Epidemiological Investigation On Parasitic Diseases of Buffaloes (Bubalus Bubalis) in Kurigram District of Bangladesh

EPIDEMIOLOGICAL INVESTIGATION ON PARASITIC
DISEASES OF BUFFALOES (Bubalus bubalis) IN

KURIGRAM DISTRICT OF BANGLADESH
A THESIS
BY
MD. ABDULLAH AL MAMUN

EXAMINATION ROLL NO. 07/VPAR/JD/08M
REGISTRATION NO. 28633
SESSION: 2001-2002
SEMESTER: JULY-DECEMBER, 2008
MASTER OF SCIENCE (MS)

IN
PARASITOLOGY
DEPARTMENT OF PARASITOLOGY
BANGLADESH AGRICULTURAL UNIVERSITY
MYMENSINGH
NOVEMBER 2008
1
A THESIS
BY

SESSION: 2001-2002
Submitted to
The Department of Parasitology
Bangladesh Agricultural University, Mymensingh
in partial fulfilment of the requirements for the degree of
MASTER OF SCIENCE (MS)

IN
PARASITOLOGY
DEPARTMENT OF PARASITOLOGY
BANGLADESH AGRICULTURAL UNIVERSITY
MYMENSINGH
NOVEMBER 2008
2
A THESIS
BY

SESSION: 2001-2002
Approved as to style and content by
(Prof. Nurjahan Begum) (Prof. Dr. Md. Motahar Hussain Mondal)

Supervisor Co-supervisor
(Prof. Dr. Md. Motahar Hussain Mondal)

Chairman
Defence Committee
and
Head, Department of Parasitology
Bangladesh Agricultural University
Mymensingh
3
NOVEMBER 2008
ACKNOWLEDGEMENT
The author always likes to bow his head to almighty Allah for his never-ending
blessing for successful completion of the work. All praises and deepest sense of
gratitude be to almighty Allah, the supreme authority of this Universe, who enabled
the author to complete this thesis.
The author wishes to express the deepest sense of gratitude, sincere appreciation,
indebtedness and best regards to his respected teacher and research supervisor
Professor Nurjahan Begum, Department of Parasitology, Bangladesh Agricultural
University, Mymensingh for his scholastic and dynamic guidance, constant
inspiration, cordial consistence, affectionate feeling, sympathetic supervision and
constructive criticism in all phases of this study and preparing the manuscript.
The author is ever grateful and immensely indebted to his honorable and respected
teacher and research co-supervisor Professor Dr. Md. Motahar Hussain Mondal,
Head, Department of Parasitology, Bangladesh Agricultural University,
Mymensingh for his valuable advice and encouragement throughout the period of
this study.
The author finds his great pleasure to express his sincere thanks and gratitude to
Professor Dr. M. Hafezur Rahman, Assistant Professor Anisuzzaman and Assistant
Professor Thahsin Farjana for their encouragement, kind and constant cooperation,
valuable advice and proper direction throughout the course of the study.
The author is grateful to his beloved parents Md. Abdus Sobur and Most.
Monowara Begum, all members of his family and relatives for their sacrifices,
blessings and encouragement throughout his study period.
The author would like to extend thanks to his friends, room-mates, hall-mates and
senior and junior brothers for their good wishes and co-operation to conduct this
research work.
The author likes to express his cordial thanks to the demonstrator Md. Jalal Uddin,
Mrs. Hosne Ara (Sr. Lab. Tech.), Md. Kamal Uddin (Lab. Tech.), Md. Joynal Abedin
(Office Attendant) and all other staffs, Department of Parasitology, Bangladesh
Agricultural University, Mymensingh for their assistance and cooperation during the
research period.
Finally, the author extends his profound thanks to Md. Anawarul Hasan Khan
(Swapan), Khan Enterprise, BAU for computer composing of the manuscript.
The Author
November 2008
4
LIST OF CONTENTS
CHAPTER TITLE PAGE NO.

1. INTRODUCTION 1
2. REVIEW OF LITERATURE 4
2.1 Buffalo rearing and its importance 4
2.2 Prevalence of endoparasites in buffaloes 7
2.2.1 Prevalence of helminths 7
2.2.2 Prevalence of enteric protozoa 13
2.3 Prevalence of ectoparasites in buffaloes 15
2.4 Prevalence of blood parasites in buffaloes 17
3. MATERIALS AND METHODS 19
3.1 Study period 19
3.2 Study area 19
3.3 Selection of buffaloes 19
3.4 Examination of buffaloes for ectoparasites 19
3.5 Collection of ectoparasites 21
3.6 Preservation of samples 21
3.7 Identification of ectoparasites 21
3.8 Collection of faecal sample 21
3.9 Examination of faecal sample 22
3.9.1 Modified Stoll’s Dilution Technique 22
3.10 Collection, staining and preservation of blood
samples 23
3.11 Blood smear Examination 23
3.12 Statistical analysis 23
4. RESULTS 24
4.1.1 Overall prevalence 24
4.1.2 Age related prevalence of endoparasites 25
4.1.3 Sex related prevalence 25
5
4.1.4 Seasonal prevalence 26
6
LIST OF CONTENTS
CHAPTER TITLE PAGE NO.

4.2 Prevalence of ectoparasites in buffaloes 27
4.2.2 Age related prevalence of ectoparasites 27
4.3.2 Age related prevalence of blood protozoa 29
5. DISCUSSION 45
5.1.2 Age related prevalence 46
5.2 Prevalence of ectoparasits in buffaloes 49
5.2.2 Age related prevalence of ectoparasites 49
5.2.4 Seasonal dynamics of ectoparasitic infestation in
buffaloes 51
5.3.2 Age related prevalence of blood protozoa 53
5.3.4 Seasonal prevalence of blood parasitic in buffaloes 53
6. CONCLUSION 55
7. REFERENCES 56
7
LIST OF TABLES
SL. NO. TITLE OF THE TABLES PAGE NO.
Table 1 Overall prevalence of endoparasites of buffaloes in

Bangladesh 31
Table 2 Age related prevalence of endoparasites of buffaloes

in Bangladesh 33
Table 3 Sex related prevalence of endoparasites of buffaloes

in Bangladesh 34
Table 4 Seasonal prevalence of endoparasites of buffaloes in

Bangladesh 35
Table 5 Overall prevalence of ectoparasites of buffaloes in

Bangladesh 36
Table 6 Age related prevalence of ectoparasites of buffaloes

in Bangladesh 37
Table 7 Sex related prevalence of ectoparasites of buffaloes in

Bangladesh 38
Table 8 Seasonal prevalence of ectoparasites of buffaloes in

Bangladesh 38
Table 9 Overall prevalence of blood parasites of buffaloes in

Bangladesh 39
Table 10 Age related prevalence of blood parasites of buffaloes

in Bangladesh 40
Table 11 Sex related prevalence of blood parasites of buffaloes

in Bangladesh 41
Table 12 Seasonal prevalence of blood parasites of buffaloes in

Bangladesh 41
8
LIST OF FIGURES
SL. NO. title of the figures PAGE NO.
Figure 1 The flow diagram-showing research out line 20
Figure 2 Overall prevalence (%) of endoparasites of buffaloes in

Bangladesh 32
Figure 3 Overall prevalence (%) of ectoparasites of buffaloes in

Bangladesh 36
Figure 4 Overall prevalence of blood parasites of buffaloes in

Bangladesh 39
Figure 5 Buffalo grazing on marshy paddy field just after

harvesting. 42
Figure 6 Preserved ectoparasites in70% alcohol. 42
Figure 7 Ova of Schistosoma spindale 42
Figure 8 Ova of Schistosoma indicum 42
Figure 9 Anaplasma marginale stained with Giems’s stain (100×) 43
Figure 10 Anaplasma marginale stained with Giems’s stain 43
Figure 11 Babesia sp. stained with Giems’s stain (100×) 43
Figure 12 Theileria sp. stained with Giems’s stain (100×) 43
Figure 13 Haematopinus tuberculatus showing sternal plate 44
Figure 14 Haematopinus tuberculatus 44
Figure 15 Boophilus microplus 44
Figure 16 Haemaphysalis bispinosa 44
9
ABSTRACT
Epidemiology of parasitic diseases of buffaloes was studied in Kurigram district of

Bangladesh from November, 2007 to October, 2008. A total of 236 buffaloes were
examined, among them 61.02%, 61.86% and 12.27% were found infected with
endoparasites, ectoparasites and blood parasites, respectively. Nine species of endoparasites
were identified, of them four species were trematodes, namely, Fasciola gigantica
(22.46%), Paramphistomum cervi (29.24%), Schistosoma indicum (1.27%), Schistosoma
spindale (0.85%); three species were nematodes namely, Toxocara vitulorum (2.54%),
Strongyles (0.85%), Strongyloides sp. (0.42%) and two species were protozoa, namely,
Eimeria sp. (3.39%) and Balantidium coli (37.29%). No cestodes were detected. Three
species of ectoparasites were identified of which, two species were arachnids, namely,
Boophilus microplus (13.98%), Haemaphysalis bispinosa (11.44%) and one species was
insect namely Haematopinus tuberculatus (51.27%). No mites were detected. During this
study, three species of blood parasites were recorded, all were protozoa, namely,
Anaplasma marginale (8.89%), Theileria sp. (2.12%) and Babesia sp. (1.69%). No filarial
worm was detected. Among the endoparasites and ectoparasites mixed infection was
common but in case of blood parasites only single infection was observed. In this
investigation, prevalence of parasites in relation to age, sex and seasonal dynamics were
also studied. Except cetoparasites, relatively higher prevalence of parasites was observed in
rainy season followed by summer and winter seasons. Significantly (p<0.01) higher
prevalence of ecto and blood parasites were recorded in female animals than males whether
males and females were almost equally (odd ratio 1.08) susceptible to endoparasitic
infection. In the age groups, young (< 2 to 5 years) were mostly susceptible to endo and
blood parasites whereas buffalo calves (0.5-2 years) were more susceptible to ectoparasites.
In the present study, EPG (Egg Per Gram of Feces) was also determined. The range of EPG
was 100-5000 for B. coli whereas it was 100-1200 for F. gigantica. Overall mean
ectoparasitic burden was 2.31±1.31 per square inch of heavily infected area. The highest
parasitic burden was recorded in case of H. tuberculatus (3.49±2.29) followed by B.
microplus (1.85±0.94) and H. bispinosa (1.59±0.69).
10
CHAPTER 1
INTRODUCTION
The world population of buffaloes (Bubalus bubalis) has been estimated at over 172
million head, more than 97% (167.6 million) of which are in Asia and the Pacific
region, mainly in India (97.7 million), Pakistan (25.5 million) and Bangladesh (0.83
million) (FAO, 2004). About 98% of buffaloes in the region are raised by small
farmers owning to less than two hectares of land and less than five buffaloes
(Mudgal, 1992). In general, the water buffalo is regarded as more productive,
healthier and more useful than the cow, especially for the poorest “backyard”
farmers in Asia (Bhat, 1999). According to the 1997 FAO report, the buffalo are
recognized as “Black Gold of Asia” (Khushk and Memon, 2004). In comparison to
research on cattle, research on water buffaloes has been much neglected (Johan,
2001-2002).
Both the swamp and river type buffaloes are found in Banglasesh (Latif, 1994).
Buffalo is considered to be a multipurpose animal and in some parts of the country,
especially in the hilly areas and low lying marshy land, the farmers are more
dependent on buffaloes than cattle. They are extensively used for agricultural land
preparation (ploughing and laddering), inter-cultural operation (racking), carting and
transportation of goods (mainly agricultural products) in rural areas, threshing and
crushing of sugarcane and oil seeds in the country (Rahman and Islam, 1992). The
buffaloes are also the source of protein. Buffalo meat is nutritionally superior than ox
meat (Ranjhan, 1992). In addition to milk, meat and draft facilities, buffaloes provide
organic wastes like dung and urine for fuel and fertilizer, dead animals and bone for
feeding of poultry and hides, horns and tallow for industrial use. The buffaloes work
slowly and without any argument. The working life of buffalo is longer than that of
11
cattle, usually more than 17 years and up to 25 years of age. Because of this
excellent draft and pulling capacity, buffaloes are called the living tractor of the East
(Cockrill, 1968).
In Bangladesh, among many causes, parasitism is thought to be a major cause that

