Use of entomopathogenic nematodes and fungi in the

Transcription

Use of entomopathogenic nematodes
and fungi in the control of pine weevil.
Chris Williams and Christine Griffin
Behavioural Ecology and Biocontrol,
Dept. of Biology,
Maynooth University,
Co. Kildare.
Ireland.
Forestry in Ireland




Currently approximately 10% of Ireland’s
surface area is forested with Coillte being
the main owner
There is an aim to increase this to 15%
over the coming years
The major plantation species are
Lodgepole pine (Pinus contorta) and
Norway Spruce (Picea abies)
The major insect pest is the large Pine
Weevil (Hylobius abietis)
Pine Weevil life cycle



Semi-volitine, usually a 2yr lifecycle
though can vary from 1-4yrs – temp
dependent
Four larval instars which develop within
the stump under the bark prior to
pupation
Clearfelled forests provide an optimum
niche for larval development and food
for adults on emergence
Summary





Biocontrol of Pine weevil
Meta-analysis of EPN efficacy
Field Trials of EPN and EPF
Susceptibility of larval and pupal
pine weevil
Effects of EPN on non-target
Coleoptera
Life cycle of Entomopathogenic Fungi
(EPF)
Conidiaspore lands on
insect
Conidiaspore germinates –
mycelium formed
Infective stage
used in
inundative
biocontrol
Under favourable
conditions - sporulation
Life cycle of entomopathogenic
nematode (EPN)
Summary





