bactrocera and species in other genera. See Appendices 3
and 4
241
HOST RANGE
Bactrocera zonata has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Arecaceae, Caricaceae, Combretaceae, Curcurbitaceae, Fabaceae,
Lecythidaceae, Malpighiaceae, Malvaceae, Myrtaceae, Punicaceae, Rosaceae, Rutaceae and
Tiliaceae (for a full list of recorded hosts see Allwood et al. 1999; Tsuruta et al. 1997).
Scientific name
Common name
Scientific name
Common name
Mangifera indica
mango
Psidium guajava
guava
Prunus persica
peach
DISTRIBUTION
Bhutan, Sri Lanka, India, Pakistan, Thailand, Vietnam, Mauritius, Egypt, UAE (Drew and Romig 2013).
REMARKS
Bactrocera zonata is a red brown species that is similar in general appearance to B. tryoni. It is easily
distinguished from B. tryoni in having the costal band interrupted beyond apex of R2+3. Bactrocera
correcta possess a similar costal band but has a black scutum and a black ‘T’ pattern on abdominal
terga III-V (Drew and Romig 2013).
PEST STATUS

Exotic


In India, Pakistan and now Egypt, it is an important fruit fly pest and causes severe damage to
peach, guava and mango
ATTRACTANT
Methyl eugenol.
242
FIGURES
Figure 105. Bactrocera zonata
Image courtesy of A. Carmichael and the International Centre for the Management of Pest Fruit Flies, Griffith
University
243
Species diagnosis: Ceratitis (Tephritidae: Dacinae:
Ceratitidini)
8.5.1 Ceratitis capitata (Wiedemann)
Common name: Mediterranean fruit fly
Trypeta capitata
DIAGNOSIS
8.5.1.1
In Australia, there are no species of Ceratitis that look similar to C. capitata. Consequently, the
following characters can be used to distinguish Ceratitis capitata from all other species of Tephritidae
occurring in Australia. Small to medium-sized, brightly coloured flies; scutellum swollen, rounded
above, shiny black with a thin sinuate yellow streak near base dorsally; scutum yellowish with
numerous black areas in a characteristic pattern; abdomen yellowish with two narrow transverse lightcoloured bands; wing relatively broad in comparison with its length, cloudy yellow, with three brown
bands on apical two-thirds, all separated from each other, and smaller dark irregular-shaped streaks
within the cells in the proximal half; cell cup with its apical extension short; males with a black
diamond-shaped expansion of the apex of the anterior orbital seta.
These characters also distinguish C. capitata from all other species in the genus wherever they may
occur worldwide. Several species of the subgenus Ceratitis closely resemble C. capitata in the
thoracic pattern, the apical expansion of cell cup, the presence of dark markings in the basal half of
the wing, and in having the anterior orbital bristle of the male modified in some way. In C. capitata, it is
black and resembles a diamond apically rather than some other shape (Foote, Blanc and Norrbom
1993).
8.5.1.2
Molecular
BsrI:
DNC
SnaBI: DNC
HhaI:
DNC
SspI:
520, 160
HinfI:
DNC
Vspl:
650, 200
Sau3AI: DNC
PCR-RFLP Test 2: DdeI and RsaI are fully diagnostic for C. capitata, although most of the others
provide additional diagnostic value.
DdeI: 1150, 270, 220, 130; RsaI: 450,380,290,260,240,110
Allozyme electrophoresis (Section 7.3.5).
244
HOST RANGE
Ceratitis capitata is a highly polyphagous species and its pattern of host relationships from region to
region appears to relate largely to what fruits are available (CABI 2007). It has been recorded on hosts
from a wide range of families. These include: Anacardiaceae, Annonaceae, Apocynaceae, Arecaceae,
Cactaceae, Caricaceae, Clusiaceae, Combretaceae, Ebenaceae, Juglandaceae, Lauraceae,
Lythraceae, Malpighiaceae, Malvaceae, Muntingiceae, Myrtaceae, Passifloraceae, Rosaceae,
Rubiaceae, Rutaceae, Santalaceae, Sapindaceae, Sapotaceae, Solanaceae and Vitaceae (for a full
list of recorded hosts see CABI 2007).
Scientific name
Common name
Scientific name
Common name
Annona cherimola
cherimoya
Malus domestica
apple
Capsicum annuum
bell pepper
Prunus spp.
stone fruit
Citrus spp.
citrus
Prunus salicina
Japanese plum
Coffea spp.
coffee
Psidium guajava
guava
Ficus carica
fig
Theobroma cacao
cocoa
DISTRIBUTION
Native to Africa, has spread to the Mediterranean region, southern Europe and Middle east, Western
Australia, Central and South America and Hawaii (PaDIL 2007). A review of the past and present
distribution of Ceratitis capitata in Australia (Dominiak and Daniels 2011). Now only found in Western
Australia, it has not been detected in any eastern State since the 1950’s.
REMARKS
The males of Ceratitis capitata are easily separated from all other members of the family by the black
pointed expansion at the apex of the anterior pair of orbital setae. The females can be separated from
most other species by the characteristic yellow wing pattern and the apical half of the scutellum being
entirely black (White and Elson-Harris 1992).
PEST STATUS

Endemic

High priority pest identified in the Mango IBP

Ceratitis capitata is an important pest in Africa and has spread to almost every other continent
to become the single most important pest species in the family Tephritidae

It is ecologically adapted to regions of Mediterranean climate and less of a problem in
subtropical and tropical areas although it can still be damaging in elevated tropical regions.
ATTRACTANT
Trimedlure/capilure and terpinyl acetate.
245
FIGURES
Figure 106. Ceratitis capitata
Image courtesy of Scott Bauer, USDA Agricultural Research Service, Bugwood.org
Figure 107. Ceratitis capitata
246
8.5.2 Ceratitis (Pterandrus) rosa (Karsch)
Common name: Natal fruit fly
Pterandrus rosa
DIAGNOSIS
8.5.2.1
Head: Anterior pair of orbital setae not modified in any way.
Thorax: Scutellum marked black and yellow, with yellow lines or areas meeting margin, such that
each apical scutellar seta is based in or adjacent to a yellow stripe; male mid-femora without stout
ventral setae; mid-tibiae with rows of stout setae along the distal half of both the anterior and posterior
edges giving a feathered appearance. Wing length 4-6 mm.
The males of most species of subgenus Pterandrus have rows of stout setae on both the anterior and
posterior edges of each mid-tibia, giving a feathered appearance. Ceratitis rosa can be separated from
most other members of this subgenus by having this feathering confined to the distal half of the tibia
and by lacking stout setae on the underside of the mid-femur. The males also lack the spatulate head
appendages of subgenus Ceratitis. Unfortunately, there is no simple method of recognizing females,
except that Pterandrus species tend to have brown wing bands and a generally brown body colour,
which contrasts with the yellow markings of C. capitata.
8.5.2.2
Molecular
species in BOLD. Low divergence clades are apparent and when used in conjunction with other
information about the specimen may be sufficient to make a confident identification
BsrI:
DNC
SnaBI: DNC
HhaI:
DNC
SspI:
570, 480
HinfI:
800, 200
Vspl:
600, 300
Sau3AI: DNC
PCR-RFLP Test 2: Fully diagnostic restriction enzymes are DdeI, HinfI, RsaI, Sau3A, SspI ad all other
enzymes have some restriction value.
DdeI: 1500, 270, 220, 170; HinfI: 780, 410,330,280,150,120; RsaI: 410, 380, 310, 290, 260, 240, 130,
110; Sau3A: 1400, 770,110; SspI: 1000, 490, 300, 240, 150
Allozyme electrophoresis: (Section 7.3.5).
247
HOST RANGE
Ceratitis rosa has been recorded on hosts from a wide range of families. These include:
Anacardiaceae, Annonaceae, Apocynaceae, Caricaceae, Clusiaceae, Combretaceae, Lauraceae,
Malvaceae, Moraceae, Myrtaceae, Oxalidaceae, Rhamnaceae, Rosaceae, Rubiaceae, Rutaceae,
Sapindaceae, Sapotaceae, Solanaceae, and Vitaceae (for a full list of recorded hosts see CABI 2007).
Scientific name
Common name
Scientific name
Common name
Coffea arabica
coffee
Prunus persica
peach
Citrus spp.
citrus
Psidium spp.
guava
DISTRIBUTION
Angola, Ethiopia, Kenya, Malawi, Mali, Mozambique, Nigeria, Republic of South Africa (KwaZulu
Natal), Rwanda, Rhodesia, Swaziland, Tanzania, Uganda, Zaire, and the islands of Mauritius and
Reunion (UF & FDACS 2009).
REMARKS
Ceratitis rosa is best recognised by its characteristic pattern of brown wing bands, the three black
areas in the apical half of the scutellum, and by the male having feathering on the mid tibiae, but no
feathering on the mid femora (White and Elson-Harris 1992). This fruit fly closely resembles the
Mediterranean fruit fly in appearance. It averages slightly larger and has characteristic picture wings
and dark black spots on the thorax. The arista of the antenna is plumose, while that of the C. capitata
bears only short pubescence. The front of the male lacks the pair of conspicuous spatulate setae
which is found on the male C. capitata. The mesothoracic tibiae of the males are clothed with dorsal
and ventral brushes of elongated bluish-black scales, lacking in the C. capitata. The ovipositor sheath
of the female is shorter than the width at its base. Length of the fly 4 to 5 mm (UF & FDACS 2009).
PEST STATUS

Exotic

High priority pest identified in the Mango IBP

Ceratitis rosa is highly polyphagous and causes damage to a very wide range of unrelated
fruit crops
ATTRACTANT
Trimedlure and terpinyl acetate.
248
FIGURES
Figure 108. Ceratitis rosa
Figure 109. Ceratitis rosa
249
Species diagnosis: Dacus (Tephritidae)
8.6.1 Dacus (Callantra) longicornis (Wiedemann)
Common Name:
See Drew & Romig (2013), p. 373.
DIAGNOSIS
8.6.1.1
A large species; face fulvous with a pair of small irregularly oval black spots; postpronotal lobes yellow
except posterodorsal corners fuscous; notopleura yellow; scutum dark red-brown without distinct dark
patterns; lateral and medial postsutural yellow vittae absent; a narrow yellow triangle running along
anterior margin of notopleural suture with base on notopleuron; mesopleural stripe narrow equal in
width to notopleuron dorsally; scutellum yellow except for broad red-brown basal band; setae: sc. 2;
prsc. absent; ia. 1; p.sa. 1; a.sa. 1; mpl. 1; npl. 2; scp. 4; legs with fore femora dark red-brown to
fuscous, mid femora dark red-brown to fuscous except fulvous on basal 1/4, hind femora dark fuscous;
fore and mid tibiae dark red-brown to fuscous, hind tibiae dark fuscous; fore tasi with all segments
dark red-brown, mid and hind tarsi with basal segment fulvous and apical four segments red-brown;
wing with cells bc and c fuscous, dense microtrichia over all of cell c and most of cell bc; a broad dark
fuscous costal band overlapping R4+5 for its entire length and sometimes becoming darker at apex;
anal streak indistinct but a broad pale fuscous area generally over cell cup and across wing margin
towards cell dm; supernumerary lobe weak; abdomen strongly petiolate, abdominal terga III-V
generally dark fuscous to black with a paler band often across posterior margin of tergum III, large
orange-brown spots posterocentrally on terga IV and V with the spot on tergum V often expanded
anteriorly into a medial longitudinal orange-brown band.
8.6.1.2
8.6.1.3
Molecular
HOST RANGE
Recorded from family Cucurbitaceae, primarily species of genus Luffa.
DISTRIBUTION
Widespread from Southern Asia to South-East Asia
250
REMARKS
Dacus longicornis possesses significant variability in colour patterns and thus some specimens may
be difficult to identify. It is distinguished by having a red-brown scutum, fuscous cells bc and c, the
costal band overlapping R4+5, face fulvous with a pair of black spots, fuscous anatergite, yellow
katatergite, postpronotal lobes mostly yellow and a narrow mesopleural stripe.
PEST STATUS
 Exotic
 A minor pest of native cucurbits, mainly Luffa species, in village situations
ATTRACTANT
Cue lure
FIGURES
Figure 110. Dacus longicornis
Image courtesy of S. Sands and the International Centre for the Management of Pest Fruit Flies,
Griffith University
251
8.6.2 Dacus (Callantra) solomonensis (Malloch)
Common Name:
Dacus (Callanatra) solomonensis Malloch
Callantra solomonensis (Malloch)
DIAGNOSIS
8.6.2.1
Large species; face with a pair of large oval black spots; postpronotal lobes yellow to yellow-brown;
notopleura yellow; scutum dark fuscous with a large irregular red-brown area on posterior region,
lateral and medial postsutural vittae absent, mesopleural stripe of medium width extending midway
between anterior margin of notopleuron and anterior npl. bristle, yellow triangle extending along
anterior margin of mesonotal suture, scutellum usually red-brown; setae: sc. 2, prsc. absent, a.sa. 1;
wing with a broad fuscous costal band and broad fulvous anal streak, costal cells orange-brown with
dense microtrichia covering both cells; abdominal terga III and IV dull black with orange-brown along
posterior margins and tergum V dark fuscous to dull black except for orange-brown medially and along
posterior margin, posterior lobe of surstylus short, abdominal sternum V with a slight concavity on
posterior margin.
8.6.2.2
8.6.2.3
Molecular
PCR-RFLP Test 2: AluI, DraI, RsaI and SspI provide diagnostic restriction patterns.
AluI: 720-770, 170, 150, 130, 120,110; DraI: 800-750, 270, 220, 180-160; RsaI: 450, 380,320,290;
SspI: 1100, 380
HOST RANGE
Reared from family Cucurbitaceae (pumpkins and cucumber)
DISTRIBUTION
Bougainville Island; Solomon Islands
REMARKS
Dacus solomonensis is a unique species in possessing a large hump on abdominal tergum V.
PEST STATUS
 Exotic
 A significant pest of some cucurbit crops in village situations in the area of distribution
252
ATTRACTANT
Cue lure
FIGURES
Figure 111. Dacus solomonensis
Image courtesy of S. Sands and the International Centre for the Management of Pest Fruit Flies,
Griffith University
253
Species diagnosis: Dirioxa (Tephritidae: Phytalmiinae)
8.7.1 Dirioxa pornia (Walker)
Common name: Island fly
Trypeta pornia
DIAGNOSIS
8.7.1.1
Head with arista plumose on dorsal surface, bare on ventral surface; thorax with scutum mostly redbrown, 6 scutellar setae; scutellum flat and bare of microsetae; legs with one strong apical spine on
mid tibiae; wing pattern as per Figure 113; abdominal terga fulvous with transverse black patterns on
terga III to V; male surstylus short and thick; female aculeus rounded and blunt at apex (pers. comm.
Drew 2010).
8.7.1.2
Molecular
BsrI:
DNC
SnaBI: DNC
HhaI:
DNC
SspI:
300, 220
HinfI:
DNC
Vspl:
DNC
Sau3AI: DNC
Allozyme electrophoresis (Section 7.3.5).
HOST RANGE
Dirioxa pornia attacks ripe, damaged and fallen fruit. It has been recorded on hosts from a wide range
of families. These include: Anacardiaceae, Annonaceae, Araucariaceae, Capparaceae, Caricaceae,
Clusiaceae, Combretaceae, Curcurbitaceae, Ebenaceae, Euphorbiaceae, Fabaceae, Lauraceae,
Lecythidaceae, Loganiaceae, Moraceae, Musaceae, Myrtaceae, Oleaceae, Oxalidaceae,
Passifloraceae, Proteaceae, Rosaceae, Rubiaceae, Rutaceae, Sapindaceae, Sapotaceae,
Solanaceae and Xanthophyllaceae (for a full list of recorded hosts see Hancock et al. 2000).
Major hosts: No major host fruits have been identified but has created occasional quarantine
problems.
DISTRIBUTION
Eastern Australia, from Iron Range, Cape York Peninsula, to southern New South Wales. Introduced
to Perth, Western Australia. (Hancock et al. 2000). Also in Northern Victoria.
254
REMARKS
Dirioxa spp. are the only tephritids with six setae on the scutellar margin, that are likely to be found in
fruit crops; the wing pattern is characteristic (White and Elson-Harris 1992).
PEST STATUS

Endemic
ATTRACTANT
Protein and citrus juice (Dominiak et aI. 2003) and is attracted to cue lure in some cases.
FIGURES
Figure 112. Dirioxa pornia
Image courtesy of Carroll et al., Pest fruit flies of the world, http://delta-intkey.com/ffa (as of 22 November 2015)
Figure 113. Dirioxa pornia
255
Species diagnosis: Rhagoletis (Tephritidae: Trypetinae:
Trypetini)
8.8.1 Rhagoletis cerasi (Linnaeus)
Common Name: European Cherry Fruit Fly
Musca cerasi Linnaeus
DIAGNOSIS
8.8.1.1
Rhagoletis cerasi possesses a unique wing colour pattern with a short broad transverse fuscous band
at base, a broad transverse fuscous band across centre from anterior margin to hind margin and
enclosing r-m crossvein, a short narrow transverse fuscous band from anterior margin to vein R4+5 and
an inverted L-shaped band at apex of wing, the stem of which encloses dm-cu crossvein.
8.8.1.2
8.8.1.3
Molecular
HOST RANGE
Recorded from family Caprifoliaceae (Lonicera species), family Rosaceae (Prunus species)
DISTRIBUTION
Northern Europe (except UK); Central Asia
REMARKS
Rhagoletis cerasi is distinguished by its unique wing colour patterns.
PEST STATUS
 Exotic
 A major pest of commercial cherries throughout its geographic range
ATTRACTANT
No known record
256
FIGURES
Figure 114. Rhagoletis cerasi
257
8.8.2 Rhagoletis cingulata (Leow)
Common name: Cherry fruit fly
Trypeta cingulata
DIAGNOSIS
8.8.2.1
Rhagoletis species are diagnosed primarily on wing patterns. The wing of R. cingulata possesses a
broad singular dark fuscous band from anterior margin to hind margin in centre of wing, an inverted
‘U’-shaped dark fuscous pattern across apex and a dark fuscous spot across apex of R4+5.
8.8.2.2
8.8.2.3
Molecular
HOST RANGE
A significant pest in commercial cherries in the eastern half of USA
DISTRIBUTION
Eastern half of USA
REMARKS
Rhagoletis cingulata is distinguished by its wing colour patterns. It is morphologically close to
Rhagoletis indifferens. See notes under R. indifferens to separate these two species.
PEST SPECIES
 Exotic
 A significant pest of cherries in USA
ATTRACTANT
No known species
258
FIGURES
Figure 115. Rhagoletis cingulata
259
8.8.3 Rhagoletis completa (Cresson)
Common name: Walnut husk fly
Rhagoletis suavis completa
Rhagoletis suavis var. completa
DIAGNOSIS
8.8.3.1
The four transverse wing bands are present and are usually distinctly separated by hyaline areas,
except in occasional specimens in which the discal and subapical bands are connected posteriorly. In
the former case, the pattern closely resembles that of R. berberis, but the host relationships of these
two species are quite different. The thorax and abdomen of R. completa are golden yellow (completely
black in R. berbeis) and the scutellum is concolorous yellow (black with a distinct yellow spot in R.
berberis) (Foote et al. 1993).
8.8.3.2
Molecular
species in BOLD. Low divergence clades may be apparent and used in conjunction with other
information about the specimen may be sufficient to make a confident identification
PCR-RFLP Test 2: AluI is fully diagnostic; DraI, HinfI, RsaI and SspI also have diagnostic value
AluI: 640, 450, 130, 120, 110
HOST RANGE
Rhagoletis completa has been recorded on hosts from two families, Juglandaceae and Rosaceae (for
a full list of recorded hosts see CABI 2007).
Scientific name
Common name
Scientific name
Common name
Juglans californica
California walnut
Juglans nigra
black walnut
Juglans hindsii
Californian black walnut
Juglans regia
walnut
DISTRIBUTION
Southern and Central USA including Mexico; adventive in Western USA since the 1920s. Also
established in Southern Europe since the early 1990s (CABI 2007).
260
REMARKS
Rhagoletis completa is an unusual economic tephritid species in that it is a major pest of walnuts, in
contrast to the soft fleshy fruit hosts of other fruit fly species. It is best diagnosed by the distinctive
wing colour patterns (Figure 116) and a red-brown thorax. It is widely distributed over Central and
Western mainland USA (pers. comm. Drew 2010).
PEST STATUS