hindering the development of livestock population (Jabber and Green, 1983). Unlike
bacterial and viral diseases, the diseases caused by parasites are of great importance.
Parasitic diseases are also emphasized for their pathogenicity and economic
importance in animals by the experts both from the government and
nongovernmental organizations. The losses due to parasites in the form of mortality,
lowered general health condition, retarded growth, lower output of work, decrease in
the production of milk and meat (Faiz, 1972). In Pakistan, parasitic diseases,
including tick born diseases, are considered a major obstacle in the health and
product performance of cattle and buffaloes. Ticks comprise a burning veterinary
problem because they transmit diseases, induce paralysis or toxicosis and cause
physical damage to livestock (Razput et al., 2005).
In developed countries, the data on epidemiology of various helminthiasis are

published in an efficient manner as an aid to combat infections more effectively. In
contrast, in developing countries, little published information exists and data on the
epidemiological aspect of helminthic infections. Helminth parasitism, especially,
gastrointestinal parasitism is one of the major health problems severely limiting the
animal productivity in dairy animals. In spite of significant production losses, which
may run into millions of rupees (Shah and Chaudhry, 1995) the problem is neglected
due to its chronic and insidious nature (Sanyal, 1998).
The diverse agroclimatic conditions, animal husbandry practices and pasture

management largely determines the incidence and severity of various parasitic
12
diseases in a region. Epidemiological pattern of the parasitic diseases in the different
agroclimatic zones of the country would provide a basis for evolving strategic and
tactical control of these diseases. The present study examines the status of parasites
in buffaloes and its management in the Bangladesh. Little information is available
regarding the epidemiology of all types of parasites in buffaloes; however, what data
are available indicate that further study of the prevalence and intensity of parasites in
buffaloes is needed.
The parasitic favour of buffaloes from different regions of Bangladesh was not
investigated thoroughly. Therefore the present study was undertaken with a view to
fulfill the following objectives-
i. To investigate the overall prevalence of parasitic diseases (ecto, endo and

blood parasites) of buffaloes in Kurigram district of Bangladesh.
ii. To study the prevalence of parasitic diseases in relation to age and sex of the
buffaloes and seasonal dynamics of the year.
13
CHAPTER 2
REVIEW OF LITERATURE
Available literatures on key factors that influence the prevalence of the parasitic
diseases of buffaloes are reviewed in this chapter. The main purpose of this chapter
is to get up-to-date information regarding the research works addressed here.
Important information related to the present study was represented below. For
obtaining the distinct evidences and key information, the whole review of literature
on the research title “Epidemiological investigation on parasitic diseases of
buffaloes (Bubalus bubalis) in Kurigram district of Bangladesh” has been
conveniently brought under following headings. These are outlined below-
 Buffalo rearing and its importance

 Prevalence of endoparasites in buffaloes
• Prevalence of helminths
• Prevalence of enteric protozoa
 Prevalence of ectoparasites in buffaloes
 Prevalence of blood parasites in buffaloes
2.1 Buffalo rearing and its importance
Sebastian et al., (1970) reported buffalo is a more efficient milk producer than an
indigenous cow in India.
Parikh (1988) reviewed that Asian civilization is primarily based on agriculture,

mostly growing rice, wheat, coarse grains, sugar cane and pulses as principal crops
and 30–50% of bovine are used for draft operations.
Agarwal and Tomar (1998) reported that in Asia, buffalo played a significant role
in overall socioeconomic development through its milk, meat, hides and draft power
14
for agricultural operations. River buffalo produces more milk than the swamp
buffalo. Average lactation yield in the best known dairy breeds, Murrah and Nili-
ravi, is around 2000 liters although elite buffalo with up to 6000 liters also exist in
India, Italy and Pakistan, which indicates its great potential for milk production.
Saadulla (1998) cited that buffalo is mostly known as Majjh or Bhains in the Indian
subcontinent. Of the 158 millions buffalo in the world, 153 millions are water
buffalo, primarily found in Asia and more than half are in India. The author also
cited that from their home in India, they were carried to Egypt by Arabic invaders in
the 9th century for meat, milk and draft power and subsequently to Europe by
pilgrims and crusaders in the middle ages.
World Bank (1998) report suggested that Bangladesh government is supporting

women's organizations to rear livestock for milk in the pattern of Swayam Krishi
Sangam; a nongovernment organization microcrediting woman in southern India to
rear buffalo. The profits from Grameen-modeled microfinance businesses in
Bangladesh increased borrower's consumption by 18% per year and the percentage
of poverty was reduced by 70% within a few years of joining.
Hoffmann (1999) suggested that approximately 20% of the value of an animal is in
the draft power it supplies. However, these outputs often don’t appear in the figures
showing the value of livestock to a country. For more than 5000 years, buffalo have
been used for draft. Buffalo dung is an excellent source of fertilizer for growing
crops and is used all over the world. Approximately 40% of the value of an animal
could be in the manure it produces.
15
Hibler (2000) reported that buffalo are particularly suited to work on wet fields with
a strong body, broad hooves, flexible pastern and fetlock joints.
Rasali (2000) observed that growth was not uniform in all buffalo rearing countries
in Asia. Although spectacular increases were observed in South Asia, countries in
eastern Asia exhibited a considerable decline in buffalo population, which is a matter
of concern.
Ali (2002) observed in Bangladesh that 22.7% and 0.7% of people surveyed use
dung energy for household and commercial purposes, respectively.
Karanth (2002) reported that buffalo dung is extensively used as fuel for cooking in
India, Pakistan, Bangladesh and in almost all countries. The first European pioneers
on the Great Plains of the USA also used dried buffalo dung, called buffalo chips, for
fuel because of the lack of wood.
Faostat (2003) reported that buffalo meat is very popular in most buffalo loving
countries. Out of 242 630 374 tons total world meat, 3 089 875 tons comes from
buffalo. India is the world's largest buffalo meat producer having approximately
14 000 buffalo slaughterhouses. Leather is another major contribution to the world
market irrespective of type. Apart from direct input in the form of leather goods, the
industry provides jobs to millions of people. Fresh buffalo hide production in the
world is 833 566 tons. The author also suggested that world buffalo population
increased by 91% between 1961 and 2001. It was 148 183 900 in 1990 and
166 419 000 in 2002, a gross increase of 12.31%.
2.2 Prevalence of endoparasites in buffaloes

2.2.1 Prevalence of helminths
16
Bhuyan (1970) recorded fasciolosis in 90.90% buffaloes, 60% cattle, 12.92% goats
and 8.34% sheep in Bangladesh by faecal sample examination.
Chowdhury (1970) recorded Neoascaeis vitulorum, Trichuris globulosa, Trichuris

tadrosi, Bunostomum phlebotomum and Bunostomum bovis from the gastro-
intestinal tract of buffaloes of different ages.
Prokopic et al. (1976) found five cestode species parasitizing ruminants for the first
time in Afghanistan viz. Moniezia benedeni, M. expansa, Avitellina
centripunctata, Stilesia globipunctata, and Thysaniezia giardi.
Dewan et al. (1979) observed the 80% mortality rate of buffalo-calves due to
Neoascaris vitulorum infestation in some areas of Mymensingh district.
Islam (1982) recorded the presence of hydatid cyst in buffaloes of different age
groups in Mymensingh, Bangladesh. The lowest percentage (8.85%) was found in 0-
3 years old but the highest percentage (68.97%) was found in 9 years old buffaloes.
Islam (1989) recorded incidence of different parasites by examination of faeces were

as follows: Fasciola gigantica 18.9%, Paramphistomes 29.5% Schistosoma indicum
1.6%, S. spindale 13.9%, S. nasalis 4.5%, Strongyloides sp. 14.8%, Capillaria sp.
8.5%, Oesophagostomum sp. 6.6%, Hookworm 8.1%, Trichostrongylus sp. 11.25%,
other Strongyles (Haemoncus and Mecistocirrus sp.) 15.20%, Toxocara vitulorum
9.7%, Eimeria zurnii 2.3% in buffaloes of Bangladesh.
Hossain (1991) carried out an investigation at Kanihari Union Mymensingh district,

Bangladesh during the period from September 1989 to June 1990 and identified
fasciolosis 12% and mixed fluke infection 10.10% in water buffaloes of 138 clinical
cases. Necropsy examination of 25 carcasses revealed hepatic fasciolosis in 24%
17
cases. By faecal sample examination of 356 animals, 12% F gigantica and 14%
amphistome infection was recorded.
Islam (1991) conducted an epidemiological study of fasciolosis in Dinajpur,

Bagerhat, Chapinawabgonj and Sirajgoni districts of Bangladesh from November
1988 to June 1991 and recorded an overall rate of infection fasciolosis in live
domestic animals 16.30% and amongst buffalo, cattle and sheep/goat the infection
rate was 20.20%, 16.80% and 13% respectively.
Islam et al. (1992) conducted an investigation to detect the presence of parasites in

water buffaloes in Bangladesh during September 1988 to August 1989 and examined
a total of 480 live buffaloes and 180 visceral samples from Dhaka, Mymensingh,
Bogra and Rajshahi areas. The authors reported an overall 18.9% F. gigantica and
29.50% Paramphistomum spp. infection by faecal examination and 46.6% F.
giganitca and 48.1% Paramphistomum spp. infection on post-mortem examination.
Motaleb (1996) studied the status of parasitic diseases in buffalo, cattle and goat in
Anwara, Chittagong and reported that 38.5% (52 to 135) buffaloes, 36.28% (119- of
328 cattle and 13.3 (6 to 45) goats were infected with Fasciola giganitca.
Alim (1997) conducted an exploratory study on the epidemio-pathology of

