ABG-186 DEC

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ABG-186 DEC
GENETIC VARIATION IN PREVALENCE OF STOMACH
WORM (Haemonchuscontortus) IN DIFFERENT
POPULATIONS OF BLACK BENGAL GOAT
MS Thesis
NABILA NOURIN RETEE
Department of Animal Breeding and Genetics
Bangladesh Agricultural University
Mymensingh
December (2014)
GENETIC VARIATION IN PREVALENCE OF STOMACH
WORM (Haemonchuscontortus) IN DIFFERENT
POPULATIONS OF BLACK BENGAL GOAT
A Thesis
Submitted to
Bangladesh Agricultural University, Mymensingh
In Partial Fulfillment of the Requirements for the Degree of
Master of Science
in
Animal Breeding and Genetics
By
NABILA NOURIN RETEE
Roll No. 13ABGJD02M
Registration No. 34379Session: 2007-08
Department of Animal Breeding and Genetics
Bangladesh Agricultural University
Mymensingh
December (2014)
GENETIC VARIATION IN PREVALENCE OF STOMACH
WORM (Haemonchuscontortus) IN DIFFERENT
POPULATIONS OF BLACK BENGAL GOAT
A Thesis
Submitted to
Bangladesh Agricultural University, Mymensingh
In Partial Fulfillment of the Requirements for the Degree of
Master of Science
in
Animal Breeding and Genetics
By
NABILA NOURIN RETEE
Appoved as to style and content by
_________________________
Prof. Dr. Md. OmarFaruque
Supervisor
________________________________
Prof. Dr. Md. MotaharHussainMondal
Co-supervisor
_________________________
Prof. Dr. Md. Omar Faruque
Chairman, Defence Committee
and
Head, Department of Animal Breeding and Genetics
December (2014)
ACKNOWLEDGEMENTS
The author has pleasure to express her greatest and deepest gratitude to the
supreme of everything Allah, for her mercy to successfully complete the thesis
work, manage each and everything soundly.
The author feels it a profound privilege to expresses her deepest sense of gratitude,
profound appreciation and heartfelt indebtedness to her research supervisor
Professor. Dr. Md. Omar Faruque, Department of Animal Breeding and Genetics,
Faculty of Animal Husbandry, Bangladesh Agricultural University, Mymensingh,
for his scholarly guidance, innovative suggestions, encouragements, constant
supervision, constructive criticism, affable behavior and untiring perseverance
throughout the tenure of this research work and preparing manuscript of this
thesis.
The author wishes to express her indebtedness with heartiest gratitude to another
research supervisor Professor Dr. Md.MotaharHussainMondal, Department of
Parasitology, Faculty of Veterinary Science, Bangladesh Agricultural University,
Mymensingh, for his outstanding inspiration, total guidance, intellectual help,
valuable criticism, keen suggestions, cordial behavior throughout this research work
and preparation of this manuscript.
The author is also thankful to the present respectable chairmanProfessor. Dr. Md.
Omar Faruque, Department of Animal Breeding and Genetics, Faculty of Animal
Husbandry, Bangladesh Agricultural University, Mymensingh, for the moral
support throughout this research work and benevolent inspiration as well as overall
his greatest kindness. The author is also indebted and grateful to all of her
honorable teachers of the Department of Animal Breeding and Genetics, Faculty of
Animal Husbandry, Bangladesh Agricultural University, Mymensingh, for their
valuable advice and help during the course of the study period.
i
The author is grateful and wants to give special thanks to the project of “Studies
on the Quantitative Traits Loci (QTL) of Economic traits in Black Bengal
goat” funded by SPGR program of NATP Phase –I and “Genetic variation on
the control of nematodes in the small ruminant in Bangladesh” funded by IAEA
(International Atomic Energy Agency, Vienna, Austria)for financial support during
the research period.
The author is very much thankful to the Sheikh HabiburRahman, Md. Jalaluddin,
Md. Mohon Mia and all field assistants for their cordial behavior and help
throughout this research work.
The author says special thanks to Desha, Pervez, Manik, Juel, Mitu, Sumi, Rupa,
Badhon, Moon and all of her friends and well-wishers for their constant company
and encouragement during this study period.
Finally, the author owes her deepest gratitude, heartfelt respects, and thanks to her
beloved parents, husband, sisters Rachi and Mugdha, brother Rakin for their
understanding, inspirations, moral support, kindness and blessings, forbearance and
endless love to complete her study.
THE AUTHOR
December, 2014
ii
GENETIC VARIATION IN PREVALENCE OF
STOMACH WORM (Haemonchuscontortus) IN
DIFFERENT POPULATIONS OF BLACK BENGAL
GOAT
NABILA NOURIN RETEE
ABSTRACT
Black Bengal goat is the only recognized breed of Livestock in
Bangladesh.A number of factors viz., reduced grazing land for goat, diseases
and parasite infection are limiting the production of Black Bengal goat in the
country nowadays. This experiment was conducted to investigate the genetic
variation in resistance to Haemonchuscontortus(a GI Nematode)in different
populations of Black Bengal goat in Bangladesh. To achieve this goal, two
studies were done. In study 1, 196 goats from different populations of Black
Bengal goat (BBW, BBC, BBH)were studied.The prevalence of
Haemonchuscontortuswas 47.52%, 46.54% and 42.85% for BBW, BBC and BBH
population respectively. So, more than 50% of Black Bengal goats under natural
grazing condition had noHaemonchuscontortusinfestation.The haematological
values (PCV and Hb) were larger in non-infectious goats than infectious goats
in all three populations. In study II, two populations of Black Bengal goat (BBC
and BBH) were studied through artificial challenge trial. 5 kids of 5 months old
were dewormed properly and infested with L3 larvae @5000/ kid. Thebody
weight (BWT),faecal egg count(FEC), haematological values (Hb and PCV) were
measured at day 0 (Day when larvae were infested artificially), day 21 (21 days
after the larvae were infested artificially) and day 28 (day after the larvae were
infested artificially). Only two kids of BBC had FEC at day 21 and day 28
thought it was low ranging from 50 to 200 only. On the other hands, no kids of
BBH had FEC even at day 28.The studies revealed that BBH had the least
prevalence and load of Haemonchuscontortusthan other two populations of Black
Bengal goat in naturally grazing condition and artificial challenge trial.This
supports
the
between
population
difference
for
resistance
to
Haemonchuscontortus inBlack Bengal goats.It might be topography, feeding habit
or genotypic differences of threepopulations of Black Bengal goats. Further
work on gene expression is needed to clarify the actual cause behind this fact.