pine weevil
Coleoptera
Factors affecting EPN efficacy against pine weevil
Meta-analysis of 22 trials in Ireland and Britain
Measures of efficacy
Year of
app.
Trial / Site Name
Location
Ballyroan trial one1
Laois, Ire.
Ballyroan trial two1
Subs.
Pine
Peat
2002Pine
Peat
Destructive
sampling mean
Cont.
Emergence
trapping mean
Cont.
93.3
126.0
140.7
84.4
% red
S.c
% red
H.d
%
para S.c
48.5
87.0
25.0
11.0
87.2
15.5
%
para
H.d
66.2
60.5
Ballybrittas1
, Ire.
2003Pine
Peat
67.2
37.1
60.0
67.7
47.0
55.2
Emo2
, Ire.
2004Pine
Peat
84.6
78.9
64.3
84.8
37.0
52.6
Donadea2
, Ire.
2005Pine
Peat
144.4
87.9
57.3
78.7
26.0
77.1
Glendine
, Ire.
2007Pine
Peat
84.3
53.2
70.7
80.6
14.0
37.0
Summerhill
, Ire.
2010Pine
Peat
80.8
260.5
53.9
n/a
71.7
n/a
Killduff
, Ire.
2011Pine
Peat
30.4
n/a
n/a
n/a
20.2
n/a
Lairg3
N. Sco.
2003Pine
Peat
n/a
33.0
70.9
n/a
75.0
n/a
Lairg3
N. Sco.
2003Pine
Peat
n/a
41.5
65.3
n/a
n/a
n/a
Ballymacshaneboy
Co. , Ire.
2007Pine
Min
34.9
55.1
13.7
75.1
9.0
34.1
Lackenrea trial 1
Co. , Ire.
2007Pine
Min
56.0
29.0
29.0
63.4
3.0
18.5
Lackenrea trial 2
Co. , Ire.
2007Spruce
Min
11.1
16.2
42.0
79.2
2.0
36.8
Annalecka4
Wicklow, Ire.
2006Spruce
Min
17.8
n/a
n/a
n/a
10.0
43.5
Glendalough4
, Ire.
2006Spruce
Min
n/a
4.9
14.8
39.1
n/a
n/a
Ballinagee4
Wicklow, Ire.
2004Spruce
Min
18.8
1.5
25.0
33.3
9.0
17.8
Oakwood4
Wicklow, Ire.
2005Spruce
Min
5.3
18.3
10.4
49.0
12.0
31.1
Deerpark
Wicklow, Ire.
2007Spruce
Min
8.6
13.4
32.2
74.2
21.0
45.5
The Rodneys
Co. , Ire.
2007Spruce
Min
n/a
1.8
31.3
n/a
n/a
n/a
Knockeen
Co. , Ire.
2007Spruce
Peat
n/a
0.5
42.9
85.7
n/a
n/a
1997Spruce
Peat
Ae Forest trial
1From
Laois, Ire.
2001
Tree
species
15
Ae Forest trial 25
, Sco.
, Sco.
1997
Spruce
Peat
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
18.3
5.2
n/a
n/a
Dillon et al. (2006), 2From Dillon et al. (2007), 3From Torr et al. (2007), 4From Dillon et al. (2008), 5From Brixey et al. (2006).
Measures of efficacy:
% weevils in stump parasitised 4 weeks after application
% reduction in number of adults emerging relative to untreated stumps
Year of
app.
Trial / Site Name
Location
Laois, Ire.
Ballyroan trial two1
Subs.
Pine
Peat
2002Pine
Peat
Destructive
sampling mean
Cont.
Emergence
trapping mean
Cont.
93.3
126.0
140.7
84.4
% red
S.c
% red
H.d
%
para S.c
48.5
87.0
25.0
11.0
87.2
15.5
%
para
H.d
66.2
60.5
Ballybrittas1
, Ire.
2003Pine
Peat
67.2
37.1
60.0
67.7
47.0
55.2
Emo2
, Ire.
2004Pine
Peat
84.6
78.9
64.3
84.8
37.0
52.6
Donadea2
, Ire.
2005Pine
Peat
144.4
87.9
57.3
78.7
26.0
77.1
Glendine
, Ire.
2007Pine
Peat
84.3
53.2
70.7
80.6
14.0
37.0
Summerhill
, Ire.
2010Pine
Peat
80.8
260.5
53.9
n/a
71.7
n/a
Killduff
, Ire.
2011Pine
Peat
30.4
n/a
n/a
n/a
20.2
n/a
Lairg3
N. Sco.
2003Pine
Peat
n/a
33.0
70.9
n/a
75.0
n/a
Lairg3
N. Sco.
2003Pine
Peat
n/a
41.5
65.3
n/a
n/a
n/a
Ballymacshaneboy
Co. , Ire.
2007Pine
Min
34.9
55.1
13.7
75.1
9.0
34.1
Lackenrea trial 1
Co. , Ire.
2007Pine
Min
56.0
29.0
29.0
63.4
3.0
18.5
Lackenrea trial 2
Co. , Ire.
2007Spruce
Min
11.1
16.2
42.0
79.2
2.0
36.8
Annalecka4
Wicklow, Ire.
2006Spruce
Min
17.8
n/a
n/a
n/a
10.0
43.5
Glendalough4
, Ire.
2006Spruce
Min
n/a
4.9
14.8
39.1
n/a
n/a
Ballinagee4
Wicklow, Ire.
2004Spruce
Min
18.8
1.5
25.0
33.3
9.0
17.8
Oakwood4
Wicklow, Ire.
2005Spruce
Min
5.3
18.3
10.4
49.0
12.0
31.1
Deerpark
Wicklow, Ire.
2007Spruce
Min
8.6
13.4
32.2
74.2
21.0
45.5
The Rodneys
Co. , Ire.
2007Spruce
Min
n/a
1.8
31.3
n/a
n/a
n/a
Knockeen
Co. , Ire.
2007Spruce
Peat
n/a
0.5
42.9
85.7
n/a
n/a
1997Spruce
Peat
Ae Forest trial
1From
Laois, Ire.
2001
Tree
species
15
Ae Forest trial 25
, Sco.
, Sco.
1997
Spruce
Peat
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
18.3
5.2
n/a
n/a
Dillon et al. (2006), 2From Dillon et al. (2007), 3From Torr et al. (2007), 4From Dillon et al. (2008), 5From Brixey et al. (2006).
Factors in meta-analysis :
Tree stump species, Soil type; Weevil population size; Nematode species
Year of
app.
Trial / Site Name
Location
Laois, Ire.
Ballyroan trial
two1
Subs.
Pine
Peat
Destructive
sampling mean
Cont.
Emergence
trapping mean
Cont.
93.3
140.7
% red
S.c
% red
H.d
%
para S.c
48.5
87.0
25.0
%
para
H.d
66.2
Laois, Ire.
2002Pine
Peat
126.0
84.4
11.0
87.2
15.5
60.5
Ballybrittas1
, Ire.
2003Pine
Peat
67.2
37.1
60.0
67.7
47.0
55.2
Emo2
, Ire.
2004Pine
Peat
84.6
78.9
64.3
84.8
37.0
52.6
Donadea2
, Ire.
2005Pine
Peat
144.4
87.9
57.3
78.7
26.0
77.1
Glendine
, Ire.
2007Pine
Peat
84.3
53.2
70.7
80.6
14.0
37.0
Summerhill
, Ire.
2010Pine
Peat
80.8
260.5
53.9
n/a
71.7
n/a
Killduff
, Ire.
2011Pine
Peat
30.4
n/a
n/a
n/a
20.2
n/a
Lairg3
N. Sco.
2003Pine
Peat
n/a
33.0
70.9
n/a
75.0
n/a
Lairg3
N. Sco.
2003Pine
Peat
n/a
41.5
65.3
n/a
n/a
n/a
Ballymacshaneboy
Co. , Ire.
2007Pine
Min
34.9
55.1
13.7
75.1
9.0
34.1
Lackenrea trial 1
Co. , Ire.
2007Pine
Min
56.0
29.0
29.0
63.4
3.0
18.5
Lackenrea trial 2
Co. , Ire.
2007Spruce
Min
11.1
16.2
42.0
79.2
2.0
36.8
Annalecka4
Wicklow, Ire.
2006Spruce
Min
17.8
n/a
n/a
n/a
10.0
43.5
Glendalough4
, Ire.
2006Spruce
Min
n/a
4.9
14.8
39.1
n/a
n/a
Ballinagee4
Wicklow, Ire.
2004Spruce
Min
18.8
1.5
25.0
33.3
9.0
17.8
Oakwood4
Wicklow, Ire.
2005Spruce
Min
5.3
18.3
10.4
49.0
12.0
31.1
Deerpark
Wicklow, Ire.
2007Spruce
Min
8.6
13.4
32.2
74.2
21.0
45.5
The Rodneys
Co. , Ire.
2007Spruce
Min
n/a
1.8
31.3
n/a
n/a
n/a
Knockeen
Co. , Ire.
2007Spruce
Peat
n/a
0.5
42.9
85.7
n/a
n/a
Ae Forest trial 15
, Sco.
1997Spruce
Peat
n/a
n/a
n/a
n/a
18.3
n/a
Ae Forest trial
1From
2001
Tree
species
25
Dillon et al. (2006),
, Sco.
2From
1997
3From
Spruce
Peat
Torr et al. (2007),
n/a
4From
n/a
n/a
5From
n/a
5.2
n/a
Brixey et al. (2006).
Nematode species highly significant: in each of two measures,
Heterorhabditis downesi more effective than Steinernema carpocapsae
80
***
70
60
***
Paired t-test,
12 sites
50
% 40
30
S. carpocapsae
S.carpocap
sae
H. downesi
20
10
0
% weevils parasitised
% reduction
of adult emergence
% reduction
of adults
Williams et al.
Site factors affecting EPN efficacy: Trials meta-analysis