Exotic

High priority pest identified in the Nuts IBP
ATTRACTANT
FIGURES
Figure 116. Rhagoletis completa
261
8.8.4 Rhagoletis fausta (Osten-Sacken)
Common name: Black cherry fruit fly
Trypeta (Acidia) fausta
Trypeta fausta
Rhagoletis fausta
Acidia fausta
DIAGNOSIS
8.8.4.1
Rhagoletis fausta is very close to R. striatella in that the posterior apical band in both species arises
from the subapical band in the vicinity of vein r-m, making an F-shaped pattern in the apical half of the
wing similar to that in the pomonella group (however, in the latter, note that the subapical band is
missing and the apical bands are connected to the discal band). In wing pattern alone, R. fausta is
unique among North American Rhagoletis in combining a very broad connection between the discal
and subapical bands in cell dm with the presence of both an anterior and posterior apical band, the
latter arising in much the same location as in R. striatella. In R. striatella, the discal and subapical
bands are separate or connected only along the posterior wing margin. In many respects, the wing
pattern of R. fausta resembles that of R. suavis, but R. fausta has both anterior and posterior apical
bands and an isolated hyaline spot in the distal half of cell cua1. Rhagoletis suavis has a yellowish
body; that of R. fausta is black and without yellowish bands at the posterior margins of the abdominal
terga (Foote et al. 1993).
8.8.4.2
Molecular
HOST RANGE
Rhagoletis fausta has been recorded on hosts the Rosaceae family (for a full list of recorded hosts see
CABI 2007).
Scientific name
Common name
Scientific name
Common name
Prunus avium
sweet cherry
Prunus cerasus
sour cherry
262
DISTRIBUTION
Widespread occurrence in western and eastern North America (United States and Canada) (CABI
2007).
REMARKS
Rhagoletis fausta is a dark coloured species with the scutum and abdominal tergites primarily black. It
also has a unique wing colour pattern (Figure 117). It is widely distributed over mainland USA where it
infests cherry varieties in the plant genus Prunus (pers. comm. Drew 2010).
PEST STATUS

Exotic

High priority pest identified in the Cherry IBP

Rhagoletis fausta is an important pest of cherries in North America
ATTRACTANT
FIGURES
Figure 117. Rhagoletis fausta
263
8.8.5 Rhagoletis indifferens (Curran)
Common name: Western cherry fruit fly
Rhagoletis cingulata indifferens
DIAGNOSIS
8.8.5.1
Rhagoletis indifferens is similar to R. cingulata in wing pattern but the anterior arm of R. indifferens is
broken to produce an apical spot in only about 5% of individuals in contrast to R. cingulata, in which
this spot is much more commonly encountered. In addition, other characters that distinguish R.
indifferens from R. cingulata are as follows:
Rhagoletis indifferens:
A. Apical yellow shading on posterior margin of tergite 5 of male lacking.
B. Black shading always present on posterior surface of fore coxa
C. Epandrium dark-coloured
Rhagoletis cingulata:
A. Apical yellow shading on posterior margin of tergite 5 of male present
B. Fore coxae concolorous yellow
C. Epandrium light-coloured
Most individuals of R. indifferens may be distinguished from those of R. chionanthi and R. osmanthi by
the differences in geographical distribution and hosts and by the generally smaller size and lesser
development of the wing bands. (Foote et al. 1993).
8.8.5.2
Molecular
264
HOST RANGE
Rhagoletis indifferens has been recorded on hosts the Rosaceae family (for a full list of recorded hosts
see CABI 2007).
Scientific name
Common name
Prunus avium
sweet cherry
DISTRIBUTION
Rhagoletis indifferens is a western North American species (Canada and United States). Adventive
populations have been found in southern Switzerland since the early 1990s (CABI 2007).
REMARKS
Rhagoletis indifferens may not be a distinct species but a colour variety of Rhagoletis cingulata. It is
distributed primarily over western regions of mainland USA where it infests wild and cultivated cherries
of the plant genus Prunus (pers. comm. Drew 2010).
PEST STATUS

Exotic

High priority pest identified in the Cherry IBP

Rhagoletis indifferens is an important pest of cherries in North America
ATTRACTANT
FIGURES
Figure 118. Rhagoletis indifferens
265
8.8.6 Rhagoletis mendax (Curran)
Common Name: Blueberry maggot
DIAGNOSIS
8.8.6.1
Rhagoletis mendax wing markings consist of broad dark fuscous basal band and an F-shaped pattern
across the entire membrane. It also lacks dark markings on the fore femora and the female aculeus is
shorter than in other species (aculeus length 0.73 – 0.89 mm).
8.8.6.2
8.8.6.3
Molecular
DNA barcoding: BOLD reference data available, but cannot be distinguished from R. pomonella
HOST RANGE
Recorded from 11 species of the genus Vaccinium (family Ericaceae). A serious pest of blueberries in
Eastern USA.
DISTRIBUTION
Eastern USA
REMARKS
Rhagoletis mendax has often been confused with R. pomonella. It is separated from R. pomonella by
the absence of dark markings on the fore femora and the shorter aculeus.
PEST STATUS
 Exotic
 A significant pest of blueberries in Eastern USA
ATTRACTANT
No known record
266
FIGURES
Figure 119. Rhagoletis mendax
Image courtesy of Carroll et al., Pest fruit flies of the world, http://delta-intkey.com/ffa (as of 14 December
2015)
267
8.8.7 Rhagoletis pomonella (Walsh)
Common name: Apple maggot
Trypeta pomonella
Trypeta (Rhagoletis) pomonella
DIAGNOSIS
8.8.7.1
Rhagoletis pomonella, together with R. zephyria, R. mendax and R. cornivora are among the most
readily recognised species of Rhagoletis by virture of their wing pattern, which consists of a slightly
oblique discal band to which the anterior and posterior apical bands are connected, forming a
characteristic F-shaped pattern in the apical half of the wing. The absence of the subapical band
distinguishes the species of the pomonella group from all other species of Rhagoletis. Rhagoletis
striatella, which also has an F-shaped apical wing pattern, is distinguished from R. pomonella by the
colour pattern of the scutellum and the additional characters given in the key to species. Rhagoletis
pomonella is separable from the other three species of the pomonella group by the presence in most
specimens of heavy black shading on the posterior surface of the fore femur, and, in specimens from
the northern part of its range, by a generally larger body size and by the longer aculeus (0.901.49mm). In the southern part of its range, specimens of R. pomonella generally are smaller than in
the north. For that reason and because of a consequently shorter aculeus, females are not separable
from those of R. mendax and R. cornivora by the use of morphological characters. Mexican specimens
of R. pomonella resemble those that occur in the United States and Canada but generally are larger
and possess a light spot near the base of the apical wing band (Foote, Blanc and Norrbom 1993).
8.8.7.2
Molecular
DNA barcoding: BOLD reference data available, but cannot be distinguished from R. mendax
PCR-RFLP Test 2: HinfI and RsaI are fully diagnostic; DraI also has some diagnostic value
HinfI: 640, 260,190,150, 110, 80, 70
HOST RANGE
Rhagoletis pomonella has been recorded on hosts from the Rosaceae family (for a full list of recorded
hosts see CABI 2007).
Scientific name
Common name
Malus domestica
apple
268
DISTRIBUTION
Canada, United States and Mexico (Carroll et al. 2002).
REMARKS
Rhagoletis pomonella possesses primarily a black scutum and abdomen and a distinctive wing colour
pattern (Figure 120). R. pomonella is difficult to distinguish from R. mendax. It has been the subject of
extensive taxonomic, ecological and pest management research in the USA and is considered the
major economic pest species within the genus Rhagoletis. It is a major pest of cultivated apples. It is
distributed over the central and north-eastern regions of mainland USA and extreme southern
Canada. In 1979 it was introduced into the western coastline of the USA and is now widespread in that
region (pers. comm. Drew 2010).
PEST STATUS

Exotic

High priority pest identified in the Apple and Pear, and Cherry IBPs

Rhagoletis pomonella, which primarily attacks apples, is the most serious fruit fly pest in North
America
ATTRACTANT
FIGURES
Figure 120. Rhagoletis pomonella
269
Species diagnosis: (Diptera: Drosophilidae)
8.9.1 Drosophila (Sophophora) suzukii
(Matsumura 1931)
Common name: Spotted Wing Drosophila (SWD)
Leucophenga suzukii
DIAGNOSIS
8.9.1.1
A factsheet (PHA 2013) and draft diagnostic identification protocol have been produced for Australia
(Blacket et al. 2015 b).
Drosophilidae are characterised by head with 1 pair of proclinate and 1 or 2 pairs of reclinate orbital
bristles; post-vertical bristles, when present, parallel or convergent; arista usually branched above and
below; mesopleuron bare; costa twice broken; auxiliary vein not reaching costal margin (Bock 1976).
Sophophora species are characterised by relatively narrow cheeks and possession (in most species)
on the abdominal tergites of dark posterior bands that are not interrupted in the mid-line (Bock 1980).
SWD has a uniform apricot coloured thorax that is free of stripes, spots or patterns, with unbroken
dorsal bands on its abdomen (Blacket et al. 2015 b).
Male SWD have a single apical wing spot (Figure 121A) and a single row of apical dark combs
running across (rather than parallel with) each of the basal two tarsal segments (Figure 121C).
Female SWD possess an ovipositor that is strongly sclerotized with robust teeth (typically with 30-36
teeth, Seljak 2013), very strongly developed along the lower half towards the ovipositor tip with a sawlike lower edge (Figure 121B), the ovipositor is 6-7 times longer than the diameter of the spermatheca
(only 2-3 times longer in most other Drosophila species) (Hauser 2011).
8.9.1.2
Molecular
DNA barcoding: BOLD reference data is available
Published molecular tests: DNA barcoding has been employed by to confirm SWD in Europe
(Calabria et al. 2012). A PCR-RFLP test has been developed by Kim et al. (2014). A High Resolution
Melt Real Time PCR test has been developed by Dhami & Kumarasinghe (2014). Species-specific
multiplex PCR primers have also been developed by Murphy et al. (2015).
270
HOST RANGE
Spotted Wing Drosophila (SWD), is one of the few species of Drosophilidae known to be a pest of
healthy, unripe fruit (Walsh et al. 2011, Atallah et al. 2014), Drosophila generally prefer to breed in fruit
that is already decaying and fermenting. SWD is a highly polyphagous pest that breeds in a wide
variety of mostly thin skinned small fruits (particularly Rosaceae) and is currently known from fifteen
plant families (Cini et al. 2012): Actinidiaceae, Adoxaceae, Caprifoliaceae, Cornaceae, Ebenaceae,
Elaeagnaceae, Ericaceae, Grossulariaceae, Moraceae, Myricaceae, Myrtaceae, Rhamnaceae,
Rosaceae, Rutaceae, Vitaceae.
Major commercial hosts (PHA 2013):
Scientific name
Common name
Scientific name
Common name
Prunus sp.
cherries & stone fruit
Fragaria ananassa
strawberry
Rubus sp.
berries
Vitis sp.
grapes
Malus sp.
apples
Pyrus sp.
pears
DISTRIBUTION
Worldwide SWD has recently rapidly spread from eastern Asia to many other regions (Walsh et al.
2011), including parts of Oceania, North America, Europe and South America. SWD is currently
unknown from Australia.
REMARKS
A large number of Drosophila species are known from Australia and many Drosophila species have
larvae that are commonly found in rotting fruit. SWD belong to the Sophophora subgenus of
Drosophila, a group that contains a large number of species (>300 worldwide), including the common
cosmopolitan D. melanogaster and D. simulans. There are twenty species currently known from
Australia (Atlas of Living Australia, http://www.ala.org.au/). Male D. melanogaster and D. simulans are
superficially similar to SWD in usually possessing dark tipped abdomens, but they differ in the
morphology of the combs on their basal tarsal segments and in having non-spotted wings (Blacket et
al. 2015 b). Female D. immigrans are superficially similar in the morphology of the ovipositor, however
the relative size of the ovipositor compared with the spermatheca differs substantially between these
two species, and in SWD the ovipositor (Figure 121B) is strongly sclerotized with robust teeth very
strongly developed along the lower half towards the ovipositor tip (Hauser 2011).
PEST STATUS

Exotic.

High level, highly invasive, pest species of a large number of soft fruits.
ATTRACTANT
Adults are known to be attracted to traps baited with a combination of wine, vinegar and soap (Landolt
et al. 2012).
271
FIGURES
Figure 121. Drosophila suzukii A. Female (left), Male (right). B. Female ovipositor. C. Male basal tarsal
segments.
Images courtesy of Deprá et al. 2014
272
9
Diagnostic resources
Key contacts and facilities
Contact
Facility
Area of expertise
Prof. RAI (Dick) Drew
International Centre for
Management of Pest Fruit Flies
Over four decades of
research experience in
taxonomy, ecology and
pest management of
tropical fruit flies in the
Subfamily Dacinae
[email protected]
Griffith University
170 Kessels Road, Nathan, Qld
4111, Australia
Phone: (07) 3735 3696
Fax:
(07) 3735 3697
Dr. David Yeates
CSIRO Entomology
Curator of Diptera
GPO Box 1700, Canberra, ACT
2601
[email protected]
Phone: (02) 6246 4001
Fax:
(02) 6246 4177
Mr. Peter S. Gillespie
Orange Agricultural Institute
Insect Collection Manager
NSW DPI
[email protected]
1447 Forest Road, Orange, NSW
2800
Phone: (02) 6391 3986
Fax:
(02) 6391 3899
Mr. Bernie Dominiak
Orange Agricultural Institute
[email protected]
NSW DPI
1447 Forest Road, Orange, NSW
2800
Surveillance, sterile insect
technique, mass rearing,
market access
Phone: (02) 6391 3986
Fax:
Leigh Pilkington
(02) 6391 3899
NSW DPI
Locked Bag 26
Gosford NSW 2250
Phone: (02) 4348 1953
Prof. Phillip Taylor
Macquarie University
Director, Biosecurity Futures Research
Centre
Department of Biological Sciences
[email protected]
Phone: (02) 9850 1311
Sydney, NSW 2109
Fax:
Dr. Anthony (Tony) R Clarke
Senior Lecturer in Ecology
[email protected]
(02) 9850 4299
Fruit fly biology and
environmentally benign
management, sterile
insect technique, lures,
monitoring, male
annihilation technique
Queensland University of
Technology
Science and Engineering Faculty
GPO Box 2434, Brisbane, Qld
4001, Australia
Phone: (07) 3138 5023
Fax:
(07) 3138 1535
273
Contact
Facility
Area of expertise
Dr Mark Schutze
Queensland University of
Technology
Fruit fly integrative
taxonomy and
systematics
[email protected]
Science and Engineering Faculty
GPO Box 2434, Brisbane, Qld
4001, Australia
Phone: (07) 3138 5023
Fax:
Ms. Jane Royer
Entomologist
[email protected]
(07) 3138 1535
Queensland Department of
Agriculture & Fisheries
Ecosciences Precinct, 41 Boggo
Rd, Dutton Park, Brisbane. QLD
4102
Phone: (07) 3255 4385
Mr. James Walker
[email protected]
Box 96
Airport Administration Centre
Cairns International Airport
Cairns, QLD 4870
Morphological
identification of most of
the 109 species of
Australian Bactrocera and
Dacus, pest species of
southeast Asia and the
Pacific and common
species trapped in PNG
Phone: (07) 4241 7845
Ms. Sally Cowan
[email protected]
Box 96
Airport Administration Centre
Cairns International Airport
Cairns, QLD 4870
Phone: (07) 4241 7857
Fax: (07) 4241 7845
Mr. Glenn Bellis
AQIS Darwin
Entomologist
1 Pederson Road, Marrara, NT
0812
[email protected]
Phone: (08) 8920 7000
Fax:
Ms. Linda Semeraro
Entomologist
[email protected]
(08) 8920 7011
Department of Economic
Development, Jobs, Transport
and Resources, Victoria
- AgriBio
5 Ring Road, Bundoora, Vic. 3083
Morphological
identification
Tephritid
molecular diagnostics
Laboratory: AgriBio, Bundoora.
Reference Collection: Victorian
Agricultural Insect Collection,
Bundoora.
Phone: (03) 9032 7000
Fax:
(03) 9032 7604
274
Contact
Facility
Area of expertise
Dr. Mali Malipatil
- AgriBio
Morphological
identification
Principal Research Scientist
[email protected]
Bundoora.
Phone: (03) 9032 7000
Fax:
Dr Mark Blacket
Entomologist
[email protected]
(03) 9032 7604
- AgriBio
Morphological
identification
Tephritid
molecular diagnostics
Bundoora.
Phone: (03) 9032 7000
Fax:
Dr. Darryl Hardie
Entomologist
[email protected]
(03) 9032 7604
Department of Agriculture and
Food Western Australia
Surveillance and fruit fly
pest management
Entomology Branch
Locked Bag 4, Bentley Delivery
Centre, WA 6983
Phone: (08) 9368 3721
Fax:
Mr. Andras Szito
Entomologist
[email protected]
(08) 9474 2405
Entomology Branch
Morphological
identification of
Tephritidae
Centre, WA 6983
Phone: (08) 9368 3721
Fax:
Mr Bill Woods
[email protected]
(08) 9474 2405
Fruit fly biology and
management
Entomology Branch
Centre, WA 6983
Phone: (08) 9368 3721
Fax:
(08) 9474 2405
Mr. Mark Adams
South Australian Museum
[email protected]
Science Centre
Morgan Thomas Lane, Adelaide,
SA 5000
Phone: (08) 8207 7305
Fax:
(08) 8207 7222
Molecular systematics of
taxonomically-problematic
groups among the
Australian fauna,
including 25 years’
experience in the rapid
genetic identification of
fruit fly larvae
275
Contact
Facility
Area of expertise
Dr. Karen Armstrong
Lincoln University
[email protected]
Bio-Protection Research Centre
Tephritid molecular
diagnostics
PO Box 85084, Lincoln University,
Canterbury 7647, New Zealand
Phone: (03) 423 0918
Fax:
(03) 325 3864
Dr. Brian Thistleton
Department of Resources
Principal Entomologist, Plant Industries
GPO Box 3000, Darwin NT 0801
[email protected]
Phone: (08) 8999 2257
Fax:
(08) 8999 2312
For international fruit fly authorities, see www.sel.barc.usda.gov/Diptera/tephriti/TephWork.htm
276
Reference collections
Collection
Location
Victorian Agricultural Insect Collection,
DEDJTR Vic.
AgriBio Building, DEDJTR Bundoora Campus, 5 Ring
Road, Bundoora, Victoria 3083.
Queensland Primary Industries Insect
Collection, DAF.
Biosecurity Queensland, DAF Ecosciences Precinct,
41 Boggo Rd, Dutton Park Brisbane QLD 4102,
Australia.
NAQS Insect Collection.
Airport Business Park, Cairns Airport Building 114,
Catalina Cresent, Cairns QLD 4870, Australia.
The Northern Territory Economic Insect
Reference Collection.
Department of Resources, Primary Industry, Berrimah
Farm, Makagon Road, Berrimah NT 0828, Australia.
Museum and Art Gallery of the Northern
Territory, NRETAS.
Conacher Street, Fannie Bay, Darwin NT 0820,
Australia.
277
Printed and electronic resources
9.3.1
Morphological keys
In practice in Australia the four paper keys that are most commonly used are:

Drew, R.A.I. (1989) The tropical fruit flies (Diptera: Tephritidae: Dacinae) of the Australian and
Oceanian regions. Memoirs of the Queensland Museum. 26: 1-521.