iii
CONTENTS
CHAPTER
TITLE
PAGE
ACKNOWLEDGEMENTS
i-ii
ABSTRACT
iii
CONTENTS
iv-v
LIST OF TABLES
vi
LIST OF PLATES
vii
LIST OF FIGURES
viii
CHAPTER
1
INTRODUCTION
1-5
CHAPTER
2
REVIEW OF LITERATURE
6-18
Importance of Black Bengal goat in
6-7
2.1
Bangladesh
2.2
Prevalence of nematode and stomach
7-9
worm in ruminant including Black Bengal
goat in Bangladesh
2.3
Haemonchuscontortus
9-10
2.4
Prevalence of Haemonchus parasite and
11-13
economic loss in ruminant including goat
CHAPTER
2.5
Prevention for Haemonchuscontortus
13-16
2.6
Research on Haemonchusresistance gene
16-18
MATERIALS AND METHODS
19-29
Study-I: Study on the prevalence of
19-25
3
3.1
parasites
3.2
Study-II: Artificial challenge trial
3.3
Study-III: Identification of population
resistance to Haemonchuscontortus
iv
25-28
29
CONTENTS
CHAPTER
CHAPTER
CHAPTER
CHAPTER
CHAPTER
TITLE
PAGE
4
RESULTS
30-41
4.1
Study-I: Prevalence of parasites
30-37
4.2
Study-II: Artificial challenge trial
38-41
4.3
Identification of population resistance to
Haemonchuscontortus
DISCUSSION
CONCLUSION
REFERENCES
5
6
7
v
41
42-44
45-46
47-59
LIST OF TABLES
TABLE
TITLE
PAGE
3.1
Schedule of artificial challenge trial
26
4.2
Prevelance of Haemonchuscontortus in BBW, BBC and BBH
30
population
4.3
ANOVA table for prevalence of Haemonchuscontortus in three
populations of Black Bengal goat
31
4.4
Prevalence of Haemonchuscontortusin different sex and age
32
gropus in Black Bengal goats
4.5
Faecal egg count (epg) and percent of animal affected by
Haemonchuscontortus
33
4.6
Variation in PCV and Hb values among the
Haemonchuscontortusinfected and non-infected goat
35
4.7
Level of significance between infectious and non-infectious
goats for PCV value by t-Test
36
4.8
Level of significance between infectious and non-infectious
goats for Hb value by t-Test
36
4.9
FEC, HB and PCV at and after artificial infestation of larvae (L3
@5000/kid)
38
4.10
Body weight of kids at and after artificial infestation of larvae
(L3 @5000/kid)
40
vi
LIST OF PLATES
TABLE
TITLE
PAGE
[
[[[
3.1
20
Different populations of Black Bengal goat. BBW: Black Bengal
goat in the western part, BBC: Black Bengal goat in the western
part, BBH: Black Bengal goat in the western part,
3.2
Black Bengal goats are under natural grazing condition. A
22
represents BBW goats, B represents BBC goats and C represents
BBH goats
[
[[
3.3
23
Clean canvass floor mat used for faeces collection
[
[[
3.4
28
Collection of Haemonchuscontortus wormfrom slaughtered
goat’sabomasums
vii
LIST OF FIGURES
TABLE
TITLE
PAGE
[[[ [
1.1
3
Dendrogram for genetic relationship of five goat
populations/breeds BBW: Bangladesh west, BBC: Bangladesh
central, BBH: Bangladesh Hilly, JAM: Exotic breeds, CRW:
Crossbred between Black Bengal and exotic breeds
[[[ [[
2.2
[
4.3
[[[
[[
Life cycle of Haemonchuscontortus
Prevelance of Haemonchuscontortus in BBW, BBC and BBH
10
[
31
population
[[
[[
4.4
34
Distribution of animals having different level of faecal egg count
of Haemonchuscontortus
[[[
[[
4.5
37
Correlation between PCV value (%) and different level of faecal
egg count of Haemonchuscontortus
4.6
37
Correlation between Hb value (gm/dl) and different level of faecal
egg count of Haemonchuscontortus
4.7
Body weight of kids at different days during artificial challenge
trial
viii
41
Chapter 1
INTRODUCTION
Goat is an important domestic animal in many parts of the world and
has served mankind for ages. It provides substance in the form of food and
clothing. This hardy ruminant can exist in harsh environment in which other
livestock species would perish. Goat grows and reproduces under extreme
conditions from rugged mountain areas where winters are bitter cold to desert
regions where it is hot and dry, and water and forage are sparse. Goats are
socio-economically important in developing countries, ensuring food and fiber
supply and providing income to small households (Lebbie, 2004; Sahlu et al.,
2004; Sahlu and Goetsch, 2005). Today, more than 90% of the goat population is
in developing countries where goat meat and meat products are considered as
one of the most important sources of income (Shrestha and Fahmy, 2007). The
goat is being recognized as a significant food source, because goats can convert
feed DM into milk as efficiently as other ruminants; 185 kg milk/100 kg of dry
OM for does compared with 162 kg for cows in temperate environment
(Spedding, 1970). Goat has been recently recognized as a tool for poverty
alleviation. Owing to increased demand for goat products, more livestock
producers are raising goats in developing countries (Sahlu and Goetsch, 2005).
Goat production provides employment for poor rural families, especially for
women and children. Especially during droughts when crops fail, goats due to
their adaption capabilities, can survive on woody browses and infrequent
watering; coupled with
their high reproductive rate and short generation
interval, goats their owners to recover quickly and economically (Lebbie, 2004;
Peacock, 2005). The special characteristics of goats that make them particularly
important in rural poor communities compared to other domestic ruminants
include: ability to graze and utilize a wide range of poor quality forages and
browse; ability to walk long distances; short generation intervals and high
1
reproductive rates; high turnover rates on investment and hence low risk on
investment; efficient utilization of marginal lands; smaller carcasses. It is
generally accepted that goats require low inputs and are easy to manage,
contributing to a sustainable economic stock for more smallholders in the
tropics and subtropics (Acharya and Battacharya, 1992). The goat is a very
important animal genetic resource in the world. The world population of goats
is 910 millions, of which 94% are found in the developing countries (FAOSTAT
2013). Africa and Asia account for about 90% of the total population in the
developing countries, including a bewildering variety of breeds. India, China,
Pakistan and Bangladesh possessed 31.2%, 29.3%, 10% and 7.5% of the total
goat of Asia, respectively. These countries together possessed about 78% of the
population of goats in Asia. Bangladesh possessed 50500 thousand goats that
represented 6.7% of the total population in the world (FAOSTAT, 2011).
More than 90% of the goat population in Bangladesh is comprised of
Black Bengal having some variants in color and size; the majority of remainder
is imported Indian breeds and their crosses (Husain, 1993). Goat Black Bengal
goat is found almost in all villages of Bangladesh. There are wide variation in
color, body size and weights of goats found in different location. Black Bengal
goats are mostly black in color which comprises 80% of the total goat
population and the rest are of white, brown, gray, mixed or spotted coat color
and any combination of those colors at any proportion (Nozawa et al., 1984). In
the livestock sector, Black Bengal goat is the only recognized breed amongst the
domestic species available throughout Bangladesh. Black Bengal goat is a dwarf
breed and famous for high fertility, prolificacy, superior chevon quality, best
quality skin, early sexual maturity, resistance against common diseases,
aseasonality, low kidding interval and very good adaptability (Hussain, 1993;
Islam et al., 2001; Devendra and Burns, 1983). They are very much adaptive to
the stressful and varied nutritional regimen. They graze a wide spectrum of
plants, grasses, shrubs and leaves of perennial trees. They can grow optimally
even without concentrate feeds.
2
Faruque (2009) classified Black Bengal goat into three sub groups
according
to
the
geographical
distribution
of
the
goats
and
DNA
polymorphisms (Fig. 1). These are Bangladesh West (BBW), Bangladesh Central
(BBC), and Bangladesh East (BBH). There is morphological and genotypic
variation among these populations. A number of exotic breeds were introduced
in the past for cross breeding purpose. As a result, many crossbred goat (cross
between Black Bengal and exotic breeds) are visible in different part of the
country. The exotic breeds and crossbreds are concentrated in the western part
of the country. In the absence of any census, it is difficult to mention the exact
population number of each population or breed. However, Black Bengal goats
are clearly predominant.
100
CRW
99
JAM
BBW
BBC
BBH
Fig 1: Dendrogram for genetic relationship of five goat populations/breeds
BBW: Bangladesh west, BBC: Bangladesh central, BBH: Bangladesh Hilly,
JAM: Exotic breeds, CRW: Crossbred between Black Bengal and exotic breeds
The Parasitism has been considered as one of the major constraints of
livestock production. Parasitism is an important limiting factor that is
responsible for deteriorating the health and productivity of livestock. Parasitic
infestations exert adverse effects on the health and productivity of animals
(Rehman et al., 2009). These effects are varied and more pronounced in goats
(Iqbal et al., 1993). Many species of parasites are found in goats and usually
include Haemonchus, Oesophagostomum, Ostertagia, Cchabertia, Nematodirus,
Trichuris, Moniezia and Fasciola. One of the most important species is
Haemonchus contortus (Husnain and Usmani, 2006).
3
Haemonchus contortus, also known as red stomach worm, wire worm or
barber's pole worm, is very common parasite and one of the most pathogenic
nematodes of ruminants. Adult worms are attached to abomasal mucosa and
feed on the blood. This parasite is responsible for anemia, bottle jaw, and death
of infected sheep and goats, mainly during summer months in warm, humid
climates. Haemonchus contortus causes helminthiasis especially parasitic gastroenteritis (PGE) which constitutes a serious health problem and limitation to the
productivity of goats and sheep throughout the world due to the associated
morbidity, mortality and cost of treatment and control measures (Silvestre et al.,
2000). It causes the animals to be unthrifty which may include the loss of
weight, low birth weight, and difficulty in kidding. It exerts the greatest
economic losses in temperate and tropical regions (Blood et al., 1979; Raza et al.,
2009; Ijaz et al., 2009).
Haemonchus contortus parasites pretenses the greatest challenge to goat
health and production in humid areas. The standard treatment to control the
challenge has been the use of anti helminthics. However, parasite resistance to
anti helminthes drug is an increasing problem. It is increasing the cost of
treatment and production. Livestock Producers and breeders are now trying to
develop breeds that have resistant to infectious diseases. In this regards, they
are trying to exploit the within and between breed genetic variation in
resistance to infectious diseases. There is well-documented evidence for within
and between breed genetic variations in resistance to infectious diseases, such
as gastrointestinal nematode infections, diseases due to mycotoxins, bacterial
diseases including foot root and mastitis, ectoparasites such as flies and lice,
and scrapie, the small ruminant transmissible spongiform encephalopathy.
