Buffalo Bulletin (September 2014) Vol.33 No.3

International Buffalo Information Center (IBIC)
BUFFALO BULLETIN
ISSN : 0125-6726
Aims
IBIC is a specialized information center on water buffalo. Established in 1981 by Kasetsart University
(Thailand) with an initial financial support from the International Development Research Center (IDRC) of
Canada. IBIC aims at being the buffalo information center of buffalo research community through out the
world.
Main Objectives
1. To be world source on buffalo information
2. To provide literature search and photocopy services
3. To disseminate information in newsletter
4. To publish occasional publications such as an inventory of ongoing research projects
Buffalo Bulletin is published quarterly in March, June, September and December. Contributions on
any aspect of research or development, progress reports of projects and news on buffalo will be considered
for publication in the bulletin. Manuscripts must be written in English and follow the instruction for authors
which describe at inside of the back cover.
Publisher
International Buffalo Information Center, Office of the University Library, Kasetsart University
Online availible
http://ibic.lib.ku.ac.th/e-Bulletin
Advisory Board
Prof. Dr. Charan Chantalakhana
Prof. Dr. John Lindsay Falvey
Prof. Dr. Metha Wanapat
Mr. Antonio Borghese
Dr. Aree Thunkijjanukij
Miss Wanphen Srijankul
Editorial Member
Dr. Pakapan Skunmun
Dr. Kalaya Bunyanuwat
Prof. Dr. Federico Infascelli
Thailand
Faculty of Veterinary and Agricultural Science, University
of Melbourne, Australia
Department of Animal Science, Faculty of Agriculture,
Khon Kaen University, Thailand
International Buffalo Federation, Italy
International Buffalo Information Center, Office of the
University Library, Kasetsart University, Thailand
Thailand
Department of Livestock Development, Thailand
Department of Veterinary Medicine and Animal Science,
University of Naples Federico II, Italy
Dr. Rafat Al Jassim

Prof. Dr. Nguyen Van Thu
Prof. K. Sarjan Rao
Prof. Dr. Masroor Ellahi Babar
Asst. Prof. Dr. Asif Nadeem
Prof. Dr. Raul Franzolin
School of Agriculture and Food Sciences, Faculty of

Science, The University of Queensland, Australia
Department of Animal Sciences, Faculty of Agriculture
and Applied Biology, Can Tho University, Vietnam
Department of Livestock Production and Management,
College of Veterinary Science, India
Virtual University of Pakistan, Pakistan
Institute of Biochemistry and Biotechnology, University of
Veterinary and Animal Sciences, Pakistan
Departamento de Zootecnia, Universidade de So Paulo,
Brazil
Editor
Dr. Sunpetch Sophon
Journal Manager
Mr. Chalermdej Taterian
Assistant Journal Manager

Miss Jirawadee Wiratto
Faculty of Veterinary
Technology, Thailand
Medicine,
Mahanakorn
of


BUFFALO BULLEITN
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Buffalo Bulletin (September 2014) Vol.33 No.3
CONTENTS
Page
Case Report
Therapeutic management of concurrent vitamin A and zinc deficiency
in buffalo calves- Case report
S. Beigh , J.S. Soodan, A. Tiko and H. Tantary................................................................................244
Successful management of a compound fracture in a buffalo using a fabricated
polyvinylchloride splint in a field setting
A. Velavan, S. Sivaraman and K. Krishnakumar..............................................................................246
Original Article
Prevalence of gastrointestinal parasites in buffaloes (Bubalus bubalis)
in and around Tirupati, India
C. Sreedevi and Md. Hafeez.............................................................................................................251
Haemato-biochemical effects of oral sub-chronic cypermethrin toxicosis in buffalo calves
Shabir Ahmad Dar, Bibhuti Ranjan and Rajdeep Kaur....................................................................256
Incidence of parthenogenetic development of buffalo (Bubalus bubalis) oocytes
Ch. Srinivasa Prasad, A. Palanisamy, S. Satheshkumar,
V.S. Gomathy and S. Rangasamy......................................................................................................267
Genetic diversity and conservation of animal genetic resources in Iraqi buffalo
using microsatellite markers
Talib Ahmed Jaayid and Maytham Abdul Kadhim Dragh...............................................................271
Cryopreservation of endometrial stromal cells of buffalo (Bubalus bubalis)
Chethan Sharma G., S.K. Singh, Jessihun Nongsiej, H.B. Rakesh,
R.P. Singh and S.K. Agarwal............................................................................................................277
Adoption level of buffalo farming practices in the arid zone of Rajasthan
M.L. Meena and Dheeraj Singh.......................................................................................................283
CONTENTS
Page
Original Article
Milk production and reproduction performance of Murrah buffaloes of Tamil Nadu, India
A.K. Thiruvenkadan, S. Panneerselvam, N. Murali, S. Selvam
and V. Ramesh Saravanakumar......................................................................................................291
Effect of genetic and non-genetic factors on morphometric traits of buffaloes
Shashi Shankar, Dhirendra Kumar and K.G. Mandal...................................................................301
Observations on buffalo sarcoptic mange in Jammu, India
T. Nazir, R. Katoch, R. Godara, Anish Yadav and Vijay Pandey...................................................308
Buffalo milk marketing by the Gujjars (Tribals) in the Jammu and Kashmir state in India
Tarunvir Singh and Sudhakar Dwivedi..........................................................................................316
Prevalence of ticks in buffaloes in the Upper Sindh Pakistan
Majid Hussain Soomro, Shahida Parveen Soomro, Muhammad Bachal Bhutto,
Zeeshan Akbar, Muhammad Yaqoob and Abdullah G. Arijo.........................................................323
Comparison of blood acid base gas parameters in venous and arterial blood
of healthy buffaloes
S.A. Hussain and S.K. Uppal.........................................................................................................328
Prevalence of Salmonella and Escherichia coli associated with diarrhea in buffalo
and cow calves
M. Anwarullah, J.A. Khan, M.S. Khan, K. Ashraf and M. Avais...................................................332
Distribution of Salmonella species in buffaloes in some middle governorates of Iraq
Afaf Abdulrahman Yousif and Ali, D.M. Al-Hashimi.....................................................................337
Case Report
THERAPEUTIC MANAGEMENT OF CONCURRENT VITAMIN A AND ZINC DEFICIENCY

IN BUFFALO CALVES- CASE REPORT
S. Beigh, J.S. Soodan*, A. Tiko and H. Tantary
donot respond to the light. The meance reflex

was almost absent but palperal and corneal reflex
was present; however, animal was not apparently
blind. The clinical examination of skin revealed
dermatophytosis with rough dry hair coat, heavy
deposition of scales, with alopecia and wrinkling
ABSTRACT
Vitamin A deficiency is either due to an
absolute deficiency of vitamin A or its precursor
carotene in the diet or secondary in which the dietary
supply of vitamin A or its precursor is adequate
but their digestion, absorption or metabolism is
interfered with to produce deficiency at the tissue
level. Secondary vitamin A deficiency may occur
in case of chronic diseases of liver or intestine or
due to deficiency of zinc (Radostits et al., 2000).
Absorption, metabolism, hepatic release, transport,
at the neck and head. Plucked hairs and scrabbed

scales were examined for fungal agent by direct
microscopy in 10% KOH and lactophenol. Samples
were inoculated on mycobiotic agar. The plates were
incubated at 28C for 2-6 weeks and examined for
colony formation. Culture examination revealed T.
verrucosum as the usual cause of dermatophytosis.
Blood samples were taken from all the animals
in heparinised vials for the estimation of vitamin
A and serum zinc concentration. Vitamin A and
serum zinc concentration were 7.12 0.34 g/dl
(reference values 15.4-32.3) and 5.12 0.26 g/L
(reference values 6-12). On the basis of history,
clinical examination and decreased serum zinc and
vitamin A concentration present, the animals were
diagnosed to be suffering from combined vitamin
A and zinc deficiency.
and tissue utilization of vitamin A may depend

on adequate zinc status while severe vitamin A
deficiency may reduce absorption and lymphatic
transport of zinc by altering synthesis of a zincdependent binding protein (Bendich, 1993).
Keyword: buffalo calves, vitamin A, zinc,
therapeutic management
CASE HISTORY AND OBSERVATION

Five buffalo calves of 1-2 years of age
were referred to the Veterinary Clinics and
Teaching Hospital of SKUAST-Jammu with the
history of sudden inappetance, reduced weight and
heavy lacrymation. Clinical examination revealed
slight clouding of the cornea, dilated pupils which
TREATMENT AND DISCUSSION

The calves were administrated orally with
zinc sulphate at the rate of 1 gm per week for 45
days with vitamin A 300,00 IU and Belamyla 3
Division of Veterinary Clinical Medicine and Jurisprudence, Faculty of Veterinary Sciences and Animal
Husbandry-R.S.Pura-181102, SKUAST- Jammu, India, *E-mail: shafullah@gmail.com
244
often results in increased prevalence of infectious

diseases like dermatophytosis in the present
case. Therefore, supplementation of zinc along
with vitamin A in vitamin A deficient animals is
beneficial .
ml intramuscularly, as reported by Anand et al.

(2005). The affected parts were topically treated
zinc oxide ointment once daily for three weeks.
Marked improvement were reported in terms of
disappearance of skin lesions and convulsions, and
the appetite returned to normal, but the clouding of
eyes remained unaffected. All other buffalo owners
were advised to supplement vitamin A and zinc to
the animals. Vitamin A has numerous functions such
as maintenance of epithelial cells, vision, immune
cell function and gene regulation (Chew, 1987).
Absorption, metabolism, hepatic release, transport,
and tissue utilization of vitamin A depends on the
adequate level of serum zinc. Deficiency of zinc
predisposes the animal for vitamin A deficiency
(Serdar et al., 2009). Zinc plays a regulatory role
on vitamin A transport mediated through protein
synthesis. Zinc deficiency depresses the synthesis
of a.. A brand of Sarabhai Zydus, Ahmedabad retinol
REFERENCES
Anand, K.J., C. L. Srinivas, Dananjay Sing, Harsha
Kumar and S.M. Dhoolapas. 2005. Zinc
deficiency in two calves. Indian Vet. J., 85:
768-769.
Bendich, A. 1993. Physiological role of antioxidant
in the immune system. J. Dairy Sci., 76:
2789-2794.
Chew, B.P. 1983. Vitamin A and -carotene in host
defence. J. Dairy Sci., 70: 2732.
Christian, P. and K.P. West. 1998. Interaction
between zinc and vitamin A: an update. Am.
J. Clin. Nutr., 68: 435-441.
Ewans, P. and Halliwell B. 2001. Micronutrients:
Oxidant/antioxidant status. Brit. J. Nutr.,
85: 57.
Naresh, R., S.K. Dwivedi, D. Swarup and S. Dey.
2001. Zinc, copper, and cobalt concentrations
in blood during inflammation of mammary
gland in dairy cows. Asian Austral. J. Anim.,
14: 564.
binding protein (RBP) in the liver and leads to lower

concentrations of RBP in the plasma (Christian
et al., 1998). Vitamin A and zinc also interact
through the ubiquitous, oxidative conversion of
retinol to retinaldehyde (retinal), a critical step in
the metabolic pathway of vitamin A that is welldescribed in the visual cycle in the retina of the
eye and requires the action of a zinc dependent
retinol dehydrogenase enzyme (Christian et al.,
1998). Zinc is also essential components of the
bodys antioxidant defense that play an important
role in the prevention of free radical-induced
damage to tissues for maintenance of health and
production (Ewans et al., 2001). Susceptibility to
inflammatory and infectious conditions increases
during zinc deficiency (Naresh et al., 2001). Along
with zinc, vitamin A is also an important defense
booster. Vitamin A increases disease resistance and
has stimulatory effects on cell-mediated immunity
(Bendich, 1993). Deficiency of vitamin A and zinc
Radostits, O.M., C.C. Gay, D.C. Blood and K.W.