fasciolosis in buffaloes from December, 1996 to November, 1997 in Bangladesh and
recorded on overall 50.65% infection on fecal sample examination. He also reported
that female animals were susceptible (52.65%) than the males (47.76%). No age
limit for infection and highest rate of infection was found in the buffaloes of above
10 years of age (65.63%).
18
Jithendran and Bhat (1999) reported that Fasciola was endemic throughout the
year, with a higher percentage infection in buffaloes than in cattle in Himachal
Pradesh, India. Other fluke and nematode infections showed a seasonal pattern in
prevalence, with a small peak in March and April followed by a high peak in July
and September. The faecal egg counts (eggs per gram) of flukes (Fasciola/
amphistomes) ranged from 50 to 300 in cattle and 50 to 400 in buffaloes, with high
loads during the rainy and post-rainy seasons. The GI nematode egg counts
(excluding Toxocara) revealed a similar trend, with the overall monthly mean epg
ranging from 85 to 1720 in cattle and 90 to 1625 in buffaloes, with a high peak
during the months of July to September. On coproculture of positive samples, the
nematode infections in order of prevalence were: Strongyloides, Trichostrongylus,
Haemonchus, Oesophagostomum, Bunostomum and Mecistocirrus.
Van et al. (2000) observed the overall prevalence of Strongyles infection was 53%
for cattle and 28% for buffaloes in Mindanao, the Philippines. The prevalence
decreased with increasing age in both animal species. Faecal egg counts were
significantly lower in buffaloes than in cattle. Adult animals had lower egg counts
than calves. This age effect was most pronounced in buffaloes. Six genera of
Strongyles nematodes were identified from the faecal cultures in both animal
species, Mecistocirrus, Haemonchus, Trichostrongylus, Cooperia, Bunostomum and
Oesophagostomum.
Azam et al. (2002) investigated the prevalence of ecto and endoparasites of buffalo
calves in 50 buffalo farms in Khadagzai area of Dir district in North West Frontier
Province, Pakistan, in April and May [year not given]. Faecal examination of calves
(n=118: age <less or =>1 year) revealed that 64.41% of the calves were positive for
internal parasites. The worm load varied significantly (P<0.05) among the farms and
was high (1600-3600 EPG) in 2%, moderate (800-1600 EPG) in 22%, low (200-800
19
EPG) in 34% and negligible (less than 200 EPG) in 42% of farms. Among the calves
examined, 50.84% had a worm burden of 200-800 EPG and 13.56% had a worm
burden of 800-1600 EPG. A high worm burden (1600-3600 EPG) was observed only
in 0.85% of the calves. Six species of nematodes and one species of trematode were
identified. No cestodes infection was encountered during the study. The incidence of
Trichostrongylus species was 21.19%, followed by Trichuris (9.32%), Haemonchus
(8.47%), Strongyloides papillosus (5.93%), Ostertagia (5.08%), Toxocara vitulorum
(1.70%), Fasciola (5.93%) and mixed infections (6.78%).
Azhar et al. (2002) reported 25.59, 26.16, 13.7 and 10.5 per cent infection rate of
fasciolosis, respectively in slaughtered buffaloes at livestock farms, veterinary
hospitals and in household buffaloes in Punjab province, Pakistan. Overall highest
(24.0%) seasonal prevalence in all types of buffaloes was recorded during autumn,
followed by spring (20.0%), winter (13.0%). While the lowest (9.0%) was recorded
during summer. It was noticed that a higher infection rate was recorded in older
buffaloes than in youngsters (below 2 years of age) where as sex showed no
significant difference. Buffaloes of either sex are equally affected.
Bachal et al. (2002) recorded the prevalence of helminths in buffalo calves as 47%.
Out of these, only 18% buffalo calves excreted eggs in their faeces that ranged from
100-400 eggs/g (epg) while 7, 13, 5.5 and 3.5% calves excreted eggs in faeces that
ranged from 401-800, 801-1200, 1201-1600, 1601 and above eggs/g, respectively.
Further, they observed that 43 and 4% buffalo calves were positive for nematodes
and trematodes, respectively, but mixed infections were observed in 1.5% calves.
Cestodes were not detected in any samples examined. Four different species of
helminths were identified, the species were: Toxocara vitulorum (33%), Ostertagia
ostertagi (8%), Trichuris ovis (2%) and Fasciola gigantica (4%). The highest egg
20
counts were recorded in 1-120 days old calves. A slightly higher prevalence
(48.30%) of helminths was found in female than in male (45.12%) calves.
Mureli et al. (2002) observed that the most common parasites of buffalo and cattle
were Coccidia, Strongyles, Trichuris, Strongyloides, Toxocara and Moniezia in
Malaysia. Moniezia infection was prevalent among buffalo calves. Farm
management factors such as rearing system, size of farm, grass and water sources,
overcrowding, drainage and manure disposal system showed a significant effect
(P<0.01) on helminthiasis level.
Morsy et al. (2005) investigated on natural infection with Fasciola species in

Tamyia Center, Egypt by stool examination and reported 20% infection in buffaloes.
Garippa (2006) reported 14.8% prevalence of Cystic Echinococcosis (CE) in cattle,

with no viable cysts recovered, and 10.5% in water buffaloes, with a fertility of
1.4%.
Haridy et al. (2006) showed that buffaloes were infected with Fasciola sp. (17.7%)
and Paramphistomum cervi (10%) in Al-Santa Center, Egypt.
Asif et al. (2007) recorded the overall prevalence of helminthiasis was 51% in cattle,
47% in buffaloes, 62% in sheep and 52% in goats, with nematodes being the most
common helminths in an irrigated area of lower Punjab, Pakistan. The prevalence of
helminths was higher in young animals compared with adults in cattle (P<0.0001),
buffaloes (P<0.0001), sheep (P<0.059) and goats (P=0.010). The prevalence of
different species of helminths also varied in different age groups, with Toxocara
vitulorum being higher in calves than adults both in cattle (P=0.017) and buffaloes
21
(P<0.0001). Sex-wise prevalence of helminths was higher in males than females for
buffaloes (P<0.0001) and sheep (P=0.014) in contrast to cattle and goats.
Condoleo1 et al. (2007) detected gastrointestinal Strongyles (33.1%), Strongyloides

spp. (3.1%), Fasciola hepatica (7.1%), Dicrocoelium dendriticum (2.4%),
Paramphistomidae (7.1%), and Moniezia spp. (2.4%) in buffaloes of central Italy
Iqball et al. (2007) recorded Toxocara vitulorum, Fasciola hepatica, Haemonchus

contortus, Bunostomum phlebotomum, Ostertagia circumcinta, Oesophagostomum
radiatum, and Trichostrongylus spp. through standard coprological examination
procedures of dairy animals of Nestle milk collection areas of Punjab (Pakistan).
Racioppi et al. (2007) reported the recent appearance of Fasciola hepatica

parasitizing different buffalo (Bubalus bubalis) populations in the province of
Corrientes. These animals share pastures with Bos taurus, Bos indicus and their
crossbreds. The author also observed 28.5% positivity and a tendency to expand to
other cattle.
Ravindran et al. (2007) reported the prevalence of visceral schistosomiasis by worm

counts from the mesentery of domestic ruminants of the hilly district of Wayanad,
located in Kerala, one of the states in South India. They found 57.3, 50, and 4.7% of
cattle, buffaloes and goats, respectively, had visceral schistosomiasis upon slaughter
at a municipal slaughter house in Kalpetta.
Shazly et al. (2007) reported the overall rates of infection of fasciolosis were
12.31%, 9.73%, 17.84% and 5.40% in cows, buffaloes, sheep and goats respectively
in Dakahlia Governorate, Egypt. The mean eggs per gram stool were 22, 13.6, 148.3
and 8.6 for cows, buffaloes, sheep and goats. The mean numbers of Fasciola
22
worms/liver/animal were 69.1, 62.7 and 208.1 for cows, buffaloes and sheep
respectively.
2.2.2 Prevalence of enteric protozoa
Fusco et al. (1997) conducted a study in 42 buffalo calves from one breeding farm in
Southern Italy infected by 7 different Eimeria species, to assess the dynamics of
oocyte elimination [date not given]. Weekly examination of each animal was
conducted from second to twelfth week of age when 100% of the subjects have shed
the oocysts. E. bareillyi and E. zuernii were detected as early as second week of age
from the faeces of 100 and 20% of the positive subjects, respectively. The other
species detected were E. ellipsoidalis and E. subspherica (third week), E.
auburnensis (fourth week), E. bovis (fifth week) and E. pellita (sixth week). None of
the subjects showed severe clinical symptoms related to coccidial infection.
NAVIDPOUR AND HOGHOOGHI (1998) REPORTED THE

INFECTION RATE OF TOXOPLASMA GONDII OF
BUFFALOES UNDER THE AGE OF A YEAR, ABOVE
A YEAR FEMALE, AND MALE WERE 10.8%, 4.7%,
12.1%, AND 5.3%, RESPECTIVELY IN IRAN TESTED
BY THE IFA TEST.
Penzhorn (2000) examined faecal specimens collected in the Kruger National Park
from 103 African buffaloes (Syncerus caffer) up to 1 year old and 283 buffaloes
older than 1 year for the presence of coccidian oocysts and nematode eggs. Most
specimens from animals older than 1 year had negative coccidian oocyst counts.
23
Positive specimens from younger animals had significantly higher coccidian oocyst
counts than those from older animals.
Azam et al. (2002) recorded that 72% of the buffalo calves were suffered from
intestinal protozoan infection in Khadagzai area of Dir district in North West
Frontier Province, Pakistan. Most of these (85%) had mixed infections of coccidia
and amoebae and the remaining 15% were infected with coccidia only. 55.93% of
the calves studied were positive for ectoparasites. The prevalence of ticks, lice, mites
and mixed infections was 5.08, 34.75, 11.86 and 4.24%, respectively.
Bastianetto et al. (2007) reported that coccidiosis is responsible for significant

economical losses in buffaloes around the world as a consequence of its high
mortality rate and interference in nutrient absorption.
Condoleo1 et al. (2007) tested for the presence of copro-antigens of C. parvum

using a commercially available ELISA in the water buffalo in central Italy. Out of
the 90 farms, 22 (24.4%) resulted positive. With respect to animals, out of the 347
faecal samples, 51 (14.7%) were found to have antigens of C. parvum. The results of
the logistic regression model showed a positive association between the positivity to
C. parvum and the high number of buffaloes on farms.
Nalbantoglu et al. (2008) first time identified eleven different Eimeria species and
one Isospora species were in 78 (75%) out of the 104 water buffaloes as follows: E.
zuernii (55.1%), E. auburnensis (44.9%), E. bovis (44.9%), E. ellipsoidalis (28.2%),
E. ankarensis (16.7%), E. subspherica (16.7%), E. alabamensis (11.5%), E.
cylindrica (10.3%), E. bareillyi (5.1%), E. canadensis (5.1%), E. brasiliensis (3.8%),
and Isospora spp. (46.2%) in the Province of Afyon, Turkey.
24
2.3 Prevalence of ectoparasites in buffaloes
Mollah et al. (1970) reported that presence of Haematopinus tuberculatus (33.5%)

in buffaloes in Dhaka and Mymensingh District of Bangladesh.
Miranpuri (1988) recorded a total of 13 ixodid tick species, Boophilus microplus,

Haemaphysalis bispinosa, Haemaphysalis cornupunctata, Haemaphysalis himalaya,
Heamaphysalis montgomeryi, Hyalomma anatolicum anatolicum, Hyalomma
dromedarii, Hyalomma marginatum isaaci, Hyalomma (Hyalommina)
brevipunctata, Hyalomma (Hyalommina) hussaini, Nosomma monstrosum,
Rhipicephalus haemaphysaloides, and Rhipicephalus turanicus from 424 buffaloes
in the northwestern states of India. Ten tick species, Amblyomma testudinarium,
Boophilus microplus, Haemaphysalis anomala, Haemaphysalis arborensis,
Haemaphysalis bispinosa, Haemaphysalis intermedia, Haemaphysalis nepalensis,
Haemaphysalis neumanni, Rhipicephalus haemaphysaloides, and Rhipicephalus
turanicus parasitising 194 buffaloes in the northeastern states of India. The author
also reported Anaplasma marginale and Babesia bigemina in 6.2 and 2.6%,
respectively, of buffaloes tested in the northeastern states, and 14.9 and 4.7%,
respectively, in the northwestern states of India.
Islam (1989) reported Haemaphysalis bispinosa (8.1%) and Haematopinus

tuberculatus (34.6%) in buffaloes of Bangladesh.
Chowdhury (1992) recorded 7 species of ectoparasites of buffaloes. The recorded

ectoparasites of buffaloes in order of predominance were Haematopinus
tuberculatus 52.85%, Boophilus microplus 21.71%, Haematobia exigua 19.42%,
Sarcoptes scabiei 17.42%, Psoroptes natalensis 7.71%, Haemaphysalis bispinosa
7.42% and Chrysomyia bezziana (Larvae infection) 4.85%. In the present study
Haematopinus tuberculatus was main pest of buffaloes in Mymensingh. The
25
incidence of ectoparasites of buffaloes were variable on the basis of age and seasons.
It was recorded that most of the pest affected much the younger hosts.
Bouattour et al. (1999) identified a total of 14 ticks (Ixodidae) species of domestic

animals in Tunisia. The author reported tick population varied in different climatic
condition and also to the species of animals.
Coronado (2001) reported Boophilus microplus as the main vector of A. mariginale.