Due to this document, within-breed variation of disease resistance in many
cases is a heritable trait. This offers the opportunity to select animals for
enhanced resistance to the disease. The feasibility of this approach has been
experimentally demonstrated and in other cases, breeding programs selecting
4
commercial animals for enhanced resistance are being successfully established,
especially for sheep as compared to goats. There are concerted efforts to find
genetic markers associated with resistance to infections, potentially allowing
selection for increased resistance in the absence of infection, especially for
scrapie and nematode parasitism (Bishop and Morris, 2004).
Black Bengal goat is the only breed of livestock in Bangladesh. Like other
ruminants, it is also infected by stomach worm (Qadir, 1967; Haq and Shaikh,
1968, Howlader et al., 1996; Karim, 2003; Shahiduzzam et al., 2003; Akanda et al,
2012; Hossen, 2012; Hassan et al., 2014) resulting in deteriorate of the health of
animal and reduction of production. It will, therefore, be worthy to develop
Black Bengal goat resistant to stomach worm. However, information or report
on within breed genetic variation for stomach worm of different populations of
Black Bengal goat is not available. This work was therefore undertaken
i.
To investigate the prevalence of Haemonchus contortus in different
populations of Black Bengal goat kept under naturally grazing condition.
ii.
To investigate the effect of artificial infestation of Haemonchus contortus in
different populations of Black Bengal goat kept under confined condition.
iii.
To identify the population of Black Bengal goat related to resistant to
Haemonchus contortus.
5
Chapter 2
REVIEW OF LITERATURE
There are a lot of work have been done on the life cycle of Haemonchus
contortus, its effect on the ruminant health and production and within and
between breed genetic resistant specially of sheep and goat in abroad and few
in the country. In the following sections, some of these reports have been
presented.
2.1 Importance of Black Bengal goat in Bangladesh
Goat is numerically and economically important and promising animal
resources in the developing countries especially in Asia and Africa (Husain,
1993). It is estimated that more than 90% of goat population consists of Black
Bengal goat (BBG). The Black Bengal goat is only breed of livestock in
Bangladesh. It is well known for high fertility, fecundity and excellent skin
quality. The Black Bengal goat is the most common and popular livestock of
Bangladesh. Among the Asian countries, Bangladesh had the 4th highest
population of goats (Amin et al., 2000). Small land requirement and adaptability
to harsh climate have made goats more suitable for rearing in our country
(Singh and Rai, 2006). Goats have been reared in Bangladesh from the time of
human settlement in this part of the earth. They are also considered a potential
genetic resource for poverty alleviation as they are the source of income for
many small and landless farmers. All the goats in Bangladesh are reared under
semi intensive system. The average number of goats per household is 2.31 and
they are mostly reared by landless, small and medium farmers (Faruque et al.,
2010). They are owned by almost all the categories of the people including
ethnic groups and poor and subsistence farmers.
Reproductive efficiency is always considered to be the most vital factor
ensuring increase in productivity to a certain environmental condition (Hossain
et al., 2004). Increased production efficiency can be obtained from goats since
6
they have a high reproductive efficiency with the potential for increased litter
size and shorter generation interval and they have a relatively higher fertility
rate in comparison to other farm animals (Williamson and Payne, 1978).
Reproductive performance of goats is a major determinant of productivity and
economic viability of commercial goat farms. The goats’ reproductive
performance is an indicator of their adaptation to the adverse conditions. Black
Bengal goat is that unique creature which has high reproductive and productive
efficiency than any other crossbred. This breed has an excellent ability to
accommodate and adapt to fluctuation in environment. The Black Bengal goat is
the most prolific of all domestic ruminants under tropical and sub-tropical
conditions and certain breeds are able to breed throughout the year.
(Devendra, 1970; Riera, 1982; Song et al., 2006).
2.2 Prevalence of nematode and stomach worm in ruminant including Black
Bengal goat in Bangladesh
In Bangladesh, parasitic infection is the major cause of hindering the
development of livestock population. The climate of Bangladesh is suitable for
the parasites, which are to great extent responsible for kid mortality in this
country. The hot and humid climatic conditions in Bangladesh are highly
conductive for the development and multiplication of parasites (Jabber and
Green, 1983).
Bangladesh is a tropical country with moderate winter and
summer. This climatic condition is suitable for the development and survival of
many parasites. Besides, in this country most of the goats are reared in rural
areas in scavenging or semi scavenging system (Devendra, 1970). In this type of
rearing, goats graze on the fields. This type of management practice plays a
vital role in the high rate of parasitic infection (Domke et al., 2011). Gastro
intestinal Nematodes are spread by orally ingesting the infective stage of the
parasite. This most commonly occurs during grazing.
A warm-wet environment of Bangladesh is most conductive to build-up of
worm population in ruminants. Amongst them, stomach worms of goat are
7
great problem (Mondal and Islam, 1994). Black Bengal goats of Bangladesh are
affected by various intestinal helminths (Qadir, 1967; Haq and Shaikh, 1968). A
number of studies carried out in different parts of the country revealed that
Haemonchus contortus parasitic infection is common in Black Bengal goat (Haq
and Shaikh, 1968; Howlader et al., 1996; Karim, 2003; Shahiduzzam et al., 2003;
Akanda et al, 2012; Hossen, 2012; Hassan et al., 2014). These studied were
conducted mainly on the naturally grazing Black Bengal goats in Chittagong,
Sylhet and Mymensingh district. These studied revealed that there was
infection of Haemonchus contortus in goats as determined by faecal egg count as
well as by the presence of worm in the abomasums; however the occurrence of
Haemonchus contortus varied with seasons of the year, and
sex and age of
animals. (Hassan et al. 2014) conducted an experiment on prevalence and
pathological effects of intestinal Helminths including Haemonchus contortus in
Black Bengal goat in Chittagong district. The study clearly suggested that Black
Bengal goats were susceptible to intestinal helminths in both winter and
summer seasons, and most of the parasites recovered were associated with the
production of variable degree of pathological lesions. Black Bengal goat is
assumed to be a disease resistance breed. In many researches, it has been shown
that the rate of occurrence of parasitic infection in Black Bengal goat is very low.
(Ratnesh et al., 2013) worked on variability of resistance in Black Bengal with
various genetic and non-genetic factors. The study found that the resistance of
kids under sire was varied significantly and it was also noticed that maternal
genetic effect has a very little impact on resistance of kids. Males (buck) were
most resistant and the kids were least resistant and the resistance of dam was in
between the male and kids population.
2.3 Haemonchus contortus
Haemonchus contortus is one of the most prevalent and pathogenic
parasite infesting the stomach of ruminants irrespective of age, gender, and
breed of the host throughout the world leading to tremendous loss in variety of
ways. So, several attempts have been made to study this parasite by a number
of researchers (Franklin, 1935; Ksull, 1939; Almedia, 1945; Silverman and
8
Patterson, 1960; Sahai and Deo, 1964; Altaif and Issa, 1983; Masud and Jamil,
1887; Gelaye and Wossene, 2003, Muzaffar, 2010, Kuchai, 2012). Adult worms
are attached to abomasal mucosa and feed on the blood. This parasite is
responsible for anemia, bottle jaw, and death of infected sheep and goats,
mainly during summer months in warm, humid climates.
The adult female worm can release between 5,000 and 10,000 eggs, which
will be passed out in the feces. Eggs then develop in moist conditions in the
feces and continue to develop into the L1 (rhabditiform), and L2 juvenile stages
by feeding on bacteria in the dung. The L1 stage usually occurs within four to
six days under the optimal conditions of 24–29 °C. The L2 rhabditiform sheds
its cuticle and then develops into the L3 filiariform infective larvae. The L3 form
has a protective cuticle, but under dry, hot conditions will not survive long. The
L3s then crawl up the blades of wet grass and wait to be ingested by a grazing
animal. Sheep, goats and other ruminants become infected when they graze and
eat grasses containing the L3 infecting larvae. The infecting larvae pass through
the first three stomachs to reach the abomasums. There, the L3s shed their
cuticles and burrow into the internal layer of the abomasums, where they
develop into L4s, usually within 48 hours, or preadult larvae. The L4 larvae
then molt and develop into the L5 adult form. The male and female adults mate
and live in the abomasum, where they feed on blood.