Drew, R.A.I. and Hancock, D.L. (1994) The Bactrocera dorsalis complex of fruit flies (Diptera:
Tephritidae: Dacinae) in Asia. Bulletin of Entomological Research. Supplementary Series 2: 168.

Drew, R. A., and Romig, M. C. (2013). Tropical Fruit Flies (Tephritidae Dacinae) of South-East
Asia: Indomalaya to North-West Australasia. CABI, Wallingford.

White, I.M. and Elson-Harris, M.M. (1992) Fruit Flies of Economic Significance: Their
Identification and Bionomics. CAB International. Oxon, UK. 601 p.
Other paper-based keys include:

Drew, R.A.I. and Raghu, S. (2002) The fruit fly fauna (Diptera: Tephritidae: Dacinae) of the
rainforest habitat of the Western Ghats, India. Raffles Bulletin of Zoology. 50, 327-352.

Drew, R. A., and Romig, M. C. (2013). Tropical Fruit Flies (Tephritidae Dacinae) of South-East
Asia: Indomalaya to North-West Australasia. CABI, Wallingford.

Drew, R.A.I., Hancock, D.L. and White, I.M. (1998) Revision of the tropical fruit flies (Diptera:
Tephritidae: Dacinae) of South-east Asia. II. Dacus Fabricius. Invertebrate Taxonomy, 12,
567-654.

Drew, R.A.I. and Romig, M.C. (2001) The fruit fly fauna (Diptera: Tephritidae: Dacinae) of
Bougainville, the Solomon Islands and Vanuatu. Australian Journal of Entomology, 40, 113150.

Drew, R.A.I., Hooper, G.H.S. and Bateman, M.A. (1982) Economic fruit flies of the South
Pacific region. Queensland Department of Primary Industries, Brisbane, Queensland. 139 pp.

White, I.M. and Elson-Harris, M.M. (1992) Fruit Flies of Economic Significance: Their
Identification and Bionomics. CAB International. Oxon, UK. 601 p.

Rohani, I. (1987) Identification of larvae of common fruit fly pest species in West Malaysia.
Journal of Plant Protection in the Tropics, 4 (2), 135-137.

Hardy, E.D. (1986) Fruit flies of the subtribe Acanthonevrina of Indonesia, New Guinea, and
the Bismarck and Solomon Islands (Diptera: Tephritidae: Trypetinae: Acanthonevrina). Pacific
Insect Monographs, No. 42. Honolulu, Hawaii. 191 p.

Hardy, E.D. (1974) The fruit flies of the Philippines (Diptera - Tephritidae). Pacific Insect
Monographs, No. 32. Honolulu, Hawaii. 266 p.

Significant information on the larvae of many Australian fruit flies, including ones not of
economic importance but that might turn up during sampling, was given in the PhD thesis of
Dr Marlene Elson-Harris lodged at the University of Queensland.
Electronic keys available include:

White, I.M. and Hancock, D.L. (2003) Fauna Malesiana [electronic key to fruit flies]. ISBN
9075000359.

White, I.M. and Hancock, D.L. (1997) Indo-Australasian Dacini Fruit Flies (CABIKEY)
International Institute of Entomology, London. CD-ROM.
278

An interactive key is also available on the Fruit Flies of the World website: http://deltaintkey.com/ffa

Key to Fruit Flies of New South Wales: http://www1.dpi.nsw.gov.au/keys/fruitfly/index.html
9.3.2
Electronic resources

Tephritid Barcoding Initiative (TBI): www.barcodeoflife.org. The TBI aims to barcode
10,000 specimens representing 2,000 species of fruit flies, including all taxa (about 350
species) of major and minor economic importance.

The Diptera Site: www.sel.barc.usda.gov/Diptera/tephriti/tephriti.htm. Contains a large
amount of biological and other information about fruit flies.

Pest Fruit Flies of the World: http://delta-intkey.com/ffa. Contains comprehensive
information and keys on fruit flies of all regions.

ANIC Anatomical atlas of flies: http://www.ento.csiro.au/biology/fly/fly.php. Great for
illustrations of every feature of acalyptrate flies.

On the fly: interactive atlas and key to Australian fly families:
www.csiro.au/resources/ps236.html.

Australian Pest and Diseases Image Library (PaDIL): www.padil.gov.au. Contains species
information as well as photos for a number of fruit fly species (endemic and exotic).

NSW government fruit fly resource:
www.agric.nsw.gov.au/Hort/ascu/fruitfly/fflyinde.htm. List of fruit fly species found in New
South Wales or believed to be present there, with links to summary information on each and
key.

International Centre for Management of Pest Fruit Flies (Griffith University and
Malaysia): http://www.icmpff.org

South Pacific fruit fly website (Pacifly): http://www.pacifly.org. Contains profiles of all
species found in the South Pacific.

Featured Creatures: http://entomology.ifas.ufl.edu/creatures/index.htm. Contains profiles
for a limited number of fruit fly species.

The Australian Plant Pest Database (APPD): http://pha.vpac.org. A national, online
database of pests and diseases of Australia's economically important plants.