Hinchcliff. 2000. Veterinary Medicine: A
Textbook of Diseases of Cattle, Sheep, Pigs,
Goats and Horses, 9th ed. WB Saunders,
Harcourt Publishers Ltd., London.
Serdar, P. and K. Funda. 2009. Serum zinc and
vitamin A concentrations in calves with
dermatophytosis. Kafkas niv. Vet. Fak.
Derg., 15: 1.
245
Case Report
SUCCESSFUL MANAGEMENT OF A COMPOUND FRACTURE IN A BUFFALO USING A

FABRICATED POLYVINYLCHLORIDE SPLINT IN A FIELD SETTING
A. Velavan*, S. Sivaraman and K. Krishnakumar
a challenging type to treat. In buffaloes repair of

fractured long bones is not commonly reported due
to their heavy body weight (Turner, 1984; Ayaz,
2000). Prognosis in adults is guarded and mainly
limited by the weight of the animal and degree of
the contamination of the wound (Mulon, 2010).
So, management of this type of fracture is not
much explored. In this paper a modified method of
utilizing a PVC (polyvinylchloride) splint for the
successful management of a compound fracture of
the metatarsal bone in a buffalo is described.
ABSTRACT
A six-year-old female Murrah buffalo was
brought with the complaint of hind limb fracture.
Clinical examination revealed compound fracture
of a metatarsal bone with an open wound on the
medial aspect. The fracture was stabilized using
a fabricated PVC splint external coaptation. The
daily dressing of wound was carried out using 5%
povidone iodine through the window provided in
the PVC splint. Inj. streptopenicillin 5 gm was given
i/m daily for 10 days. The animal started partial
weight bearing on the fractured limb on day 40. On
day 90, the animal was bearing its complete weight
on the fractured limb. There was no complication
reported thereafter.
HISTORY AND CLINICAL SIGNS

A six-year-old female buffalo weighing 450
kilograms was reported to the Teaching Veterinary
Clinical Complex, Veterinary College and
Research Institute, Orathanadu with the complaint
of breaking a hind limb while jumping across a
feed manger. The animal was already treated by a
local vet using plaster of paris for a week. Clinical
examination of the animal revealed that it was a
compound fracture of right metatarsal bone (Figure
1). The open wound was located on the medial
aspect of metatarsal bone (Figure 2). Fractured
bone fragment was visible through the wound site.
Pus discharge was noticed at the fractured site.
Keywords: compound fracture, metatarsal bone,

buffalo, fabricated, PVC splint
INTRODUCTION
In farm animals, limb fractures are
common and occur subsequent to trauma during
handling or dystocia (Anderson and Jean, 2008).
Limb fractures are classified depending on the
anatomical location, presence of external wound,
extent of bone damage and direction of the fracture
line. Among these types, the compound fracture is
Department of Teaching Veterinary Clinical Complex, Veterinary College and Research Institute, Orathanadu,
Thanjavur-614625, India, *E-mail: velvetdr2003@gmail.com
246
RESULTS AND DISCUSSION
amount of soft tissues covering over the metatarsal

and metacarpal bones, these fractures are frequently
converted into compound fractures resulting from
the penetration of the bone within (Ayaz, 2000).
Due to the lack of satisfactory immobilizing devices
with open dressing facilities, compound fractures
usually do not respond to the treatment and develop
such complications that only amputation will save
the animal (Nayak and Samantara, 2010). Since,
the compound fracture needs daily dressing of the
wound, external skeletal fixation is the right choice.
Economic considerations and non-availability of
orthopaedic implants for large animals at the field
level makes compound fractures non-treatable.
Dealing with the compound fracture will always
remain a challenge to the clinician (Mulon, 2011).
Moreover, radiological assessment is almost always
impracticable at the field level, for there is no x-ray
infrastructure facility.
Considering all these factors, we fabricated
the PVC splint with window provision for daily
dressing of wound. Common antibiotics used in
orthopaedic infection are penicillin, cephalosporin,
fluoroquinolone and trimethoprim-sulpha as it has
been established they reach a tissue concentration
above MIC in bones. The control of the infection
is the main target to reach the ultimate goal of
fracture healing (Mulon, 2010). In this present
case I organisms were identified in the wound and
inj. streptopenicillin was used. Administration of
antioxidant vitamins A, E and C could accelerate
bone healing after long bone fixative surgery
(Sandukji et al., 2011). In this present case apart
from calcium and phosphorus supplementation,
administration of vitamins AD3E would help in
The wound at the medial site was dressed

using 5% povidone iodine and Ringers lactate
solution and bandaged. Cotton padding was
provided from stifle to fetlock joint except at the
wound area. The fracture was stabilized using a
fabricated PVC splint. Fabrication of PVC splint
was done as per the following procedure. A 10
mm thickness, 4 diameter PVC pipe was cut at
75 cm length. Then the PVC pipe was divided
into two halves and a 5 cm rectangular piece was
removed from the medial side of the splint. This
window provision was done for the daily dressing
of the wound. Then the splint was heated at fire
and fabricated according to the shape of the hind
limb from stifle to hoof. The fabricated PVC splint
is shown in Figure 3. Over the cotton padding
the fabricated PVC splint was applied. Fractured
fragments were aligned anatomically and adhesive
tape was applied over the fabricated PVC splint and
bandaged. The bandage was covered with parceling
tape to keep the bandage dry despite urine spillage.
Daily wound dressing was carried out using 5%
povidone iodine and bandaged for 40 days. Inj.
streptopenicillin 5 gm i/m was given for 10 days.
Supplements like Inj.vitamin AD3E 10 ml i/m
and Inj.phosphorus 10 ml i/m were administered
once weekly for 4 weeks . Oral calcium 60 ml
was advised for 40 days. Animal was kept under
complete rest. On day 40, the wound had healed
completely (Figure 4). The animal started mild
weight bearing from day 20. Partial weight bearing
was observed on day 40. Complete weight bearing
of the fractured limb was noticed on day 90.The
fabricated PVC splint removed on day 90.
The compound fracture is one type of
fracture wherein there is communication between
the fracture site and outer skin wound. Due to limited
earlier bone healing. So, compound fracture in

buffaloes can be successfully treated using the
above protocols.
247
Figure 1. Buffalo lying down with fractured hind limb before treatment.
Figure 2. Fractured hind limb (white arrow) indicates wound at the medial side of fractured area.
248
Figure 3. Fabricated PVC splint (white arrow indicates window provision for daily wound dressing).
Figure 4. Status of the animal after 40 days of treatment with partial weight bearing.
249
REFERENCES
Anderson, D.E. and G. St. Jean. 2008. Management
of fractures in field settings. Vet. Clin. North
Am. Food Anim. Pract., 24: 567-582.
Ayaz, M.M. 2000. Successful repair of metacarpal
fracture in a buffalo using a novel gadget.
Pak. Vet. J., 20: 49-50.
Mulon, P.Y. 2011. Management of open fractures
in
cattle.
www.acvs.org/symposium/
proceedings 2011/ data/papers/222/pdf:
672-673.
Nayak, S. and S. Samantara. 2010. Management
of open facture and dislocation in bovines
without apmuation. www.vetscan.co.in., 5:
68.
Sandukji, A., H. Al-Sawaf, A. Mohamadin,
Y. Alrashidi and S.A. Sheweita. 2011.
Oxidative stress and bone markers in plasma
of patients with long-bone fixative surgery:
role of antioxidents. Hum. Exp. Toxicol., 30:
435-442.
Turner, A.S. 1984. Large animal orthopaedics,
p. 816-821. In Jennings, Jr. P.B. (ed). The
Practice of Large Animal Surgery, Vol 2.
W.B. Saunders Co, Philadelphia. USA.
250
Original Article
PREVALENCE OF GASTROINTESTINAL PARASITES IN

BUFFALOES (BUBALUS BUBALIS) IN AND AROUND TIRUPATI, INDIA
C. Sreedevi1 and Md. Hafeez2
ABSTRACT
gastrointestinal parasitic infections in buffaloes in

and around Tirupati.
The prevalence of gastro-intestinal

parasites of buffaloes was studied in and around
Tirupati of Andra Pradesh for a period of one
year through coprological examination. A total
of 694 buffaloes were examined; among them,
279 (40.20%) buffaloes were found infected with
one or more species of gastro-intestinal parasites.
Ten species of gastro-intestinal parasites were
identified; of them, seven species were helminthes,
namely, Amphistome, Fasciola, Strongyles,
Strongyloides, Toxocara, Trichuris, Moniezia spp.
and two species were protozoa, namely, Buxtonella,
Eimeria and Entamoeba spp. Amphistomes were
predominant (15.42%) followed by Strongyles
(6.19%). Among the gastro-intestinal parasites
mixed infection was common (3.17%). In this
study, prevalence of parasites in relation to age and
seasonal dynamics was also studied. Significantly
(p<0.05) higher prevalence of gastro-intestinal
parasites was observed in the rainy season
followed by the summer and winter seasons. In
the age groups, adults (above 1 year) were most
(p<0.05) susceptible to gastro-intestinal parasites.
Keywords: gastrointestinal parasites, buffalo,

prevalence, age, season
INTRODUCTION
Bubalus bubalis (buffalo) is one of the
most important species of domestic livestock as a
source of dairy, meat, manure and drought power
and plays an important role in Indian rural economy.
In India, the majority of small and marginal farmers
are more dependent on buffaloes than cattle for their
livelihood as they also serve as an insurance against
the risk of crop failure due to natural calamities
(Dhanda, 2004). The Food and Agriculture
Organization (FAO, 2000) has termed the buffalo
as an important but an asset undervalued. Buffalo
diseases have been considered as one of the major
constraints for the development of the industry in the
developing countries causing substantial economic
loss to poor subsistence farmers (Jithendran and
Bhat, 1999). The parasitic diseases, gastro-intestinal
helminthiasis, coccidiosis, fasciolosis and mange are
not less important in buffaloes than other infectious
diseases (Griffiths, 1974). Epidemiological survey
Based on the results on the prevalence of gastrointestinal parasites in buffaloes it is obligatory to