9
Fig. 2: Life cycle of Haemonchus contortus
There are significant observable genetic and phenotypic divergences
among Haemonchus contortus species. Microsatellite markers have been used to
investigate genetic divergence, whilst phenotypic divergence has been
considered through individual worm morphology, isolate life history traits and
the effect of isolates upon the host. In an experiment genetic and phenotypic
divergence between isolates of Haemonchus contortus in Australia was observed
(Lello et al., 2004). They conclude that there is significant observable genetic
divergence between isolates of H. contortus in Australia. Phenotypic divergence
is also observed, and potentially has significant implications for both economic
losses due to haemonchosis on individual properties and for decisions
regarding the regulation of stock movements in Australia.
10
2.4 Prevalence of Haemonchus parasite and economic loss in ruminant
including goat
In the tropics, the most important nematode species affecting ruminants
are Haemonchus contortus, Trichostrongylus species, Nematodirus species Cooperia
species, Bunostomum species Oesophagostomum species. Heamonchus contortus
commonly known as the twisted stomach worm is a blood sucking nematode
parasite, primarily occurring in the abomasum of small ruminants, notably
sheep and goats. It has been ranked as the most important parasite of ruminants
in all regions across the tropics and subtropics and causes an insidious drain on
production, weight losses and even mortality in young animals as reported by
(Bhat et al., 2010).
Gastrointestinal parasites not only affect the health but also affect the
productive and reproductive performance of the cattle and buffalo.
Gastrointestinal worms are recognized as by for the most significant part of
diseases in livestock sector (Waller 1997). It has been established that parasitic
infestation results in considerable losses in milk production in cattle and buffalo
(Hayat et al., 1984). Among the predisposing factors of internal parasites
infection are climates, nutritional deficiency, grazing habits, immunological
status, pasture management, presence of intermediate host and vector and the
number of infective larvae and eggs in the environment. Damages inflicted to
the health and productivity includes loss in body weight, poor reproductive
performance, digestive disturbance, and emaciation for longer period (Radostits
et al., 1994). Other economic losses are poor work performance, involuntary
culling, lower milk production, treatment costs, and mortality in heavily
parasitized animals (Lebbie et al., 1994). So it is important to control internal
parasites through better management as in developed countries, and
knowledge on prevalence of these parasites is mandatory.
11
In most sheep production areas, internal or gastro-intestinal parasites
(i.e. worms) are usually the primary disease affecting sheep and lambs. Sheep
are more susceptible to internal parasites than most other types of farm
livestock for several reasons. Their small fecal pellets disintegrate very easily
thus releasing the worm larvae onto pastures. Nematode parasites are the major
animal health constraint in sheep production on pasture. Chemical control
using anthelmintic drenches has been a reliable means of nematode control for
the last 40 years but increasingly nematodes are becoming resistant to
anthelmintics. Nematodes resistant to all major classes of anthelmintics have
now been documented throughout the world for the three major sheep
nematode species – Haemonchus contortus, Teladorsagia circumcincta and
Trichostrongylus colubriformis. Multiply drug-resistant H. contortus is now
making small ruminant production, in some areas of the tropics, unsustainable.
(Vanimisetti et al., 2004) describes the Inheritance of fecal egg count and packed
cell volume and their relationship with production traits in sheep infected with
Haemonchus contortus. They concluded that ewes did not lose weight and lambs
continued to grow throughout the measurement period, indicating no major
negative effects of infection on BW in this production environment. There was
considerable seasonal variation in patterns of FEC over time in ewes. Generally,
autumn-lambing ewes showed an early increase in FEC, whereas springlambing ewes showed a later increase in FEC.
Goat rearing is hindered by various problems, among them parasitism is
an important limiting factor. Haemonchus contortus is a highly pathogenic and
economically important gastrointestinal parasite of goats. These parasites are
common blood feeders that cause anemia, reduced productivity and can lead to
death in heavily infected animals. The results of an experiment revealed that
43.10% (459) goats were infected with different species of nematodes including
Haemonchus contortus (14.65%), Trichuris ovis (8.17%), Trichostrongylus axei
(7.61%), Trichostrongylus colubriformis (6.76%), Oesphagostomum columbianum
(5.35%), Ostertagia circumcincta (5.35%), Chabertia ovina (4.79%) and Strongyloides
12
papillosus (4.51%). This experiment was carried out by (Akhter et al., 2011) in
Hyderabad and Adjoining Areas. But there are some variations of resistance in
goats infected with Haemonchus contortus. (Costa et al., 2000) studied the
variability between and within breeds with respect to nematode egg counts
(EPG), packed cell volume (PCV) and hemoglobin (Hb) in yearling female goats
of the Caninde(15), Bhuj (6) and Anglo-Nubian (15) breeds,
exposed to
Haemonchus contortus. There are many associated risk factors influencing the
prevalence of gastrointestinal helminthes including age, sex, weather condition
and husbandry or management practices (Miller et al., 1998; Khan et al., 2009).
Parasitism and gastrointestinal nematode parasitism is arguably the most
serious constraint affecting small ruminant production world-wide especially
goat population. Economic losses are caused by decreased production, cost of
prevention, cost of treatment, and the death of infected animals. Problems with
nematode parasitism are often classified as production disease (i.e. chronic
subclinical condition affecting productivity such as weight loss, reduced weight
gain, reproductive inefficiency, etc.). It should be noted here that is more when
goats are managed as grazers. When goats are managed as browsers, exposure
to nematode parasites is reduced and subsequently the effects are not as severe
(Miller et al., 2005).
2.5 Prevention for Haemonchus contortus
Internal parasites are a significant threat facing today’s small ruminant
producer. Problems associated with parasites, particularly those of the
gastrointestinal tract of goats can cause irreversible damage or even death to the
animal, reduced performance and economic loss for the producer. Animals that
are overburdened with parasites can be hindered in their reproductive
performance, experience reduced growth rates, and become less productive
overall, whether their purpose be meat, fiber, or milk. Control of internal
parasites, especially gastrointestinal nematodes including Haemonchus contortus
13
(barberpole worm, stomach worm), is a primary concern for many goat
producers and is particularly challenging in humid regions.
Anthelmintics are drugs that either kill egg laying adults, or kill larvae
before they become adults and become capable of laying eggs. Current control
practices rely heavily upon the frequent use of anthelmintics which leads to the
development of resistance to the anthelmintics (Pandey and Sivaraj, 1994).
There are various types of anthelmintics (albendazole, fenbendazole,
mebendazole,
oxfendazole,
ivermectin,
moxidecti
etc.).
While
some
anthelmintics are effective in killing Haemonchus, there are two major problems
that arise when using anthelmintics. The first problem that affects mainly sheep
and goat producers is that there are very few anthelmintics that are actually
approved by the FDA for use in small ruminants. The second problem is the
resistance that parasites have developed to many anthelmintics. Resistance
occurs when a drug is overused and the parasites develop a tolerance to the
drug, making it no longer effective in killing them. Resistance makes it very
difficult to effectively control Haemonchus because it lowers the number of
options available to treat the parasite, especially since resistance to one drug
often means that a parasite will be resistant to all drugs in that compound class.
Prevention and control of the parasites that infect sheep and goats are
becoming increasingly difficult due to generations of overuse and improper use
of the available anthelmintic dewormers, which results in increasing resistance
by parasites to common anthelmintics. In the past, control of parasites relied
almost exclusively on multiple and regular anthelmintic drenching, with the
aim to maximize livestock productivity and profitability. However, as it has
been established that the use of anthelmintics has several drawbacks such as:
the
increasing
incidence
of
parasite
resistance
against
the
available
anthelmintics; consumer concerns regarding drug residues in food products
and in the environment; the negative effect of preventive treatments on the
development of natural immunity against helminthes (Ketzis et al., 2006).
14
Pasture management is one of the best outcomes of parasite control
program is the reduction of number of parasites that the goats are exposed. This
can be accomplished by managing pastures in a way that will reduce the
parasitic load. Use control grazing practices to optimize pasture production.
This is a better practice than continuous grazing on the same pasture because
goats will return to the same areas where their favorite plants are growing.
Those areas will then become heavily infected by gastrointestinal parasite
larvae. Put goats in a browse area, such as a woodlot, when hot, humid
environmental conditions favor the rapid life cycle of gastrointestinal parasites.
By browsing, goats will not consume forage close to the ground where the
parasite larvae are located. In addition, many browse plants have the additional
benefit of harboring high tannin concentrations. Tannins have been shown to
reduce fecal egg counts and possible gastrointestinal parasite larvae numbers
(Kate, 2010).
The another way of parasite control is the pasture rotation or rotational
grazing which have the ability to break the parasite cycle has been tossed
around for years. The main reason to use pasture rotation is not for parasite
control but to provide the most nutritious forage for growth and development
of goat. But pasture management have the same limitations that it cannot fully
prevent the parasitic infection of goat.