Atlas of Living Australia: http://www.ala.org.au/
279
Supplier details
Supplier
Address
Contact details
Life Technologies
5 Caribbean Drive
Ph: 1 300 735 292
(for PCR)
Scoresby, VIC 3179
Fax: 1 800 067 639
http://www.thermofisher.com
Astral Scientific
PO Box 232
Ph: 1800 221 280
Gymea, NSW 2227
Fax: (02) 9540 2051
http://astralscientific.com.au/
Bio-Rad Laboratories Pty. Ltd.
Level 5, 446 Victoria Road
Gladesville, NSW 2111
GENESEARCH
14 Technology Drive
(agents for New England
Biolabs)
Arundel, QLD 4214
Interpath services
1/46 Sheehan Rd
Heidelberg West, VIC 3081
Ph: (02) 9914 2800 or 1800 224
354
Fax: (02) 9914 2888
Ph: 1800 074 278 or (07) 5594
0299
www.genesearch.com.au
Ph: (03) 9457 6277 or 1800 626
369
Fax: (03) 9458 4010
http://www.interpath.com.au/
Invitrogen
PO Box 4296
Ph: 1800 331 627
(for primer synthesis)
Mulgrave, VIC 3170
Fax: (03) 9562 7773
Mirella Research Pty. Ltd.
PO Box 365
Ph: (03) 9388 1088 or 1800 640
444
Brunswick, VIC 3056
Promega Corporation
PO Box 7417
(for Molecular weight marker)
Alexandria, NSW 2015
Fax: (03) 9388 0456
Ph: (02) 8338 3800 or 1800 225
123
Fax: (02) 8338 3855 or 1800 626
017
www.promega.com
Qiagen Pty Ltd
PO Box 25
Ph: (03) 9489 3666
(for DNA extraction)
Clifton Hill, VIC 3068
Fax: (03) 9489 3888
www.qiagen.com
Roche Diagnostics Australia
Pty. Ltd.
31 Victoria Avenue
Ph: (02) 9899 7999
Castle Hill, NSW 2154
Fax: (02) 9634 2949
Sigma-Aldrich Pty. Ltd.
PO Box 970
(for chemicals)
Castle Hill, NSW 1765
Ph: 1800 800 097 or (02) 9841
0555
Fax: 1800 800 096 or (02) 9841
0500
www.sigmaaldrich.com
280
10 References
Allwood, A.J., Chinajariyawong, A., Drew, R.A.I., Hamacek, E.L. and Hancock, D.L. (1999). Host plant
records for fruit flies (Diptera: Tephritidae) in South East Asia. Raffles Bulletin of Zoology Supplement
No. 7: 1-92.
Anon. (1996). Code of Practice for the management of Queensland Fruit Fly. Standing Committee on
Agriculture and Resource Management, Department of Primary Industries, Canberra.
Armstrong K.F. and Cameron, C.M. (1998). Molecular Kit for Species Identification Fruit Flies
(Tephritidae), Lincoln University.
Armstrong K.F., and Ball S.L. (2005). DNA barcodes for biosecurity: invasive species identification.
Philosophical Transactions of the Royal Society of London B: Biological Sciences. 360: 1813–1823.
Armstrong, K. F., Cameron, C. M., & Frampton, E. R. (1997). Fruit fly (Diptera: Tephritidae) species
identification: a rapid molecular diagnostic technique for quarantine application. Bulletin of
Entomological Research, 87(02): 111-118. doi: doi:10.1017/S0007485300027243
Atallah J, Teixeira L, Salazar R, Zaragoza G, & Kopp A. (2014). The making of a pest: the evolution of
a fruit-penetrating ovipositor in Drosophila suzukii and related species. Proceedings of the Royal
Society B: Biological Sciences 281, 20132840.
Barr N., Ruiz-Arce R., and Armstrong K. (2014). Using Molecules to Identify the Source of Fruit Fly
Invasions. pp 321-378 In: Trapping and the Detection, Control, and Regulation of Tephritid Fruit Flies,
Springer, Netherlands.
Bergsten J., Bilton D.T., Fujisawa T., Elliott M., Monaghan M.T., Balke M., Hendrich L., Geijer J.,
Herrmann J., Foster G.N., Ribera I., Nilsson A.N., Barraclough T.G. and Vogler A.P. (2012). The Effect
of Geographical Scale of Sampling on DNA Barcoding. Systematic Biology 61: 851–869
Blacket M.J. (2011). Identification of Queensland Fruit Flies using molecular markers: Mitochondrial
DNA sequence variation in Southern QFF. Department of Primary Industries Victoria Report (June
2011).
Blacket M.J., Rice A.D., Semeraro L., and Malipatil M.B. (2015 a). DNA-based identifications reveal
multiple introductions of the vegetable leafminer Liriomyza sativae (Diptera: Agromyzidae) into the
Torres Strait Islands and Papua New Guinea. Bulletin of Entomological Research. 105(05): 533-544.
Blacket M.J., Semeraro L, & Malipatil MB. (2015 b). Draft diagnostic protocol for Spotted Wing
Drosophila (Drosophila suzukii). Submitted to the Australian Grape and Wine Authority (AGWA) and
the Subcommittee on Plant Health Diagnostics (SPHD).
Blacket M.J., Semeraro L., and Malipatil M.B. (2012). Barcoding Queensland Fruit Flies (Bactrocera
tryoni): impediments and improvements. Molecular Ecology Resources, 12: 428–436.
Bock IR. (1976). Drosophilidae Australia. I. Drosophila (Insecta:Diptera) Australian Journal of Zoology
Supplementary Series 40, 1–105.
Boykin L.M., Armstrong K., Kubatko L. and De Barro P. (2012). DNA barcoding invasive insects:
database roadblocks. Invertebrate Systematics. 26: 506–514.
Burgher-MacLellan, K.L., Gaul, S., MacKenzie, K. and Vincent, C. (2009). The use of real-time PCR to
identify blueberry maggot (Diptera: Tephritidae, Rhagoletis mendax) from other Rhagoletis species
and in lowbush blueberry fruit (Vaccinium angustifolium). Acta Horticulturae 810, 265-274 DOI:
10.17660/ActaHortic.2009.810.34 http://dx.doi.org/10.17660/ActaHortic.2009.810.34
281
CABI (2007) Crop Protection Compendium, 2007 Edition. CAB International, Wallingford, UK. Online
version: http://www.cabi.org/compendia/cpc/ [accessed September 2015].
Calabria G, Maca J, Bachli G, Serra L, & Pascual M. (2012). First records of the potential pest species
Drosophila suzukii (Diptera: Drosophilidae) in Europe. Journal of Applied Entomology 136, 139-147.
Cameron, E.C., Sved, J.A., and Gilchrist, A.S. (2010). Pest fruit fly (Diptera: Tephritidae) in
northwestern Australia: one species or two? Bulletin of Entomological Research. 100(2):197-206.
Carroll, L.E., I.M. White, A. Freidberg, A.L. Norrbom, M.J. Dallwitz, and F.C. Thompson. (2002
onwards). Pest fruit flies of the world. Version: 8th December 2006. http://delta-intkey.com [accessed
May 2010].
Casana-Giner V, Oliver JE, Jang EB & Carvalho LA. (2003). Synthesesand behavioral evaluations of
fluorinated and silylated analogs ofraspberry ketone as attractants for melon fly, Bactrocera cucurbitae
(Coquillett). Journal of Entomological Science 38: 111–119.
Clarke, A.R., Armstrong, K.F., Carmichael, A.E., Milne, J.R., Raghu, S., Roderick, G.K. and Yeates,
D.K. (2005). Invasive phytophagous pests arising through a recent tropical evolutionary radiation: The
Bactrocera dorsalis complex of fruit flies. Annual Review of Entomology 50: 293-319.
Clarke, A. R., Powell, K. S., Weldon, C. W., & Taylor, P. W. (2011). The ecology of Bactrocera tryoni
(Diptera: Tephritidae): what do we know to assist pest management? Annals of Applied Biology
158(1): 26-54.
Cowley, J.M. (1990). A new system of fruit fly surveillance trapping in New Zealand. New Zealand
Entomologist 13: 81-84.
Cowley, J.M., Page, F.D., Nimmo, P.R. and Cowley, D.R. (1990). Comparison of the effectiveness of
two traps for Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) and implication for quarantine
surveillance systems. Journal of the Australian Entomological Society 29: 171-176.
Dean M.D. and Ballard J.W. (2001). Factors affecting mitochondrial DNA quality from museum
preserved Drosophila simulans. Entomologia Experimentalis et Applicata 98: 279–283
Deprá M, Poppe JL, Schmitz HJ, De Toni DC, & Valente VLS. (2014). The first records of the invasive
pest Drosophila suzukii in the South American continent. Journal of Pest Science 87, 379-383.
Dhami, M., Gunawardana, D. N., Voice D. & Kumarasinghe L. (2015). A real-time PCR toolbox for
accurate identification of invasive fruit fly species. Journal of Applied Entomology. In press
Dhami MK & Kumarasinghe L. (2014). A HRM real-time PCR assay for rapid and specific identification
of the emerging pest Spotted-Wing Drosophila (Drosophila suzukii). PLoS ONE 9, e98934.
Dominiak, B.C., Gilmour, A.R., Kerruish, B., and Whitehead, D. (2003). Detecting low populations of
Queensland fruit fly Bactrocera tryoni (Froggatt) with McPhail and Lynefield traps. General and
Applied Entomology 32: 49-54.
Dominiak, B.C. and Nicol, H.I. (2010). Field performance of McPhail and Lynefield traps for monitoring
male and female sterile Bactrocera tryoni (Froggatt) and wild Dacus newmani (Perkins). Pest
Management Science 66: 741-744.
Dominiak, B. C. and Daniels, D. (2011). Review of the past and present distribution of Mediterranean
fruit fly (Ceratitis capitata Wiedemann) and Queensland fruit fly (Bactrocera tryoni Froggatt) in
Australia. Australian Journal of Entomology 51: 104-115.
Dominiak, B.C., Kerruish, B., Barchia, I., Pradhan, U., Gilchrist, A.S., and Nicol, H.I. (2011). The
influence of mixtures of parapheromone lures on trapping of fruit fly in New South Wales, Australia.
Plant Protection Quarterly 26(4): 136-140.
282
Dominiak, B.C., Campbell, A.J., Jang, E.B., Ramsey, A. and Fanson, B.G. (2015a). Field evaluation of
melolure, a formate analogue of cuelure, and reassessment of fruit fly species trapped in Sydney, New
South Wales, Australia. Journal of Economic Entomology 108: 1176-1181.
Dominiak, B.C., Fay, H.A.C. and Fanson, B.G. (2015b). Field evaluation of the efficacy of zingerone
and other chemical lures on seven species of fruit fly in Sydney, New South Wales, Australia. Plant
Protection Quarterly 30: 67-71.
Dominiak, B.C. and Senior, L. (in prep.). Review of the biology, distribution and control of Cucumber
Fruit Fly, Bactrocera cucumis (French) (Diptera: Dacinae) in Australia.
Drew, R.A.I. (1989). The tropical fruit flies (Diptera: Tephritidae: Dacinae) of the Australian and
Oceanian regions. Memoirs of the Queensland Museum. 26: 1-521.
Drew, R.A.I. (2004). Biogeography and speciation in the Dacini (Diptera: Tephritidae: Dacinae).
Bishop Museum Bulletin in Entomology 12: 165-178.
Drew, R.A.I. and Hancock, D.L. (1994). The Bactrocera dorsalis complex of fruit flies (Diptera:
Tephritidae: Dacinae) in Asia. Bulletin of Entomological Research. Supplementary Series 2: 1-68.
Drew, R.A.I., G.H.S. Hooper and M.A. Bateman. (1982). Economic Fruit Flies of the South Pacific
Region. Queensland Department of Primary Industries.
Drew, R.A.I. and Lambert, D.M. (1986). On the specific status of Dacus aquilonis and Dacus tryoni
Diptera Tephritidae. Annals of the Entomological Society of America. 79(6): 870-878.
Drew, R.A.I., J. Ma, S. Smith, and J. M. Hughes. (2011). The taxonomy and phylogenetic relationships
of species in the Bactrocera musae complex of fruit flies (Diptera: Tephritidae: Dacinae) in Papua New
Guinea. Raffles Bull. Zool. 59:145–162.
Drew, R. A. and Romig, M. C. (2007). Records of fruit flies and new species of Dacus (Diptera:
Tephritidae) in Bhutan. Raffles Bulleting of Zoology 55: 1-21.
Drew, R. A., and Romig, M. C. (2013). Tropical Fruit Flies (Tephritidae Dacinae) of South-East Asia:
Indomalaya to North-West Australasia. CABI.
Fay, H.A.C. (2012). A highly effective and selective male lure for Bactrocera jarvisi (Tryon) (Diptera:
Tephritidae). Australian Journal of Entomology 51: 189–197.
Flath, R.A., Cunningham, R.T., Liquido, N.J. and McGovern, T.P. (1994). Alpha-Ionol as Attractant for
Trapping Bactrocera latifrons (Diptera: Tephritidae). Journal of Economic Entomology 87(6):14701476.
Floyd, R., Lima, J., deWaard, J., Humble, L., and Hanner, R. (2010). Common goals: policy
implications of DNA barcoding as a protocol for identification of arthropod pests. Biological Invasions.
12: 2947-2954.
Folmer, O., Black, M., Hoeh, W., Lutz, R. and Vrijenhoek, R. (1994). DNA primers for amplification of
mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular marine
biology and biotechnology 3: 294-299.
Foote, R.H., Blanc, F.L. and Norrbom, A.L. (1993). Handbook of the Fruit Flies (Diptera: Tephritidae)
of America North of Mexico. Comstock, Ithaca, N.Y.
Gilbert MTP, Moore W, Melchior L, Worobey M (2007). DNA Extraction from Dry Museum Beetles
without Conferring External Morphological Damage. PLoS ONE 2(3): e272.
doi:10.1371/journal.pone.0000272
Gilbert, M.T.P., Moore, W., Melchior, L., and Worobey, M. (2007). DNA extraction from dry museum
beetles without conferring external morphological damage. PLoS ONE 2: e272.
283
Gillespie, P. (2003). Observations on fruit flies (Diptera: Tephritidae) in New South Wales. General
and Applied Entomology 32: 41-47.
Hall B. (2011). How to send samples for diagnosis in Australia: Plant disease and Insect identification.
CRC Plant Biosecurity Publication, http://plantbiosecuritydiagnostics.net.au/wordpress/wpcontent/uploads/2012/12/CRC-packaging-brochure.pdf
Hancock, D.L., Hamacek, E.L., Lloyd, A.C. and Elson-Harris, M.M. (2000). The distribution and host
plants of fruit flies (Diptera: Tephritidae) in Australia. Queensland Department of Primary Industry.
Information series Q199067. Brisbane, Queensland. 75pp.
Hauser M. (2011). A historic account of the invasion of Drosophila suzukii (Matsumura) (Diptera:
Drosophilidae) in the continental United States, with remarks on their identification. Pest Management.
Science 67, 1352–1357.
[IAEA] International Atomic Energy Agency. (2003). Trapping guidelines for area-wide fruit fly
programmes. IAEA/FAO-TG/FFP. IAEA, Vienna, Austria.
Jiang F., Jin Q., Liang L., Zhang A.B. and Li Z.H. (2014). Existence of Species Complex Largely
Reduced Barcoding Success for Invasive Species of Tephritidae: A Case Study in Bactrocera spp.
Molecular Ecology Resources. 14: 1114–1128.
Jinbo U., Kato T. and Ito M. (2011). Current progress in DNA barcoding and future implications for
entomology. Entomological Science 14: 107–124
Kim SS, Tripodi AD, Johnson DT, & Szalanski AL. (2014). Molecular Diagnostics of Drosophila suzukii
(Diptera: Drosophilidae) using PCR-RFLP. Journal of Economic Entomology 107, 1292-1294.
Krosch, M. N., Schutze, M. K., Armstrong, K. F., Graham, G. C., Yeates, D. K., & Clarke, A. R. (2012).
A molecular phylogeny for the Tribe Dacini (Diptera: Tephritidae): systematic and biogeographic
implications. Molecular Phylogenetics and Evolution 64(3): 513-523. doi:
10.1016/j.ympev.2012.05.006
Landolt PJ, Adams T, Davis TS, & Rogg H. (2012). Spotted Wing Drosophila, Drosophila suzukii
(Diptera: Drosophilidae), Trapped with Combinations of Wines and Vinegars. Florida Entomologist 95,
326-332.
Lawson, A.E., McGuire, D.J., Yeates, D.K., Drew, R.A.I. and Clarke, A.R. (2003). Dorsalis: an
interactive identification tool to fruit flies of the Bactrocera dorsalis complex. Griffith University.
Brisbane, Australia. [CD-ROM]
Leblanc L, Vueti ET, Drew RAI & Allwood AJ. (2012). Host plant records for fruit flies (Diptera:
Tephritidae: Dacini) in the Pacific islands. Proceedings of the Hawaiian Society 44: 11-53.
Leblanc L, William J & Allwood AJ. (2004). Host Fruit of Mango Fly (Bactrocera frauenfeldi (Schiner))
(Diptera: Tephritidae) in the Federated States of Micronesia. Micronesica 37: 21-31.
Leblanc, L; Balagawi, S; Mararuai, A; Putulan, D; Tenakanai, D; Clarke, A R. (2001). Fruit flies in
Papua New Guinea. Pest Advisory Leaflet, Secretariat of the Pacific Community. Plant Protection
Service; 37.
Lou M., and Golding G.B. (2010). Assigning sequences to species in the absence of large interspecific
differences. Molecular Phylogenetics and Evolution 56: 187–194
Manchenko, G.P. (1994). Handbook of detection of enzymes on electrophoretic gels. CRC Press,
Boca Raton, Florida.
Mantel BL & Upton MS. (2010). Methods for Collecting, Preserving and Studying Insects and Other
Terrestrial Arthropods, 5th edition. The Australian Entomological Society Miscellaneous Publication
no. 3. pp. 81.
284
McKenzie, C., Gillispie, P. and Hailstones, D. (2004). Draft molecular diagnostic standard for fruit flies
of economic significance to Australia. Plant Health Australia and NSW Department of Primary
Industries.
McKenzie, L., Dransfield, L., Driver, F. and Curran, J. (1999). Molecular protocols for species
identification of tephritid fruit flies. Report to AQIS. CSIRO, Canberra.
McQuate GT & Peck SL. (2001). Enhancement of attraction of alpha ionol to male Bactrocera latifrons
(Diptera: Tephritidae) by addition of a synergist cade oil. Journal of Economic Entomology 94: 39–46.
McQuate GT, Jang EB & Siderhurst M. (2013). Detection/Monitoring of Bactrocera latifrons (Diptera:
Tephritidae): Assessing the potential of prospective new lures. Proceedings of the Hawaiian
Entomological Society 45: 69-81
Metcalf, R.L., Mitchell, W.C., and Metcalf, E.R. (1983). Olfactory receptors in the melon fly Dacus
cucurbitae and the oriental fruit fly Dacus dorsalis. Proceedings of the National Academy of Sciences
of the United States of America 80: 3143-3147.
Moreau C.S., Wray B.D., Czekanski-Moir J.E. and Rubin B.E.R. (2013). DNA preservation: a test of
commonly used preservatives for insects. Invertebrate Systematics 27: 81–86
http://dx.doi.org/10.1071/IS12067
Murphy KA, Unruh TR, Zhou LM, Zalom FG, Shearer PW, Beers EH, Walton VM, Miller B, & Chiu JC.
(2015). Using comparative genomics to develop a molecular diagnostic for the identification of an
emerging pest Drosophila suzukii. Bulletin of Entomological Research 105, 364-372.
Norrbom, A.L. (2003). The Diptera site. Systematic Entomology Laboratory, ARS, USDA and
Department of Entomology, NMNH, SI. www.sel.barc.usda.gov/diptera/tephriti/tephriti.htm [accessed
May 2010].
O’Loughlin, G.T., East, R.W., Kenna, G.B. and Harding, E. (1983). Comparison of the effectiveness of
traps for the Queensland fruit fly, Dacus tryoni (Froggatt) (Diptera: Tephritidae). General and Applied
Entomology 15: 3-6.
Osborne, R., Meats, A., Frommer, M., Sved, J.A., Drew, R.A.I., and Robson, M.K. (1997). Australian
Distribution of 17 Species of Fruit Flies (Diptera: Tephritidae) Caught in Cue Lure Traps in February
1994. Australian Journal of Entomology 36: 45-50.
PaDIL (2007). Pest and disease image library. An Australian Government initiative. www.PaDIL.gov.au
[accessed May 2010].
PHA. (2013). Fact Sheet: Spotted-winged drosophila.
http://www.planthealthaustralia.com.au/pests/spotted-winged-drosophila/
Queensland Department of Primary Industries and Fisheries (2002). OP-03: Fruit fly identification
guidelines. Operational Procedures. August, 2002.
Ratnasingham S., and Hebert P.D.N. (2013) DNA-Based Registry for All Animal Species: The Barcode
Index Number (BIN) System. PLoS ONE 8(8): e66213. doi:10.1371/journal.pone.0066213
Richardson, B. J., Baverstock, P. R., and Adams, M. (1986). Allozyme electrophoresis. A handbook for
animal systematics and population studies. Academic Press, Sydney, Australia.
Royer JE & Hancock DL. (2012). New distribution and lure records of Dacinae (Diptera: Tephritidae)
from Queensland, Australia, and a description of a new species of Dacus Fabricius. Australian Journal
of Entomology 51: 239–247.
Royer JE, DeFaveri S & Lowe G. (2014). Cucumber volatile blend, a promising female-biased lure for
Bactrocera cucumis (French 1907) (Diptera: Tephritidae: Dacinae), a pest fruit fly that does not
respond to male attractants. Australian Journal of Entomology 53: 347–352
285
Royer JE, Wright CW & Hancock DL. (2015). Bactrocera frauenfeldi (Schiner) (Diptera: Tephritidae),
an invasive fruit fly in Australia that may have reached the extent of its spread due to environmental
variables. Australian Journal of Entomology accepted, in press.
Royer JE. (2015). Responses of fruit flies (Tephritidae: Dacinae) to novel male attractants in north
Queensland, Australia, and improved lures for some pest species. Australian Journal of Entomology.
early view available at: http://onlinelibrary.wiley.com/doi/10.1111/aen.12141/pdf
Schutze M.K., Aketarawong N., Amornsak W., Armstrong K.F., Augustinos A.A., Barr N., Bo W.,
Bourtzis K., Boykin L.M., Cáceres C., Cameron S.L., Chapman T.A., Chinvinijkul S., Chomič A., De
Meyer M., Drosopoulou E., Englezou A., Ekesi S., Gariou-Papalexiou A.,Geib S.M., Hailstones D.,
Hasanuzzaman M., Haymer D., Hee A.K.W., Hendrichs J., Jessup A., Ji Q., Khamis F.M., Krosch
M.N., Leblanc L., Mahmood K., Malacrida A.R., Mavragani-Tsipidou P., Mwatawala M., Nishida R.,
Ono H., Reyes J., Rubinoff D., San Jose M., Shelly T.E., Srikachar S., Tan K.H., Thanaphum S., Haq
I., Vijaysegaran S., Wee S.L., Yesmin F., Zacharopoulou A., & Clarke A.R. (2015) Synonymization of
key pest species within the Bactrocera dorsalis species complex (Diptera: Tephritidae): taxonomic
changes based on 20 years of integrative morphological, genetic, behavioural, and physiological data.
Systematic Entomology 40(2): 456-471.
Secretariat of the Pacific Community (2006). Pacific Fruit Fly Web. www.spc.int/pacifly [accessed
May 2010]
Seljak G. (2013). PM 7/115 (1) Drosophila suzukii. Bulletin OEPP/EPPO Bulletin 43, 417–424
Semeraro, L. and Malipatil, M.B., (2005). Molecular diagnostic test for Queensland fruit fly larvae
(Bactrocera tryoni Froggatt).
Smit J., Reijnen B., and Stokvis F. (2013). Half of the European fruit fly species barcoded (Diptera,
Tephritidae); a feasibility test for molecular identification. ZooKeys 365: 279–305.
Smith, E.S.C. and Chin D. (1987). Current fruit fly research in the Northern Territory. Proceedings of
the 4th Workshop in Tropical Entomology. Darwin, NT 10-15 May 1987, pp. 128-135.
Smith, E.S.C., Chin, D., Allwood, A.J., and Collins, S.G. (1988). A revised host list of fruit flies (Diptera:
Tephritidae) from the Northern Territory of Australia. Queensland Journal of Agricultural and Animal
Sciences 45(1): 19-28.
Tan KH & Nishida R. (2000). Mutual reproductive benefits between a wild orchid, Bulbophyllum
patens, and Bactrocera fruit flies via a floral synomone. Journal of Chemical Ecology 26: 533–546.
Tsuruta, K., White, I.M., Bandara, H.M.J., Rajapakse, H., Sundaraperuma, S.A.H., Kahawatta,
S.B.M.U.C., and Rajapakse, G.B.J.P. (1997). A preliminary notes on the host plants of fruit flies of the
tribe Dacini (Diptera: Tephritidae) in Sri Lanka. Esakia 37: 149-160.
University of Florida and the Florida Department of Agriculture and Consumer Services (2009).
Featured Creatures. http://entomology.ifas.ufl.edu/creatures/index.htm [accessed May 2010].
Van Velzen R, Weitschek E, Felici G, Bakker FT (2012). DNA Barcoding of Recently Diverged
Species: Relative Performance of Matching Methods. PLoS ONE 7(1): e30490.
doi:10.1371/journal.pone.0030490
Vargas, R.I., Pinero, J.C. and Leblanc, L. 2015. An overview of Pest Species of Bactrocera Fruit Flies
(Diptera: Tephritidae) and the Integration of Biopesticides with other biological approaches for their
management with a focus on the Pacific Region. Insects 6: 297-318.
Vink C.J., Thomas S.M., Paquin P., Hayashi C.Y., and Hedin M. (2005). The effected of preservatives
and temperature on arachnid DNA. Invertebrate Systematics 19: 99-104.
Virgilio M., Jordaens K., Breman F.C., Backeljau T., De Meyer M. (2012). Identifying Insects with
Incomplete DNA Barcode Libraries, African Fruit Flies (Diptera: Tephritidae) as a Test Case. PLoS
ONE 7: e31581.
286
Virgilio, M., Jordaens, K., Verwimp, C., White, I. M., & De Meyer, M. (2015). Higher phylogeny of
frugivorous flies (Diptera, Tephritidae, Dacini): Localised partition conflicts and a novel generic
classification. Molecular phylogenetics and evolution 85: 171-179.
Walsh DB, Bolda, MP, Goodhue, RE, Dreves, AJ, Lee, J et al. (2011). Drosophila suzukii (Diptera:
Drosophilidae): Invasive Pest of Ripening Soft Fruit Expanding its Geographic Range and Damage
Potential. Journal of Integrated Pest Management 2, 1-7.
Wang X-C., Liu C., Huang L., Bengtsson-Palme J., Chen H., Zhang J-H., Cai D. and Li J-Q. (2015).
ITS1: a DNA barcode better than ITS2 in eukaryotes? Molecular Ecology Resources 15: 573–586
Wang, Y., Yu, H., Raphael, K., & Gilchrist, A. S. (2003). Genetic delineation of sibling species of the
pest fruit fly Bactocera (Diptera: Tephritidae) using microsatellites. Bulletin of Entomological Research
93(04): 351-360. doi: doi:10.1079/BER2003249
Waterhouse, D. F. (1993). The Major Arthropod Pests and Weeds of Agriculture in Southeast Asia.
Canberra, Aust.: ACIAR. Monograph 21: 141.
White, I.M. and Elson-Harris, M.M. (1992). Fruit Flies of Economic Significance: Their Identification
and Bionomics. CAB International. Oxon, UK. 601 p.
Yu, D., Chen, Z., Zhang, R., & Yin, W. (2005). Real-time qualitative PCR for the inspection and
identification of Bactrocera philippinensis and Bactrocera occipitalis (Diptera: Tephritidae) using SYBR
Green assay. The Raffles Bulletin of Zoology 53(1): 73-78.
Yu, D., Zhang, G., Chen, Z., Zhang, R., & Yin, W. (2004). Rapid identification of Bactrocera latifrons
(Dipt., Tephritidae) by real‐time PCR using SYBR Green chemistry. Journal of Applied Entomology
128(9‐10): 670-676.
287
11 Appendices
288
APPENDIX 1
Rearing fruit fly larvae to adults for the purpose of identification – basic
method
Pupae need to develop in a dry medium such as sand or sawdust. Once flies start to emerge they
need to be provided with access to water and sugar for survival and for colour development. After
about four days they may be collected, killed and prepared for study (Lawson et al. 2003).
You will need:
 a clear plastic tub with a well fitted lid with a large hole cut in it
 mesh to put between the tub and the lid
 river sand or untreated sawdust as a pupation media
 sugar cubes, water, chux type sponge, inactive torula yeast (protein source) - to feed the flies

Process:
 Put about 1cm of sand or sawdust in the bottom of the tub.
 Put infested fruit on the sawdust.
 If the fruit is large, wet and sloppy you may want to put the fruit on another smaller container
with a mesh lid, inside the other container, so it drains into that rather than the media.
 Leave the tub somewhere that is roughly the same temperature as outside, away from ants
and direct sunlight. If it is cooler and you have access to a controlled temperature room, you
may want to keep the fruit at warmer temperatures e.g. 27OC and 70% RH.
 Allow about 5+ days for the larvae to pupate. In cooler conditions or with harder fruit such as
apples this may take up to 30 days.
 After 5 days check for pupae that have emerged as adults. Don’t open the lid as any adults
may escape. Check for adults every day after this.
 Once you see adults they will need the food and water. Put a sugar cube on top of the mesh;
put a 3cm slice of the chux in a small container of water and have one end resting on the
gauze so the flies can drink from the wicking chux; mix the yeast with enough water to make a
paste put a dob of this on top of the gauze.
 Wait 5-7 days after flies have emerged to kill them. They need this amount of time to fully
develop and colour up so they are identifiable.
 You can kill the flies by putting the whole rearing tub in a large freezer.
Note: if you have fruit that may be a quarantine intercept, rearing should only be undertaken in an
appropriately accredited quarantine containment laboratory. You may have an exotic and don’t want to
breed it and have them escape!
Suppliers:
Fermex Inactive Yeast from Lesaffre. See website for contacts in your state
http://www.fermex.com.au/products/?id=7 (~$70/15kg)
http://www.fermex.com.au/contact_us/
Inactive torula yeast in smaller quantities can also be purchased from health food stores.
APPENDIX 2
Composition and preparation of reagents
5X TBE buffer
Combine:
 450 mM Tris base
 450 mM Boric acid
 10 mM EDTA (pH 8.0)
 Store at room temperature
Dilute to 1X TBE with millipore water prior to use.
1% Agarose gel
1. 1 g of DNA grade agarose per 100 mL of 1X TBE.
2. Melt in a microwave.
3. Pour into a prepared gel tray when agarose has cooled sufficiently.
4. Allow the gel to set at room temperature for at least 30 min before use.
6 x Loading dye
Combine:
 1X TBE buffer
 0.25% (w/v) Bromophenol Blue
 0.25% (w/v) Xylene cyanol FF
 30 % (v/v) Glycerol
Store at room temperature.
10X PBS
Combine:
 1.37 M NaCl
 27 mM KCl
 43 mM Na2HPO4.7H2O
 14 mM KH2PO4
Autoclave and store stock at room temperature.
Dilute to 1X PBS with sterile water for use. Store at room temperature in sterile bottle.
Preparing PCR primers, 10 μM
1. Add nuclease free water to the freeze-dried primer stock: to calculate quantity of water to add,
find the nmol reading for each primer and move decimal place forward (to right) of the nmol
reading eg. 51.7 nmol = 517 μL of nuclease free water to be added to primer stock.
2. Prepare a 1 in 10 dilution of primer for final stock of 1 μM, e.g. if preparing 500 μL, then add 450
μL of nuclease free water and 50 μL of primer from 10 μM stock.
Preparing dNTP’s
These can be purchased premixed, or be contained within a PCR kit. A cheaper alternative is to prepare
by combining individual nucleotide stocks, each with initial concentration of 100 mM. To prepare a 400
μl dNTP stock, final concentration of 2.5 mM:
 Prepare Eppendorf tubes (1.5 ml) for final stock
 Add 360 μl nuclease free water to each Eppendorf tube