They also observed intrauterine infection of fetus with A. marginale.
Islam et al. (2006) recorded five species of ixodid ticks namely, Boophilus
microplus (56.3%), Haemaphysalis bispinosa (11.3%), Rhipicephalus sanguineus
(14.7%), Hyalomma anatolicum anatolicum (15.0%) and Amblyomma testudinarium
(2.8%) of domestic animals in Bangladesh. The data showed that Boophilus
microplus occurred predominantly on cattle (42.4%). The other hosts involved were
buffaloes (12.5%), goats (25.5%) and pigs (8.2%). The population density of these
ticks was significantly (p < 0.01) influenced by the changing of seasons.
Veneziano1 et al. (2007) found 11.0% (14/127) of the farms and in the 4.5%
(34/762) of the animals were infected with Haematopinus tuberculatus in water
buffalo farms in central Italy. The presence Haematopinus tuberculatus should be
routinely considered because it is a cause of serious health, production and economic
damages in intensive breeding buffaloes.
2.4 Prevalence of blood parasites in buffaloes
Lalchandani (2001) reported 39.21 percent buffaloes infected with different blood
protozoa (Anaplasma, Babesia and Theileria). The prevalence of anaplasmosis,
babesiosis, theileriosis, babesiosis + anaplasmosis, anaplasmosis + theileriosis,
Babesiosis + theileriosis and anaplasmosis + babesiosis + theileriosis was 20.11,
5.97, 2.39, 6.77, 1.19, 1.39 and 1.39 percent respectively.
26
Ahmad et al. (2002) found that 6.78% and 10.45 of the cattle and buffaloes
respectively were positive for T. evansi in Malaysia. Several factors such as location,
herd management and vector incidence have a bearing on the prevalence of T.
evansi infection.
Latif et al. (2002) reported the infection rate with Theileria parasites in ticks
collected from buffalo-grazed pastures was high and produced fatal theileriosis in
susceptible cattle in the Highveld of Zimbabwe. Similarly, adult R. appendiculatus
ticks artificially fed as nymphs on the buffaloes produced fatal infections in
susceptible cattle.
Singla et al. (2002) reported babesiosis in a Murrah buffalo, 3 years of age, in India
in the 4th month of its 4th lactation presented with high rise of temperature
(103.50F), haemoglobinuria and decrease in milk yield from 12 kg to 4 kg daily. The
animal was infested with Boophilus microplus.
Daiz et al. (2003) carried out a study to determine the prevalence of A. marginale in
Venezuela by indirect immunofluorescence test (IFA) and buffy coat examination.
Through the IFA technique, a prevalence rate of 95.4% was observed; whereas by
the buffy coat examination, 56.9% positive to A. marginale. There were no
significant difference between the presence of A. marginale and the sex and age of
the animals.
Rajput et al. (2005) conducted a comparative study on the prevalence of Anaplasma

parasite on ticks carrying buffaloes and cattle. Five hundred blood samples of both
animals (250 of each) were collected. The author revealed that 205 (41%) animals
had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma
marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes
and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in
buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were
found positive in buffaloes and cattle respectively. Comparative study revealed that
27
the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing
percentage in female of both animals was slightly higher than that of the male.
Maharjan and Mishra (2006) observed the prevalence of trypanosomiasis in 16 of

240 (6.67%) in domestic animals of Makawanpur district of Nepal out of which 9 of
105 were (8.57%) cattle; 5 of 75 (6.67%) buffalos, and 2 of 15 (13.3%) dogs, while
none of the goats and pigs acquired infection. The disease was found maximum
during rainy season 9 of 82 (10.98%) with higher prevalence among cross breeds
than that of local breeds.
Tamasaukas et al. (2006) concluded that trypanosomiasis due to T. vivax in

buffaloes had a high seroprevalence in dual purpose systems at Guarico state,
Venezuela. The average seroprevalence was 29.5%, with a distribution of 75% in
adult animals. Seroprevalence rates were 75 and 25% in female and male buffaloes,
respectively.
28
CHAPTER 3
MATERIALS AND METHODS
3.1 Study period
The investigation was carried out during the period from November, 2007 to
October, 2008. Although there are six seasons in Bangladesh but only three seasons
are prominent such as summer (March-June), rainy (July-October) and winter
(November-February). So, the experimental period was divided into these three
seasons for the convenience of the study.
3.2 Study area
Samples were collected from the different areas of Kurigram district for the
convenience of the study and availability of the buffaloes. Morphological
examination was conducted in the Department of Parasitology, Bangladesh
Agricultural University, Mymensingh.
3.3 Selection of buffaloes
Two hundred and thirty six buffaloes were selected randomly. The age of the
buffaloes were 6 months and above. During collection of samples the age, sex,
breed, place of farming and season of the year were carefully recorded.
3.4 Examination of buffaloes for ectoparasites
The selected buffaloes were thoroughly investigated by close inspection for the
detection of ectoparasites and clinical manifestations relevant to ectoparasitic
infestation.
29
Experimental design
Selection of Buffaloes
Collection of Collection of Collection of blood Collection of data from

feces ectoparasites from ear vein buffalo owners
Preservation of Preservation and Preparation of slides

feces in 10% preparation of stained with Giemsa’s
formalin permanent slides stain
Examination of feces Examination under Examination under

using Stoll’s method microscope and microscope and
under microscope identification identification
Statistical analysis
Figure 1. The flow diagram-showing research out line
30
3.5 Collection of ectoparasites
Ticks and lice were collected from the different parts of the body of the individual
buffalo by hand picking. When required, small camel hair brush dipped in ethanol
was used for the collection of ticks. The point of attachment was smeared with
ethanol. Adequate precautions were taken to preserve the mouth parts and
appendages of the ectoparasites during collection.
3.6 Preservation of samples
Ticks and lice were preserved in 70% alcohol in clean, well-stopped glass vials and
labeled properly.
3.7 Identification of ectoparasites
Presumptive identifications were made while preserved in ethyl alcohol under

stereoscopic microscope and final identifications were made under compound
microscope. For this, collected arthropods were processed for permanent mounting
using methods suggested by Cable (1967). Lice were identified following the
methods suggested by Herms and James (1961). Ticks were identified as described
by Soulsby (1982).
3.8 Collection of faecal sample
The buffaloes were selected randomly irrespective of age, sex, condition/ nutritional
status and level of husbandry from the buffalo reared areas as mentioned above.
Most of the buffaloes in the rural areas are reared in free range grazing system. The
age of the buffaloes was determined from the official record book, or by
interrogating the farmers, or by examining the teeth. After taking all the relevant
information, the faecal samples were collected directly from the rectum of the
animals. Before collection, the animals were restrained properly and all possible
hygienic measures including wearing of apron, hand gloves and gumboot were taken
31
to avoid contamination. Fresh faecal samples were also collected from the ground
when the animals were found in the act of defecation. A total of 236 samples were
collected. About 20-25 grams of faeces were collected from each buffalo. Each
sample was kept in separate polythene bag, tied carefully and numbered properly and
the samples were preserved in 10% formalin. The correctly labeled and properly
numbered polythene bags containing the faecal samples with all required information
were brought to the laboratory and examined.
3.9 Examination of faecal sample
The fecal samples were examined by Modified Stoll’s Dilution Technique as

described by Soulsby (1982).
3.9.1 Modified Stoll’s Dilution Technique
This technique was following for total egg count per gram of faeces (EPG) as
described by Soulsby, (1982). The technique was as follows:
i. The faecal sample was first well mixed and then 3 grams of faces were
weighed with the help of a balance and weight box and put in 100 ml
graduated beaker.
ii. The beaker was then filled with water up to 42 ml mark.

iii. Some small glass beads were added.
iv. The feces was thoroughly mixed with water by magnetic stirrer.
v. The mixture was then strained with a coffee strainer.
vi. The strained mixture was again shaken and 0.15ml of mixture was taken with
a 1 ml special pipette and put on a glass slide and covered with a cover slide.
Care was taken to avoid bubble formation
vii. The slide was then placed under a microscope and the eggs were identified
and counted. The total number of eggs of parasites found in the slide was
multiplied by 100 to get the Eggs Per Gram of Feces (EPG).
32
3.10 Collection, staining and preservation of blood samples
Blood samples were collected from ear vein of the suspected buffaloes and thin
smears were prepared on clean glass slides. To prepare a thin smear, a small drop of
blood was placed on the side of a properly clean, dry, grease free glass slide and thin
smear was made with the help of another even edged glass slide by applying even
pressure. The smears were then air dried for 10 minutes and fixed with absolute
acetone free methyl alcohol, stained with Giemsa’s stain and air dried (Cable, 1967).
3.11 Blood smear examination
The stained slides were examined under compound microscope in higher

magnification (40× and 100×) with oil emersion for detection of protozoa as
described by Cable (1967). Identification was based on the morphology as described
by Levine (1985) and Soulsby (1982). Anaplasma sp. was identified according to the
morphological description given by Soulsby (1982).
3.12 Statistical analysis
Statistical analyses were carried out by Statistical Package for Social Science (SPSS)
using F test. To compare the prevalence of parasites in both sexes, data were
analysed by using paired sample t-test (Mostafa, 1989). Odd ratio was calculated
according to the formula given by Schlesselman (1982).
33
CHAPTER 4
RESULTS
4.1.1 Overall prevalence of endoparasites
During this study (November 2007 to October, 2008) a total of 236 buffaloes were
examined through fecal sample examination, of which 144 (61.02%), were found
infected with one or more species of endoparasites. A total of nine species of
endoparasites (ova/eggs) were identified, of them four species were trematode,
namely Fasciola gigantica (22.46%), Paramphistomum cervi (29.24%),
Schistosoma indicum (1.27%), Schistosoma spindale (0.85%), three species were
nematode namely Toxocara vitulorum (2.54%), Strongyles (0.85%), Strongyloides
sp. (0.42%) and two species were protozoa namely Eimeria sp. (3.39%) and
Balantidium coli (37.29%). No cestodes were detected. From this experiment, it was
observed that, prevalence of B. coli. (37.29%) was the highest whereas
Strongyloides sp. (0.42%) was the least (Table 1, Figure 2).
In this study, EPG (Egg Per Gram of Feces) was also determined. The range of EPG
varies among the parasites. EPG count was the highest in case of B. Coli infection
(100-5000) followed by Fasciola gigantica (100-1200), Paramphistomum cervi
(100-500), Eimeria sp. (100-300). Toxocara vitulorum (100-200) and the other
parasites had the same EPG (100). Mean EPG count was also high in case of B. coli
(452.27±359.22) followed by that of Eimeria sp. (250.00±130.93); F. gigantica
(201.88±142.18), Paramphistomum cervi (186.95±119.95) and Toxocara sp.
(116.67±40.83). A low parasitic burden was found in case of Strongyles species,
Strongyloides sp., S. indicum and S. spindale (100.00±0.00).
34
4.1.2 Age related prevalence of endoparasites
Age of the host had an effect on the prevalence of endoparasites of buffaloes.