But individual goats vary in their degree of susceptibility to Haemonchus
and other parasites. Some animals, by means of their genetics, are much more
resistant or resilient to parasitic infections, and can survive parasite levels
without showing any symptoms while another animal may be killed by that
level of infestation. This genetic resistance can be used when selecting breeding
stock, since a herd that exhibits more innate resistance to parasites will cost less
to maintain and will ultimately be more profitable to the producer. So it should
be the main to develop parasite resistance goat breed.
15
The increasing prevalence of anthelmintics resistance during the recent
years and also great concern about drug residues in the food chain have
necessitated the search for an effective alternative strategy to control
roundworm (especially H. contortus). This can be resolved by utilizing principle
of breeding for genetic resistance which is a component of integrated parasite
management (IPM) programmes (Woolaston and Windon, 2001; Bishop and
Morris, 2004). (Albers et al., 1987) reported that resistance to H. contortus was
moderately heritable; indicating that selection and breeding goat for increased
resistance is possible. Therefore shifting to resistant variety or designing cross
breeding programmes involving resistant goat breeds are thus most promising
application in combating nematode infection. In view of the above facts, the
present work has been undertaken on Black Bengal goat to study the variability
of resistance in Black Bengal goat naturally infected with H. contortus (Ratnesh
et al., 2013).
2.6 Research on Haemonchus resistance gene
The farm animal genetic materials are a rich collection of genetic
variations with either useful or harmful consequences on health and
productivity. These variations are usually remain in the form of single
nucleotide polymorphisms (SNPs), deletions of nucleotides or whole genes,
gene or whole chromosomal rearrangements, gene duplications, copy number
polymorphisms, copy number variations and presence/absence of transposable
elements.
Selection for increased resistance exploits the genetic variability in
resistance observed in most commercial sheep breeds and in some goat breeds
(Baker and Gray, 2004). One approach is to select resistant animals using
phenotypic traits such as the faecal egg count (FEC), PCV, eosinophil count,
body weight (BW) and immunoglobulin A and E (IgE and IgA) activity which
are indirect measures of resistance. Selection for phenotypic traits has been
16
successfully used in Australia and New Zealand (Karlsson et al., 1991; Kemper
et al., 2010); however, it is costly and time-consuming as it requires animals to
be challenged with parasites. Considering the cost and time, for goat breeding
and genetics, MAS is now considered the optimal choice (Weller and Ron,
2011). Unfortunately, little information about the candidate genes affecting
diseases resistance traits is available. For this reason, excavating the suitable
candidate DNA markers that correlate with growth and diseases resistance
traits in goat breeds has become a major objective.
Exploration of genetic variation either within specific regions of genome
or more specifically in candidate genes involved in innate and adaptive
immune pathways may help to identify a set of DNA markers significantly
associated with parasite resistance characteristics. The former approach in
terms of quantitative trait locus (QTL) analysis is a powerful method to
understand genotype-phenotype relationship. Several QTL studies on parasite
resistance characteristics have been reported in sheep. Very few works have
been done on goat in the whole world.
The major histocompatibility complex (MHC) plays a key role in the
immune response. Clear evidence has been reported of genetic markers in the
class I and class II regions of the MHC on sheep chromosome 20 (OAR20) being
associated with resistance to nematodes. For example, the DRB gene on OAR20
(a part of class II of MHC) and the interferon gamma gene (IFNG) (Schwaiger et
al., 1995; Buitkamp et al., 1996; Paterson et al., 1998) both play a role in
regulation of immune responses to infections. (Schwaiger et al., 1995) found in
Scottish Blackface sheep that one allele (DRB1) of the DRB locus within the
ovine MHC II on OAR20 was associated with 98% reduction in worm egg
count. From 23 polymorphic alleles identified at the DRB locus in Polish Heath
sheep, two alleles (482 bp and 530 bp) demonstrated a significant association
with resistance to GIN, whereas another allele (568 bp) conferred increased
susceptibility to parasites (Charon et al., 2002). (Stear et al., 1996) studied in
17
Scottish Blackface lambs infected with Ostertagia circumcincta and showed that
class I alleles were in close linkage disequilibrium with the DRB1 alleles and
they were associated with a 10-fold reduction in FEC. In another study, the
MHC class I microsatellite (DYA) had a significant and moderately strong
(eightfold) association with FEC (Buitkamp et al., 1996). These authors stated
that the disease susceptibility locus is closer to the class II region than to the
class I region, and both of these host defence mechanisms (MHC I and II) are
likely to involve a type 2 helper T (TH2)-cells. The identification of QTL for
parasite resistance may vary depending on the nematode species, the
challenging regime and the indicator trait measured (Dominik, 2005).
18
Chapter 3
MATERIALS AND METHODS
This experiment was conducted in the Department of Animal Breeding and
Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh,
Gurudaspur and Baraigram Upazila of Natore district and Bandarban Hill
district on the Black Bengal goats from 15th July, 2013 to 30th September, 2014.
Animal Genetics Laboratory under the Department of Animal Breeding and
Genetics and Parasitology Laboratory under the Department of Parasitology
were used for analysis of faeces and blood. To achieve the goal, overall
activities were divided into the following three studies:
 To study the prevalence of Gastro-intestinal nematode including
Haemonchus contortus in different populations of Black Bengal goats kept
under natural grazing condition.
 To investigate the effect of artificial infestation of Haemonchus contortus in
different populations of Black Bengal goat kept under confined condition.
 To identify the population related to resistant to Haemonchus contortus.
Study –I
3.1 Study on the prevalence of parasites
3.1.1 Experimental site
The experiment was carried out from August, 2013 to November, 2013 covering
the rainy in three areas of Bangladesh. These were Chapainawabganj and
Rajshahi (western part of Bangladesh), Mymensingh (the central part of
Bangladesh) and Bandarban Hill district (eastern part of Bangladesh). The three
areas represent three topographical conditions and three different ecologies.
Chapainababgonj and Rajshahi district was dry area, Mymensingh was flood
fed area. Bandarban Hill district was hilly and forest area.
19
3.1.2 Experimental animal
Geographically and genetically, Black Bengal goat can be divided into three
populations: BBW (Black Bengal goat in the western part), BBC (Black Bengal
goat in the central part), BBH (Black Bengal goat in the eastern part and hilly
region) (Faruque, 2009). No work had been done on the naturally grazing Black
Bengal goats in hilly region like Bandarban Hill district (BBH) and dry region
like Chapainawabganj and Rajshahi (BBW). Present study represents three
genetically representative populations of Black Bengal goats and also wider
geographical areas of Bangladesh covering these populations.
200 goats of various ages (4-18 months) were randomly selected from the three
areas. Among them, 106 were female and 94 were male goats. The Black Bengal
goat population of Chapainawabganj and Rajshahi has been termed as BBW
(Black Bengal goat in the western part), the Black Bengal goat population of
Mymensingh has been termed as BBC (Black Bengal goat in the central part),
and the Black Bengal goat population of Bandarban Hill district has been
termed as BBH (Black Bengal goat in the eastern part) also represent goat of
hilly region.
20
BBC
BBW
BBH
Plate -1: Different populations of Black Bengal goat. BBW: Black Bengal goat
in the western part, BBC: Black Bengal goat in the western part, BBH: Black
Bengal goat in the western part
3.1.3 Management of the animals
All the goats under this study were reared in semi intensive system. This
system allowed the goats to graze freely in the natural pasture in the day time
about 6 to 8 hours and kept in confinement during night time. The BBW goats
obtained leaves of fruit tress (Mango and Jack fruit) in addition to grass as the
region was rich in fruit trees. The BBC goats lived on only grass of fellow land
as the region was flood fed area and lack of fruit trees. The BBH goats browsed
mostly on leaves of herbs in the forest. In all the cases, goats did not receive any
anti helmintic drugs as the owners did not practice any deworming program for
their goats.
21
A
B
C
Plate -2: Black Bengal goats are under natural grazing condition. A represents
BBW goats, B represents BBC goats and C represents BBH goats
3.1.4 Sample collection
Fresh fecal sample (near to 5 gm) was collected from each goat. Precaution was
taken so that there was no dust during the faeces collection using clean canvass
floor mat in most cases. The goat under study was kept confined on this floor
mat until it discharged the faeces. The faeces was preserved in 5% formalin and
carried to laboratory. Blood was also collected from the same goat when it was
allowed by the owners for estimating Haemoglobin value (Hb) and Packed cell
volume (PCV %). Blood was collected in EDTA coated venoject tube, carried in
ice box and kept in refrigerator at 40C until analyzed.