Add 10 μl of each dNTP to each final stock eg. 10 μl of A, 10 μl of C, 10 μl of G, 10 μl of T, to
each tube. Mix well
This should make a 1 in 10 dilution = 2.5 mM.
Preparing molecular weight marker (100 bp ladder)
For 250 μg mL-1 concentration of marker:
 400 μL marker
 200 μL loading dye
 600 μL double-distilled H2O
Proportion is 2:1:3 respectively
Preparing an agarose gel - basic equipment and materials
 agarose powder
 electrolytic buffer (TAE or TBE)
 SYBR® Safe DNA gel stain (or ethidium bromide if alternative is not available)
 loading dye (X6)
 large glass flask and plastic jar (250 mL)
 molecular weight marker (100 bp ladder)
 microwave
 plate and combs
 pipette tips - non-aerosol
 pipettes
Preparation of gel:
1. Gel size may be 50 mL, 100 mL, or 200 mL.
2. Use a 1.5% agarose gel for PCR product resolution and a 2-3 % gel for restriction digest
products
3. According to gel size and concentration, add TBE to the appropriate weight of agarose powder
4. Microwave until boiling point and solution is clear; care not to superheat and cause solution
erruption
5. Pour agarose into a plastic beaker and add SYBR® Safe to solution once the agarose is heated.
Add 1 μL of SYBR® Safe to 10 ml of agarose solution. If using ethidium bromide, add directly
to glass beaker and use the same quantity per mL, but make sure to use a 1 in 10 ethidium
bromide solution.
6. Pour gel onto plate and allow to cool for 30 min or until gel is opaque.
7. Run electrophoresis machine. Time will vary depending on size and density of gel eg. 250 mL
gel can run at 90 V for 3-4 h. A 50 mL gel may run for 30 min to 1 h at 100 V.
Viewing Results:
1. For gels with SYBR Safe, place onto the Safe Imager Transilluminator (with amber filter unit)
and cover with camera box for viewing image on screen. If using ethidium bromide place gel
onto a UV light trans-illuminator and cover with camera box.
2. Save gel image electronically or print.
3. For PCR products, a single bright band at around 500-1000 bp (Test 1) or 1500 bp (Test 2)
indicates successful PCR amplification; any samples showing multiple bands should not be
used for RFLP, and either the source of contamination (two DNA’s in the sample) or PCR
optimisation needs to be considered
4. For restriction digest products, use a 100 bp ladder to determine size of fragments and compare
with species fragment size chart in Armstrong & Cameron 1998; results of all enzymes used
need to agree for diagnosis.
APPENDIX 3
Restriction Enzyme Haplotype Chart
(adapted from Armstrong & Cameron 1998)
Restriction enzyme pattern types (see photographs in Appendix 4) are represented by different letters. Same
letters with different numbers are similar patterns, but distinguishable within a pattern type. A minimum of four
enzymes is recommended, to include one or more of those highlighted where possible.
= unique pattern,
Genus
Subgenus
Anastrepha
Bactrocera
Ceratitis
Dacus
Rhagoletis
Afrodacus
Austrodacus
Bactrocera
Daculus
Notodacus
Zeugodacus
Ceratitis
Callantra
= unique within a pattern type,
Species2
Abv.3 Alu I
Cfo I
A. bistrigata
A. fraterculus
A. grandis
aA. ludens
A. obliqua
A. serpentina
A. sororcula
A. striata
aA.suspensa
A. zenildae
B. jarvisi
B. cucumis
bB. aquilonis
B. carambolae
B. curvipennis
cB. dorsalis
dB. facialis
eB. frauenfeldi
B. kirki4
B. latifrons
B. melanotus
B. musae
bB. neohumeralis
cB. papayae18
dB. passiflorae
cB. philippinensis18
B. psidii
B. quadriestosa
eB. trilineola
B. trivialis
bB. tryoni
B. umbrosa
B. zonata
B. oleae
B. xanthodes
B. cucurbitae
C. capitata
C rosa
D. solominensis
R. completa
R. pomonella
Ab
Af
Ag
Al
Ao
As
Aso
Ast
Asp
Az
Bj
Bc
Ba
Bcm
Bcp
Bd
Bf
Bfr
Bk
Bl
Bm
Bmu
Bn
Bpy
Bp
Bph
Bpd
Bq
Btl
Btr
Bt
Bu
Bz
Bo
Bx
Bcb
Cc
Cr
Ds
Rc
Rp
A2/A3
A1
A2
A1
A2
A3
A1
A2
A1
A1
B2
B4
B2
B1
B2
B2
B2
B1
B1
B3
B2
C
B2
B2
B2
B2
B2
B2
B1
B2
B2
B3
B1
B4
A2
B5
A4
A5
A1
A3
B3
A2
A1
A2
A1
A2
B
A1
A2
A1
A1
C1
D1
C3
C5
C1
C1
C2
C5
C5
C3
C1
C1
C3
C1
C2
C1
C1
C1
C5
C3
C3
C4
C5/C1
C6
C7
D2
D3
D4
E
F
D1
= unique to group of indistinguishable species.
Restriction Enzyme Pattern Types1
Dde I
Dra I
Hinf I
Nco I
A1
A1
B
A1
A1
C
A1
A1
A1
A1
A5
A3
A5
A6
A5
A5
A4
A6
A6
A4
A5
A5
A5
A5
A4
A5
A5
A5
A6
A5
A5
A4
A6/A5
A2
A2
A1
D
E
A1
A3
A4
A1/A2
A1
A3
A1
A1
A2
A1
A2
A1
A1
E1
B1
E1
D1
E1
D2
D3
E2
E2
E3
E1
D2
E1
D2
D3
D2
E1
E1
E2
E1
E1
F
D1/D2
G1
C
B2
D1
E4
B3
B4
G2
B
A1
B
C
A2
B
A1
B
C
D
E1
F1
E1
E2
E1
E1
E3
E1
E1
E3
E1
E1
E1
E1
E3
E1
E1
E1
E1
E1
E1
E3
E1/E2
E4
E4
F2
E2
H
F1
A3
G
A1
A1
A1
A1
A1
A2
A1
A1
A1
A1
A2
A1
A2
B1
A2
A2
A2
A2
A2
B2
A2
B3
A2
A2
A2
A2
A2
A2
A2
A2
A2
A2
B1/A2
A1
A1
A1
A3
A4
A1
A2
A2
Rsa I
Sau 3A Ssp I
B4
A
B1
A
A
B2
A
B3
A
A
C1
F1
C1
D
C2
C1
E1
D
D
E2
C1
C1
C1
C1
E1
C1
C1
C1
D
C1
C1
G
D/C1
B5
I
J
K
N
L
F2
M
A1/A2
A1
B
A1
C1
C2
C3
A2
A1
A1
D1
A3
D1
D3
D1
E1
D2
D3
D3
F
D1
E1
D1
E1
D2
E1
D1
D1
D3
D1
D1
D2
D3/E1
A2
A1
A4
E2
G
A1
A2
A3
Some species show two patterns correlating with different populations, given as Type 1/Type 2 respectively.
Species in bold are listed on the Biosecurity (Notifiable Organisms) Amendment Order (1997); species prefaced with the same
superscript letter are as yet indistinguishable using the current list of restriction enzymes.
3
Abbreviated species names assigned for routine use as in Appendix 7.
4
B. kirki can be distinguished from B. frauenfeldi and B. trilineola using an additional restriction enzyme Acc I, see Addendum.
1
2
18
Synonymised with B. dorsalis
D2
A1
B1
A1
D1
A2
A1
D2
A1
A1
E
F1
G1
D3
G3
D3
D3
H
H
A2
H
D3
G1
D3
D3
D3
I
A2
H
D3
G1
D3
D3
B3
C
F3
G2
J
B2
F2
I
APPENDIX 4
Diagnostic restriction patterns
(adapted from Armstrong & Cameron, 1988)
The restriction patterns are summarised in the following photographs (overpage). Aim to use four
enzymes to identify unknown specimens. At least one example of each species is presented; see
Appendix 3 for species names in full.
The tables of fragments provide only approximate sizes (bp) based on the limitations of estimating from
electrophoresis gels. However, comparative estimations are still diagnostic.
mw, molecular weight marker (100bp ladder, Gibco BRL) where highlighted bands indicate 2000 bp
(top), 600 bp (middle) and 100 bp (bottom).

B. correcta, not included in the report or elsewhere in that Manual.
Alu I (AG/CT)
D1 F E D3 D2 C7 C6 C4
C3 C5 C1 C1 C2 C3 C1 C1 C3
C5 C5 C2 C1 C1 C5 A1 D1 C1 A1 A1 A2 A1 B A2 A1 A2 A1
Rp Rc Ds Cc Bcb Bx Bo Bu
Bt Btl Bq Bpd Bp Bn Bmu Bm Bl
Bk Bfr Bf Bd Bcp Bcm Az Bc Bj
mw
mw
D1 D3
D4
Rp Cc ____Cr____
mw
mw
C1 C5 C1 D2 C3 C3 C3 C3
C3 C3
Bpy Bcm Bd Bcb Btr Bt Bn Ba
Btr Bl Bcr Bcm Bd Bz1 Bz2
mw mw
C5C5C1
C5 C1
mw mw
Haplotype
Az Asp As tAso As Ao Al Ag Af
mw
A1 A1 A2 A1 B A2 A1 A2 A1 A2 A2
Az Asp Ast AsoAsAo Al Ag AfAb2Ab1
mw mw
mw
Fragment sizes (bp)
A1
1080-1050
240
120
A2
1150-1100
240
120
240
120
B
800-790*
340-330
C1
790-770
C2
770-760
C3
780-770
C4
770
C5
780-760
C6
D4
240-230*
250
330-310
240
700-660*
200
180-170
130
120
110
170
130
120
110
170
130
120
110
170
130
120
110
170
130
120
110
170
130
120
110
170
130
120
110
130
120
110
130
120
110
130
120
110
130
120
110
130
120
110
130
120
110
1200-1180
D2
D3
270
680
C7
D1
290-280
980
1300
1700
E
720-700*
F
640
450
170 150
______________________________________________________________________________________
*Sometimes double band
Cfo I (GCG/C)
B3 A3 A1 A4 B5 A2 B4 B3
B2 B1 B2 B2 B2 B2 C B2 B3
B1 B1 B2 B2 B2 B1 A1 B4 B2
A1 A1 A2 A1 A3 A2 A1 A2 A1
Bt Btl Bq Bpd Bp Bn BmuBm Bl
Az Asp Ast AsoAs Ao Al Ag Af
mw
B3 A4
mw
A5
Rp Cc ___Cr___
mw
mw
B2 B1 B2 B5 B2 B2 B2 B2
B2 B3 B1 B1 B2 B1 B1
A1 A1 A3 A1 A3A2A1 A2 A1 A3 A2
Az Asp Ast AsoAsAoAl Ag Af Ab2 Ab1
mw mw
mw mw
mw mw
Note: refer to original photograph to check for consistency with subtle variation between B group.
Haplotype
Fragment size (bp)
A1
980-940
A2
A5
B1
B2
530
1000
A3
A4
mw
530
1050-1020
530
1250
530
1600
530
1010-990
980-930*
B3
B4
B5
C
900
870-820
530
180-170
530
180
530
180-170
530
170
720-700* 530
900-880*
530
170
240
___________________________________________________________
* Sometimes double band
Dde I (C/TNAG)
A4 A3 A1 D A1 A2 A2 A4
A5 A6 A5 A5 A4 A5 A5 A5 A4
Rp Rc Ds Cc BcbBx Bo Bu
mw
A6 A6 A4 A5 A5 A6 A1 A3
Bt Btl Bq Bpd Bp BnBmuBm Bl
mw
A4 D
E
Rp Cc ___Cr___
A5 A1 A1 A1 A1 C A1 A1 B A1
Bk Bfr Bf Bd Bcp Bcm Az Bc
mw
Bj Az AspAstAso As Ao Al Ag Af
mw
mw
A5 A6 A5 A1 A5 A5 A5 A5
A5 A4 A6 A6 A5 A6 A5
A1 A1 A1 A1 C A1 A1 B A1 A2 A1
Az Asp Ast Aso AsAo Al AgAf Ab2 Ab1
mw mw
mw mw
Haplotype
mw mw
mw
Fragment size (bp)
A1
800-750 270
220
180-160
A2
860-830
270
220
170
A3
900
270
220
160-150
A4
950-930
270
220
160
270
220
170-160
270
220
170-160
270
220
270
220 210
270
220
270
220
A5
1000-980*
A6
1030
B
850
C
D
E
830-800*
1150
1500
120
130
170
___________________________________________________________________________________
Dra I (TTT/AAA)
G2 B4 B3 D1B2 C G1 F
E1 E2 E1 E1 D3 E1 D2E1E3 E2 E2 D3 D2 E1 D1A1 B1 E1
A1 A1 A2 A1 A2 A1 A1 A3 A1
Rp Rc Ds Cc BcbBx Bo Bu Bt Btl Bq Bpd BpBnBmuBmBl Bk Bfr Bf Bd Bcp Bcm Az Bc Bj Az AspAstAsoAsAo Al Ag Af
mw
mw
G2 D1
E4
Rp Cc ___Cr___
mw
mw
A1 A1 A2 A1 A2 A1 A1 A3 A1 A1 A2
Bt Bn Ba
Btr Bl Bcr Bcm Bd Bz1Bz2
Az Asp Ast Aso As Ao Al Ag Af Ab2 Ab1
mw
mw mw
mw mw
mw
Fragment size
A1
1150-1080
A2
1150-1100 400
A3
370
1180
320
1250
B2
1100
B3
B4
mw
E1 E3 D3 D1 D2 D1 D2
Haplotype
B1
mw
E1 E1 E1
200
120
200
120
1120
180
1250
C
1150-1100*
D1
1200
D2
1220-1200
D3
1200
150
180
130
250-240*
70?
360-350
110-100
320-310
110
290
E1
1030-990*
E2
1020
E3
1020
E4
1040
F
1020
G1
1120
G2
1120
110
320-310
180-160
110-100
360-350
180-150*
280
160
410
110
(160)x2
280
160
170
200
_________________________________________________________________________________
*Sometimes double band
110
110 [90,70?]
150
130
140 120
100
Hinf I (G/ANTC)
G A3 F1 E2 F2 E4 E4 E3
E1 E1 E1 E1 E3 E1 E1E1E3
E1 E1 E3 E1 E1 E2 D F1 E1
D C B A1 B A2 C B A1
Rp Rc Ds Cc Bcb BxBoBu
Bt Btl Bq Bpd Bp BnBmuBmBl
Bk Bfr Bf Bd BcpBcmAz BcBj
AzAspAst Aso As Ao Al Ag Af
mw
mw
G E2
H
Rp Cc ____Cr_____
mw
mw
mw
mw
E1 E3E1Φ E2 E1 E2 E1
D C B A1 B A2 C B A1 B B
Btr Bt Bn Ba
Btr Bl Bcr BcmBd Bz1 Bz2
AzAspAstAsoAsAo Al Ag Af Ab2 Ab1
mw
mw mw
Haplotype
mw mw
mw
Frgagment size (bp)
A1
580-570
A2
610
A3
B
mw
E1 E1 E1 E1
630
280
260 240 150
110 80
270 240 150
110 80
260
1200-1100
C
590-580 490-480
D
380
70
150
110 80
70?
150
110 80
70
150
110 80
70
270 240 200
150
110 80
70
E1
1010-970*
260
150
110 80
70
E2
1050
260
150
110 80
70
260
150
110 80
70
260
E3
920
E4
880-860
F1
900-850
F2
780
G
H
640
780
150
110 80
70
370
150
80
70
370
150
80
70
150
110 80
70
260
410 330 280
190
150 120
________________________________________________________________________________
NcoI (C/CATGG)
A2 A2 A1 A3 A1 A1 A1 A2
A2 A2 A2 A2 A2 A2B3A2 B2
A2 A2 A2 A2 A2 B1 A1 A1 A2
A1 A1 A1 A1 A2 A1 A1 A1 A1
Rp Rc Ds Cc Bcb BxBoBu
Bt Btl Bq BpdBp BnBmuBm Bl
Az AspAstAso As Ao Al Ag Af
mw
A3
mw
A4
Cc ____Cr____
mw
mw
mw
mw
A2 B1 A2 A1 A2 A2 A2 A2
A2 B2A2Φ B1 A2 B1 A2
Btr Bl Bcr Bcm Bd Bz1 Bz2 Az Asp Ast Aso As Ao Al Ag Af Ab2 Ab1
mw mw
mw mw
Haplotype
m w mw
A3
mw
Fragment size
A1
A2
A1 A1 A1 A1 A2 A1 A1 A1 A1 A1 A1
1600-1500 (uncut)
1700 (uncut)
1800 (uncut)
B1
1050
B2
1050
B3
1050
680
__________________________________________________
590
630
Rsa I (GT/AC)
M F2 L K
J
I B5 G
mw
C1 D C1 C1 E1 C1 C1 C1 E2
D D E1 C1 C2 D
Bt Btl Bq Bpd Bp Bn Bmu Bm Bl
mw
I
K
N
mw
A A B3 A B2 A A B1 A
Az Asp Ast Aso As Ao Al Ag Af
mw
BpyBcm Bd Bcb Btr Bt Bn Ba
mw mw
mw
C1 E2 DΦ D C1 D C1
C1 D C1 J C1 C1 C1 C1
Bx Cc ___Cr____
A F1 C1
Az Asp Ast Aso As Ao Al AgAfAb2Ab1
mw mw
Haplotye
mw
A A B3 A B2 A A B1 A B4 B4
mw mw
mw
Fragment size
A
380-340∆
B1
440 430
B2
480
B3
480-450
B5
430
C1
530-500*
C2
490
360
290
380 370
290
(370-360)x2
B4
290
290
410
380 370
290
400
370
290
460-440*
410
290
460
410
290
410
290
D
520-500
E1
500
420 410
290
E2
500
(410)x2
290
F1
570
F2
570
G
I
490
410
380
510-500*
300
290
300
290
410
750
290 260
150
370 360
J
310
290
K
450
410 400
380
290 260 240 110
L
450
380
320
M
580
380
330
___________________________________________________________________________________
∆
Several patterns, combinations of 3 bands; * sometimes double band
290
230
Sau 3A (/GATC)
A3 A2 A1 E2 A4 A1 A2 D2 D1 D1 D3 D1 E1 D1 D2 D1 E1 D1
F D3 D3 D2 E1 D1 D3 D1 A3 D1 A1 A1 A2 C3 C2 C1 A1 B A1
Rp Rc Ds Cc Bcb Bx Bo Bu Btr Bt Btl Bq Bpy Bpd Bp Bn Bmu Bm
Bl Bk Bfr Bf Bd Bcp Bcm Ba Bc Bj Az Asp Ast Aso As Ao Al Ag Af
mw
mw
A3 E2
G
Rp Cc ____Cr____
mw
mw
E1 D3 E1 A4 D1 D1 D1 D1
D1 F
mw mw
D3
D3 E1 D3 E1
mw mw
A1 A1 A2 C3 C2 C1 A1
B
A1 A2 A1
Az Asp Ast Aso As Ao Al Ag Af Ab2 Ab1
mw mw
Haplotype
mw
Fragment size
A1
770
A2
770
A3
770 700
A4
770
530
110
B
770
530
110 80
C1
770
C2
770
C3
770
D1
770
D2
770
D3
770
E1
E2
800
880
620-580*
110
670-660*
110
110
450
520/500/480/400**
440
200-170
110
200-190
110
140-130
410-400 390-360*
380 360
450-440* 380-370
110
110
110
110
770
110
770
110
F
760 730
110
____________________________________________________________________________________________