follow integrated strategies and measures to control
Department of Veterinary Parasitology, NTR College of Veterinary Science, Gannavaram. Andhra Pradesh,
India, E-mail: sdevichennuru@rediffmail.com
2
Department of Veterinary Parasitology, College of Veterinary Science, Sri Venkateswara Veterinary
University, Tirupati, Andhra Pradesh, India
251
of parasitic infection is an important aid to combat

infections more effectively and in controlling
economic losses by adopting effective control
measures. The incidence of G.I. parasites in cattle
and buffaloes from different parts of India has been
published from time to time for this purpose (Sanyal
et al., 1992; Pal et al., 2001; Muraleedharan, 2005;
Ashutosh Wadhwa et al., 2011.). The present study
was undertaken to study the prevalence of common
The prevalence of various gastrointestinal

parasitic infections in buffaloes is summarized
in Table 1. Faecal examination of 694 buffaloes
revealed 40.20 percent incidence of parasitic
infections. Jagannath et al. (1988) and Hirani
et al. (1999) also reported 42.12 percent and
38.86 percent of incidence of gastrointestinal
helminthiasis in buffaloes in Karnataka and Gujarat,
respectively. Whereas Muraleedharan (2005)
and Ashutosh Wadhwa et al. (2011) recorded the
lower percentages of 20.45 percent in Karnataka
and 13 percent in Rajastan, respectively. Slightly
higher prevalence was observed by Anish yadav et
al. (2004) and Mamun et al. (2011) who recorded
60.51% and 61.02% in Jammu and Bangladesh.
Low and high prevalence of gastrointestinal
helminthiasis from different parts of India could
be due to the deworming of buffaloes and the
managemental practices followed in the particular
area. A significant (P<0.05) relationship between
age group and incidence of parasitism was observed
in the present study. The results indicated higher
prevalence rate in adults than young buffaloes
(Anish yadav et al., 2004); this could be due to
the grazing of adult animals in and around marshy
lands that were infested with infective stages of
parasites. Contrary to this Samanta and Santra
(2009) reported high prevalence rate in the belowone-year age group.
Amphistome, Fasciola sp., Strongyles,
Strongyloides sp., Toxocara sp., Trichuris sp.,
Moniezia sp., Buxtonella sp., Eimeria sp., and
Entamoeba sp. were recorded in the studied
geographical area. The helminthic infection was
30.54 percent followed by protozoa (6.48%). The
prevalence of amphistomes recorded was higher
(15.42%) than other common parasitic infections.
G. I. parasites in and round Tirupathi, Andhra

Pradesh, India.
MATERIALS AND METHODS

Six hundredninety-four buffaloes from six
surrounding villages and organized dairy farms
located in and around Tirupati were randomly
selected for the study. The faecal samples were
collected as per standard procedure throughout
the year for a one-year period for observing the
G. I. parasites in different seasons viz. summer
(March-June), monsoon or rainy (July-October)
and winter (November-February). The faecal
samples were examined by direct microscopic
method and concentration methods (sedimentation
and floatation) and samples positive for helminth
eggs and protozoan cysts/oocysts were specifically
identified and counted (Soulsby, 1982). Samples
positive for coccidian oocysts were preserved in
2.5% potassium dichromate solution for species
specific identification. The quantum of infection
among the animals was derived in terms of
percentage positive of the total samples examined.
Data obtained were classified according to age and
season and were analyzed as per standard statistical
techniques (Snedecor and Cochran, 1980).
252
253
Grand
total
Summer
Winter
Monsoon
Season
76
117
198
391
20
24
87
131
19
63
90
172
115
204
375
694
< 1 yr
1-3 yr
3 yr
Total
< 1 yr
1-3 yr
3 yr
Total
< 1 yr
1-3 yr
3 yr
Total
< 1 yr
1-3 yr
3 yr
Total
%
Infection
No. of
samples
examined
Age
group
Amphistome
15.42
1
24
45
70
3
12
15
6
16
22
1
33
73
107
Fasciola
2.02
2
6
8
2
2
2
2
4
4
10
14
Strongyles
6.19
10
18
28
1
2
3
6
1
4
4
9
2
16
25
43
Strongyloides sp.
2.59
2
4
9
15
1
1
1
3
3
7
4
2
12
18
1.44
8
8
1
1
1
1
10
10
Toxocara sp.
Trichuris sp.
1.58
3
4
7
1
1
1
2
3
1
6
4
11
Moniezia sp.
11
44
83
138
3
7
17
27
4
18
25
47
18
69
125
212
Total
0.57 30.54
1
1
2
1
1
1
1
3
1
4
Balantidium sp.
4.89
8
13
21
1
5
6
3
4
7
12
22
34
Protozoa
1.15
4
1
5
1
1
2
1
1
6
2
8
Eimeria sp.
Helminthes
0.43
1
1
2
1
1
1
2
3
Entamoeba sp.
Table 1. Seasonal and age wise prevalence of gastrointestinal parasites of buffaloes.
Total
6.48
5
10
13
28
1
3
5
9
1
3
4
8
7
16
22
45
3.17
3
1
4
8
1
2
5
8
2
1
4
6
6
4
12
22
Mixed
40.20
19
55
100
174
5
12
27
44
7
22
32
61
31
89
159
279
Total
positive
samples
Similar findings on the prevalence of higher

percentages of amphistome infections in bovines
have been recorded from Gujarat, Kashmir and
Bangladesh (Hirani et al., 1999; Anish yadav et al.,
2004; Mamun et al., 2011). Buffaloes are usually
exposed to a higher risk of infection with snailborne helminthes due to their tendency to seek
rivers, pools or swamps for wallowing (Cockrill,
1974).
stages on pasture. Sanyal and Singh (1995) also

indicated an increased parasitic burden in hosts
and pastures during the rainy season, based on
nationwide survey on parasitic epidemiology in
dairy animals in seven different agro climatic zones
of India. With the onset of winter, the infection
rate gradually decreased, agreeing with the report
of Samanta and Santra (2009). The results of the
present study revealed that the prevalence of gastrointestinal parasites in buffaloes is very common
and quite severe. Keeping in view these factors,
strategic treatment and control programme may
be formulated to control gastrointestinal parasitic
infections in buffaloes in Tirupati and elsewhere in
Andra Pradesh.
Mixed infections of Toxocara vitulorum,

Strongyloides papillosus and Eimeria sp. were
observed in the below-one-year age buffaloes
as pointed out by Bharkad et al. (1999), but ova
of Amphistomes and cysts of Buxtonella were
observed only in animals above one year of age.
Mixed infection with ova of Toxocara sp. and
Strongyloides sp. could be due to their common
prenatal and transmammary route of infection. In
addition Toxocara vitulorum ova were exclusively
identified in below-one-year age buffaloes as this
parasite is spontaneously expelled at the age of up
to 4 months (Usharani Devi et al., 2000). Absence
of toxocarosis in aboveoneyear age animals might
be due to the arrested larval development resulting
in non-patent infection. The infection rates of
Moniezia sp. (0.57%) and Eimeria sp. (1.15%)
were similar to the report of Muraleedharan (2005)
and were quite low compared to the report from
ACKNOWLEDGEMENT
The authors are thankful to the Associate
Dean, College of Veterinary Science, Tirupati for
the facilities provided.
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Gastrointestinal parasitic infestation profile
of bovines at R.S. Pura, Jammu. J. Vet.
Parasitol., 18(2): 167-169.
Ashutosh Wadhwa, R.K., Tanwar, L.D. Singla, S.
Eda, Naveen Kumar and Yogesh Kumar.
2011. Prevalence of gastrointestinal
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Bharkad, G.P., P.D. Deshpande and B.W. Narladkar.
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Seasonal effect on prevalence of G. I.
parasites in the present study revealed that the
infection rate was significantly higher (P<0.05)
during the rainy season (44.50%) followed by the
summer (35.46%) and winter (33.58%) seasons
similar to the observations reported by Mamun et
al. (2011). The environmental conditions of this
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and buffalo-calves in and around Ranch,
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Hansani, L.G. Kathiria and P.V. Patel. 1999.
Prevalence of gastrointestinal parasitic
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Rahman. 1988. Gastrointestinal parasites
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West Humid Himalayan Region of India
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(Bubalus bubalis) in Kurigram district of
255
Original Article
HAEMATO-BIOCHEMICAL EFFECTS OF ORAL SUB-CHRONIC CYPERMETHRIN

TOXICOSIS IN BUFFALO CALVES
Shabir Ahmad Dar, Bibhuti Ranjan and Rajdeep Kaur*
due to their high efficacy against target species,

their relatively low mammalian toxicity and rapid
biodegradability. Synthetic pyrethroid pesticides
account for over 30% of the global pesticide use
and these are now being preferentially used in
place of organophosphates and organochlorines
ABSTRACT
Cypermethrin, a type II synthetic
pyrethroid insecticide, at a dose rate of 0.5 mg/kg/
day for 14 consecutive weeks, produced mild signs
of toxicity in buffalo calves. Repeated exposure to
cypermethrin produced a significant increase in the
plasma levels of lactate dehydrogenase (15.4%),
gamma-glutamyl transpeptidase (18.7%), aspartate
aminotransferase (13.5%), blood urea nitrogen
(75%) and plasma creatinine (33.9%). It also
produced significant decrease in the levels of total
proteins (4.9%), haemoglobin concentration (9.5%),
packed cell volume (8.9%) and total erythrocytic
count (14.3%), but, there was a significant increase
in the erythrocyte sedimentation rate (6.6%).
(El-Tawil and Abdel-Rahman, 2001).