22
Plate 3. Clean canvass floor mat used for faeces collection
3.1.5 Faecal egg count (FEC)
Feacal egg was counted in the laboratory of Parasitology. A modified McMaster
technique was used to determine faecal egg counts and to identify different ova
and oocyst of nematode species following Cable (1957). In this method, 4gm of
faeces was thoroughly suspended in 56ml saturated salt solution. The
suspension was stirred through a 150 mm mesh sieve to remove the course
particles. A portion of the suspension was withdrawn with the help of Pasteur
pipette and allowed to run into the chambers of the McMaster slide. The slide
was allowed to stand for 5 minutes to float the eggs. The eggs in the two
chambers were counted using low power objectives (10X magnification). The
number of eggs per gram of feces was calculated by using the following
formula:
Number of egg in two chambers
Number of egg in one gm =
x dilution factor
0.3
23
Total volume of suspension in ml
Dilution factor =
Total volume of feces
In this study, a sensitivity saws 50 i.e. each egg represented 50 epg
3.1.6 Estimation of Packed cell volume (PCV)
Packed cell volume was measured from blood in the laboratory of Animal
Genetics. Packed cell volume is the fraction of whole blood volume that consists
of red blood cells (RBC). The procedure of PCV estimation has been given
below:
1. Well mixed blood sample was drawn by the special loading pipette.
2. The loading pipette was placed at the bottom of the hematocrit tube.
3. The hematocrit was filled with blood through the little pressure on the
rubber bulb and simultaneously removing the loading pipette up to 0 or
10 marks.
4. Blood above 0 or 10 mark should be removed with the help of cotton.
5. Then all of the hematocrit tubes were placed in the centrifuge machine
and centrifuged for 30 min at 3000 rpm.
6. After centrifugation the tube was removed from the machine and packed
cell volume or hematocrit was recorded from the bottom of the
graduated scale.
7. The result was expressed in percentage (%).
24
3.1.7 Estimation of Haemoglobin value (Hb)
Haemoglobin value was measured from blood in the laboratory of Animal
Genetics. Haemoglobin value was measured by acid hematin method with the
help of Haemometer. The procedure for measuring Hb has been described
below:
1. 2% HCl solution was taken in the diluting tube up to 2 marks.
2. Well mixed blood sample was drawn by the shale pipette up to 20 mm
mark.
3. The blood from the pipette was rinsed for 2-3 times.
4. The content was mixed thoroughly and left for five minutes in the
comparator.
5. After five minutes distilled water was added drop wise and mixed with
the help of stirrer.
6. The mixing was continued until and unless the color in the diluting tube
matched with the color of the comparator.
7. After matching the tube was removed and result was recorded from the
graduated scale.
8. The result was expressed in g/dl
Study –II
3.2 Artificial challenge trial
In order to determine the relative resistant of different populations of
Black Bengal goats to Haemonchus contortus, an artificial challenge trial was
conducted based on the result of prevalence of Haemonchus contortus in different
regions and populations of goat. The prevalence of Haemonchus contortus was
maximum in BBW (section 4.1) and minimum in BBH.
Faruque (2013)
conducted artificial challenge trial on BBW and BBH. So, BBH and BBC
populations were selected in the present for artificial challenge trial. The
artificial challenge trial was done as per the schedule shown below:
25
Table 1: Schedule of artificial challenge trial
Date
Activitity
Day
Transfer of BBC and BBH to individual goat
pan of AI shed
01.05.14
Quarantine & acclimination
1 week
Parasite culture, H.contortus
22.04.14
Faeces examination and Deworming
08.05.14
Faeces examination
16.05.14
Artificial challenge, infestation of L3
18.05.14
Day 0
FEC, PVC, Hb, BW
08.06.14
Day21
FEC, PVC, Hb, BW , Slaughter of animals for
15.06.14
Day 28
5000 L3 per kid
FEC, PVC, Hb, BW
tissue collection (liver)
FEC= Faecal egg count, PCV = Packed cell volume, Hb = Haemoglobin,
BW = Body weight
3.2.1 Experimental site
This work was carried out in AI Centre at Bangladesh Agricultural
University, Mymensingh from 1st May to 15th June, 2014.
3.2.2 Experimental animal
Ten Black Bengal female kids of 5 months age were used for the study.
Among them, 5 kids were procured from Mymensingh and 5 kids were
procured from Bandarban Hill district.
26
3.2.3 Management of animals
After procurement of kids, the feaces were collected from all the kids and
examined for Haemonchus contortus ova. Haemonchus contortus ova were found
in some kids of BBC group. So, all the kids were treated with anti-helminthes
(Ivermectin @ 1 ml per kid). Feaces were again examined after 7 days for the
presence of Haemonchus contortus ova. No Haemonchus contortus ova were found.
So the kids were ready for artificial challenge trial. During that seven days
prepatory period and the artificial challenge trial, all the kids were kept in
individual kid pen and stall fed with grass and concentrate mixture.
3.2.4 Production of L3 larvae of Haemonchus contortus
To produce L3 larvae of Haemonchus contortus, guts of a number of goats
were collected from slaughter house of Mymensingh, abomasun was dissected
and mature female stomach worms were collected. All the female worms were
transferred to a pastel motor and mashed. PBS (Phosphate Buffer Solution) was
added to it. Then the solution was filtrated with a sieve. One microscope slide
was examined at very low powers initially 10x of microscope. It was done to
know the concentration of eggs in the sample. The standard concentration of
eggs in a sample was 200-300/drop. The sample, if found positive, was
transferred to Petridis and kept at 250C in the lab. The sample was checked
every day to find out the growth of larvae until third stage larval were
observed. The culture was then ready for infesting the kids artificially. The
culture was prepared in the laboratory of Parasitology.
27
Plate 4. Collection of Haemonchus contortus worm from slaughtered goat’s
abomasums
3.2.5 Artificial challenge trial (infestation of larvae artificially)
Larvae, cultured in the laboratory, were infested artificially to all 10 kids
of two populations. Care was taken so that kid swallowed the larvae. Body
weight of kids was taken before infestation of larvae. Faeces and blood was
collected to estimate FEC, Hb and PCV. Date on which larvae were infested to
kids as counted day 0. Body of weight kids were again taken on day 21 and 28.
Faeces and blood was also collected to estimate FEC, Hb and PCV on day 21
and day 28. The artificial trail ended on day 28 as animals were slaughtered.
FEC, PVC and Hb were used as indicator for resistance to parasite.
28
3.3 Identification of population resistance to Haemonchus contortus
The individuals, kept under naturally grazing condition and having no
Haemonchus contortus ova in the faeces, and higher PCV and Hb values, were
considered resistance to Haemonchus contortus. Similarly the individuals, kept
under artificial challenge trial, and having no Haemonchus contortus ova in the
faeces, and higher PCV and Hb values, were also considered resistance to
Haemonchus contortus. If most of the individuals in population had no parasitic
or few load in the faeces (less than 300 epg), that population was considered
resistance to Haemonchus contortus. However, all the data obtained in study I
and study II was analyzed statistically to find out the difference in parasitic
load of within and between populations of Black Bengal goat. For between
population difference, data were analyzed following Completely Randomize
Design after Snedecor and Cochran (1980). For within population difference,
data was analyzed following student t-test after Snedecor and Cochran (1980).
29
Chapter 4
RESULTS
Study –I
4.1 Prevalence of Haemonchus contortus
Haemonchus contortus was found in all the three populations of Black
Bengal goat populations. However, parasitic load varied within and between
populations. The prevalence of Haemonchus contortus was 47.52%, 46.54% and
42.85% for BBW, BBC and BBH population respectively (Table 2 and Fig. 3). So,
more than 50% of Black Bengal goats under natural grazing condition remained
non-infected by Haemonchus contortus.
Table 2: Prevalence of Haemonchus contortus in BBW, BBC and BBH
population
Population
Total number of
goat studied
(n)
No. goat infected
with Haemonchus
contortus
(n)
*Prevalence of
Haemonchus
contortus
(%)
BBW
101
48
47.52c
BBC
39
18
46.54b
BBH
56
24
42.85a
*Values with different superscript within column differed significantly, at
p<0.05.
30
70
Prevalence of H. contortus %
60
50
40
30
20
10
0
BBW
BBC
BBH
Different populations of Black Bengal goat
Fig. 3: Prevalence of Haemonchus contortus in BBW, BBC and BBH
population
Table 3: ANOVA table for prevalence of Haemonchus contortus in three
populations of Black Bengal goat
Sources of
Level of
Variation
d.f.
S.S.
M.S.
F
significance
Treatment
2
5880128
2940064
16.9
0.01
Error
87
15135122
173966.9141
Total
89
The prevalence of Haemonchus contortus differ significantly at 1% (P < 0.01)
among three populations of Black Bengal goat.