Diagnostic 80 bp and 100 bp bands not visible here, see photograph above; * sometimes double band.
** any combination of two bands
Ssp I (AAT/ATT)
D3 A2 I
D3 H A2 H H H D3 D3
D3 D3 D3 G1 G1 G1 E F3 C B3 F1
D2 A1 A1 D2 A2 D1 A1 A1 B1 A1 A1
Btr Bq Bpd BmuBm Bl Btl Bk Bfr Bp Bf
Bcm Bpy Bd Bt Bn Ba Bj Bcb Bx Bo Bc
Ab Az Aso Ast As Ao Asp Al Ag Af Af
mw
mw
B2 G2 J
J
G1 G1 G1 D3 I
Ds Cc Cr Cr
mw
mw
A2 E G3
Bt Bn Ba Bp Bpd Bq Bj Bcp
mw mw
D3
D3 A2
D3 D3 D3 D3
Rp Rc Ds Cc
Bu
mw mw
mw
Fragment size
A1
1500 (uncut)
1700 (uncut)
B1
1050
B2
370
1100
B3
350
1200
D1
(60)
380
1150
C
210
110
1350-1300
150-140
1650-1600
100
E
880
F1
100
650
900 640
F3
880
G1
500
1000
G2
1020
G3
1020
H
640
880
F2
550
100
520
580
100
100
1450
I
1350
250
_____________________________________________________________________________________________________