The toxicity of pyrethroid insecticides to
mammalian animals has received much attention
in recent years because animals exposed to these
insecticides exhibit changes in their physiological
activities besides other pathological features.
Cypermethrin is a synthetic type II pyrethroid.
It is widely used as an insecticide in developing
countries for controlling pests and in various
agricultural practices (Usmani and Knowles, 2001).
Due to the central role played by the liver in the
detoxification of cypermethrin, there is a tendency
for its accumulation and subsequent toxicity to the
liver, disrupting the normal hepatic functioning.
Low levels of chronic exposure to agricultural
chemicals may not have clinically recognizable
symptoms but could produce subtle cumulative
effects that eventually affect the health of an
organism. The potential hazard due to pesticide
residues on the health of livestock is a growing
concern.
Although extensive research work is being
done on various aspects of synthetic pyrethroids,
including their metabolism, pharmacological
Keywords: buffalo calves, cypermethrin, oxidative

stress, insecticide, toxicity
INTRODUCTION
In the present world scenario, insect
pest management has gained impedus, leading
to constant evolution of pesticide technology
as a result of which newer and safer compounds
are being developed for various agricultural and
veterinary practices. Pyrethroid pesticides have
gained popularity over other conventional pesticides
Department of Veterinary Pharmacology and Toxicology, College of Veterinary Science, Guru Angad Dev
Veterinary and Animal Sciences University, Ludhiana - 141 004, India, *E-mail: rajdeepkaur@gadvasu.in
256
(LDH), plasma gamma-glutamyl transpeptidase

(GGT), aspartate aminotransferase (AST), alanine
aminotransferase (ALT), blood urea nitrogen (BUN),
plasma creatinine and total proteins were estimated
using Bayer Autopak kits on a Photometer 5010
(Nicholas Piramal). In addition, the haematological
parameters like erythrocyte sedimentation rate
(ESR), packed cell volume (PCV), haemoglobin
concentration, total erythrocyte count (TEC), total
leucocyte count (TLC), differential leucocyte count
(DLC), mean corpuscular volume (MCV), mean
corpuscular haemoglobin concentration (MCHC)
and mean corpuscular haemoglobin (MCH) were
estimated by the methods described by Benjamin
(1985). Statistical analysis was done using an
SPSS 16.0 software package.
characteristics, ecotoxicity and detection of residues,

little attention has been paid to their biochemical
effects in nontarget species. Although some work
on the toxicity of cypermethrin has been done in
different animals, there is limited information
available regarding its toxic mechanisms in buffalo
species. So, in order to better understand the
mechanisms involved in cypermethrin toxicity, it
was thought pertinent to investigate its ability to
modulate various biochemical and haematological
parameters in buffalo calves.

The experiments were performed on eight
healthy male buffalo calves of 6-12 months of
age and weighing between 60-120 kg, procured
from the University Dairy Farm and local market.
The animals were acclimatized in the animal
shed of department under uniform conditions for
2 weeks prior to the commencement of study.
The animals were dewormed, fed seasonal green
fodder and wheat straw and had access to water
ad libitum. Permission to conduct the experiment
was duly obtained from the University Animal
Ethics Committee. The animals were randomly
divided into two groups of four animals, each.
Animals of Group 1 served as healthy control
whereas Group 2 animals were orally administered
cypermethrin at a dose rate of 0.5 mg/kg/day for 14
consecutive weeks. Blood samples were collected
in heparinized vials via jugular venipuncture at
weekly intervals during the treatment period and on
2nd week after the withdrawal of treatment. Levels
of cypermethrin were analyzed in the plasma of
animals by gas chromatography using an electron
capture detector (Gill et al., 1996). Various
parameters viz. plasma lactate dehydrogenase
RESULTS
Oral administration of cypermethrin at the
dose rate of 0.5 mg/kg/day for 14 consecutive weeks
resulted in an increase in plasma cypermethrin
concentration for the initial four weeks, and
thereafter, the cypermethrin concentration declined
slowly (Figure 1). The initial incline could be
due to continuous exposure of cypermethrin,
and thereafter, the decline in cypermethrin
concentration may be attributed to the induction of
the metabolizing enzymes or decrease of plasma
protein levels as observed in the present study.
Cypermethrin
exposure
produced
significant elevation in the enzymatic activity
of lactate dehydrogenase and gamma-glutamyl
transpeptidase from the 12th week onwards (Table
1). This elevation in LDH and GGT was observed
to be 16.3% and 23.6%, respectively, after 14 weeks
of insecticide exposure. Similarly, the enzymic
activity of aspartate aminotransferase increased
257
significantly from the 12th week onwards, elevating
weeks post treatment. These findings are in league

with those reported by Padma and Ashok (2010) in
rats, Nagarjuna et al. (2008) in rabbits and Khan et
al. (2009) in goats, exposed to pyrethroids.
Diseased or damaged kidneys cause an
elevated BUN because the kidneys are less able
to clear urea from the bloodstream. Elevated
blood urea is correlated with an increased protein
catabolism in the mammalian body (Murray et
al., 1990). Increase in plasma creatinine and
BUN levels probably indicate renal damage,
which may be attributed to urinary obstruction,
which potentiates decreased secretion of urea
from the body. Decrease in protein levels of
plasma are usually seen in chronic liver diseases
due to impaired synthesis and in renal diseases
due to loss of proteins. Hypoproteinemia is seen
in inflammations and is due to increased protein
catabolism during stress. Leakage of proteins is
also observed during glomerulonephritis. The
decrease in protein levels of plasma in the present
investigation could be attributed to impairment of
protein synthesis following chronic exposure to
cypermethrin.
Long term oral exposure to cypermethrin
(0.5 mg/kg; 14 weeks) resulted in significant changes
in the blood profile of exposed animals. This was
manifested by a significant decrease in haemoglobin
(18.7%) and packed cell volume (11.7%) by the
14th week of treatment, as depicted in Table 3. The
by 14.9% by the end of treatment period (Table

1). On the other hand, no significant changes were
observed in the activity of alanine aminotransferase
in cypermethrin exposed animals (Table 1). These
findings are consistent with those reported by Aslam
et al. (2010), Khan et al. (2009) and Remya et al.
(2010) in poultry, goats and rats, respectively.
The increase in plasma LDH activity in
present investigation reflects damage to a range of
tissues including skeletal, cardiac muscles, kidney
and liver. Although the exact cause of increased
GGT level in the present study could not be
ascertained, yet cholestatic disorders of all species
are associated with increased GGT activity (Braun
et al., 1987). Aspartate aminotransferase is a not
an organ specific enzyme; however, as the cardiac
and skeletal muscles have high concentrations of
aspartate aminotransferase (Brar et al., 2000), its
elevation in present investigation suggests marked
muscular damage. Alanine aminotransferase is
employed as a marker of hepatocellular damage in
various species and is usually elevated in disorders
of inflammatory, toxic or degenerative origin (Dial
1995). Leakage of this enzyme into the blood occurs
when there is damage to hepatocytes, so its level
increases in plasma. In large domestic species, the
activity of ALT in the liver is low and hence during
liver injury, the ALT is not remarkably elevated
(Tennant 1997).
Cypermethrin
produced
significant
increase in the levels of blood urea nitrogen
and plasma creatinine to the extent of 75% and
33.9%, respectively by the 12th week of insecticide
levels however returned to normal within 2 weeks

post-treatment. Similarly, a significant decline in
TEC was observed in insecticide exposed animals
from the 10th week onwards, declining by 16.9%
by the 14th week of treatment (Table 3). In contrast,
there was a significant increase in ESR from the
8th week onwards reaching a maximum of 8.02%
by the 14th week of cypermethrin exposure (Table
3). However, the values returned to normal during
exposure (Table 2). However, chronic cypermethrin

exposure resulted in a significant decline in the
total plasma proteins, to the extent of 4.9% after
the 14th week of cypermethrin exposure (Table 2).
The levels however returned to normal within 2
258
259
Treatment
1215.6 35.1
1227.0 46.4
1235.0 48.4
1256.0 55.6
1271.0 51.6
1302.4 52.4
1334.4 53.6
1344.8 64.1
1366.4 59.7
1370.2 55.9
1423.056.5**
1410.6 45.8*
1413.651.2**
1291.4 40.9
1193.449.6
1136.247.9
1174.644.6
1203.456.6
1189.356.3
1216.644.2
1235.764.6a
1214.063.8a
1256.748.9a
1236.064.5a
1233.053.6a
1224.364.3a
1228.757.6a
1222.749.5a
Lactate dehydrogenase
Control
Treatment
15.70.9a
15.01.1
14.00.8
12.80.8
14.00.8
15.71.0
15.20.7
15.00.7a
15.11.0a
14.91.0a
15.40.8a
15.00.7a
15.30.8a
15.00.9a
15.20.7
14.40.8
15.00.8
13.80.8
14.80.7
14.80.8
16.40.7
16.60.8
16.40.8
16.60.8
17.00.8
17.40.7*
17.40.7*
17.80.8*
Gamma-glutamyl
transpeptidase
Control
Treatment
123.76.5a
119.66.3
122.66.4
132.44.0
131.05.9
133.04.9
119.86.4
131.06.1a
125.010.7a
122.79.7a
124.78.2a
127.76.5a
126.37.6a
127.05.8a
134.84.9
123.28.0
121.46.3
121.48.3
125.010.2
124.08.8
133.09.2
131.29.1
139.27.2
135.47.2
137.06.1
145.07.6**
143.06.6**
141.66.8**
Aspartate aminotransferase
Control
Values given are expressed as U/L and represent the Mean S.E. of 5 animals unless stated.
a= Mean S.E. of 4 animals.
Values with superscript in a given row differs significantly from each other. (*P<0.05 and **P<0.01).
0
2
3
4
5
6
7
8
9
10
12
13
14
Post treatment
2
Treatment
Time
(weeks)
Treatment
35.03.1a
30.21.2
29.42.6
30.05.4
33.84.8
35.04.6
33.22.6
35.83.1a
34.06.3a
37.05.7a
35.35.8a
34.76.0a
34.04.3a
33.05.6a
37.23.3
31.21.6
31.03.0
33.63.0
39.05.1
39.25.1
33.05.3
38.63.4
41.03.4
44.63.9
43.46.2
45.25.6
42.25.2
37.45.1
Alanine aminotransferase
Control
Table 1. Effect of repeated oral administration of cypermethrin 0.5 mg/kg/day on plasma lactate dehydrogenase, gamma-glutamyl transpeptidase,
aspartate aminotransferase and alanine aminotransferase in buffalo calves.
260
1.270.08
1.190.08a
4.00.6a
5.40.3
Treatment
Creatinine
1.140.06
1.200.05
1.200.07
1.130.08
1.130.05
1.220.09
1.180.06
1.230.11
1.110.06
1.270.12
1.240.06
1.230.08
a
1.160.06
1.270.06
a
1.250.08
1.300.10
a
1.150.10
1.410.11
a
1.170.09
1.450.10**
1.120.10a
1.500.12**
1.210.08a
1.490.11**
1.160.10a
1.500.12**
Control
Control
Treatment
Blood urea nitrogen
3.20.4
3.60.7
4.20.8
3.80.4
3.20.7
3.20.5
3.60.5
3.20.4
2.80.3
2.60.3
3.20.4
3.80.4
a
2.50.6
3.40.9
a
4.30.3
5.41.6
a
4.31.5
6.70.4**
4.70.7a
7.20.6**
4.00.6a
7.00.5**
4.30.3a
7.40.5*
4.00.6a
7.11.0**
Values given are expressed as mg/dl and represent the Mean S.E. of 5 animals unless stated.
Time (weeks)
Treatment
0
2
3
4
5
6
7
8
9
10
12
13
14
Post-treatment
2
7.110.09a
7.110.10
Control
Treatment
Total proteins
7.210.18
7.170.19
7.110.17
7.060.17
7.110.21
7.140.22
7.160.18
7.090.18
7.050.15
7.010.17
7.070.16
7.080.15
a
7.020.22
6.980.11
a
7.100.20
6.910.13
a
7.040.19
6.900.25
a
7.030.17
6. 810.12
a
7.030.15
6.760.15
a
7.080.16
6.750.11*
7.080.14a
6.730.13*
Table 2. Effect of repeated oral administration of cypermethrin 0.5 mg/kg/day on blood urea nitrogen, creatinine and total proteins in buffalo calves.
261
37.620.64
38.051.08a
8.790.25a
8.850.27
Control
Treatment
Packed cell volume
38.461.19
38.550.56
38.290.78
38.211.12
37.820.34
37.870.46
37.690.42
37.540.43
36.660.81
36.560.68
37.490.45
36.720.60
a
37.030.83
35.860.54
a
36.830.51
34.820.79**
37.400.85a
34.380.95**
37.100.62a
34.311.0 **
37.871.07a
34.330.87**
37.371.20a
34.040.95**
37.580.95a
34.511.03*
Control
Treatment
Haemoglobin
8.960.29
9.100.31
8.940.40
9.120.21
8.750.31
9.030.32
8.840.21
9.010.24
8.900.28
8.840.35
8.900.34
8.710.31
a
8.990.30
8.830.30
a
9.150.26
8.360.27
a
8.860.28
8.240.31
a
8.880.25
8.150.30**
8.870.23a
8.070.22**
8.650.28a
7.940.25**
8.730.23a
7.900.31**
131.71.5a