31
The prevalence of Haemonchus contortus also varied with age and sex of
the Black Bengal goats (Table 4). From Table 4, it appeared that the prevalence
of Haemonchus contortus was more in female goats than male goats. The
prevalence increased with the increase in age of the goats.
Table 4. Prevalence of Haemonchus contortus in different sex and age gropus
in
Black Bengal goats
Parameters
No. of animals
examined
(n)
Prevalence
No of animal
with positive
faecal egg count
% +ve
(n)
Sex
Age
Male
91
34
37.36
Female
107
52
48.59
4m-8m
76
34
44.73
9m-18m
90
47
52.22
19m-3y
32
21
65.62
The maximum and minimum faecal egg count (epg) of Haemonchus contortus
were 2200 and 0, 1500 and 0, 200 and 0 for BBW, BBC and BBH population
respectively. The mean faecal egg counts were 663.54 + 69.77, 647.22 + 112.11
and 81.25 + 7. 86 epg for BBW, BBC and BBH population respectively. Goats,
which were infected by Haemonchus contortus, had different level of parasitic
load as revealed by faecal egg count. This has been presented in Table 5 and Fig.
4.
32
Table 5. Faecal egg count (epg) and percent of animal affected by
Haemonchus contortus
Population of goat
Faecal egg count( egg per gram)
% of animal
affected
BBW
>200
8.33
200-450
39.59
500-950
27.08
1000-1450
16.67
1500-2000
6.25
<2000
2.08
100.00
BBC
>200
5.55
200-450
44.45
500-950
16.67
1000-1450
27.78
1500-2000
5.55
<2000
0.00
100.00
BBH
>200
95.84
200-450
4.16
500-950
0.00
1000-1450
0.00
1500-2000
0.00
<2000
0.00
100.00
33
100
90
% of animal having parasitic load
80
70
60
50
BBW
40
BBC
30
BBH
20
10
0
Faecal egg count (egg per gram)
Fig. 4: Distribution of animals having different level of faecal egg count of
Haemonchus contortus
From the tables and figs, it appears that BBH had the least prevalence
and load of Haemonchus contortus among three populations of Black Bengal goat.
Other two populations had almost similar prevalence and load of Haemonchus
contortus.
4.1.1 Haematological values
The maximum and minimum PCV (%) and Hb (g/dl) value were 48 and 10.5,
36 and 8.5, 24 and 8.5 for BBW, BBC and BBH population respectively that were
infected with Haemonchus contortus. The maximum and minimum PCV (%) and
Hb (g/dl) value were 59 and 9.8, 44 and 8.4, 31 and 9.6 for BBW, BBC and BBH
population respectively that were not infected with Haemonchus contortus. The
34
mean and standard error of PCV and Hb value of BBW, BBC and BBH
populations are shown in Table 6.
Table 6. Variation in PCV and Hb values among the Haemonchus contortus
Infected and non-infected goat
Population
Goat infected with Haemonchus
contortus
PCV (%)
BBW
Hb (g/dl)
SE(±)
Mean
SE(±)
Mean
SE(±)
Mean
SE(±)
35.7
1.86
7.46
0.30
43.19
3.02
8.9
0.23
29.67
(20)
2.40
6.53
(8)
0.40
(5)
BBH
PCV (%)
Mean
(20)
BBC
Hb (g/dl)
Goat not infected with
Haemonchus contortus
24.00
(1)
36.38
(8)
2.71
(4)
00
8.50
(1)
00
24.43
(7)
7.80
(4)
1.27
8.00
(7)
*Number of animal studied has been indicated in the Parenthesis
From the above table the infectious and non-infectious goats show different
PCV and Hb value. So we measured the level of significance between infectious
and non-infectious goats for PCV and Hb value by t-Test. The results are shown
in the Table 7 and 8.
35
0.32
0.55
Table 7. Level of significance between infectious and non-infectious goats for
PCV value by t-Test
Population
df
t-calculated
t-tabulated
Level of
value
value
significance
BBW
13
2.11
1.77
**
BBC
7
1.84
1.82
NS
BBH
0
0.34
-
NS
** Significant at 5% (P < 0.05); NS = Non-significant
Table 8. Level of significance between infectious and non-infectious goats for
Hb value by t-Test
Population
Df
t-calculated
t-tabulated
Level of
value
value
significance
BBW
25
3.81
1.71
**
BBC
8
2.44
1.86
**
BBH
0
0.90
-
NS
** Significant at 5% (P < 0.05); NS = Non-significant
The correlation between parasitic load and Heamatological values has been
presented in Fig. 5 and Fig. 6. From the figures it appears that parasitic load had
negative effects on the haematological values of goats.
36
60
y = -0.0157x + 42.387
r = 0.689**
50
PCV value (%)
40
30
20
10
0
0
200
400
600
800
1000
1200
1400
No. of Ova
Fig. 5: Correlation between PCV value (%) and different level of faecal egg
count of Haemonchus contortus
12
y = -0.0022x + 8.4903
r = 0.612**
10
Hb value (gm/dl)
8
6
4
2
0
0
200
400
600
800
1000
1200
1400
1600
No. of Ova
Fig. 6: Correlation between Hb value (gm/dl) and different level of faecal egg
count of Haemonchus contortus
Study –II
37
1600
4.2 Artificial infestation of Haemonchus contortus
From the information from literatures and experiment I, two populations of
Black Bengal goat viz., BBC and BBH were selected for artificial trial. The result
of FEC, Hb and PCV at day 0 (Day when larvae were infested artificially), day
21 (21 days after the larvae were infested artificially) and day 28 (day after the
larvae were infested artificially) has been presented in Table 9.
Table 9. FEC, HB and PCV at and after artificial infestation of larvae
(L3 @5000/kid)
Population
BBC
BBH
Kid
No
Day 0
Day 21
Day 28
FEC
Hb
PCV
FEC Hb
PCV FEC Hb
Epg
g/dl
%
epg
g/dl %
epg
g/dl %
BBC1
0
12.6
38
0
12.6
39
0
12.6
39
BBC2
0
10
28
50
9.8
28
200
9.5
26
BBC3
0
13
30
50
13
29
150
12.6
28
BBC4
0
10.6
34
0
10.6
33
0
10.6
34
BBC5
0
10
28
0
10
28
0
10
28
BBH1
0
10.8
30
0
10.8
30
0
10.8
30
BBH2
0
10.8
27
0
10.8
28
0
10.8
28
BBH3
0
10
24
0
10
25
0
10
24
BBH4
0
10
26
0
10
26
0
10
26
BBH5
0
11
28
0
11
28
0
11
29
FEC – Faecal egg count, Hb – Haemoglobin, PCV- Packed cell volume,
epg - egg per gram
38
PCV
Table 9 reveals that there was no egg in the faeces of Kids in BBH
population indicating there was no Haemonchus contortus development. On the
other hand, only two kids of BBC population had eggs in the faeces indicating
there was Haemonchus contortus development. However, the total FEC was low
ranging from 50 to 200 only. The other three kids had no egg in the faeces. The
corresponding Hb and PCV values decreased slightly in those two kids. The
other kids that had no egg of Haemonchus contortus even after artificial
infestation of larvae showed almost constant level of HB and PCV. This
indicates that BBH population was resistant to Haemonchus contortus whereas
BBC was susceptible to Haemonchus contortus. This also reflected in body weight
change of the kids after the artificial infestation of larvae. The result of body
weight of kids at day 0 (Day when larvae were infested artificially), day 21 (21
days after the larvae were infested artificially) and day 28 (day after the larvae
were infested artificially) has been presented in Table 9.
39
Table 10. Body weight of kids at and after artificial infestation of larvae
(L3 @5000/kid)
Population
BBC
BBH
Kid No
Day 0
Day 21
Day 28
Body weight (Kg)
Body weight
(Kg)
Body weight (Kg)
BBC1
7.5
8.0
8.2
BBC2
7.0
7.6
7.75
BBC3
7.5
7.9
8.0
BBC4
7.0
7.5
7.6
BBC5
7.0
7.7
8.1
BBH1
8.0
8.7
8.9
BBH2
8.0
8.65
8.9
BBH3
7.5
8.0
8.2
BBH4
7.5
8.1
8.35
BBH5
9.0
9.6
9.8
When this data was placed in the graphs, there appeared that Kid2 and kid3
gained almost no body weight (Fig. 7).