Nematode species: significant
(H. d > S. c.)
Analysis (GLM) for each species separately
 Soil type
 Tree (stump) species: spruce or pine
 Weevil population size (density dependent
effects)
Williams et al.
Soil type, but not tree species or weevil population affects efficacy
by each nematode species for each of our two measures
Nematode
Response
Factor
F ratio P
variable
S. carpocapsae
% parasitism
% reduction
H. downesi
% parasitism
% reduction
Tree species
0.134
0.721
Soil type
3.433
0.089
No. weevils
0.093
0.765
Tree species
0.158
0.697
Soil type
5.913
0.029
Tree *Soil
0.685
0.422
Tree species
0.694
0.424
Soil type
9.874
0.010
Tree species
0.242
0.633
Soil type
4.720
0.055
Tree *Soil
0.950
0.353
Adjust Model
ed R2
P
0.667
0.001
0.882
<0.001
0.932
<0.001
0.961
<0.001
Williams et al.
Forest soils: “peat” or “mineral”
Peat: 44% of Irish forests
planted on drained /cut
peat bog
Mineral: podsols, gleys,
brown earths
Peat soil favours parasitism by both EPN species
70
% weevils parasitised
60
Peat
50
Mineral
40
30
20
10
0
Steinernema carpocapsae
Heterorhabditis downesi
Williams et al.
Peat soil favours reduction in adult emergence by both EPN species
80
70
60
50
40
30
Peat
90
Mean percentage reduction in emergence
% reduction in emergence
90
Peat
80
Mineral
Mineral
70
60
50
40
30
20
10
0
20
Nematode species
10
0
Williams et al.
Why are peat soils suitable for EPN in forest ecosystem?