133.81.4
Control
Treatment
Erythrocyte sedimentation rate
130.21.8
129.61.2
131.01.4
131.21.9
131.82.7
129.62.9
130.01.6
132.21.7
131.61.4
133.21.8
132.21.6
135.62.1
a
131.02.3
134.21.6
a
133.02.1
135.21.4
a
131.31.8
137.41.7**
133.01.7a
139.01.7**
131.72.6a
139.82.3**
132.02.6a
139.02.2**
131.32.9a
140.02.5*
Values given are expressed as g/dl and represent the Mean S.E. of 5 animals unless stated.
Time (weeks)
Treatment
0
2
3
4
5
6
7
8
9
10
12
13
14
Post-treatment
2
5.050.24a
5.110.26
Control
Treatment
Total erythrocytic count
5.080.29
5.210.12
5.080.13
5.180.29
5.050.21
5.220.25
5.060.23
5.140.22
5.030.16
5.240.25
5.100.22
5.030.27
a
5.110.20
4.910.26
a
5.090.17
4.740.24
a
5.050.21
4.670.23
a
5.130.18
4.670.18*
5.000.22a
4.490.25*
5.030.19a
4.310.19**
5.030.21a
4.380.21**
Table 3. Effect of repeated oral administration of cypermethrin 0.5 mg/kg/day on haemoglobin, packed cell volume, erythrocyte sedimentation rate and
total erythrocytic count in buffalo calves.
Treatment
Treatment
262
75.934.74a
10580852.3
75.537.26
74.344.55
73.715.02
72.526.59
73.013.14
71.632.56
72.603.49
71.823.84
70.711.88
71.532.20
71.550.72
74.053.34
73.928.17
74.293.29
Treatment
17.671.47a
17.630.90
17.590.58
17.321.56
17.661.33
17.680.93
17.441.12
17.920.56a
17.360.59a
17.551.08a
17.330.62a
17.740.87a
17.190.66a
17.340.53a
17.610.89
17.291.94
17.590.85
17.590.44
17.270.57
17.330.57
17.210.62
17.420.95
16.980.43
17.151.00
17.141.21
17.410.59
17.391.78
17.010.41
Mean corpuscular
haemoglobin
Control
Treatment
23.220.53a
23.150.85
23.351.63
23.142.35
23.721.08
24.280.61
23.741.14
24.721.14a
23.90.81a
23.681.07a
23.940.02a
23.441.09a
23.150.19a
23.280.67a
23.591.65
23.251.074
23.861.52
23.862.13
23.561.83
24.190.66
23.711.15
24.250.80
24.021.18
23.971.25
23.951.58
23.511.23
23.530.37
22.901.20
Mean corpuscular
haemoglobin concentration
Control

Values given are expressed as 106/mm3 for total erythrocytic count and as mm3 and represent the Mean S.E. of 5 animals unless stated.
76.152.23
75.313.00
74.833.27
74.453.03
72.823.10
73.462.12
72.521.81a
72.360.79a
74.111.51a
72.382.63a
75.682.10a
74.252.55a
74.933.16a
Mean corpuscular
volume
Control
10420671.1
10820674.1
9944752.3
11680687.0
10560933.1
10320772.9
10760855.9
10560605.5
9240640.8
9080336.8
10930913.5
11170592.5
9560613.8
Total leukocytic count
Control
0
10590613.8
2
11000528.0
3
10700637.3
4
10760568.7
5
10920862.0
6
10600911.7
7
11700810.7a
8
11733712.7a
9
8850859.8a
10
8953535.6a
12
12366856.8a
13
11666529.2a
14
9266581.2a
Post-treatment
2
10625632.5a
Treatment
Time
(weeks)
Table 4. Effect of repeated oral administration of cypermethrin 0.5 mg/kg/day on total leukocytic count, mean corpuscular volume, mean corpuscular
haemoglobin and mean corpuscular haemoglobin concentration in buffalo calves.
263
5.20.7
4.80.5
4.60.4
3.80.4
3.81.0
3.80.7
3.50.6a
3.50.3a
3.70.9a
3.30.3a
6.30.9a
4.01.0a
50.6a
4.30.9a
69.47.9
63.25.8
63.47.1
66.02.4
59.86.2
62.24.1
61.53.3a
64.52.3a
62. 73.2a
64.31.9a
65.013a
72.74.3a
67.02.1a
65. 72.4a
Control
M
1. 0.3a
2.20.6
3.00.4
2.80.6
1.80.4
1.80.8
1.80.6
1.80.3a
1.50.3a
1.70.3a
1.70.3a
2.30.0a
0.00.3a
2.0 0.3
0. 70.3a
1.00.3
0.80.2
1.20.4
0.60.24
0.40.24
0.60.24
0.50.3a
0.50.5a
0.00.0a
0.70.3a
0.00.0a
0.0.0.0a
0.00.0a
26.61.1
23.26.1
202. 6
21.82.8
22.83.7
24.83.4
26.42.3
24.81.7
27.21.6
23.84.4
25.42.9
23.26.1
27.41.3
22.61.8
67.21.5
68.66.0
70.61.9
71.41.8
70.43.4
68.83.9
66.62.7
68.42.4
65.81.9
69.84.7
67.82.9
69.25.9
68.01.4
71.01.8
4.60.4
4.80.6
4.80.4
3.80.6
4.20.4
4.00.3
4.40.6
4.20.6
4.20.4
4.60.7
4.6.2
4.40.5
2.80.5
4.20.6
Treatment
M
1.60.2
2.80.4
3.60.5
2.60.7
2.00.3
2.00.5
2.00.4
2.20.4
2.00.3
1.40.2
2.00.3
2.60.4
1.40.2
2.00.3
0.00.0
0.60.2
1.00.4
0.40.2
0.60.2
0.40.2
0.60.4
0.40.2
0.80.4
0.40.2
0.20.2
0.60.2
0.40.2
0.20.2
Values given are expressed as % and represent the Mean S.E. of 5 animals unless stated, N, L, M, E, and B stads for neutrophils, lymphocytes,
monocytes, eosinophils and basophils respectively.
Treatment
0
22.27.5
2
28.25.8
3
28.07.1
4
27.83.2
5
34.26.1
6
31.63.9
7
32.73.2a
8
30.02.2a
9
32.02.1a
10
30.01.0a
12
26.39.7a
13
22.73.6a
14
25.71.6a
Post-treatment
2
27.72.1a
Time
(weeks)
Table 5. Effect of repeated oral administration of cypermethrin 0.5 mg/kg/day on differential leukocyte count in buffalo calves.
Figure 1. Serum cypermethrin concentration in buffalo calves after oral administration of cypermethrin 0.5
mg/kg/day.
two weeks post-treatment. However, cypermethrin
exposure failed to produce any significant changes
in total leukocytic count (TLC) and differential
leukocytic count (DLC) (Table 4 and Table 5). Table
4 further illustrates that cypermethrin produced
no significant changes in any of the erythrocytic
indices i.e. mean corpuscular haemoglobin,
mean corpuscular volume and mean corpuscular
haemoglobin concentration. These observations
are in agreement with those reported by Remya et
al. (2010) in rats, Summaira et al. (2010) in broiler
chicks, Khan et al. (2009) in goats and Yousef et al.
(2003) in rabbits intoxicated with cypermethrin.
Significant reduction of haemoglobin
content during the later period of the investigation
could be related to decreased synthesis of red
blood cells in bone marrow (Mandal et al., 1986),
or reduced biosynthesis of heme in bone marrow
(Khan et al., 2009) or as a result of increased rate
of erythrocyte destruction. This could probably

be explained by the effect of cypermethrin on
erythropoiesis. The subsequent recovery of
haemoglobin after termination of cypermethrin
treatment indicates that the damage produced was
of reversible nature. Consonantly, the decrease in
PCV could be attributed to the decrease in TEC.
The decrease in TEC, Hb concentration and PCV
observed in this study could be the result of disruptive
action of cypermethrin on the erythropoietic tissue
as a result of which the viability of the cells might
have been affected.
Therefore, on the basis of the observations
of the present investigation, it can be concluded
that chronic exposure to cypermethrin induces
significant biochemical and haematological
alterations in buffalo calves.
264
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Rodwell. 1990. Harpers Biochemistry, 22nd
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and P. Doss. 2008. Alteration in protein
metabolic profiles in gastrocnemius muscle
tissue of rats during cypermethrin toxicosis.
Journal of the Indian Society of Toxicology,
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Padma, S. and K.S. Ashok. 2010. Cypermethrin
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Remya, R. Nair, Mammen J. Abraham, C.R.
Lalithakunjamma, N.D. Nair and C.M.
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Summaira, S., A. Khan, M.Z. Khan, F. Aslam,
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Pharmacol. Res., 44: 33-40.
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Development of a method for the analysis
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Khan, Ahrar, Hafiz A.M. Faridi, Muhammad Ali,
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266
Original Article
INCIDENCE OF PARTHENOGENETIC DEVELOPMENT OF