40
12
10
8
BWT at D0
6
BWT at D21
BWT at D28
4
2
0
BBC1 BBC2 BBC3 BBC4 BBC5 BBH1 BBH2 BBH3 BBH4 BBH5
Fig. 7: Body weight of kids at different days during artificial challenge trial
4.3 Identification of Haemonchus contortus resistant population
The results of artificial challenge trial reveled that BBH population was
resistance to Haemonchus contortus while BBC population is susceptible to
Haemonchus contortus. The results of prevalence of Haemonchus contortus
revealed that most of individuals of BBH had no parasitic load. On the other
hands, individuals having parasitic load in BBH population had very low level
of parasitic load (Table 2 and Table 4). These results support the between
population difference in resistance to Haemonchus contortus in Black Bengal
goats. Table 4 also supports and provides evidence of within-population
resistance to Haemonchus contortus in Black Bengal goats.
41
Chapter 5
DISCUSSION
Black Bengal goat is the only recognized breed of Livestock in Bangladesh.
It is also one of most important ruminant livestock found almost every
household in rural area of the country. Its contribution is reflected in supplying
nutritious meat, valuable leather, income generation and poverty alleviation. It
has also cultural and religious role in the society. However, a number of factors
are limiting the production of Black Bengal goat in the country now a day. The
reducing grazing land for goat, diseases and parasite infection are also limiting
factors for the production of Black Bengal goat. Among the gastrointestinal
nematodes, Haemonchus contortus is the most frequent in Black Bengal goat of
Bangladesh as reported by a number of investigators viz., Qadir (1967), Haq
and Shaikh (1968), Howlader et al., (1996), Karim (2003), Shahiduzzam et al.
(2003), Akanda et al. (2012), Hossen (2012) and Hassan et al. (2014). Most of
those investigators worked on the Black Bengal goats in Mymensingh district
using slaughter house samples while few investigators worked on the Black
Bengal goats in Chittagong district. It may be mentioned here that Mymensingh
is considered as flood fed area which is also suitable area for harboring the
gastrointestinal nematodes like Haemonchus contortus. The Black Bengal goats of
Mymensingh and adjacent districts have been term as BBC in the present study.
So it can be said those studies represent the prevalence of Haemonchus contortus
in BBC. None of them worked on the naturally grazing Black Bengal goats in
hilly region like Bandarban Hill district and dry region like Chapainawabganj
and Rajshahi. The Black Bengal goats of these two regions have been termed as
BBW and BBH respectively in the present study. It may therefore be said that
the present study represents wider geographical areas of Bangladesh and
includes naturally grazing goats of three majors Black Bengal populations viz,
BBW, BBC and BBH.
42
In the present study, the prevalence of Haemonchus contortus was more in
BBC and BBW populations than BBH (Table 1). The prevalence of Haemonchus
contortus were 47.52%, 46.54% and 42.85% in BBW, BBC and BBH population
respectively. Faecal egg count of individual goat, if occurred, was also very high
in BBW and BBC (up to 2200) than BBH (up to 450). This is evidence from the
faecal egg count as presented in Table 5. One reason might be the soil structure
and vegetation pattern of Bandarban Hill district as well as feeding behavior of
BBH goats there. The soil of Bandarban Hill district is loose sandy soil covered
mostly by forest. There are a lot of herbs and shrubs there. BBH goat browses
the leaves of these herbs and shrubs. That might assist in reducing
gastrointestinal parasitic burden. On the other hand, BBC and BBW goats live in
flood fed area and eat grasses in most cases. The topography as well as feeding
habits might assist in increasing the parasitic burdens in these two populations.
Karim (2003), Shahiduzzam et al. (2003), Akanda et al. (2012), Hossen (2012) and
Hassan et al. (2014) reported slight higher prevalence of Haemonchus contortus in
BBC than the present study. This might be use of abomasums as a source of
parasite in their study while the present study is based on faecal egg count.
There is no other investigation yet now except this about the prevalence of
Haemonchus contortus in BBW and BBH population. Faruque (2009) reported
differences in the genotypes of BBW, BBC and BBH goats in Bangladesh. There
may be genetic difference in resistance to Haemonchus contortus in BBH
population. However, this should be confirmed by gene expression study.
In addition to population variation in prevalence of Haemonchus contortus,
there was also variation in age and sex within the population for prevalence of
Haemonchus contortus (Table 2). The prevalence of parasite increased with the
age and sex. The older and female goats had higher percent of prevalence of
Haemonchus contortus. This finding of present study is in agreement with the
findings of Howlader et al. (2002), Shahiduzzam et al. (2003), Akanda et al.
(2012), Hossen (2012) and Hassan et al. (2014).
43
The haematological values presented in Table 6, Fig. 5 and Fig. 6 revealed
that goats free of Haemonchus contortus had higher haematological values than
goats infested with Haemonchus contortus. This is supported by investigation
made on goat and sheep by other scientists in abroad (Buvanendran, et al., 1981;
Karlsson, et l., 1991; Yadav et al., 2006). The BBH population had significantly
higher PCV and Hb values as this population had less Haemonchus contortus
load.
The artificial challenge trial also corresponded with the above findings.
There was no egg or ova in the faeces at day 28 and 21 after the infestation of
larvae parasite in BBH. On the other hand, two kids BBC2 and BBC3 had some
eggs in the faeces at day 21 and day 28 after the infestation of larvae parasite.
This study indicates that BBH is more resistance to Haemonchus contortus than
BBC. In an artificial challenge trial done on BBH and BBW by Faruque (2013),
BBH was found more resistance to Haemonchus contortus than BBW.
The discussion made so far indicates that BBH goats are more resistance to
Haemonchus contortus than BBC and BBW goats. There is also variation in
resistance to Haemonchus contortus within BBH goat population. Further studies
on the gene expression should be conducted to confirm these findings.
The between and within population variation of resistance to Haemonchus
contortus in different populations of Black Bengal goats will assist in future to
develop Haemonchus contortus resistant Black Bengal goat in the country.
44
CHAPTER 6
CONCLUSION
Black Bengal goat is the only recognized breed of Livestock in
Bangladesh. a number of factors viz., reduced grazing land for goat, diseases
and parasite infection are limiting the production of Black Bengal goat in the
country nowadays. This experiment was conducted to investigate the genetic
variation in resistance to Haemonchus contortus (a GI Nematode) in different
populations of Black Bengal goat in Bangladesh. To achieve this goal, two
studies were done. Study I was carried out to find out the incidence of
Haemonchus contortus in different populations of Black Bengal goat kept under
naturally grazing condition. 196 goats from different populations of Black
Bengal goat (BBW, BBC, BBH) were studied. The prevalence of Haemonchus
contortus was 47.52%, 46.54% and 42.85% for BBW, BBC and BBH population
respectively. So, more than 50% of Black Bengal goats under natural grazing
condition remained non-infected by Haemonchus contortus. The mean faecal egg
counts were 663.54 + 69.77, 647.22 + 112.11 and 81.25 + 7. 86 epg for BBW, BBC
and BBH population respectively. The study revealed that BBH had the least
prevalence and load of Haemonchus contortus among three populations of Black
Bengal goat. The maximum and minimum PCV (%) and Hb (g/dl) value were
48 and 10.5, 36 and 8.5, 24 and 8.5 for BBW, BBC and BBH population
respectively that were infected with Haemonchus contortus. The maximum and
minimum PCV (%) and Hb (g/dl) value were 59 and 9.8, 44 and 8.4, 31 and 9.6
for BBW, BBC and BBH population respectively that were not infected with
Haemonchus
contortus.
The
correlation
between
parasitic
load
and
Heamatological values were estimated and it seemed that parasitic load had
negative effects on the haematological values of goats. In study II, two
populations of Black Bengal goat (BBC and BBH) were studied for artificial
challenge trial. 5 kids of 5 months old were dewormed and infested with L3
larvae @5000/ kid. The body weight, FEC, Hb and PCV were measured at day 0
(Day when larvae were infested artificially), day 21 (21 days after the larvae
45
were infested artificially) and day 28 (day after the larvae were infested
artificially). Only two kids of BBC had FEC at day 21 and day 28 thought it was
low ranging from 50 to 200 only. On the other hands, no kids of BBH had FEC
even at day 28.
The studies revealed that BBH had the least prevalence and load of
Haemonchus contortus than other two populations of Black Bengal goat in
naturally grazing condition and artificial challenge trial. This supports the
between population difference for resistance to Haemonchus contortus in Black
Bengal goats. It might be topography, feeding habit or genotypic differences of
three populations of Black Bengal goats. Further work on gene expression is
needed to clarify the actual cause behind this fact.
The between and within population variation of resistance to Haemonchus
contortus in different populations of Black Bengal goats will assist in future to
develop Haemonchus contortus resistant Black Bengal goat in the country.
46
Chapter 7
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