Direct effect of open texture: facilitates
movement (active and passive) of EPN
Possible indirect effects:
moisture content
 temperature, pH
 antagonists
 transmission of host-associated signals
etc

Tree stump species does not affect EPN efficacy either as
% parasitism or % reduction relative to untreated;
BUT: the number of weevils emerging per untreated stump
is much higher in pine than spruce, making population
reduction by EPN difficult in pine stumps
Number of
pine weevils
emerging
per
untreated
stump
Pine
Pinus contorta
Spruce
Picea sitchensis
Williams et al.
Summary





pine weevil
Coleoptera
Field trials of EPN
and EPF
-
All sites are planted with Pinus contorta
Control 500ml water
B. bassiana 109 spores per stump
S. carpocapsae 3.5x106 IJs per stump
B. bassiana + S. carpocapsae ½ dose
10 stumps per treatment sampled four
weeks after application
10 stumps per treatment sampled with
emergence traps throughout the summer
Emergence trap and destructively sampled
stump
Overview of field trials
Year
app
Site
Er /
pro
Cont
2010
Hortland
Pro
X
2011
Balivor
Pro
X
2010
Summerhill Er
2011
Kilduff
Er
2012
Rossnagad
Er
Er = Eradicant; Pro = prophylactic
Nem
(S.c)
Fung
(B.b)
Mix
(Sc
+mix)
Other
X
Results Eradicant 1
(Summerhill)
Proportion of Individuals dead
Summerhill 2010 Destructive sampling
Proportion of individuals dead
1.0
a
0.8
a
0.6
0.4
0.2
b
c
0
Control Nema
(S.c)
Fungi
(B.b)
Mixed
(S.c + B.b)
Infection status of individuals
Dead (unknown cause)
250
No. of Pine Weevils
Fungus-killed
Nematode-killed
200
Alive
150
100
50
0
Control
Nematodes
Fungus
Mixed
Number of adults emerging Summerhill
(2010)
Number of adults emerging
300
c
250
b
200
150
a, b
a
100
50
0
Control Nema
(S.c)
Fungi Mixed
(B.b) (S.c + B.b)
Number of adults emerging
Number of adults emerging Summerhill
(2011)
12.5
10.0
7.5
5.0
2.5
0
Control Nema
(S.c)
Fungi
(B.b)
Mixed
(S.c + B.b)
Summerhill results


All treatments significantly better than
control (2010) with Nematode (S.c.)
and mixed (S.c. + B.b. at half dose)
applications being the most promising
Emergence trapping in the second year
(2011) showed overall reduced
populations with the same numerical
trends in weevil population reductions
Kilduff



A large experiment was conducted at
Kilduff (2011) to test the efficacy of EPN
(Steinernema carpocapsae) & EPF
(Beauveria bassiana, Metarhizium
anisopliae (Met 52)) alone, in
combination and at different dose rates
Overall there were 10 treatments
Assessments were made by both
destructive sampling and emergence
trapping
Results – Eradicant 2
(Kilduff)
Destructive sampling Kilduff (2011)
Proportion of individuals dead
Proportion of Individuals dead
Treatments including EPN
0.5
b
b
0.4
ab
0.3
ab
0.2
0.1
0
a
a
a
a
a
a
EPF only treatments and control
c
fS
+
+
et
e
ria
52
Sc
uv
e
ea
fM
ha
l
fB
ha
l
c
do
s
Sc
er
ia
er
ia
et
52
fd
os
e
ll
Fu
52
et
52
Be
au
v
al
H
do
se
M
M
Be
au
v
do
se
do
se
fS
al
H
al
H
al
f
Fu
ll
H
Fu
ll
do
se
100
M
et
120
al
f
or
e
sp
co
nt
ro
l
140
H
ry
D
No. of Pine Weevils
Infection status of individuals
Nematode-killed with Fungus
Dead (unknown cause)
Fungus-killed
Nematode-killed
Alive
80
60
40
20
0
Results Kilduff