Prevalence of endoparasites were significantly (p<0.01) higher (65.85%) in young
animals aged > 2-5 years than in buffalo calves aged 0.5 to 2 years (63.16%) and in
older animals aged > 5 years (59.66%). Calculated odd ratio implied that young
animals were 1.30 and 1.12 times more susceptible to infection than older animals
and calves, respectively. On the other hand, calves and older animals were almost
equally (odd ratio 1.16) susceptible to endoparasitic infection. Older animals were
infected by highest 8 species of parasites followed by young (7 species) and calves
(5 species) of parasites but F. gigantica, Paramphistomum cervi, B. coli, Eimeria sp.
and T. vitulorum were common among the age groups (Table 2).
In calves (0.5 to 2 years), prevalence was relatively higher in case of B. coli

(38.89%), followed by that of Fasciola gigantica (22.22%), T. vitulorum (16.67%),
Paromphistomum sp. (11.11%) and Eimeria sp. (5.56%). In young animals (> 2 to 5
years), B. coli was the main endoparasite recorded (41.46%) followed by
Paramphistomum cervi (36.59%) and F. gigantica (17.07%). In older animals (>5
years), prevalence was also higher in case of B. coli (35.59%), followed by
Paramphistomum cervi (29.38%); F. gigantica (23.73%); Eimeria sp. (2.82%);
Toxocara vitulorum, Strongyles and S. indicum (1.13%) and Strongyloides sp.
(0.56%).
35
4.1.3 Sex related prevalence of endoparasites
In this study, it was detected that prevalence of endoparasites were slightly higher
(p<0.01) is male (61.34%) than the female (59.52%) animals. Male buffaloes were
1.08 times more vulnerable to endoparasitic infection than females. It was interesting
that, S. spindale was only detected in female animals. In both the sexes, prevalence
was the highest in case of B. coli infection (37.11% in male and 35.71% in female)
followed by Paramphistomum cervi (29.38% in male and 28.57% in female), F.
gigantica (24.23% in male and 14.29% in female), Eimeria sp. (2.58% in male and
7.14% in female). The other parasites were found in negligible percentages (Table
3).
4.1.4 Seasonal prevalence of endoparasites
In the present study, it was observed that seasons of the year had a profound effect
on the prevalence of endoparasites in buffaloes. Prevalence of endoparasites was
significantly (p<0.01) higher in rainy season (71.70%) followed by summer
(58.90%) and winter (57.27%) seasons. In this study, it was also revealed that
buffaloes were 1.77 and 1.89 times more susceptible to endoparasitic infection in
rainy season than summer and winter seasons, respectively. On the other hand, in
summer and winter season buffaloes were almost equally (odds ratio 1.07) prone to
endoparasitic infection (Table 4).
In rainy season, prevalence were relatively higher in case of B. coli (54.72%),

followed by that of Paramphistomum cervi (45.28%); F. gigantica (13.21%); S.
spindale (3.77%); T. vitulorum, S. indicum and Strongyloides sp. (1.89%). Eimeria
sp. and Strongyles species were not found in rainy season. Interestingly, S. spindale
was only detected in the rainy season. In summer season, prevalence was higher in
case of B. coli (30.14%) followed by Paramphistomum cervi (28.77%), F. gigantica
36
(26.03%), Eimeria sp. (5.48%), T. vitulorum (2.74%) and both S. indicum and
Strongyles species (1.37%). No Strongyloides sp. was detected. In winter season,
prevalence was higher in case of B. coli (33.64%) followed by F. gigantica
(24.55%), Paramphistomum cervi (21.82%), Eimeria sp. (3.64%), T. vitulorum
(2.73%). Stronglyes and S. indicum were the least (0.91%).
4.2 Prevalence of ectoparasites in buffaloes
4.2.1 Overall prevalence of ectoparasites
During this study, a total of 236 buffaloes were examined, of which 61.86% were
found infested with one of more species of ectoparasites. A total of 3 species of
ectoparasites were identified, of which 2 species were arachnids namely
Haemaphysalis bispinosa (11.44%) and Boophilus microplus (13.98%) and 1 species
was insect namely Haematopinus tuberculatus (51.27%) (Table 5, Figure 3).
In this study, mean parasitic burden was also determined. The range of parasitic
burden in case of H. tuberculatus was 1-13 per square inch of heavily infested area
followed by that of B. microplus (1-4) and H. bispinos (1-3). Mean parasitic burden
was also high in case of H. tuberculatus (3.49±2.29) followed by that of B.
microplus (1.85±0.94) and H. bispinosa (1.59±0.69). From this experiment, it was
found that prevalence of H. tuberculatus (51.27%) was the highest whereas H.
bispinusa (11.44%) was the least.
4.2.2 Age related prevalence of ectoparasites
Age of the host had an effect on the prevalence of ectoparasites of buffaloes.

Prevalence of ectoparasites was significantly (p<0.01) lower (58.52%) in older
animals aged > 5 years than in young aged > 2 to 5 years (70.73%) and in calves
aged 0.5 to 2 years (73.68%). Calculated odd ratio implied that young animals were
37
1.71 times more susceptible to ectoparasitic infestation than older animals. On the
other hand, buffalo calves were 1.16 and 1.98 times more vulnerable to ectoparasitic
infestation than young and older, respectively (Table 6).
In all the age groups, prevalence of H. tuberculatus was the highest. In calves (0.5 to
2 years), prevalence was relatively higher in case of H. tuberculatus (68.42%)
followed by that of H. bispinosa (15.79%) and B. microplus (5.26%). In young
animals (> 2 to 5 years), prevalence was also higher in case of H. tuberculatus
(63.41%) followed by that of H. bispinosa (17.07%) and B. microplus (14.63%). In
older animals (> 5 years), prevalence was higher in case of H. tuberculatus (46.59%)
followed by B. microplus (14.77%) and H. bispinosa (9.66%).
4.2.3 Sex related prevalence of ectoparasites
From this study, it was revealed that prevalence of ectoparasites were comparatively
higher (p<0.01) in female (85.71%) than the male (56.70%) buffaloes. Female
buffaloes were 4.58 times more susceptible to ectoparasitic infestations than the male
buffaloes. In male buffaloes, prevalence was higher in case of H. tuberculatus
(46.39%) followed by B. microplus (12.89%) and H. bispinosa (10.82%). In female
buffaloes, prevalence was also higher in case of H. tuberculatus (73.81%) followed
by H. bispinosa (26.19%) and B. microplus (19.05%) (Table 7).
4.2.4 Seasonal prevalence of ectoparasites
In the present study, it was observed that season of the year had a significant
(p<0.01) effect on the prevalence of ectoparasites in buffaloes. Prevalence of
ectoparasites was the highest in winter season (80.00%) followed by summer
(50.68%) and rainy (39.62%) seasons. In this study, it was also revealed that
buffaloes were 6.09 times more susceptible to ectoparasitic infestations in the winter
38
season than in rainy season. But in summer season, buffaloes were 1.57 times more
vulnerable to cetoparasitic infestations than rainy seasons. On the other hand, in the
winter season buffaloes were 3.89 times more prone to ectoparasitic infestations than
in summer season (Table 8).
In all the seasons, prevalence was the highest in case of H. tuberculatus but varies
significantly among the seasons. In rainy season, prevalence was 28.30%, 13.21%
and 9.43% in H. tuberculatus, H. bispinosa and in B. microplus, respectively. In the
summer season, the rate of infestation was 38.96%, 17.81% and 8.22% in H.
tuberculatus, B. microplus and H. bispinosa, respectively. On the other in winter
season, the prevalence of H. tuberculatus, H. bispinosa and B. microplus was
70.91%, 17.27% and 13.64%, respectively.
4.3.1 Overall prevalence of blood parasites
During this study (November, 2007 to October, 2008), a total of 236 buffaloes were
examined for blood parasites, of which 12.71% were found positive. A total of 3
species of blood parasites were detected, all were protozoa namely Anaplasma
marginale (8.89%), Theileria sp. (2.12%) and Babesia sp. (1.69%). No filarial
worms were detected. Only single infection was detected, no mixed infection was
observed (Table 9, Figure 4).
4.3.2 Age related prevalence of blood parasites
From this study, it was revealed that age of the host had a significant (p<0.01) role
on epidemiology of blood parasites. Prevalence of blood parasites was lower
(5.26%) in calves aged 0.5 to 2 years than older animals aged > 5 years (12.50%)
and in young animals aged > 2 to 5 years (17.07%). Calculated odd ratio implied that
young were 3.70 times more susceptible to blood parasites than calves. But the older
39
animals were 2.57 times more vulnerable to infection than calves. On the other hand,
young animals were 1.44 times more prone to infection than older animals (Table
10).
In all the age groups, prevalence of anaplasmosis was the highest. In buffalo calves
(0.5 to 2 years), A. marginale (5.26%) was the only organism detected. In young
animals (> 2 to 5 year), the rate of infection of A. marginale, Theileria sp. and
Babesia sp. were 12.20%, 2.44% and 2.44%, respectively. In older animals (> 5
years) 8.52%, 2.27% and 1.70% infection of A. marginale, Theileria sp. and Babesia
sp., respectively were recorded.
4.3.3 Sex related prevalence of blood parasites
In this study, it was revealed that prevalence of blood parasites was comparatively
higher in female (23.81%) than the male (10.31%) buffaloes. Female animals were
2.72 times more susceptible to blood protozoan infection than males. In both the
sexes prevalence of Anaplasma marginale was the highest whether prevalence of
Babesia sp. was the least. 7.73%, 1.55% and 1.03% infection of A. marginale,
Theileria sp. and Babesia sp., respectively were recorded in male animals. On the
other hand, the rate of infection was 14.28%, 4.76% and 4.76%, respectively for A.
marginale, Theileria sp. and Babesia sp. in females (Table 11)
4.3.4 Seasonal prevalence of blood parasites
In the present study, it was observed that seasons of the year had a profound effect
on the prevalence of blood parasites. In buffaloes, prevalence of blood parasites was
the higher (p<0.01) in rainy season (16.98%), followed by summer (12.33%) and in
winter (10.91%) seasons. In this study, it was also revealed that buffaloes were 1.45
and 1.67 times more susceptible to blood parasitic infection in rainy season than in
40
summer and winter seasons, respectively. On the other hand, buffaloes were almost
equally (odds ratio 1.15) susceptible to infection in summer and winter seasons
(Table 12).
In all the seasons, prevalence of anaplasmosis was the highest. In rainy season, the
prevalence of A. marginale, Theileria sp. and Babesia sp. were 11.32%, 3.77% and
1.89%, respectively. On the other hand in summer season, the prevalence of
anaplasmosis was 9.59% and the prevalence of Theileria sp. and Babesia sp. was the
same (1.37%). In winter season, buffaloes were equally susceptible to Theileria sp.
and Babesia sp. (1.82%) whether the prevalence of A. marginale was 7.27%.
Table 1. Overall prevalence of endoparasites of buffaloes in Kurigram, Bangladesh
Name of parasites No of animals Percentage Egg Per Gram of Feces (EPG)