80
1400
D2
D3
F2 B2 G2
mw mw
Haplotype
A2
I
mw
D3
Fragment not always visible
150
100
APPENDIX 5
Dominiak, Daniels, and Mapson, (2011). Review of the outbreak
threshold for Queensland fruit fly (Bactrocera tryoni Froggatt). Plant
Protection Quarterly, 26(4), 141.
This paper has been reproduced with permission.
Plant Protection Quarterly Vol.26(4) 2011 141
Review of the outbreak threshold for Queensland
fruit fly (Bactrocera tryoni Froggatt)
i.
Bernard C. DominiakA, David DanielsB and Richard MapsonC
Department of Primary Industry NSW, Locked Bag 21, Orange, New South
Wales 2800, Australia and the Department of Biological Sciences, Macquarie
University, New South Wales 2109, Australia.
B
Department of Agriculture, Fisheries and Forestry, PO Box 858, Canberra,
ACT 2601, Australia.
C
Department of Primary Industries Victoria, 621 Burwood Highway,
Knoxfield, Victoria 3180, Australia.
A
ii.
Abstract
Fruit flies cause losses in horticultural
produce across the world and are a major
quarantine concern for most countries.
Queensland fruit fly (Qfly) is a native to
Australia and is also present in a small
number of Pacific Island countries. The
detection of Qfly in recognized pest
free areas triggers quarantine restrictions from domestic and international
markets. In Australia, the detection of
five male flies has been taken to indicate an outbreak (i.e. unacceptable risk).
Matching the domestic standard, many
countries have accepted the 5-fly limit as
a quarantine threshold. But some other
countries have set the detection of two
male flies, or even a single fly, as the
threshold for an outbreak. This different
standard creates an administrative complexity for exporters and trade regulators.
In this paper, we review the published
science covering the impediments to pest
establishment. Outbreak data from Victoria and New South Wales during 2007
and 2009 are reviewed in relation to the
2-fly and 5-fly thresholds. Large volumes
of fruit have been traded within Australia and internationally based on the 5-fly
threshold without incident and there is
no evidence that the 2-fly threshold is
more appropriate. While Qfly is recognized as being capable of longer distance
dispersal than some other fruit fly species, it is also recognized as a poor colonizer. The 5-fly threshold is proposed as
the most appropriate threshold for imposition of quarantine restrictions and
is recommended as a universal standard
for harmonization of quarantine regulations.
iii.
Introduction
There are about 4500 species of fruit flies
worldwide. In the Pacific area alone, there
are 350 species of which at least 25 species
are regarded as being of major economic
importance (Allwood 2000). The genus
Bactrocera contains over 400 species, distributed primarily though the Asia-Pacific
area including Australia (Drew 1974).
Tephritid fruit flies cause direct losses to
many fresh fruit and some vegetable industries, resulting in adverse impacts on
trade and economies of many countries
(Li et al. 2010, Stephenson et al. 2003).
With the increasing globalization of trade
(Stanaway et al. 2001, Plant Health Australia 2010), fruit flies pose a major quarantine concern that is currently monitored
through regional surveillance programs
(International Atomic Energy Agency
2003, Stephenson et al. 2003, Oliver 2007).
The Queensland fruit fly Bactrocera
tryoni (Froggatt) (Diptera: Tephritidae)
(Qfly) is a major fruit fly pest of Australian horticulture, attacking most fruit and
many vegetable crops (e.g. stone fruit, citrus, coffee, tomato, capsicum, pome fruit)
(Bateman 1991, Anon. 1996, Hancock et al.
2000). Qfly is an Australian native and is
currently only found in Australia and on
some Pacific islands (Drew 1989, White
and Elson-Harris 1992). Given its pest status within Australia, Qfly is also a significant quarantine concern for many trading
partners. Markets trading in commodities
that may be subject to Qfly infestation
require assurance of reliable monitoring
grids, evidence-based outbreak thresholds and appropriate quarantine measures
(Bateman 1991, Anon. 1996, Clarke et al.
2011).
In the early 1990s, Bateman (1991) reviewed existing domestic trade conditions
and recommended a uniform agreement
among the Australian states for the management of and trade in Qfly host commodities. In response, the Code of Practice for the Management for Queensland
Fruit Fly (Anon. 1996) was published, with
particular emphasis on managing the TriState Fruit Fly Exclusion Zone (FFEZ) so
that fruit could be traded domestically
with increased efficiency. The FFEZ production area is managed as a pest free
area and is recognized by all Australian
states as being free from economic fruit
flies. Strict quarantine measures are in
place to prevent entry of fruit flies and any
incursions invoke a rapid and thorough
eradication response. Within the FFEZ,
four separate pest free areas have been
established to facilitate trade into international markets. These include the Riverina
area of New South Wales, the Sunraysia
region of Victoria/New South Wales, the
Riverland area of South Australia, and the
Shepparton Irrigation Region of Victoria.
Under some circumstances, Qfly do enter
the FFEZ and are detected in monitoring
traps (Dominiak et al. 2003a, Dominiak
and Coombes 2009). Single-fly detections
are almost always isolated incursions
that do not indicate breeding populations
(Meats et al. 2003).
For domestic trade (Anon. 1996), an
outbreak is declared following one (or
more) of three thresholds. These thresholds are the detection of:
(1) five male flies within 1 km within 14
days, or
(2) one mated female, or
(3) one or more larvae in fruit grown in the
area.
The quarantine distance around any outbreak is 15 km. This domestic trade agreement (Anon. 1996) was broadly adopted
in principle by 19 countries as the basis
of international trade. However some
key components of this agreement, such
as the outbreak threshold, have not been
accepted by some importing countries. In
1996, the outbreak threshold varied from
1, 2 and 5 male flies for 1, 14, and 3
countries respectively (Robert McGahy
personal communication). The threshold
of two male flies and five flies (hereafter
referred to as 2-fly and 5-fly thresholds)
are the most commonly used quarantine
thresholds. The 2-fly threshold is based
on detections within 400 m while the 5-fly
threshold is based on detections within
1 km. By 2009, with increased international acceptance of the 5-fly threshold,
this position had changed with 1, 11 and 9
countries accepting 1, 2 and 5 male flies respectively as outbreak thresholds (David
Daniels personal communication). These
different outbreak thresholds lack a robust
scientific basis and create complex administration procedures for trade regulators.
An agreed evidence-based Qfly outbreak
threshold would harmonize market requirements and thereby facilitate domestic and international trade (Clarke et al.
2011). A universal outbreak threshold
would have major implications for trade,
quarantine and the minimization of pesticides in the environment as part of eradication programs (cover and bait sprays).
There is a geometric expansion of areas
requiring disinfestation unnecessarily by
each kilometre of quarantine radius for
outbreaks triggered by a low threshold
(Clarke et al. 2011).
The purpose of this paper is to review
the data from February 2007 to April 2009
for 2-fly and 5-fly thresholds for fruit fly
outbreaks in Victoria and New South
142 Plant Protection Quarterly Vol.26(4) 2011
Wales, and to examine the published scientific evidence since 1996 regarding incursions, survival, breeding populations
and the resultant outbreak thresholds.
This review will focus only on male flies
as most outbreaks are triggered by the detection of male flies.
iv.
Impediments
establishment
Founding propagules
to
pest
It has been shown that the introduction
of fruit flies into pest free areas is most
often the result of illegal transportation
into and the inappropriate disposal of infested host material within the pest free
area (Bateman 1991, Dominiak et al. 2000,
Dominiak and Coombes 2009). This indicates that relatively small parcels of fruit
flies are the source of most Qfly detections. Qfly dispersal from these points of
introduction is limited by lifespan and the
ability to find food to sustain the effort of
longer or frequent short flights, survive
adverse weather and avoid predation
(Meats and Smallridge 2007, Meats and
Edgerton 2008, Gilchrist and Meats 2011,
Weldon and Taylor 2011). Immature fruit
flies disperse for about two weeks in random directions and do not travel in pairs
(Fletcher 1974a). Following the introduction of small numbers of Qfly into fruit fly
free areas, the chances of a sexually mature
male and female occurring in the same
tree or group of trees after many days of
dispersal is extremely low (Fletcher 1974a,
Bateman 1977, Meats 1998, Weldon 2003,
2007, Weldon and Meats 2010). Following
the introduction of propagules of infested
fruit into fruit fly free areas, Meats (1998)
and Meats et al. (2003) proposed that flies
disperse into a mate-free void and self extinguish, as it becomes increasingly unlikely that they will participate in mating
and will therefore not establish a breeding
population.
Nutrition
Nutrition is key for Qfly survival, dispersal, reproduction and establishment
of new populations. Wild flies must find
sugar, minerals, water and protein from
products such as bird faeces, honeydew
and fruit juice (Bateman 1972, Drew et al.
1984, Dalby-Ball and Meats 2000). In dry
environments, these products are difficult
to find. The average lifespan of Qfly without food and water is approximately 45
hours (Weldon and Taylor 2010, Dominiak unpublished). Qfly require a balanced
diet, as diets with too much or too little
protein and carbohydrate result in adverse effects on either longevity or reproduction (Prabhu et al. 2008, Fanson et al.
2009). Protein feeding by post-teneral Qfly
has been consistently reported to enhance
sexual performance (Perez-Staples et al.
2007, 2008, Prabhu et al. 2008). Bacteria
on the surfaces of leaves and fruit appear
to be a key food source for Qfly (Drew
1987, Drew and Lloyd 1987, Fletcher 1987).
However in fruit fly free regions of southern Australia, populations of these bacteria may be infrequent and erratic owing to
unfavourable climate (Drew et al. 1984,
Courtice and Drew 1984). In the absence
of these bacteria, Qfly must find protein
from alternative ephemeral sources (Perez-Staples et al. 2007, Weldon and Taylor
2011) and therefore a large proportion of
flies may not reach sexual maturity and
contribute to population growth.
Climate in the FFEZ is normally dry
and crops require irrigation. The combination of low humidity and starvation are
considerably more punitive for Qfly survival than starvation alone (Weldon and
Taylor 2010). Desiccation resistance is generally lower for females than males and resistance also declines with age. Therefore,
the lack of available food resources in the
environment diminishes the chance of survival to maturity and the chance to compete for a mating. In summary, the FFEZ
usually presents as a hostile environment
and affords very limited resources for the
establishment and spread of Qfly.
Dispersal before mating
Qfly actually spend very little time flying.
Fletcher (1973, 1989) noted that flies spend
most of their time making trivial flights or
walking within the tree canopy. In response to higher fruit abundance, both
male and female Qfly visit more leaves
and hence spend more time in trees containing more fruit (Dalby-Ball and Meats
2000). Flies move around the canopy primarily by walking, and when they do fly,
it is usually over distances of less than 50
mm in an upward direction. In laboratory
observations, wild Qfly spend only about
0.6% of their time in flight with walking
(67.5%), inactivity (18.0%) and grooming
(14%) taking up the remainder of their
time (Weldon et al. 2010). In the field, Ero
et al. (2011) reported that resting was the
most commonly observed behaviour for
Qfly while feeding was rarely observed.
The flight activity patterns and shortrange dispersal patterns of emerged adults
are similar for male and female Qfly (Weldon and Meats 2007, Weldon et al. 2010).
Clarke and Dominiak (2010) found a high
correlation between male and female trap
catches and suggested that changes in
male distribution also reflect the distribution of female Qfly. Fletcher (1973) reported that the weekly declines of released
Qfly were similar for males and females.
Meats (1998) also assumed that males and
females had similar dispersal. Therefore
the trapping of male flies is likely to reflect
a similar number of female flies in the environment.
Mating after dispersal
Male Qfly use pheromones and acoustic signals to attract sexually receptive
females, and mate only during a brief period of about 30 minutes at dusk (Tychsen
and Fletcher 1971). Males gather on the
upwind side of trees, where they release
pheromone and fan their wings, directing
the pheromone stream through the foliage
(Tychsen 1977). Male calling is energetically expensive and calling in aggregations
maximizes their chances of mating success
(Weldon 2007). Males downwind of an aggregation might fly upwind in response to
pheromone being released by calling
males. There is a period of only about ten
minutes during which males could fly to
join the flying swarm (Tychsen 1977), and
only enough time to mate once at each
dusk, although males may mate in many
dusk periods over their lifetime (Fay and
Meats 1983, Radhakrishnan and Taylor
2008, Radhakrishnan et al. 2009). Males do
not mate when temperatures at dusk are
below 15°C with 50% of males mating at
20°C or higher temperatures (Meats and
Fay 2000). Qfly have a relatively poor capacity to locate an odour source and it has
been suggested that pheromones operate
mainly within a single tree canopy (Meats
and Hartland 1999, Weldon 2007). Acoustic cues are only effective over a short distance of about 50 cm (Mankin et al. 2004,
2008, Sivinski personal communication).
Female Qfly move directly towards the
males from up to 50 cm away (Tychsen
1977).
Odour plumes carried by light winds in
trees usually become chaotic within a few
centimetres of their source and provide
few cues as to the direction of the source
(Griffiths and Brady 1995). Qfly compensate for the diffused odour by making a
series of short flights or walks (Meats and
Hartland 1999) or by using large visual
cues such as foliage to locate the source
of odours (Dalby-Ball and Meats 2000).
Female Qfly visit single male Qfly less frequently than aggregations (Weldon 2007).
If the Qfly population is sparse, these limitations therefore result in single males being unlikely to attract a female and mate.
Meats (1998) estimated the chance of
a successful mating between two Qfly on
the same tree of 5 m × 5 m to be about 0.1%.
Even in small cages, the chance of mating
was only 0.8% (Fay and Meats 1983). A
male Qfly has about a one in 400 chance of
being in the right place at the right time if
the density of males in the area was only
one per hectare. Meats (1998) estimated
that a single mating was probable when
there were six male and six female Qfly
present per hectare.
Current outbreak thresholds
Following the detection of small numbers
of male Qfly (the number depends on the
importing market), trading partners may
fear that fruit harvested for trade could
contain larvae that might establish populations in areas currently free from this
pest. In Anon. (1996), a breeding population is considered to have three indicators. Two are direct indicators; larvae detected in fruit harvested within the area
or a mated female detected in monitoring
traps. In fruit fly free areas, larval searches
are not routinely undertaken by regulatory authorities at times when no fruit
flies are detected, although they are sometimes conducted to meet some importing
country requirements. If present, larvae
are generally detected and reported by
the public but these are rare events in the
FFEZ. Because of inefficiency and difficulty of detecting larvae, a monitoring grid or
array has been established to provide an
early warning of incursions by adult Qfly.
Qfly populations are known to occur
naturally in about a 50:50 male:female ratio (Dominiak et al. 2008, Clarke and Dominiak 2010). In the FFEZ, female Qfly are
poorly attracted to monitoring traps (Dominiak et al. 2003a, Dominiak 2006, Dominiak and Nicol 2010). However, these
traps and lures may be more successful
in tropical regions (Clarke and Dominiak
2010). Due to the lack of reliable female
lures, the monitoring array relies primarily on the trapping of male flies and this
is a common situation in most countries
(International Atomic Energy Agency
2003). In Australia, Willison discovered
that male Qfly are attracted to raspberry
ketone and subsequently experimented
with a related chemical, cuelure (Allman
1958). Cuelure breaks down into raspberry ketone and this process is accelerated
in the presence of moisture (Metcalf 1990).
Sexually mature male Qfly are attracted to
raspberry ketone in nature (Tan and Nishida 1995). While male flies trapped may be
sexually mature, there is no current technology which can indicate if a Qfly male
has mated and therefore that a breeding
population exists. In the absence of this
technology, Bateman (1991) proposed that
five male flies are an indicator of a breeding population and this is later supported
by Meats (1998).
male flies are detected within one kilometre of each other within 14 days, 31 supplementary traps must be deployed within
200 metres (the outbreak zone) of the 2-fly
detection and fruit must be checked for
larvae. Supplementary traps must stay
in place for nine weeks and be inspected
twice weekly. If fewer than five male Qfly
are trapped within 1 km within any 14
day period, an outbreak is not declared. In
essence, it is deemed that a breeding
population does not exist. If a total of five
or more Qfly are detected within any 14
day period, an outbreak is declared for all
domestic and international markets. After the outbreak declaration, no produce
within the outbreak zone (within 200 m of
the detection point) can be traded. All
produce between 200 m and 15 km (the
suspension area) must be treated with an
approved disinfestation protocol before
being transported into or sold in fruit fly
sensitive markets (Jessup et al. 1998, De
Lima et al. 2007).
The detection date of the last fly
trapped is used to determine the reinstatement of area freedom based on generation tables in Anon. (1996). For some
countries, these reinstatement periods
vary from one generation plus 28 days, 12
weeks, three generations and one year.
However apart from noting these differing standards, these reinstatement periods
will not be discussed in detail further in
this paper. Some countries have adopted
the 2-fly threshold (within 400 m) as the
outbreak threshold rather than the 5-fly
threshold (within 1 km). For Australian
exporters and regulators, the different outbreak thresholds result in disrupted trade
and an administration burden. Moreover,
the disparity in outbreak thresholds and
reinstatement periods places regulatory
authorities in a difficult position, needing to impose movement controls on host
commodities destined for markets with
different requirements.
Conditions under the current code
Australian states and territories have
agreed to the 5-fly threshold as an outbreak threshold. This agreement allows
susceptible produce to be traded based
on the specified conditions before or after an outbreak is declared. What happens
when a trading partner requires a different threshold?
In the Australian response, the detection of two flies requires the deployment
of supplementary traps and fruit searches.
However since the Australian 5-fly outbreak threshold is not reached, no movement controls are imposed and fruit may
move unrestricted from a 2-fly zone to
any part of the pest free area or the rest
of Australia. Further, no chemical control
measures are deployed. This contrasts
with countries that are more risk averse
Bateman (1991) and subsequently Anon.
(1996) recommended that five male flies
trapped within 1 km of each other within
a 14 day period was an appropriate outbreak threshold, or in essence indicated
unacceptable risk of a breeding population. This standard has been accepted
for domestic trade within Australia and
by many international trading partners.
However, some countries choose lower
outbreak thresholds. Presumably, these
lower standards are thought to provide a
higher level of assurance, but there have
been no empirical studies to support this.
As part of the 5-fly standard in Anon.
(1996), there is an intermediate step, presumably to further investigate for the presence of a breeding population. When two
v.
Implications for different
outbreak thresholds
and use a 2-fly threshold. A fruit fly outbreak in any country normally requires
an eradication response and movement
controls. Since Australia does not deploy
these responses for a 2-fly threshold, the
interpretation by a 2-fly importing country is that potentially infested produce can
move from the area immediately around
the 2-fly threshold to any other district.
What is the Australian response to these
mixed thresholds? Australia only imposes
eradication or movement controls after a
5-fly threshold and therefore countries
using the 2-fly threshold may deem the
entire or part of the pest free area infested.
Trade in fruit fly host commodities under area freedom arrangements into 2-fly
sensitive markets is likely to cease for the
entire or part of the pest free area. Costly
phytosanitary treatments are usually required for these 2-fly markets. The alternative is that Australia aligns its trade standard with the 2-fly threshold, and moves to
a lower universal outbreak threshold. This
action would decrease fruit fly free trade
because the 2-fly threshold is reached
more frequently than the 5-fly threshold.
Due to the difficulties in servicing markets with different outbreak thresholds,
would markets currently accepting the 5fly threshold then also align with the2fly threshold? This possible change in
outbreak threshold results in potentially
all countries accepting the lowest outbreak
threshold. One country is even more risk
averse, requiring a 1-fly threshold for
Qfly. If this strategy was adopted internationally by all countries for all species, the
1-fly threshold would become an unreasonable burden on all international trade.
This strategy would significantly increase
pesticide use in field eradication programs
and cause most fruit to be unnecessarily
treated with undesirable impact on the
environment; some chemicals such as methyl bromide are green house gases. There
would be significant benefits in harmonizing outbreak thresholds, but empirical evidence is required to support a preferred
universal threshold.
vi.
New information published
since the early 1990s.
Bateman’s (1991) report was the basis for
the current thresholds for outbreaks and
these were adopted as a code of practice
(Anon. 1996). More data of Qfly outbreaks
have been published since Bateman (1991)
and Anon. (1996), and these more recent
publications may prove instructive in assessing the relative merits of the 5-fly and
2-fly thresholds. The monitoring grid is
either a 400 m array in towns or a 1000
m array in orchards (Anon. 1996, Meats
1998). Fruit flies are reported to rarely disperse as far as one kilometre over their
lifetime (Maelzer 1990, Bateman 1991,
Meats 1996, Dominiak et al. 2003b, Meats
et al. 2003, 2006, Meats and Edgerton 2008,
Weldon and Meats 2010, Gilchrist and
Meats 2011). Given the large size of the
FFEZ, we can then surmise that introductions of Qfly usually result from the
carriage by humans of infested produce,
and this is supported by assessment at
roadblocks (Bateman 1972, Dominiak et
al. 2000, Sved et al. 2003, Maelzer et al. 2004,
Dominiak and Coombes 2009). Clift and
Meats (2005) used Bayesian scenario analysis to show that introductions by local
inhabitants contributed more to outbreaks
than passing travellers. Most humans reside in urban areas and therefore the more
intense monitoring array (400 m) in towns
is a reflection of the greater risk (Meats
1998, Maelzer et al. 2004). Townships
also provide better environments for survival and development of fruit flies than
the surrounding rural areas (Yonow and
Sutherst 1998, Raghu et al. 2000, Dominiak
et al. 2006). Backyard environments are
typically well watered and contain both
sheltered microclimates and host fruit
trees. Larger urban areas have an urban
heat island which further minimizes the
adverse effects of cold weather (Torok et
al. 2001, Dominiak et al. 2006). The one kilometre grid is used in lower risk rural and
orchard areas. These relatively sparsely
populated rural areas are unlikely to be
the first point of introduction of infested
fruit and if they are, rural areas generally
provide less favourable environments for
fruit fly survival (Dominiak et al. 2006).
Meats (1998) suggests that a detection
of two male flies within a two week period on the one kilometre grid represents
a density between 2.1 and 6.57 flies per
hectare within the outbreak zone (200 m
radius from the discovery point). The upper estimate of 6.57 flies per hectare represents the most extreme situation in which
the source of the incursion is directly in the
centre of four adjacent traps in a grid, maximizing its distance from any trap. Meats
(1998) proposed that when the density of
flies within the outbreak zone exceeded
six flies per hectare (of each sex), there was
potential (albeit a very low risk) for one
pair to mate. Superficially, the upper estimate of 6.57 flies per hectare appears to
exceed the minimum density required for
a mating to occur by 0.57 flies per hectare.
However the theoretical minimum breeding density proposed by Meats (1998) of
six male flies is an extremely conservative
estimate and is essentially only a ‘best
guess’ based on the information available
at that time. Several critical factors used to
obtain this theoretical minimum breeding
estimate remain poorly understood. Meats
(1998) estimated that the probability of a
successful mating in the field was less than
0.1 although in calculating the minimum
breeding density, the model assumed that
it was equal to 0.1. This estimate of 0.1 was
based on unpublished observations and
has not been substantiated with data or
confirmed experimentally in the field. The
model also assumes that there are ten dusk
periods available for mating and that mating can occur each and every dusk period.
Tychsen and Fletcher (1971) concluded
that mating only occurs within a 30 minute
period each day so that sexually mature
flies must be in close proximity at this time
for mating to occur. Meats (1998) acknowledges that his estimate of ten dusk periods
is also too high as it does not take into account adverse weather, the inhospitable
environment, and other factors unfavourable to fruit flies. In reality, mating will
only occur under favourable conditions
and in the presence of an adequate population. Another factor included in the estimate was dispersal behaviour observed by
Fletcher (1973, 1974a, 1974b) in a commercial orchard at Wilton, New South Wales.
Fletcher’s conclusions are specific to the
coastal environment where his study was
conducted and cannot be directly applied
to inland pest free areas that are much less
favourable to fruit flies (Dominiak et al.
2006). Meats (1998) also acknowledged in
his closing remarks that verification of the
models is still required and to date this issue remains unresolved. Meats (1998) recognized that his interpretation of trapping
rates on the 1 km grid is conservative, and
accordingly did not recommend that the
detection of two flies should be the threshold for quarantine precautions, but rather
a threshold to intensify the grid.
vii.
Data for 2007–2009 period
The period from February 2007 to April
2009 was chosen as a base to compare 2fly and 5-fly thresholds. Information was
provided by the state departments of
agriculture in Victoria and New South
Wales; there were no outbreaks in the