BUFFALO (BUBALUS BUBALIS) OOCYTES
Ch. Srinivasa Prasad1, A. Palanisamy2, S. Satheshkumar2, V.S. Gomathy2 and S. Rangasamy3
ABSTRACT
rate is much lower in buffaloes, due to their inherent

poor developmental competence resulting in poor
cleavage and embryo development. Recent reports
suggested that in vitro matured buffalo oocytes had
better inherent developmental competence. Oocyte
maturation can be defined as those events associated
with the initiation of germinal vesicle breakdown
(GVBD) and completion of the first meiotic division
(Leibfried-Rutledge et al., 1987). Maturation allows
the oocyte to express its developmental potential
after fertilization and is not merely confined
to nuclear events or the ability to be fertilized
(Gordon, 2003). Maturation media supplemented
with hormones, serum, growth factors etc. improve
the in vitro developmental competence of oocytes.
Oocyte maturation triggered by hormone signaling
alters membrane permeability, causes ion currents
and increases cytosolic free calcium [Ca2+]i. At the
The incidence of parthenogenetic

development of oocytes in routine IVF practices i.e.
without any chemical stimuli, was assessed. After
24 h of maturation while washing and transferring
to fertilization medium nine (1.39%) oocytes out
of 646 oocytes were found cleaved. All those
parthenotes (cleaved oocytes) were transferred to
embryo culture medium instead of fertilization
medium for observing further development. It was
found that three (33.33%) out of nine were arrested
at the four-cell stage and the remaining (66.67%)
were arrested at the two-cell stage itself.
Keywords:
buffaloes,
parthenogenesis, oocytes
Bubalus
bubalis,
end of maturation, the oocyte reaches an adequate

Ca2+ store.
INTRODUCTION
In buffalo the overall in vitro embryo
production efficiency is lower than in cattle, mainly
due to the lower cleavage rate (Gasparrini, 2002). In
vitro fertilization procedures in water buffalo were
derived from those in cattle. However, the success

Ovaries from sexually mature buffaloes
(Bubalus bubalis) were collected irrespective of age,
Department of Veterinary Physiology, Madras Veterinary College, Chennai - 600 007, India, E-mail:
spchigurupati@rediffmail.com
2
Centralized Embryo Biotechnology Unit, Department of Animal Biotechnology, Madras Veterinary
College, Tamilnadu Veterinary and animal Sciences University, Chennai - 600 007, India
3
Department of Animal Reproduction, Gynaecology and Obstetrics, Madras Veterinary College, Chennai 600 007, India
267
percent of oocytes were classified as Grades A, B,

C, D and E (expanded oocytes) respectively. After
to fertilization medium nine (1.39%) oocytes out
of 646 oocytes were found cleaved. All those
parthenotes (cleaved oocytes) were transferred to
embryo culture medium instead of fertilization
medium for observing further development. It was
found that three (33.33%) out of nine were arrested
at the four-cell stage and the remaining (66.67%)
were arrested at the two-cell stage itself. It was also
found that all the parthenotes were derived from
culture grade oocytes (A and B grade oocytes).
Chemical activation of in vitro matured
buffalo oocytes successfully supported development
to the blastocyst stage (Gasparrini, 2003). Sperm
penetration provides the natural signal for oocyte
activation. The sperm-oocyte interaction results
in the release of Ca2+ from intracellular stores,
in the form of repetitive waves or spikes that are
responsible for meiotic progression and early
development (Bootman and Berridge, 1995). All
methods of chemical activation mimic this process
by providing adequate but nontoxic calcium
signals to the MII arrested oocytes with different
mechanisms.
Ion currents and cytosolic free calcium
([Ca2+]i) elevations are crucial events in triggering
the complex machinery involved in both gamete
maturation and fertilization. Oocyte maturation is
triggered by hormone signaling which causes ion
currents and [Ca2+]i increase. Because extracellular
Ca2+ is required for in vitro GVBD (DeFelici and
Siracusa., 1982) and for first meiotic division (Paleos
and Powers, 1981), Ca2+ ion transport through the
plasma membrane seems to play a functional role
in maturation. As a consequence of a net uptake
of Ca2+ through the plasma membrane, at the end
of maturation, the oocyte reaches an adequate
body condition, stage of oestrous cycle and season

from the Chennai Corporation abattoir and utilized
in this study. The ovaries were removed within
30 minutes of slaughter and washed in phosphate
buffered saline (PBS) supplemented with 50 g/ml
gentamicin sulphate to remove blood and extraneous
material. The washed ovaries were transported at 37oC
in a thermos flask in the same media to the laboratory
within 30 minutes. The extra-ovarian tissues were
trimmed off and the ovaries were washed with
PBS to remove blood clots and superficial bacterial
contamination. The washed ovaries were kept in a
sterile beaker containing PBS supplemented with 50
g/ml gentamycin until oocyte retrieval by aspiration.
The oocytes were screened under a stereozoom
microscope, washed thrice in 35 mm petridishes and
graded based on their cumulus mass investment and
homogeneity of ooplasm as described by Nandi et al.
(1998).
All the oocytes aspirated were subjected
to in vitro maturation in TCM-199 supplemented
with FCS and hormones FSH, LH and oestardiol.
The cumulus oocyte complexes (COCs) were
rinsed three times in maturation medium and were
transferred to 100 l of IVM droplets (15- 20 COCs
per droplet). The oocytes were allowed to mature in
these droplets at 38.5oC in an atmosphere of 5 percent
CO2 in air for 24 h in a CO2 incubator. Observation
for cleavage of oocytes, if any, was made after
to fertilization medium.

By aspiration technique, 646 oocytes were
retrieved from 344 buffalo ovaries, with recovery
rate of 1.87 oocytes per ovary. 10.52 (68), 33.74
(218), 29.25 (189), 16.40 (106) and 10.06 (65)
268
Figure 1. Parthenogenetic oocyte (2-cell) with

intact cumulus mass ( X 200).
Figure 2. Parthenogenetic oocyte (2-cell) with out

cumulus mass ( X 200).
Figure 3. Parthenogenetic oocyte (4-cell) ( X 200).
Figure 4. Degenerating parthenote (X 200).
Ca2+ store. This storage may occur via either gap

junctions or specific channels. In the oocyte at
fertilization also a dramatic ion flux occurs together
with a temporally abrupt and/or spatially confined
intracytoplasmic calcium [Ca2+]i) increase (Boni et
al., 2007). Sperm-induced calcium (Ca) oscillations
at the time of fertilization are a required signal for
activation in most mammalian oocytes. A clear
relationship between electrical properties of the
oocyte plasma membrane and [Ca2+]i modifications
has also been recorded following fertilization, as
well as following chemical oocyte activation or
after exposure to specific Ca2+ releasers (Tosti et
al., 2002).
Based on the results it was assumed that in
vitro maturation might have induced calcium (Ca)

oscillations due to altered electrical properties of
the oocyte plasma membrane as that of chemical
oocyte activation by calcium ionophore resulting
in cleavage, but to a lesser extent indicating that
buffalo oocytes had better inherent developmental
competence and that the poor cleavage and embryo
development following IVF may be due partly to
the poor quality of frozen/thawed sperm, improper
sperm capacitation and/or fertilization as opined
by Mishra et al. (2006). From this study, it was
concluded that cleavage alone is not a reliable
indicator of fertilized oocytes because oocytes
matured in vitro may undergo parthenogenic
development to the four-eight cell stage (Shioya et
269
Leibfried-Rutledge, M.L., E.S. Crister, W.H.

Eyestone and D.L. Northey. 1987.
Developmental potential of bovine oocytes
matured in vivo and in vitro. Biol. Reprod.,
36: 376-383.
Mishra, V., A.K. Misra and R. Sharma. 2006.
A comparative study of parthenogenic
activation and in vitro fertilization of
bubaline oocytes. Anim. Reprod. Sci.,
103(3-4): 249-259.
Nandi, S., M.S. Chauhan and P. Palta. 1998. Influence
of cumulus cells and sperm concentration
on cleavage rate and subsequent embryonic
development of buffalo (Bubalus bubalis)
oocytes matured and fertilized in vitro.
Theriogenology, 50(8): 1251-1262.
Paleos, G.A. and R.D. Powers. 1981.The effect of
calcium on the first meiotic division of the
mammalian oocyte. J. Exp. Zool., 217: 409416.
Shioya, Y., M. Kuwayama, M. Fukushima, S.
Iwasaki and A. Hanada. 1988. In vitro
fertilization and cleavage capability
of bovine follicular oocytes classified
by cumulus cells and matured in vitro.
Theriogenology, 30(3): 489-496.
Tosti, E., R. Boni and A. Cuomo. 2002. Fertilization
and activation currents in bovine oocytes.
Reproduction, 124: 835-846.
al., 1988), where in vitro developmental block may

or may not occur.
ACKNOWLEDGEMENT
The work was carried out in a scheme on
Establishment of Buffalo ES-Cell Line funded
by the Department of Biotechnology, Government
of India. The authors are thankful to the funding
agency.
REFERENCES
Boni, R., R. Gualtieri, R. Talevi and Elisabetta
Tosti. 2007. Calcium and other ion dynamics
during gamete maturation and fertilization.
Theriogenology, 68S: S156-S164
Bootman, M.D. and M.J. Berridge. 1995. The
elemental principles of calcium signaling.
Cell, 83: 675-678.
DeFelici, M. and G. Siracusa. 1982. Survival of
isolated, fully grown mouse ovarian oocytes
is strictly dependent on external Ca2+. Dev.
Biol., 92: 539-543
Gasparrini, B. 2002. In vitro embryo production
in buffalo species: state of the art.
Theriogenology, 57(1): 237-256
Gasparrini, B., V. Caracciolo di Brienza, L.
Boccia, R. Di Palo and G. Neglia Discizia.
2003. The parthenogenetic development
of buffalo (Bubalus bubalis) oocytes after
chemical stimulation. In Proceedings of
15th Congress A.S.P.A., Parma, Iyaly. Ital. J.
Anim. Sci., 2(Suppl. 1): 139-141.
Gordon, I. 2003. Laboratory production of cattle
embryos. Biotechnology in Agriculture
No.27 , 2nd ed. p. 119.
270
Original Article
GENETIC DIVERSITY AND CONSERVATION OF ANIMAL GENETIC RESOURCES IN

IRAQI BUFFALO USING MICROSATELLITE MARKERS
Talib Ahmed Jaayid1 and Maytham Abdul Kadhim Dragh2
ABSTRACT
INTRODUCTION
In our study, conducted in Iraq and

Huazhong University, China, divided Iraq into three
main regions: a southern area including Basrah,
Missan, and Dhi-Qar, a middle area including AlQadisiyah, Babil, Karbala and Baghdad, and a
northern area including Diyala, Kirkuk and Mosul.
The aim of the study was to measure the genetic
diversity, polymorphism and heterozygosity in
The buffalo contributes effectively in the

agricultural economy and food security in the
countries of the Indian Subcontinent and South East
Asia, through meat, milk, leather and labor. It is
well known that the buffalo was domesticated very
early in history, but when and where is unknown
(Cockrill, 1974). The water buffalo emerged in East
Asia (Potts 1996) and mainland South East Asia. It
spread north and west to China and to the Indian
subcontinent (Lau, 1998). The buffalo has been
present in the valley of the Indus River in the Indian
subcontinent since about 4000-5000 years ago, but
there are areas of independent domestication of
water buffalo in Mesopotamia and China earlier
than this-about 2500-7000 BC (Haynes et al., 1991;
Payne, 1991 and Bradley, 2006). Macgregor (1939)
classified buffalo into two types according to formal
criteria and behavior: the river buffalo in the Indian
subcontinent and westerly to the Balkan region and
the swamp buffalo in Southeast Asia, India, Nepal
and northeast to the valley of the Yangtze River in
China. Barker et al. (1997) was the first to analyze
the genetic diversity of buffalo breeds in South
Asia using protein coding spatial microsatellites.
They noted a clear distinction between these two
types: river and marsh buffaloes, the latter being
raised mainly in the swamps of Southeast Asia,
and this distinctiojn was confirmed later by Zhang
Iraqi buffaloes using microsatellite techniques.