Destructive sampling indicated that
those treatments that included S.c
were the best
Evidence that nematode treatments
may be as effective at half doses
No significant difference in emergence
numbers between any treatments
Distinguishing between applied and
native EPF


Applied EPF (Beauveria bassiana)
was distinguished from native EPF
(Beauveria caledonica) using RFLPs.
This was done at a field trial site
where only Beauveria bassiana was
used
Distinguishing between native and
applied EPF using RFLPs
RFLP analysis of Fungus-infected
insects recovered at Rossnagad



RFLP’s were used to identify fungusinfected weevils recovered at destructive
sampling at Rossnagad
Most of the infected weevils were infected
with the native Beauveria caledonica, but
some, particularly from the treated
stumps were B. bassiana infected
This shows that soil and bark are not
necessarily barriers to conidia movement
to insects
Applied and native EPF isolated from
infected weevils from Rossnagad
Conclusions of Field trials




Variable success of biological control agents, which appear
to be site-specific
There appears to be persistent effects into the summer after
application
Evidence of efficacy of nematodes as an eradicant
treatment, but no evidence of fungus efficacy as a
prophylactic
Some evidence that nematodes efficacy can be as effective
at half dose, or in combination with fungus, as at full dose
Summary





pine weevil
Coleoptera
“Differential susceptibility of pine
weevil larvae and pupae to
entomopathogenic nematodes and
death of adults infected as pupae”
Mean no. Insects dead/24
Larvae
30
25
20
15
Control
10
5
0
0
5
10
15
20
Time (days)
Pupae
Hd 50
Hd 250
Mean no. insects dead/24
30
25
Sc 50
20
15
10
Sc 250
5
0
-5 0
5
10
Time (days)
15
20
Percentage of dead and infected
insects as either pupae or adults (N =
160)
Summary





pine weevil
Coleoptera
Effects of entomopathogenic nematodes on
non-target beetle communities





Non-target beetles were monitored in 14 trials
(10 sites)
4 trials were assessed in the months immediately
following nematode application
10 trials were assessed in the year after
application
Whereas site and tree species appeared to
significantly affect abundance, diversity, species
richness and community composition
Nematode treatment had no effect on any of
these variables
Results
MRPP Results
Effect of Location
Effect of Tree Species
No effect of treatment with EPN
References





Williams C.D., Dillon A.B., Girling R. and Griffin C.T. (2013) “Organic
soils promote the efficacy of entomopathogenic nematdoes, with
different foraging strategies, in the control of a major forest pest: a
meta-analysis of studies to date” Biological Control 65(3): 357-364.
Williams C.D., Dillon A.B., Hennessy R., Mc Namara L., Harvey C.D.
and Griffin C.T. (2013) “Control of a major pest of forestry, Hylobius
abietis, with entomopathogenic nematodes and fungi using eradicant
and prophylactic strategies” Forest Ecology and Management 305:
212-222.
Williams C.D., Dillon A.B., Ennis D., Hennessy R. and Griffin C.T.
(2015) "Differential susceptibility of pine weevil (Hylobius abietis)
larvae and pupae to entomopathogenic nematodes and death of
adults infected as pupae.” BioControl. DOI: 10.1007/s10526-0159658-3
Dillon A., Foster A., Williams C.D. and Griffin C. (2012)
“Environmental safety of entomopathogenic nematodes – effects on
abundance, diversity and community structure of non-target beetles
in a forest ecosystem” Biological Control 63(2): 107-114.
All papers available at:
http://nuim.academia.edu/WilliamsChristopher
Acknowledgements







Aoife Dillon (Coillte)
Minshad Ansari and Tariq Butt
John Dunbar, Rose Waldron, Alicia Bétry, Karen Morris,
Karen Shevlin, Roseanne Hennessy and Alan Hildebrand for
help with destructive and emergence trap collections.
Johan Van Vlaenderen and Louise McNamara (MCOP,
DAFM)
Chris Harvey and Conor Meade
Becker Underwood for supplying nematodes and fungus.
Work was funded by the EU INTERREG (Ireland-Wales
programme) and DAFM.

Use of entomopathogenic nematodes and fungi in the

Transcription

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