affected (%) Range Mean±SD
(N=236)
F. gigantica 53 22.46 100-1200 201.88±142.18
B. coli 88 37.29 100-5000 452.27±359.22
Paramphistomum cervi 69 29.24 100-500 186.95±119.95
Eimeria sp. 8 3.39 100-300 250.00±130.93
T. vitulorum 6 2.54 100-200 116.67±40.83
S. indicum 3 1.27 100-100 100.00±0.00
S. spindale 2 0.85 100-100 100.00±0.00
Strongyles 2 0.85 100-100 100.00±0.00
Strongyloides sp. 1 0.42 100-100 100.00±0.00
Total 144* 61.02 100-5000 178.57±88.12
* = Total no. of animals affected is less than the summation of individual infection because
same animal was infected by more than one type of endoparasites
N = Total animals examined
41
70
60
50
Percentage (%)
40
30
20
10
0
F.
B.
Pa
Ei
T.
S.
St
St
To
S.
m
ro
ro
g
vi
ta
co
ra
ind
s
ig
er
ng
n
pi
tu
l
m
li
gy
a
ia
icu
n
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ph
yl
nt
da
lo
e
sp
um
m
ica
ist
ide
el
.
om
ss
um
p.
sp
.
Name of parasites
Figure 2. Overall prevalence (%) of endoparasites of buffaloes in Kurigram, Bangladesh
42
Table 2. Age related prevalence of endoparasites of buffaloes in Kurigram, Bangladesh
Age Name of parasites No. of Percentage Egg Per Gram of Feces Odds ratio
recovered animals (%) (EPG)
affected Range Mean±SD
F. gigantica 4 22.22 100-200 125.00±50.00
Buffalo B. coli 7 38.89 100-900 347.06±264.85

Calves (0.5- Paramphistomum cervi 2 11.11 100-400 193.33±138.70 Young vs
2years) Calves =
Eimeria sp. 1 5.56 500-0.00 500±0.00 1.12
n=19 T. vitulorum 3 16.67 100-200 133.33±57.74
Subtotal= 12* 63.16 100-900 259.74±102.26
F. gigantica 7 17.07 100-1400 300±286.48
B. coli 17 41.46 100-1000 347.06±264.85
Young (>2-5 Young vs
Eimeria sp. 2 4.88 100-200 150.00±70.71
years) Older =
T. vitulorum 1 2.44 100-100 100.00±0.00 1.30
n=41
S.indicum 1 2.44 100-100 100.00±0.00
S.spindale 2 4.88 100-100 100.00±0.00
Subtotal= 27* 65.85 100-1400 184.34±108.68
F. gigantica 42 23.73 100-1200 193.04±192.85
B. coli 63 35.59 100-5000 347.06±264.85
Older (>5 Eimeria sp. 5 2.82 100-300 150.00±70.71 Calves vs
years) T. vitulorum 2 1.13 100-100 100.00±0.00 Older =
n=176 Strongyles 2 1.13 100-100 100±0.00 1.16
S. indicum 2 1.13 100-100 100.00±0.00

Strongyloides sp. 1 0.56 100-100 100±0.00
Subtotal= 105* 59.66 100-5000 160.43±83.39
Level of significance (0.0002)**
n = Total animals examined

** = Means p<0.01
43
Table 3. Sex related prevalence of endoparasites of buffaloes in Kurigram, Bangladesh
Parameters Name of parasites No. of Percentage Egg Per Gram of Feces Odds ratio
animals % (EPG)
affecte Range Mean±SD
d
F. gigantica 47 24.23 100-1300 242.18±201.89
B. coli 72 37.11 100-5000 622.60±454.02
Eimeria sp. 5 2.58 100-500 130.93±75.00

Male
T. vitulorum 4 2.06 100-200 40.82±116.67
n=194
Strongyles 2 1.03 100-100 100.00±0.00
S. indicum 2 1.03 100-100 100.00±0.00
Subtotal= 119* 61.34 100-5000 182.06±120.56

Male vs
F. gigantica 6 14.29 100-1400 206.00±148.00 Female =
1.08
B .coli 15 35.71 100-5000 450.00±431.03
Eimeria sp. 3 7.14 100-500 250.00±130.93
Female T. vitulorum 2 4.76 100-200 116.67±40.82

N=42 S. spindale 2 4.76 100-100 100.00±0.00
Strongyles 0 0.00 100-100 100.00±0.00
S. indicum 1 2.38 100-100 100.00±0.00
Subtotal= 25* 59.52 100-5000 178.97±96.89

** = Means p<0.01
44
Table 4. Seasonal prevalence of endoparasites of buffaloes in Kurigram, Bangladesh
Season Name of parasite No. of Percentage Egg Per Gram of Feces (EPG) Odds
animals (%) Range Mean±SD ratio
affecte
d
F. gigantica 7 13.21 100-1200 201.88±112.18
B. coli 29 54.72 100-5000 452.27±405.63
Rainy S. spindale 2 3.77 100-100 100.00±0.00 Rainy vs

Summer
n=53 T. vitulorum 1 1.89 100-200 116.67±40.82 = 1.77
S. indicum 1 1.89 100-100 100.00±0.00
Subtotal= 38* 71.70 100-5000 192.96±113.09
F. gigantica 19 26.03 100-1200 217.39±256.74
B. coli 22 30.14 100-5000 471.18±332.51

Summer
Summer Eimeria sp. 4 5.48 100-300 250.00±130.93 vs
n=73 T. vitulorum 2 2.74 100-200 120.00±44.72 Winter
= 1.07
Strongyles 1 1.37 100-100 100.00±0.00
S. indicum 1 1.37 100-100 100.00±0.00
Subtotal= 43* 58.90 100-5000 204.87±126.54
F. gigantica 27 24.55 100-1200 192.59±123.80
B. coli 37 33.64 100-5000 302.70±236.27
Winter Eimeria sp. 4 3.64 100-300 275.00±170.78 Rainy vs

Winter
n=110 T. vitulorum 3 2.73 100-200 133.33±57.74 = 1.89
S. spindale 1 0.91 100-100 100.00±0.00
Strongyles 1 0.91 100-100 100.00±0.00
Subtotal 63* 57.27 100-5000 183.21±102.44

** = Means p<0.01
45
Table 5. Overall prevalence of ectoparasites of buffaloes in Kurigram, Bangladesh
Name of parasites No. of animals affected Percentage (%) Parasitic burden

N= 236 Range Mean±SD
H. tuberculatus 121 51.27 1-13 3.49±2.29
B. microplus 33 13.98 1-4 1.85±0.94
H. bispinosa 27 11.44 1-3 1.59±0.69
Total= 146* 61.86 1-13 2.31±1.31

* = Total no. of animals affected is less than the summation of individual infestation because
same animal was infested by more than one type of ectoparasites
70
60
50
Percentage (%)
40
30
20
10
0
H.tuberculatus B.microplus H.bispinosa Total infected
Name of parasites
Figure 3. Overall prevalence (%) of ectoparasites of buffaloes in Kurigram, Bangladesh
46
Table 6. Age related prevalence of ectoparasites of buffaloes in Kurigram, Bangladesh
Age of Name of No. of Percentage Parasitic burden Odds ratio

animals parasites animals (%) Range Mean±SD
recovered affected
H. tuberculatus 13 68.42 1-13 3.48±2.30
Buffalo
calves (0.5- B. microplus 1 5.26 1-4 1.85±0.94 Calves vs
2years) Young =
H. bispinosa 3 15.79 1-3 1.59±0.69 1.16
n=19
Subtotal= 14* 73.68 1-13 2.31±1.31
H. tuberculatus 26 63.41 1-13 3.49±2.30

Young (> 2-
B. microplus 6 14.63 1-4 1.83±0.95 Young vs
5 years)
H. bispinosa 7 17.07 1-3 1.61±0.67 Older = 1.71
n=41
Subtotal= 29* 70.73 1-13 2.33±1.31
H .tuberculatus 82 46.59 1-13 3.65±2.36

Older (>5
B. microplus 26 14.77 1-4 1.85±0.97 Calves vs
years)
H. bispinosa 17 9.66 1-3 1.41±0.62 Older = 1.98
n=176
Subtotal= 103* 58.52 1-13 2.31±1.32
Level of significance P=0.0014**

** = Means p<0.01
47
Table 7. Sex related prevalence of ectoparasites of buffaloes in Kurigram, Bangladesh
Parameters Name of parasites No. of Percentage Parasitic burden Odds ratio

animals (%) Range Mean±SD
affected
H. tuberculatus 90 46.39 1-13 3.37±2.40
Male B. microplus 25 12.89 1-4 1.80±0.91

n=194 H. bispinosa 21 10.82 1-2 1.29±0.47
Subtotal= 110* 56.70 1-13 2.15±1.26 Female vs

Male =
H. tuberculatus 31 73.81 1-8 3.81±1.94 4.58
Female B. microplus 8 19.05 1-4 2.00±1.07
n=42 H. bispinosa 11 26.19 1-3 2.10±0.74
Subtotal= 36* 85.71 1-8 2.64±1.25
Level of significance 0.0071**

* Total no. of animals affected is less than the summation of individual infestation because
** = Means p<0.01
Table 8. Seasonal prevalence of ectoparasites of buffaloes in Kurigram, Bangladesh
Season Name of No. of Percentage Parasitic burden Odds ratio
parasites animals (%) Range Mean±SD
affected
H. tuberculatus 15 28.30 1-5 2.87±1.25
Rainy B. microplus 5 9.43 1-2 1.40±0.55 Summer vs

N=53 H. bispinosa 7 13.21 1-3 1.57±0.79 Rainy = 1.57
Subtotal= 21* 39.62 1-5 1.95±0.87
H .tuberculatus 28 38.36 1-8 3.18±1.83
Summer B. microplus 13 17.81 1-4 2.00±1.00 Winter vs

Summer =
N=73 H .bispinosa 6 8.22 1-2 1.25±0.50 3.89
Subtotal= 37* 50.68 1-8 2.14±1.11
H .tuberculatus 78 70.91 1-13 3.71±2.56
Winter B. microplus 15 13.64 1-4 1.87±0.99 Winter vs

n=110 H. bispinosa 19 17.27 1-3 1.69±0.70 Rainy =6.09
Subtotal= 88* 80.00 1-13 2.42±1.42
Level of significance 0.0001**

** = Means p<0.01
48
Table 9. Overall prevalence of blood parasites of buffaloes in Kurigram, Bangladesh
Name of parasites No. of animals affected Percentage (%)