Sixty-nine blood samples were collected from
unrelated animals. Six microsatellite markers were
used (ETH125, CSSM060, BM1706, ETH02,
ETH225 and INRA005). The polymerase chain
reaction (PCR) was done using specific bovine
primers and a genetic analyzer (ABI-3730). Our
results revealed that all the six markers amplified
the DNA. The marker INRA005 did not show high
polymorphism; it only revealed three alleles (137141 bp). The marker ETH152 showed the highest
level of polymorphism; it has sixteen alleles ranged
between 192-217 bp. This study showed that there
are three main clusters: the first one included Basrah,
Baghdad and Al-Qadisiyah, the second consisted
of Kirkuk and Missan, while the third consisted of
Babil and Mosul.
Keywords: Iraqi buffalo, microsatellites, PCR,
genetic diversity
1
2
Production Department, College of Agriculture, Basrah University, Iraq, E-mail: taleb1968@yahoo.com

Department of Animal Production, College of Agriculture, Basrah University, Basrah, Iraq
271
et al. (2007). Today, almost all breeds have been

described according to geographical location and
genetic phenomena. Using the new biotechnology
in national research centers, local breeds have
been identified in China, India and Pakistan, the
Philippines, and Vietnam.In China, four types of
buffaloes were identified in accordance with the
geographical distribution (Wenping et al., 1998).
Using molecular data, Zhang et al., 2007 classified
Chinese buffalo breeds into four groupsthough
these are not commensurate with the classification
proposed by Wenping (1998). Therefore, there is a
need to determine the breeds genetically in order
to obtain correspondence between geographic and
genetic classifications of the Chinese breeds and
the Indian (Kumar et al., 2006). The discovery of
the existence of genetic differences at the level of
DNA can bring about a revolutionary strengthening
of programs of genetic improvement of this
animal. Current DNA technology can cover most
of the requirements for this purpose. Several recent
studies have been publishedon the domestic buffalo
using microsatellite markers. These include studies
of the swamp buffalo in south-east Asia (Barker et
al., 1997), in India (Kumar, 2006), and in China
(Zhang, 2007) and of the river buffalo in Greece
and Italy (Moioli, 2001). While many researchers
have investigated genetic polymorphism in the
Blood samples: Blood samples were

collected from 96 individuals from the three main
regions in Iraq (approximate 24 from the south
and 24 individuals from the north of the country
and 48 from the middle region of Iraq) all the
blood samples were from unrelated animals. Six
microsatellite markers recommended by FAO and
ISAG for domestic buffalo diversity studies were
used (Hoffmann et al., 2004). Forward primers
were end-labeled with fluorescent dyes (6-FAM,
TET and HEX). Genotypes for each marker were
determined using ABI 3730 DNA Sequencer
(Applied Biosystems) with the internal size
standard Gene-Scan (Applied Biosystems).
Data analysis
Allele frequency, the number of alleles per
locus, observed heterozygosity (HO) and expected
heterozygosity (HE) were calculated using
microsatellite toolkit. To remove the bias of sample
size, a corrected mean number of alleles (MNA)
value for each locus was obtained by computing
the means of 96 random samples of 24 individuals
for each population except buffaloes of the
middle area, which was 48 individuals. Wilcoxon
signed rank tests were conducted to examine the
significant differences on MNA and HE between
each pair of populations. Exact tests for deviations
from Hardy-Weinberg equilibrium (HWE) were
performed for each populationlocus combination
using GENEPOP version 3.4 with the P-values
obtained by the Markov Chain randomization
test. FST values per pair of populations (Weir &
Cockerham, 1984) were computed and tested
using the FSTAT program (Goudet, 2002). Two
approaches were employed to investigate the
genetic relationships among the populations. First,
buffalo, in Iraq, this issue has not been studied

enough. The present study is to fill the gap in this
important aspect and to throw light on the genetic
situation in Iraq. Further scientific objectives are
development of biodiversity restoration approaches
in agricultural biodiversity; managing agricultural
biodiversity to improve productivity and conserve
diversity and making the first stage of animal
genetic resources conservation.
272
0.869 in the ETH152 marker, while the observed

heterozygosity varied from 0.145 in the INRA005
marker to 0.916 in the CSSM060 marker. Most
loci showed heterozygosity rates above 0.5, which
means these markers are highly informative
markers (Table 2).
Relationships
among
populations:
A neighbor-joining tree on the basis of genetic
distances was constructed (Figure 1), and for the
first time in Iraq, three main clusters have been
detected in this genetic tree map. The first of these
clusters includes Basrah, Baghdad, and Qadisseah
provinces, the second consisted of the buffalo of
both Kirkuk and Missan provinces, while the third
cluster included both Babil and Mosul provinces.
The least genetic distance was between the buffalo
of Baghdad and Qadisseah provinces, which was
0.083, while the highest distance was between the
buffalo of Mosul and Babil and between the buffalo
of Mosul and Kirkuk provences, which was about
0.458.
Neis DAgenetic distances (Nei et al., 1983) were

calculated and then used to construct the neighborjoining tree using MEGA software. Secondly,
principal component analysis (PCA) with gene
frequency was conducted.
RESULTS
Genetic variability: In all 70 alleles were
detected across the six microsatellite loci .The
total number of alleles per locus (TNA) varied
from three (INRA005) to 16 (ETH152). The MNA
across thesix loci in Iraqi indigenous buffaloes was
5.98. The results of the present study showed that
all the loci used in this study amplified the DNA
of the Iraqi buffalo and showed a high level of
polymorphism except INRA005 (Table 1).
Heterozygosity: From the results of the
present study, all the six loci showed variable rate
of heterozygosity The expected heterozygosity
ranged from 0.134 in the INRA005 marker to
POP 1
POP 5
POP 6
POP 3
POP 7
POP 2
POP 4
POP 8
0.05
0.04
0.03
0.02
0.01
0.00
Figure 1. Genetic linkage map of the Iraqi buffalo.

273
Table 1. Polymorphism of 6 microsatellite loci in 3 Iraqi indigenous buffalo populations.
Locus
ETH152
CSSM060
BM1706
ETH02
ETH225
INRA005
Mean
Total number of alleles (TNA)

Southern area
Middle area
Northern area
8
9
7.5
6.5
7.5
6.5
6.5
8
8.5
6.5
6
6.5
4.5
4.75
4.5
2.5
2
2.5
5.75
6.20
6
Mean number of
alleles (MNA)
8.16
6.83
7.66
6.33
4.58
2.33
5.98
Table 2. Mean of heterozygosity of six loci in the Iraqi buffaloes.

Locus
ETH152
CSSM060
ETH02
BM1706
ETH225
INRA005
Ho and He *
Ho
He
Ho
He
Ho
He
Ho
He
Ho
He
Ho
He
Southern area
0.875
0.865
0.916
0.782
0.75
0.740
0.75
0.786
0.625
0.577
0.208
0.197
*HO: Observed heterozygosity, HE: Expected Heterozygosity.
274
Meddle area
0.729
0.865
0.833
0.827
0.770
0.753
0.791
0.833
0.770
0.628
0.145
0.134
Northern area
0.75
0.869
0.791
0.824
0.791
0.766
0.708
0.085
0.75
0.693
0.458
0.365
DISCUSSION
CONCLUSION
Most of the microsatellite loci used here

were highly informative in the Iraqi buffalo. In the
present study, a high level of genetic variability
was revealed in Iraqi buffalo. The mean expected
heterozygosity of eight populations varied between
0.197 in the INRA005 to 0.865 in the ETH152 loci.
The mean observed heterozygosity was 0.208 in
the INRA005 and 0.916 in the CSSM060 loci. Our
results were supported by many studies conducted
in other countries, all of which showed the
weakness of the INRA005 marker to the degree of
the absence of polymorphism like in Iranian buffalo
We have presented the first study of

genetic diversity of Iraqi domestic buffalo using
microsatellite markers recommended by FAO and
ISAG. Results indicate distinct genetic variation,
high levels of genetic differentiation and genetic
structure with three major clusters. These findings
could provide an objective basis for classification
and conservation of indigenous buffalo resources.
(Mirhoseinei et al., 2005) or weak polymorphism

in the Egyptian buffalo (Zeinab, 2005), with some
exceptions as in the French buffalo which showed
more than three allelesand in the Italian buffalo, in
which it reached six alleles (Ciampolini et al., 1995).
These differences may be due to the differences in the
breed or cluster included in the study.
Among the six microsatellite loci used in
this study, the ETH152 marker showed the presence
of 16 alleles, and this can be considered the highest
number of alleles of this marker that has ever
reported. In a study on microsatellite markers in
the Indian buffalo, this marker showed only three
alleles (Kale, 2010), while in the Iranian buffalo
it has between five and sevenalleles only (Seyed
et al., 2005). In Egyptian buffalo the number of
alleles for this marker was eight (Zeinab, 2005).
All of these studies and many other studies did not
reach the number in the present study-the marker
ETH152 had 16 alleles only in Iraqi buffalo. This
fact supports our theory that the Iraqi buffalo
originated in Iraq and was not imported from
India.
The authors are thankful to Professor Dr.