South Australian portion of the FFEZ
during this period. Climatically, autumn
2007 experienced near neutral values for
the Southern Oscillation Index (SOI) with
most parts of New South Wales and Victoria receiving average rainfall (Braganza
2008). The study area received slightly
above average rainfall in spring and summer of 2007 followed by dry conditions
in autumn, winter and spring 2008 (Duell
2009, Qi 2009). Average to below average
rainfall occurred in the FFEZ in summer
2008–2009 and autumn 2008 however several exceptional heatwaves occurred in
February 2009 (Mullen 2009). In this period, there were 27 outbreaks and these
were allocated to one of two categories.
Category A was a response after detection of two flies, where 31 supplementary
traps were deployed and larval searches
undertaken according to Anon. (1996). No
eradication or product movement controls
were imposed. Trade to countries using
the 2-fly threshold would have been suspended for that area. Trade was reinstated only after no flies were trapped for a
period of one generation plus 28 days.
There was no restriction of trade with any
Australian states or any 5-fly markets.
There were 19 outbreaks in this category
(Victoria: Invergorden 18 March 2008; Cobram 12 March 2008; Barooga 13 March
2008; Shepparton 10 April 2008; Bunbartha
11 April 2008; Katunga 14 April 2008; Numurkah 15 April 2008; Cobram East 2 June
2008; Echuca 18 September 2008; Irymple
24 March 2009. New South Wales: Yenda
11 April 2007; Darlington Point 26 April
2007; Yanco 29 May 2007; Lake Wyangan
12 March 2008; Hillston town 15 April
2008; Yenda 16 April 2008; Yanco 16 September 2008; Leeton town 16 September
2008; Hillston orchard 22 September 2008).
Category B was based on a 5-fly threshold. Subsequent procedures were according to Anon. (1996); supplementary traps
and larval searches were conducted,
eradication programs and product movement controls were initiated, and a 15 km
suspension zone was established. Trade in
fruit fly free produce to all domestic and
international markets (including countries
using the 2-fly threshold) was suspended
for all host commodities grown within the
suspension zone until there were no flies
trapped for one generation plus 28 days.
There were eight outbreaks in this category (Victoria: Koonoomoo 2 February
2007; Invergordon 20 March 2008; Bunbartha 22 April 2008; Katunga 13 May 2008;
Cobram East 19 June 2008; Shepparton 3
April 2009. New South Wales; Narrandera
23 May 2007; Yanco 28 October 2008.)
Of the 27 outbreaks, 19 Category A
outbreaks (70.4% of all outbreaks) did
not progress to a Category B outbreak
despite supplementary trapping and larval searches. Even with the low level of
progression to the 5-fly threshold (29.6%),
all susceptible host produce from the pest
free area required disinfestation before
being exported to markets requiring any
threshold other than the 5-fly threshold.
Meats et al. (2003) found 71% of single Qfly
detections did not lead to 5-fly outbreaks
and self extinguished without any eradication response. The 2007–2009 data for
the 2-fly threshold of 70.4% is consistent
with Meats et al. (2003).
Riverina trade volume since 1996
There has been considerable trade in host
produce from the FFEZ since 1996 using
the 5-fly threshold without any reports of
larvae found in produce. This confirms
that area freedom certification procedures
for Australia’s pest free area are robust
given that consumers are highly likely to
report and return damaged fruit to retailers. The volume of produce varies from
year to year. Australian Bureau of Statistics (2008) reported that, for the statistic
local areas of Carrathool, Griffith, Leeton
and Murrumbidgee, 8586, 166 689 and
172 387 tonnes of stone fruit, oranges and
other citrus respectively was produced.
These combined industries are valued at
$86.492 M (Australian Bureau of Statistics
2008). Given the volume and value of fruit
traded annually, if the 2-fly threshold was
an accurate indicator of crop infestation, it
is likely that Qfly would have been detected in consignments in domestic or international market during the past 15 years.
viii.
Closing comments
ix.
Acknowledgments
x.
References
Qfly is recognized as a poor colonizer in
fruit fly free areas such as the FFEZ, owing
to hostile conditions for survival and reproduction (Bateman 1972, 1977, Fletcher
1987, Edge et al. 2001, Meats et al. 2003,
Weldon 2007). Even introduction by human activity (jump dispersal) very rarely
results in establishment (Maelzer et al.
2004, Meats and Edgerton 2008). Given the
large volume of produce traded without
incident, the 5-fly threshold has a proven
track record of success in providing highly
effective phytosanitary assurance. Based
on the evaluation of outbreak data, there is
no indication that the 2-fly threshold provides any additional assurance. On this
basis, we recommend that international
trading partners adopt the 5-fly threshold
as a universal threshold that provides a
high level of assurance and also enables
increased trading opportunity.
The authors would like to thank the many
trap inspectors who inspect and report
on fly detections in Victoria and New
South Wales. The identification services
in both states are also greatly appreciated.
The administration and Information and
Communications Technology services in
state Departments also make a significant
contribution to maintaining and reporting
of databases which provided the information in this paper. Earlier versions of this
manuscript were improved by comments
from Satendra Kumar, Sarah Sullivan, Lionel Hill and Associate Prof Phil Taylor.
Allman, S.L. (1958). Queensland fruit fly.
New South Wales Department of Agriculture. Conference of Commonwealth
and State Entomologists. Brisbane, May
1957. Commonwealth Scientific and
Industrial Research Organisation, pp.
68-70.
Allwood, A. (2000). Regional approach to
the management of fruit flies in the
Pacific Island countries and territories.
In ‘Area-wide control of fruit flies and
other insect pests’, ed. K.H. Tan, pp.
439-48. (Penerbit Universiti Sains Malaysia, Penang).
Anon. (1996). Code of practice for the
management of Queensland fruit fly.
Standing Committee on Agriculture
and Resource Management, Department of Primary Industries, Canberra.
Australian Bureau of Statistics (2008).
Agricultural commodities: small area
data, Australia, 2005–2006. Report
71250DO003.
Bateman, M.A. (1972). The ecology of fruit
flies. Annual Review of Entomology 17,
493-518.
Bateman, M.A. (1977). Dispersal and species interactions as factors in the establishment and success of tropical fruit
flies in new areas. Proceedings of the Ecological Society of Australia 10, 106-12.
Bateman, M.A. (1991). The impact of fruit
flies on Australian horticulture. Horticultural Policy Council Report No. 3.
ISBN 0642161100.
Braganza, K. (2008). Seasonal climate summary southern hemisphere (autumn
2007): La Nina emerges as a distinct
possibility in 2007. Australian Meteorological Magazine 57, 65-75.
Clarke, A.R. and Dominiak, B.C. (2010).
Positive correlation of male and female
Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) catches in orange-ammonia
traps. General and Applied Entomology
39, 9-13.
Clarke, A.R., Powell, L.S., Weldon, C.W.
and Taylor, P.W. (2011). The ecology of
Bactrocera tryoni (Diptera: Tephritidae): what do we know to assist pest
management? Annals of Applied Biology
158, 26-54.
Clift, A. and Meats, A. (2005). Use of a
Bayesian Belief Network to identify situations that favour fruit fly incursions
in inland SE Australia. International
Congress on Modelling and Simulation
MODSIM 2005, eds A. Zerger and R.M.
Argent, pp. 170-6. (Modelling and Simulation Society of Australia and New
Zealand).
Courtice, A.C. and Drew, R.A.I. (1984).
Bacterial regulation of abundance in
tropical fruit flies (Diptera: Trephritidae). Australian Zoology 21, 251-68.
Dalby-Ball, G. and Meats, A. (2000). Influence of the odour of fruit, yeast and
cuelure on the flight activity of the
Queensland fruit flies, Bactrocera tryoni
(Froggatt) (Diptera: Tephritidae). Australian Journal of Entomology 39, 195-200.
De Lima, C.P.F., Jessup, A.J., Cruickshank, L., Walsh, C.J. and Mansfield,
E.R. (2007). Cold disinfestation of citrus
(Citrus spp.) for Mediterranean fruit fly
(Ceratitis capitata) and Queensland fruit
fly (Bactrocera tryoni) (Diptera: Tephritidae). New Zealand Journal of Crop and
Horticultural Science 35, 39-50.
Dominiak, B.C. (2006). Review of the use
of protein food based lures in McPhail
traps for monitoring Queensland fruit
fly Bactrocera tryoni (Froggatt) (Diptera:
Tephritidae). General and Applied Entomology 35, 7-12.
Dominiak, B.C., Campbell, M., Cameron, G. and Nicol, H. (2000). Review of
vehicle inspection historical data as
a tool to monitor the entry of hosts of
Queensland fruit fly Bactrocera tryoni
(Froggatt) (Diptera: Tephritidae) into a
fruit fly free area. Australian Journal of
Experimental Agriculture 40, 763-71.
Dominiak, B.C. and Coombes, N.M.
(2009). Fruit carrying characteristics
of travellers into a quarantine zone in
New South Wales in 1999/2000. Plant
Protection Quarterly 24, 14-19.
Dominiak, B.C., Gilmour, A.R., Kerruish,
B. and Whitehead, D. (2003a). Detecting low populations of Queensland
fruit fly Bactrocera tryoni (Froggatt) with
McPhail and Lynfield traps. General and
Applied Entomology 32, 49-53.
Dominiak, B.C., Mavi, H.S. and Nicol,
H.I. (2006). Effect of town microclimate
on the Queensland fruit fly Bactrocera
tryoni. Australian Journal of Experimental
Agriculture 46, 1239-49.
Dominiak, B.C., McLeod, L.J. and Landon, R. (2003b). Further development of
a low-cost release method for sterile
Queensland fruit fly, Bactrocera tryoni
(Froggatt) in rural New South Wales.
Australian Journal of Experimental Agriculture 43, 407-17.
Dominiak, B.C. and Nicol, H.I. (2010).
Field performance of Lynfield and
McPhail traps for monitoring male and
female sterile (Bactrocera tryoni Froggatt) and wild Dacus newmani (Perkins).
Pest Management Science 66, 741-4.
Dominiak, B.C., Sundaralingham, S., Jiang,
L., Jessup, A.J. and Barchia, I.M. (2008).
Production levels and life history traits
of mass reared Queensland fruit fly
Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) during 1999/2002 in Australia. Plant Protection Quarterly 23, 131-5.
Drew, R.A.I. (1974). The responses of fruit
fly species (Diptera: Tephritidae) in the
South Pacific area to male attractants.
Journal of the Australian Entomological
Society 13, 267-70.
Drew, R.A.I. (1987). Behavioural strategies
of fruit flies of the genus Dacus (Diptera: Tephritidae) significant in mating
and host-plant relationships. Bulletin of
Entomological Research 77, 73-81.
Drew, R.A.I. (1989). The tropical fruit flies
(Diptera: Tephritidae) of the Australasian and Oceanian regions. Memoirs of
the Queensland Museum. 26, 1-521.
Drew, R.A.I. and Lloyd, A.C. (1987). Relationship of fruit flies (Diptera: Tephritidae) and their bacteria to host plants.
Annals of the Entomological Society of
America 80, 629-36.
Drew, R.A.I., Zalucki, M.P. and Hooper,
H.H.S. (1984). Ecological studies of
Eastern Australia fruit flies (Diptera:
Tephritidae) in their endemic habitat.
I. Temporal variation in abundance.
Oecologia (Berlin) 64, 267-72.
Duell, R. (2009). Seasonal climate summary southern hemisphere (autumn 2008):
a return to neutral ENSO conditions
in the tropical Pacific. Australian Meteorological and Oceanographic Journal 58,
63-77.
Edge, V., Gardner, R., Green, A., Cock, K.
and Cornish, J. (2001). Technical review
of the Tri-State Strategy for Queensland
Fruit Fly. Report to Standing Committee on Agriculture and Resource Management. January 2001, 62 pp.
Ero, M.M., Hamacek, E. and Clarke, A.R.
(2011). Foraging behaviours of Diachasmimorpha kraussi (Fullaway) (Hymenoptera: Braconidae) and its host
Bactrocera tryoni (Froggatt) (Diptera:
Tephritidae) in a nectarine (Prunus persica (L.) Batsch var. nectarina (Aiton)
Maxim) orchard. Australian Journal of
Entomology 50, 234-40.
Fanson, B.G., Weldon, C.W., Perez-Staples, D., Simpson, S.J. and Taylor, P.W.
(2009). Nutrients, not caloric restriction,
extend lifespan in Queensland fruit flies
(Bactrocera tryoni). Aging Cell 8, 1-10.
Fay, H.A.C. and Meats, A. (1983). The effects of age, ambient temperature, thermal history and mating history on mating frequency in males of the Queensland fruit fly, Dacus tryoni. Entomologia
Experimentalis et Applicata 35, 273-6.
Fletcher, B.S. (1973). The ecology of a natural population of the Queensland fruit
fly, Dacus tryoni IV. The immigration
and emigration of adults. Australian
Journal of Zoology 21, 541-65.
Fletcher, B.S. (1974a). The ecology of a
natural population of the Queensland
fruit fly, Dacus tryoni V. The dispersal
of adults. Australian Journal of Zoology
22, 189-202.
Fletcher, B.S. (1974b). The ecology of a natural population of the Queensland fruit
fly, Dacus tryoni VI. Seasonal changes in
fruit fly numbers in the areas surrounding the orchard. Australian Journal of Zoology 22, 353-63.
Fletcher, B.S. (1987). The biology of dacine
fruit flies. Annual Review of Entomology
32, 115-44.
Fletcher, B.S. (1989). Movements of tephritid fruit flies. In ‘Fruit flies, their
biology, natural enemies and control’
Vol. 3B, eds A.S. Robinson and G.
Hooper, pp. 209-19. (Elsevier, Amsterdam).
Gilchrist, A.S. and Meats, A.W. (2011).
Factors affecting the dispersal of largescale releases of the Queensland fruit
fly, Bactrocera tryoni. Journal of Applied
Entomology 37, 186-8.
Griffiths, N. and Brady, J. (1995). Wind
structure in relation to odour plumes in
tsetse fly habitats. Physiological Entomology 20, 286-92.
Hancock, D.L., Hamacek, E.L., Lloyd, A.C.
and Elson-Harris, M.M. (2000). The distribution and host plants of fruit flies
(Diptera: Tephritidae) in Australia. DPI
Publications, Brisbane, Australia. ISSN
0727-6273.
International Atomic Energy Agency
(2003). Trapping guidelines for areawide fruit fly programmes. International Atomic Energy Agency, Vienna.
pp. 47.
Jessup, A.J., Carswell, I.F. and Dalton, S.P.
(1998). Disinfestation of fresh fruits
from Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) with combination
mild heat and modified atmosphere
packaging. Australian Journal of Entomology 37, 186-8.
Li, B., Ma, J., Hu, X., Liu, H., Wu, J., Chen,
H. and Zhang, R. (2010). Risk of introducing exotic fruit flies, Ceratitis capitata, Ceratitis cosyra and Ceratitis rosa
(Diptera: Tephritidae), into southern
China. Journal of Economic Entomology
103, 1100-11.
Maelzer, D.A. (1990). Fruit fly outbreaks in
Adelaide, S.A., from 1948–49 to 1986–
87. I. Demarcation, frequency and temporal patterns of outbreak. Australian
Journal of Zoology 38, 439-52.
Maelzer, D.A., Bailey, P.T. and Perepelicia, N. (2004). Factors supporting the
non-persistence of fruit fly populations
in South Australia. Australian Journal of
Experimental Agriculture 44, 109-26.
Mankin, R.W., Anderson, J.B., Mizrach, A.,
Epsky, N.D., Shuman, D., Heath, R.R.,
Mazor, M., Hetzroni, A., Grinshpun, J.,
Taylor, P.W. and Garrett, S.L. (2004).
Broadcasts of wing-fanning vibrations
recorded from calling male Ceratitis
capitata (Diptera: Tephritidae) increases
captures of females in traps. Journal of
Economic Entomology 97,1299-309.
Mankin, R.W., Lemon, M., Harmer,
A.M.T., Evans, C.S. and Taylor, P.W.
(2008). Time-pattern and frequency
analyses of sounds produced by irradiated and untreated male Bactrocera
tryoni (Froggatt) (Diptera: Tephritidae)
during mating behaviour. Annals of the
American Entomological Society 101, 66474.
Meats, A. and Fay, F.A.C. (2000). Distribution of mating frequency among males
of the Queensland fruit fly, Bactrocera
tryoni (Froggatt), in relation to temperature, acclimation and change. General
and Applied Entomology 29, 27-30.
Meats, A. and Hartland, C.L. (1999). Upwind anemotaxis in response to cuelure by the Queensland fruit fly Bactrocera tryoni. Physiological Entomology
24, 90-7.
Meats, A. and Smallridge, C.J. (2007).
Short and long range dispersal of Medfly, Ceratitis capitata (Diptera: Tephritidae) and its invasive potential. Journal
of Applied Entomology 8, 518-23.
Meats, A., Smallridge, C.J. and Dominiak, B.C. (2006). Dispersion theory and
the sterile insect technique: application to two species of fruit fly. Entomologia Experimentalis et Applicata 119,
247-54.
Meats, A.M. (1996). Demographic analysis of sterile insect trials with Queensland fruit fly Bactrocera tryoni (Froggatt)
(Diptera: Tephritidae). General and Applied Entomology 27, 2-12.
Meats, A.M. (1998). The power of trapping grids for detecting and estimating
the size of invading propagules of the
(Froggatt) (Diptera: Tephritidae) risks
of subsequent infestation. General and
Applied Entomology 28, 47-55.
Meats, A.M., Clift, A.D. and Robson, M.K.
(2003). Incipient founder populations
of Mediterranean and Queensland fruit
flies in Australia: the relation of trap
catch to infestation radius and models
for quarantine radius. Australian Journal
of Experimental Agriculture 43, 397-406.
Meats, A.M. and Edgerton, J.E. (2008).
Short- and long-distance dispersal of
Queensland fruit fly, Bactrocera tryoni
and its relevance to invasive potential,
sterile insect technique and surveillance trapping. Australian Journal of Experimental Agriculture 48, 1237-45.
Metcalf, R.L. (1990). Chemical ecology of
Dacine fruit flies (Diptera: Tephritidae). Annals of the Entomological Society
of America 83, 1017-30.
Mullen, C. (2009). Seasonal climate summary southern hemisphere (summer
2008-09): a weak, brief La Nina returns.
Bumper wet season in tropical Australia: exceptional heatwaves in southeastern Australia. Australian Meteorological
and Oceanographic Journal 58, 275-84.
Oliver, J. (2007). Summary of national fruit
fly-related activities released. Plant
Protection News. February 2007. Department of Agriculture, Fisheries and
Forestry.
Perez-Staples, D., Harmer, A.M.T., Collins, S.R. and Taylor, P.W. (2008). Potential for pre-release diet supplements
to increase the sexual performance and
longevity of male Queensland fruit
flies. Agricultural and Forest Entomology
10, 255-62.
Perez-Staples, D., Prabhu, V. and Taylor,
P.W. (2007). Post-teneral protein feeding enhances sexual performance of
Queensland fruit flies. Physiological Entomology 32, 127-35.
Plant Health Australia (2010). National
Plant Biosecurity Strategy. Plant Health
Australia, Deakin, Australian Capital
Territory.
Prabhu, V., Pérez-Staples, D. and Taylor,
P.W. (2008). Protein: carbohydrate ratios promoting sexual activity and longevity of male Queensland fruit flies.
Journal of Applied Entomology 132, 57582.
Qi, L. (2009). Seasonal climate summary
southern hemisphere (spring 2008): La
Nina pattern returning across equatorial Pacific. Australian Meteorological and
Oceanographic Journal 58, 199-208.
Radhakrishnan, P., Perez-Staples, D.,
Weldon, C.W. and Taylor, P.W. (2009).
Multiple mating and sperm depletion
in male Queensland fruit flies: effects
on female remating behaviour. Animal
Behaviour 78, 839-46.
Radhakrishnan, P. and Taylor, P.W. (2008).
Ability of male Queensland fruit flies
to inhibit receptivity in multiple mates,
and the associated recovery of accessory glands. Journal of Insect Physiology
54, 421-8.
Raghu, S., Clarke, A.R., Drew, R.A.I. and
Hulsman, K. (2000). Impact of habitat
modification on the distribution and
abundance of fruit flies (Diptera: Tephritidae) in southeast Queensland.
Population Ecology 42, 153-60.
Stanaway, M.A., Zalucki, M., Gillespie,
P.S., Rodriguez, C.M. and Maynard,
G.V. (2001). Pest risk assessment of insects in sea cargo containers. Australian
Journal of Entomology 40, 180-92.
Sved, J.A., Yu, H., Dominiak, B. and Gilchrist, A.S. (2003). Inferring modes of
colonization for pest species using heterozygosity comparisons and sharedallele test. Genetics 163, 823-31.
Stephenson, B.P., Gill, G.S.C., Randall, J.L.
and Wilson, J.A. (2003). Biosecurity approaches to surveillance and response
for new plant pest species. New Zealand
Plant Protection 56, 5-9.
Tan, K.H. and Nishida, R. (1995). Incorporation of raspberry ketone in the male
rectal glands of the Queensland fruit fly
Bactrocera tryoni Froggat (Diptera:
Tephritidae). Applied Entomology and
Zoology 30, 494-7.
Torok, S.J., Morris, J.G., Skinner, C. and
Plummer, N. (2001). Urban heat island
features of southeast Australian towns.
Australian Meteorological Magazine 50, 113.
Tychsen, P.H. (1977). Mating behaviour of
the Queensland fruit fly, Dacus tryoni
(Diptera: Tephritidae), in field cages.
Journal of the Australian Entomological
Society 16, 459-65.
Tychsen, P.H. and Fletcher, B.S. (1971).
Studies on the rhythm of mating in the
Queensland fruit fly, Dacus tryoni. Journal of Insect Physiology 17, 2139-56.
Weldon, C. (2003). Effectiveness of coloured unbaited sticky traps for monitoring dispersal of gamma-irradiated
(Froggatt) (Diptera: Tephritidae). General and Applied Entomology 32, 55-60.
Weldon, C. (2007). Influence of male aggregation size on female visitation in
Bactrocera tryoni (Froggatt) (Diptera:
Tephritidae). Australian Journal of Entomology 46, 29-34.
Weldon, C. and Meats, A. (2007). Shortrange dispersal of recently emerged
males and females of Bactrocera tryoni
(Froggatt) (Diptera: Tephritidae) monitored by sticky sphere traps baited with
protein and Lynfield traps baited with
cue-lure. Australian Journal of Entomology 46, 160-6.
Weldon, C. and Meats, A. (2010). Dispersal
of mass-reared sterile, laboratory-domesticated and wild male Queensland
fruit flies. Journal of Applied Entomology
134, 16-25.
Weldon, C.W., Prenter, J. and Taylor, P.W.
(2010). Activity patterns of Queensland
fruit flies (Bactrocera tryoni) are affected
by both mass-rearing and sterilization.
Physiological Entomology 35, 148-53.
Desiccation resistance of adult Queensland fruit flies Bactrocera tryoni decreases with age. Physiological Entomology 35,
385-90.
Sexual development of wild and massreared male Queensland fruit flies in
response to natural food sources. Entomologia Experimentalis et Applicata 139,
17-24.
White, I.M. and Elson-Harris, M.M. (1992).
‘Fruit flies of economic significance:
their identification and bionomics’.
(CAB International, Oxon, UK).
Yonow, T. and Sutherst, R.W. (1998).
The geographical distribution of the
Queensland fruit fly, Bactrocera (Dacus)
tryoni, in relation to climate. Australian
Journal of Agricultural Research 49, 93553.
APPENDIX 6
Real-time PCR 18S internal control for PCR competency
All DNA extractions need to be tested for PCR competency before performing the real-time PCR assay.
This can be done by using the 18S internal control TaqMan real-time PCR assay (Applied Biosystems)
or conventional PCR using LCO1490 and HCO2198 primers (Folmer et al., 1994).
The 18S real-time PCR assay is conducted using TaqMan Ribosomal RNA Control Reagent kit (VIC
probe) according to the Manufacturer’ instructions with modification. The kit is supplied with primers,
probe and control material. The PCR assay should be run in duplicate for each sample, and with 20 µl
or 10 µl volumes. The PCR compositions are listed in Table S1 and the PCR cycles are listed below
the table.
Table S1 TaqMan Internal Control of qPCR for 18S ribosomal RNA (20µl*)
PCR reagent mix
Reaction volume (1 x)
Master mix (10 x)
Sterile distilled water
7.7
77
2X SsoAdvanced Universal Probe
supermix
10
100
10 µM forward primer (Rib for)
0.1
1.0
10 µM reverse primer (Rib rev)
0.1
1.0
40 µM 18S probe (Rib pr)
0.1
1.0
DNA template
2.0
*For 10 µl volume, halve all reagents.
PCR cycling parameter
1 x cycle
95 °C, 2 min
40 x cycles
95 °C, 5 sec
60 °C, 5 sec
Plate read after each cycle
For the analysis of the real-time PCR, see section 7.3.4.3. In this assay, VIC is the fluorophore
attached to the 18S probe.
For the interpretation of the results, Cq values of 30 is the cut-off point:
 If the Cq values ≤30 cycles, the DNA extraction is PCR competent, and the DNA can be used
for real-time PCR assay for B. tryoni complex.
 If the Cq values >30 cycle, re-extraction is required.
APPENDIX 7
Additional information on the validation of the real-time PCR assay for B.
tryoni complex
a. Specificity test for Tephritidae specimens in the real-time PCR assay
Specificity testing showed that the real-time PCR assay could detect the targeted species: B. tryoni, B.
aquilonis, and B. neohumeralis (Table S2), but did not cross-react with the non-target species, including
B. correcta, B. cucurbitae, B. dorsalis, B. papaya, B. invadens, B. kandiensis, B. jarvisi, B. zonata,
Ceratitis capitata, C. cosyra, C. rosa and Dirioxa pornia (Dhami et al., 2015).
Different body parts were used for DNA extraction (Table S2). All B. tryoni complex samples tested
gave Cq value under 25 cycles except the DNA samples from the empty pupal case (Table S2).
Table S2 Tephritidae samples tested in the real-time PCR assay for B. tryoni, showing parts of the
samples tested.
Sample Reference
Morphological identification
Mean Cq
Method
BQ166
life stage, body part
B. tryoni
18.62
QG
Adult abdomen × 1
BQ136
B. tryoni
13.81
QG
Adult abdomen × 1
T15_00083
Tephritidae
19.21
QG
Larva pieces
T15_00500a
Tephritidae
15.95
QG
Larva pieces
T14_00158
B. tryoni
21.54
QG
Adult leg x 1
T14_01221
B. tryoni
23.01
QG
Adult leg x 1
BQ14
B. aquilonis
11.53
QG
Abdomen x 1
NM11
B. neohumeralis
20.48
NM
Abdomen x 1
NM12
B. neohumeralis
17.37
NM
Head/thorax
NM13
B. neohumeralis
18.79
NM
Abdomen x 1
NM14
B. neohumeralis
17.71
NM
Head/thorax
NM15
B. neohumeralis
21.33
NM
Head/thorax
T15_6386/33
B. tryoni
26.67
QG
Empty pupal case
T15_6386/34
B. tryoni
27.21
QG
Empty pupal case
Note: All samples were tested in duplicate or triplicate and all Cq values provided are means of the repeats. DNA extraction
method refers to Qiagen Blood and Tissue kit (QG) and NucleoMag 96 Tissue Kit (NM). Empty pupal case samples were
extracted using an overnight incubation step.
b. Real-time PCR assay for B. tryoni and B. curvipennis interceptions
B. curvipennis and B. tryoni samples were tested using the real-time PCR assay against B. tryoni
complex. One leg from each individual adult specimen and part of larvae intercepted in 2015 was used
for DNA extraction (method see section 7.3.1). The B. curvipennis adult samples were from a laboratory
colony in New Caledonia 2013 and are dry mounted specimens in the PHEL collection; one leg from
each specimen was used for DNA extraction. All the DNA samples were tested with 18S internal control
PCR and B. tryoni real-time PCR assays.
All DNA extractions were PCR competent in the 18S internal control assay with Cq less than 30 cycles
(Table S3 and Figure S1). The B. tryoni complex real-time PCR assay gave Cq <25 cycles for B. tryoni
samples and >27 cycles for B. curvipennis (Table S3 and Figure S1).
Table S3 Cq values of B. tryoni and B. curvipennis samples using 18S internal controls and B. tryoni
real-time PCR assays
Species IDa
Life stage
Year collected
Cq (18S)
Cq (B. tryoni)
B. tryoni
larva
2015
23.07
15.21
B. tryoni
larva
2015
21.26
14.78
B. tryoni
adult
2015
23.07
20.59
B. tryoni
adult
2015
23.81
20.62
B. curvipennis
larva
2015
25.9
27.85
B. curvipennis
larva
2015
21.63
27.72
B. curvipennis
adult
2013
24.51
34.81
B. curvipennis
adult
2013
27.56
28.47
Note: aSpecies ID are confirmed by morphology and/ or COI sequences. DNA was extracted using a piece of a larva or an adult
leg. Note: B. curvipennis adult samples were over two-year-old dried mounted specimens, conventional PCR of the DNA with
LCO1490 and HCO2198 primers produced faint PCR products (not shown), indicating DNA degradation in the samples.
18S (VIC)
B. tryoni
B. curvipennis
B. tryoni (FAM)
B. tryoni
B. curvipennis
Figure S1 Amplification curves for 18S internal control (top) and B. tryoni assays (bottom).
Top (18S internal control assay): the orange colour curves shows amplification of B. curvipennis
samples and the dark green of B. tryoni samples.
Bottom (B. tryoni assay): the orange colour curves show amplification of B. curvipennis samples and
blue curves B tryoni samples
Plant Health Australia
1/1 Phipps Close
DEAKIN ACT 2600
Phone: +61 2 6215 7700
Email:[email protected]
PHA 15-025
www.planthealthaustralia.com.au

Australian Handbook for the Identification of Fruit Flies

Transcription

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