N=236
A. marginale 21 8.89
Theileria sp. 5 2.12
Babesia sp. 4 1.69
Total= 30 12.71
14
12
10
Percentage (%)
0
A. marginale Theileria sp. Babesia sp. Total infected
Name of parasites
Figure 4. Overall prevalence of blood parasites of buffaloes in Kurigram, Bangladesh
49
Table 10. Age related prevalence of blood parasites of buffaloes in Kurigram, Bangladesh
Age of animals Name of parasites No. of animals Percentage Odds ratio

identified affected (%)
Buffalo Calves A. marginale 1 5.26
(0.5-2years)
Theileria sp. 0 0.00 Young vs Calves
n=19 =3.70
Babesia sp. 0 0.00
Subtotal= 1 5.26
Young (>2-5 years) A. marginale 5 12.20

n=41 Theileria sp. 1 2.44 Young vs Older
Babesia sp. 1 2.44 =1.44
Subtotal= 7 17.07
Older (>5 year) A. marginale 15 8.52

n=176 Theileria sp. 4 2.27 Older vs Calves =
Babesia sp. 3 1.70 2.57
Subtotal= 22 12.50
Level of significance p=0.0017**

** = Mean P<0.01
50
Table 11. Sex related prevalence of blood parasites of buffaloes in Kurigram, Bangladesh
Parameters Name of parasites No. of animals Percentage (%) Odds ratio

identified affected
Male Theileria sp. 3 1.55

n= 194 Babesia sp. 2 1.03
Subtotal= 20 10.31 Female vs Male =

A. marginale 6 14.28 2.72
Female Theileria sp. 2 4.76

n= 42 Babesia sp. 2 4.76
Subtotal= 10 23.81

** = Mean P<0.01
Table 12. Seasonal prevalence of blood parasites of buffaloes in Kurigram, Bangladesh
Seasons Name of parasites No. of animals Percentage Odds ratio

identified affected (%)
Rainy Theileria sp. 2 3.77 Rainy vs Summer =

n=53 Babesia sp. 1 1.89 1.45
Subtotal= 9 16.98
A. marginale 7 9.59
Summer Theileria sp. 1 1.37 Summer vs Winter =

n=73 Babesia sp. 1 1.37 1.15
Subtotal= 9 12.33
A. marginale 8 7.27
Winter Theileria sp. 2 1.82

Rainy vs Winter = 1.67
n=110 Babesia sp. 2 1.82
Subtotal= 12 10.91

** = Mean P<0.01
51
Figure 5. Buffalo grazing on marshy Figure 6. Preserved ectoparasites in70%
paddy field just after harvesting. alcohol.
Figure 7. Ova of Schistosoma spindale Figure 8. Ova of Schistosoma indicum
52
Figure 9. Anaplasma marginale stained Figure 10. Anaplasma marginale stained
with Giems’s stain (833×) with Giems’s stain
Figure 11. Babesia sp. stained with Figure 12. Theileria sp. stained with
Giems’s stain (833×) Giems’s stain (833×)
53
Figure 13. Haematopinus tuberculatus showing Figure 14. Haematopinus tuberculatus
sternal plate
Figure 15. Boophilus microplus Figure 16. Haemaphysalis bispinosa
54
CHAPTER 5
DISCUSSION
5.1.1 Overall prevalence of endoparasites
The research work indicates that, the water buffaloes are very much susceptible to
endoparasitic infection. About 61.02% buffaloes were found to be infected with
endoparasites, of them prevalence of B. coli (37.29%), Paramphistomum cervi
(29.24%), F. gigantica (22.46%), Eimeria sp. (3.39%), T. vitulorum (2.54%), S.
indicum (1.27%), S. spindale (0.85%), Strongyles (0.85%) and the least
Strongyloides sp. (0.42%) were recorded. Similar studies were conducted by some
other scientists in Bangladesh and in abroad. Similar findings were reported by
Azam et al. (2002) who revealed that 64.41% of the buffaloes were positive for
internal parasites in Pakistan. Slightly lower prevalence were observed by Motaleb
(1996) and Bachal et al. (2002) who recorded 38.5% and 47% buffaloes suffered
from different types of helminths in Bangladesh and in Pakistan, respectively.
The present finding is in agreement with the earlier findings of Hossain (1991),
Morsy et al. (2005) and Shazly et al. (2007) who recorded 24%, 20% and 9.73%
infection of fasciolosis in Bangladesh and Egypt, respectively. Islam (1991) reported
that Fasciola infection in buffaloes varies from place to place and recorded 12.5%,
28.8% and 17.3% infection in Dinajpur, Bagerhat and Chapainawabgonj district,
respectively. Bhuyan (1970) recorded fasciolosis in 90.90% buffaloes, which is
much higher than the present finding.
Islam et al. (1992) reported an overall 18.9% F. gigantica and 29.50%

Paramphistomum spp. infection by fecal sample examination in buffaloes, which
supported the present findings. Hossain et al. (1991) reported slightly lower
55
prevalence of F. gigantica (12%) and amphistome (14%) infection in buffaloes of
Bangladesh by fecal sample examination. Alim (1999) reported that overall 50.65%
Fasciola infection in buffaloes in different parts of Bangladesh which was much
higher than the present finding. Van et al. (2000) observed that overall prevalence of
Strongyles infection were 53% for buffaloes in Mindanao, the Philippines. These
finding is also higher than the current findings. Haridy et al. (2006) reported 17.7%
Fasciola and 10% Paramphistomum infection in buffaloes in Al-Sanla centre, Egypt.
Condoleol et al. (2007) detected 33.1% Strongyles, 3.1% Strogyloides sp. and 7.1%
Paramphistomum cervi in buffaloes in central Italy. Ravindran et al. (2007) reported
50% buffaloes having visceral schistosomiasis in India. This finding is much higher
than the present findings.
In this experiment, no cestodes were detected which supported the findings of Azam
et al. (2002) and Bachal et al. (2002) but Condoleol et al. (2007) and Prokopic et al.
(1976) reported the buffalo cestodes in Italy and Afganistan, respectively. Azam et
al. (2002) reported that 72% of the buffalo calves were suffered from intestinal
protozoan infection in Pakistan which is much higher than the present finding.
Nalbantoglu et al. (2008) reported 75% of water buffaloes were suffered from
different Eimeria spp. in Turkey. The variations among the findings might be due to
the difference in the sample size, selection of samples, techniques of sample
collection, period and place of study, environmental factors, breed of the animals etc.
5.1.2 Age related prevalence of endoparasites
During the study, it was revealed that, age of the buffaloes had a significant (p<0.01)
effect on endoparasitic infection. Young animals (65.85%) were more susceptible to
infection than calves (63.16%) and older animals (59.66%), which supported the
findings of Asif et al. (2007) who reported the higher prevalence of helminths
56
infection in young animals compared to adults in buffaloes of Pakistan. The present
finding is also in agreement with the earlier report of Azhar et al. (2002) who noticed
that higher infection rate was recorded in buffaloes aged > 2 years of age than the
buffaloes aged < 2 years. But the present finding is in contrast to the previous reports
of Alim (1997) who observed that infection rate of fasciolosis increased with the
increase of age. The cause of this variation in the prevalence of infection in different
age groups of buffaloes is difficult to explain but it might be due to an
immunological phenomenon as it was stated by Baily (1971) who suggested that the
fasciolosis is not as self limiting in the buffaloes as this in the cattle.
5.1.3 Sex related prevalence of endoparasites
There was significant (p<0.01) difference in the rate of infection in between the male
and female buffaloes. It was observed that, the prevalence of endoparasites was
slightly higher in male (61.34%) than in female animals (59.52%) which supported
the findings of Asif et al. (2007) who reported the higher prevalence of endoparasites
in males than females for buffaloes in Pakistan. But this report is in contrast to the
previous report of Bachal et al. (2002) who reported a slightly higher prevalence
(48.30%) of helminths in female than in male (45.12%) in buffalo calves. Alim
(1997) also reported that females (52.65%) were more susceptible to Fasciola
infection than male (47.76%). On the other hand, Azhar et al. (2002) reported no sex
variation. He noticed that buffaloes of either sex were equally affected. This
disparity among the findings can not be explained exactly but it might be assumed
that hormonal influence may be associated with this. Methods of study, selection of
samples and breed of buffaloes may also be associated with this.
5.1.4 Seasonal prevalence of endoparasites
Seasonal fluctuation of the year had a significant (p<0.01) effect on the prevalence of
endoparasitic infection in buffaloes. A relatively higher infection with endoparasites
57
were observed in rainy season (71.70%), followed by summer (58.90%) and winter
(52.27%) seasons. The present finding is much higher than the previous reports of
Azhar et al. (2002) who reported the overall highest (24.0%) seasonal prevalence in
all types of buffaloes during autumn, followed by spring (20.0%), winter (13.0%),
while the lowest (9.0%) was recorded during summer in Pakistan. The present
finding is in agreement with the earlier reports of Islam (1989) who reported the
prevalence of fasciolosis by feces examination was 28.6%, 18.7% and 11.7%,
respectively, during winter, summer and rainy season. But the author reported the
much higher prevalence of Strongyloides sp. during winter (17.1%), summer (20%)
and in rainy season (8.3%). The contrast in between the present and earlier findings
can be explained by the fact of variation in the geographical location of the
experimental area and also the methods of study. Moreover, in this study, year was
divided into three seasons but in other parts of the world there were four seasons. So,
this difference in the division of seasons had made some over lapping of months and
seasons. Therefore, that might have created some contradictions. However, the
highest prevalence in rainy season may be due to high humidity and heavy rainfall
which favors the parasitic growth and multiplication.
58
5.2 Prevalence of ectoparasits in buffaloes
5.2.1 Overall prevalence of ectoparasits
The research work revealed that the buffaloes were very much susceptible to
ectoparasitic infestation. About 61.86% buffaloes were found to be infested with
ectoparasites, of them prevalence of H. tuberculatas was the highest (51.27%)
followed by B. microplus (13.98%) and H. bispinosa (11.44%). Similar findings
were reported by some other scientist in Bangladesh. Mollah et al. (1970), Islam
(1989) and Chowdhury (1992) reported 33.5%, 34.6% and 52.85% prevalence of H.
tuberculatus in Bangladesh, respectively. On the other hand, Islam (1989),
Chowdhury (1992) and Islam et al. (1992) reported 8.1%, 7.42% and 11.3%
prevalence of H. bispinosa, respectively in the country which exactly supported the
present findings. But the present findings is in contrast to the previous findings of
Chowdhury (1992) and Islam et al. (2006) who reported 21.71% and 56.3%
prevalence of B. microplus, respectively in the country which is much higher than
the present findings. During this experiment no mite was detected but Chowdhury
(1992) detected 17.42% and 7.71% prevalence of Sarcoptes scabei and Psoroptes
natalensis, respectively in Bangladesh. This difference in the present and past
findings might be due to the methods of study and selection of samples.
5.2.2 Age related prevalence of ectoparasites
From this study, it was revealed that age of the host had a significant (p<0.01) effect
on ectoparasitic infestation. Buffalo calves were comparatively more susceptible
(73.68%) than young (70.73%) and older animals (58.52%). It was observed that,
susceptibility decreased with the increase of age. Similar studies were conducted by
some other scientist in Bangladesh. Islam (1989) reported that prevalence of H.
tuberculatus was the highest (65.8%) in the age groups less than < 2 years of age
which supported the present findings. During this experiment it was observed that
59
prevalence of H. bispinosa was the highest (17.07%) in the young (> 2 to 5 years).
This finding is in agreement with the earlier reports of Islam (1989) who recorded
highest (15.4%) prevalence of H. bispinosa in the buffaloes aged between 2-4 years.
But the author did not report in any case of B. microplus infestation. The present
finding is in contrast with the earlier reports of Chowdhury (1992) who recorded the
higher (60%) prevalence of H. tuberculatus in the young (2-4 years) whether highest
(8%) prevalence of H. bispinosa in the calves (< 1 year). Chowdhury (1992) also
reported the higher (24%) prevalence of B. microplus in the buffaloes aged between
1-2 years which is much higher than the present findings. These variations among
the present and previous studies might be due to the place and method of study,
geographical locations, selection of samples etc. It is very difficult to explain exactly
the frequent occurrence of ectoparasitic infestation in calves. But it may be assumed
that the less developed immune system of the calves may be responsible for the
higher prevalence of ectoparasitic infestation. On the other hand, buffaloes
developed immunity with the increase of age, so susceptibility decreased with the
increase of age.
5.2.3 Sex related prevalence of ectoparasites
From this study, it was found that the prevalence of ectoparasites were significantly
(p<0.01) higher in female (85.71%) animals than the male (56.70%). But the
prevalence of endoparasites was slightly higher in males than the female. This
disparity might be due to the methods of study, selection of samples and parasitic
species variation. In both the male and female groups, H. tuberculatus was the main
pest. Although the exact cause of higher prevalence of ectoparasitic infestations in
female buffaloes can be explained but it can be hypothesized that some hormonal
influences may be associated with this phenomenon. Lloyd (1983) reported that
higher level of prolactin and progesterone hormones make the individual more
60
susceptible to any infection. Moreover, stresses of production, such as, pregnancy
and lactation make the female animals more susceptible to any infection.
5.2.4 Seasonal dynamics of ectoparasitic infestation in buffaloes
ectoparasitic infestations in buffaloes. A relatively lower infestation with
ectoparasites was observed in rainy season (39.62%) followed by summer (50.68%)
and winter (80.00%) season. The present finding is supported by the findings of
Islam (1989) who reported the higher incidence of H. tuberculatus and H. bispinosa
in winter season. Chowdhury et al. (1992) also supported the present findings. He
also recorded the higher incidence of H. tuberculatus in winter season. During this
study, it was observed that the higher prevalence of B. microplus and H. bispinosa
was observed during winter and summer, respectively, which is in contrast to the
findings of Chowdhury (1992) who recorded the higher prevalence of B. microplus
and H. bispinosa in summer and winter, respectively. The contrast in between the
present and earlier finding can be explained by the fact of variation in the
geographical location of the experimental area, topography and composition of soil
type temperature and humidly. However, the highest prevalence in winter season
might be attributed to favourable climatic condition in winter supplemented by little
hygienic care like washing, grooming etc.
61
5.3.1 Overall prevalence of blood parasites
The research work highlights that, the buffaloes of Bangladesh are susceptible to
blood parasitic infection. About 12.71% buffaloes examined were found to be
infected with blood parasites, of them prevalence of A. marginale was the highest
(8.89%) followed by Theileria sp. (2.12%) and Babesia sp. (1.69%). No mixed
infection was recorded. The present findings is much lower than that of Razput et al.
(2005) and Buriro et al. (1990) who recorded 30% buffaloes were infected with
Anaplasma sp. in Pakistan. Lalchandani (2001) also reported that 39.22% buffaloes
infected with different types of blood protozoa (Anaplasma, Babesia and Theileria).
In Bangladesh, Islam (1992) only reported the presence of Trypanosoma theiliri
(0.4%). This difference may be attributed to the differences in climatic condition and
intensity of tick infestation in the area. However, methods of study may also be
responsible for the variation. During this experiment, samples were randomly
collected from the animals whether the animals were suffered from tick infestations
or not or either the animals showed any clinical signs. In this study, it was observed
that infestations of ticks were so low. But Tabanus sp, was found in most of the
buffalo sheds although their prevalence was not determined. Ristic (1968) reported
that Tabanus sp. can transmit the Anaplasma organism. So, in this study, it was
suggested that Tabanus might play a vital role in transmission of Anaplasma
organism. Moreover, the study area was in the border area and the border is not strict
enough. So, the free entrances of Indian buffaloes are more common, which may
also be the source of blood parasites.
62
5.3.2 Age related prevalence of blood parasites
During the study, it was revealed that, age of the buffaloes had a significant (p<0.01)
effect with blood protozoan infection. Young (> 2 to years) were mostly susceptible
(17.07%) followed by older (> 5 years) (12.50%) and buffalo calves (0.5-2 years)
(5.26%). Buffalo calves were less susceptible to the infection due to phenomenon
"Inverse Age Resistance” (Soulsby, 1982). Prevalence was peak in young animals
and gradually decreased with the increase of age as older animals develop immunity
against the organisms.
5.3.3 Sex related prevalence of blood parasites
From this study, it was observed that prevalence of blood parasites were significantly
(p<0.01) higher in female animals (23.81%) than the male (10.31%). Rajput et al.
(2005) also reported the higher prevalence of anaplasmosis in female (30.28%) than
in male (29.33%). Although the exact cause of higher prevalence of blood parasitic
infection in female buffaloes can not be explained but it can be by hypothesized that
some hormonal influence may be associated with this phenomenon. Lloyd (1983)
reported that higher level of prolaction and progesterone hormones make the
individual more susceptible to any infection. Moreover, stresses of production such
as, pregnancy and lactation make the individual more susceptible to any infection.
However, methods of study may also be associated with this variation.
5.3.4 Seasonal prevalence of blood parasites in buffaloes
blood parasitic infections. A relatively higher infection with blood parasites were
observed in rainy season (16.98%) followed by summer (12.33%) and winter season
(10.91%). It was surprised that prevalence of blood parasites was higher in rainy
season although the vector species were found lowest in rainy season. This
63
difference might be due to the methods of study and selection of samples. It might be
happened that, Tabanus sp. was responsible for this high prevalence rate as during
this experiment Tabanus sp. was found more during the rainy season. Moreover,
highest prevalence in rainy season may be due to high humidity and heavy rainfall
which created the favourable condition for the organisms.
64
CHAPTER 6
CONCLUSION
In this study, epidemiology of parasitic diseases of buffaloes was studied. Prevalence
of endoparasites was studied by examination of feces only. But it was difficult to
species wise identification of the parasite (like strongyles) through ova detection. So,
it would be better to examine the suspected organs for the detection of whole
parasites. During this study, prevalence of ectoparasites was also determined. Only
two species of ticks and one species of lice were determined but no mites and flies
were recorded. Further studies should be conducted to identify these species of
parasites. Prevalence of blood parasites were also determined in the present study by
blood smear examination. But it would be better, if a sero-surveillance could be
conducted. Result of sero-surveillance (detection of antibody against blood parasites)
would give an idea about the endemic stability. Besides, in this study, actual losses
due to parasitic diseases in buffaloes in terms of mortality, morbidity and treatment
cost have to be determined. But it would help to justify the necessity of control
program against the parasitic diseases. So, further study should also be conducted to
assess the economic losses due to parasitic diseases of buffaloes and to find out
effective control strategies against it.
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Epidemiological Investigation On Parasitic Diseases of Buffaloes (Bubalus Bubalis) in Kurigram District of Bangladesh

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EPIDEMIOLOGICAL INVESTIGATION ON PARASITIC

DISEASES OF BUFFALOES (Bubalus bubalis) IN

MD. ABDULLAH AL MAMUN

MASTER OF SCIENCE (MS)

MD. ABDULLAH AL MAMUN

MASTER OF SCIENCE (MS)

MD. ABDULLAH AL MAMUN

Approved as to style and content by

(Prof. Nurjahan Begum) (Prof. Dr. Md. Motahar Hussain Mondal)

(Prof. Dr. Md. Motahar Hussain Mondal)

CHAPTER TITLE PAGE NO.

CHAPTER TITLE PAGE NO.

SL. NO. TITLE OF THE TABLES PAGE NO.

Table 1 Overall prevalence of endoparasites of buffaloes in

Table 2 Age related prevalence of endoparasites of buffaloes

Table 3 Sex related prevalence of endoparasites of buffaloes

Table 4 Seasonal prevalence of endoparasites of buffaloes in

Table 5 Overall prevalence of ectoparasites of buffaloes in

Table 6 Age related prevalence of ectoparasites of buffaloes

Table 7 Sex related prevalence of ectoparasites of buffaloes in

Table 8 Seasonal prevalence of ectoparasites of buffaloes in

Table 9 Overall prevalence of blood parasites of buffaloes in

Table 10 Age related prevalence of blood parasites of buffaloes

Table 11 Sex related prevalence of blood parasites of buffaloes

Table 12 Seasonal prevalence of blood parasites of buffaloes in

SL. NO. title of the figures PAGE NO.

Figure 1 The flow diagram-showing research out line 20

Figure 2 Overall prevalence (%) of endoparasites of buffaloes in

Figure 3 Overall prevalence (%) of ectoparasites of buffaloes in

Figure 4 Overall prevalence of blood parasites of buffaloes in

Figure 5 Buffalo grazing on marshy paddy field just after

Figure 6 Preserved ectoparasites in70% alcohol. 42

Figure 7 Ova of Schistosoma spindale 42

Figure 8 Ova of Schistosoma indicum 42

Figure 9 Anaplasma marginale stained with Giems’s stain (100×) 43

Figure 10 Anaplasma marginale stained with Giems’s stain 43

Figure 11 Babesia sp. stained with Giems’s stain (100×) 43

Figure 12 Theileria sp. stained with Giems’s stain (100×) 43

Figure 13 Haematopinus tuberculatus showing sternal plate 44

Figure 14 Haematopinus tuberculatus 44

Figure 15 Boophilus microplus 44

Figure 16 Haemaphysalis bispinosa 44

Epidemiology of parasitic diseases of buffaloes was studied in Kurigram district of

In Bangladesh, among many causes, parasitism is thought to be a major cause that

In developed countries, the data on epidemiology of various helminthiasis are

The diverse agroclimatic conditions, animal husbandry practices and pasture

i. To investigate the overall prevalence of parasitic diseases (ecto, endo and

 Buffalo rearing and its importance

2.1 Buffalo rearing and its importance

Parikh (1988) reviewed that Asian civilization is primarily based on agriculture,

World Bank (1998) report suggested that Bangladesh government is supporting

Hoffmann (1999) suggested that approximately 20% of the value of an animal is in

could be in the manure it produces.

dung energy for household and commercial purposes, respectively.

2.2 Prevalence of endoparasites in buffaloes

Chowdhury (1970) recorded Neoascaeis vitulorum, Trichuris globulosa, Trichuris

Islam (1989) recorded incidence of different parasites by examination of faeces were

Hossain (1991) carried out an investigation at Kanihari Union Mymensingh district,

Islam (1991) conducted an epidemiological study of fasciolosis in Dinajpur,

Islam et al. (1992) conducted an investigation to detect the presence of parasites in

Alim (1997) conducted an exploratory study on the epidemio-pathology of

Morsy et al. (2005) investigated on natural infection with Fasciola species in