Chao from Huazhong University in China for
scientific support for our work.
ACKNOWLEDGEMENTS
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Tan and K. Byrne. 1997. Genetic diversity
of Asian water buffalo (Bubalus bubalis)
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276
Original Article
CRYOPRESERVATION OF ENDOMETRIAL STROMAL CELLS

OF BUFFALO (BUBALUS BUBALIS)
Chethan Sharma G., S.K. Singh*, Jessihun Nongsiej, H.B. Rakesh,
R.P. Singh and S.K. Agarwal
ABSTRACT
cryopreserved without any major impairment in

their morphological and functional integrity.
The aim of the present experiment was

to cryopreserve buffalo endometrial stromal cells
and to study their morphological and physiological
characteristics. Stromal cells isolated by enzymatic
digestion from the buffalo uterus of day 1-5 of
the estrous cycle, were cultured and frozen at
-80oC using DMSO as a cryoprotectant. Frozen
Keywords: cryopreservation, buffalo, stromal

cells, culture, prostaglandins
INTRODUCTION
Prostaglandins (PGE2 & PGF2) are
important regulators of reproductive events viz.,
luteolysis, ovulation, implantation and parturition
in farm animals. PGF2 plays a major role in
regulation of cyclicity, on the other hand, PGE2
exerts action opposite to PGF2 and favours
establishment of pregnancy by its luteoprotective
action. The endometrium is a complex tissue
containing mainly epithelial and stromal cells.
Both types of cells produce isoform of PG, but
have different morphological and physiological
properties (Fortier et al., 1988; Asselin et al.,
1997). Endometrial epithelial cells mainly secrete
PGF2 whereas stromal cells are the principal
source of PGE2. Identification of a particular cell
type responsible for observed effect and interaction
between different cell types is difficult to study with
mixed cell culture. An in vitro system for stromal
cell culture, therefore, constitutes a good model to
study regulation of prostaglandin synthesis (Parent
et al., 2003; Guzeloglu et al., 2004), which avoids
cells were thawed at 37oC and recultured. The

culture media was collected when both unfrozen
(control) and frozen cells reached confluence for
estimation of PGE2 and PGF2 using enzymelinked immunosorbent assay. Frozen stored buffalo
endometrial stromal cells immediately after thawing
appeared as single or in small clumps, glistening
and rounded which changed their morphology from
rounded to flat spindle shaped immediately 24 h
after seeding retaining their normal morphological
characteristics. The viability of frozen thawed
stromal cells at the time of seeding estimated by
Trypan blue dye exclusion was 63.3%. There was
no significant difference in the basal production of
PGE2 and PGF2 by the frozen-thawed and unfrozen
buffalo stromal cells (289.38 4.39, 3.28 0.16
pg/g protein vs. 305.97 3.20, 4.26 0.17 pg/g
protein, respectively) on day 7 of culture indicating
no adverse effect of cryopreservation on their
functional characteristics. The results demonstrated
that buffalo endometrial stromal cells can be
Division of Animal Reproduction, Indian Veterinary Research Institute, Izatnagar, Bareilly (U.P.), 243 122,
India *E-mail: singhsk2003@yahoo.co.in
277
antibiotics and transported to the laboratory on ice

immediately after slaughter. White side test (Popov,
1969) and uterine cytology (Kasimanickam et al.,
2004) were done to rule out infection.
Buffalo endometrial stromal cells were
isolated by enzymatic digestion as per Gupta et
al. (2010) with suitable modifications. Briefly, the
myometrial layers were dissected and the horn was
everted to expose the uterine epithelium and placed
in a beaker containing 0.3% trypsin III in HBSS
and digested for 2 h at 5% CO2, 95% humidified
air at 37C. After incubation the digested horn
was further placed for second digestion containing
0.064% trypsin III and collagenase type II and
0.032% DNase I for 1 h. All scrapings and washings
were combined with the digested cells and fetal
calf serum was added to a final concentration of
10% to block the action of trypsin followed by
centrifugation at 1200 rpm for 10 minutes. The
pellets were pooled and washed thrice with HBSS
and finally suspended in 1 ml RPMI-1640 medium
with 10% FCS, 50 g/ml gentamicin and 0.25
g/ml Amphotericin-B. Cells were counted using
haemocytometer and viability was determined by
trypan blue dye exclusion test (Freshney, 2000).
Cell concentration was adjusted to 6.25X105 cells/
animal handling, ethics problems and variability of

results too. Recently, stromal cell culture (Gupta et
al., 2010) has also been established in buffalo in
our laboratory, as a model to study the regulation
and modulation of PGF2 and PGE2.
To study the regulatory mechanism
involved in PG synthesis related to either luteolysis
or establishment of pregnancy, there is a need to
cryopreserve uterine endometrial cells in order
to conduct in vitro experiment at any point of
time. Cryopreservation has most commonly been
used for the long term storage of biological cells.
Freezing and storage of endometrial cells allows
accessibility of cells to conduct experiment at any
point of time, reduces repeated procurement of
genital tracts for isolation of cells and reduction in
variability of results as well. Cryopreservation of
cells derived from reproductive tracts e.g. bovine
endometrial epithelial and stromal cells (Murakami
et al., 2003), bovine endothelial cells from the
corpus luteum (Acosta et al., 2007), porcine
endometrial epithelial cells (Kim et al., 2010) and
equine endometrial epithelial and stromal cells
(Szostek et al., 2012) have been established. Till
date, information on cryopreservation of stromal
cells in buffalo is lacking. The present experiment,
therefore, was designed with the objective to study
the effect of cryopreservation on stromal cell
morphology and physiology.
ml in RPMI-1640 media plated on 6 well plates

and cultured at 37C in an atmosphere of 5% CO2,
95% humidified air for 18 h to allow attachment
of stromal cells. Media were changed every
2 days until monolayer. Further, stromal cells were
subjected to indirect immuno-fluorescent staining
for specific marker i.e. fibronectin (stromal
cells). Cells stained positive giving green colored
fluorescence indicating presence of specific stromal
cells (Figure 1-D).
Cells were harvested upon reaching
monolayer using 0.05% trypsin and 0.02% EDTA
for 5-10 minutes at 37oC and washed two times

Isolation of endometrial stromal cells
Buffalo uteri along with intact ovaries
having corpus hemmorrhagicum (approximately
day 1-5 of the estrous cycle) apparently free of
infection were procured from the local abattoir,
placed in sterile HBSS supplemented with
278
unfrozen cells.
with HBSS. Final cell pellet was suspended in

chilled freezing media (70% culture media + 10%
DMSO + 20% serum) to adjust cell concentration
at the density of 1X106 cells/ml after counting and
viability assessment. One ml of cell suspension was
dispensed into 2 ml cryovial and subjected to slow
freezing in cryo 1oC freezing container (isopropanol
bath) and kept at -80oC in a deep freezer. Cells were
thawed by plunging cryovials into a water bath
maintained at 37oC. Contents were centrifuged and
the cell pellet was suspended in RPMI 1640 media
with 10% FCS supplemented with antibiotics and
counted using haemocytometer, the quantitative
survivability of the cells was determined by
staining the frozen thawed cells with 0.4%Trypan
blue (Freshney, 2000). Growth characteristics of
the frozen thawed stromal cells were evaluated by
re-culturing the cells in RPMI-1640 at a density of
1X105 viable cells/ml in 6 well plate. Media were

Frozen stored buffalo endometrial stromal
cells immediately after thawing appeared as single
or in small clumps, glistening and rounded which
changed their morphology from rounded to flat
spindle shaped immediately 24 h after seeding
retaining their normal morphological characteristics
and reached confluence or monolayer by day 7
culture (Figure 1-A,B,C). The viability of frozen
thawed stromal cells at the time of seeding estimated
by Trypan blue dye exclusion was 63.3%. The
loss of viability may be due to stress of freezingthawing process causing injury to the plasma
membrane, a primary target of cryo-damage (Gao
and Crister, 2000). Production of PGE2 is a useful
changed every 2 days till confluence/ monolayer.

The supernatant were collected after
reaching monolayer on day 7 for PGE2 and PGF2
estimation using an ELISA Kit (Neogen, U.S.A).
Cells were harvested using 0.05% trypsin and
0.02% EDTA for 5-10 minutes at 37oC. The cell
pellets were dissolved with 0.3 M NaOH and 1%
sodium lauryl sulfate and the protein concentration
of cell lysates was determined by the Bradford
method (Bradford, 1976) using BSA as standard
(Banglore Genei, Banglore). Total PGE2 and PGF2
concentration per well were expressed as picograms
per microgram of total cellular protein.
parameter for determining the functional status of

bovine endometrial stromal cells. PGE2 has been
reported as one of the principal prostaglandin
secreted by endometrial stromal cells in cattle and
buffalo (Parent et al., 2003; Gupta et al., 2010).
There was no significant difference in the basal
production of PGE2 and PGF2 by the frozenthawed and unfrozen buffalo stromal cells (289.38
4.39, 3.28 0.16 pg/g protein vs. 305.97
3.20, 4.26 0.17 pg/g protein, respectively) on
day 7 of culture indicating no adverse effect of
cryopreservation on their functional characteristics.
No difference in the production of PGE2 and PGF2
by frozen thawed stromal cells as compared to
unfrozen cells indicated resistance of stromal cells
to the freezing and thawing process (Murakami
et al., 2003). The results also corroborated with
the finding of Szostek et al. (2012) in equines.
In the present study successful cryopreservation
of buffalo endometrial stromal cells without any
Statistical analysis
The data are shown as mean SEM of
values obtained in three separate experiments, each
performed in triplicate. The independent t test
was used to show the statistical significance among
basal production of PGE2 and PGF2 in frozen and
279
Table 1. Basal prostaglandin production (pg/g protein) by frozen thawed buffalo endometrial stromal cells.
Groups
Frozen stromal cells
Unfrozen stromal cells (Control)
t value
PGE2
289.38 4.39
305.97 3.20
3.05 NS
PGF2
3.28 0.16
4.26 0.17
4.06 NS
(B)
(A)
(C)
(D)
Figure 1. Frozen-thawed buffalo endometrial stromal cell immediately after seeding (A), 24 h
after seeding (B), at confluence/ monolayer (C) Immunofluorescent staining of buffalo
endometrial stromal cells (stained positive with anti-fibronectin) (D).
280
major impairment in morphological and functional

integrity was achieved and may serve as useful
model to study cell biology and physiology of the
endometrium particularly in buffalos.
Freshney, R.I. 2000. Culture of Animal Cells: A

Manual of Basic Technique, 4th ed. WileyLiss Inc., New York, USA. p. 361.
Gao, D. and J.K. Crister. 2000. Mechanism of
cryoinjury in living cells, ILAR J., 41: 187196.
Gupta, C., S.K. Singh, R.K. Baithalu, K.R. Anuj,
A. Saxena, R.P. Singh and S.K. Agarwal,
2010. Morphological and functional
characterization of endometrial stromal
cells in buffalo, p. 119. In International
Symposium on Biotechnologies for
Optimization of Reproductive Efficiency of
Farm and Companion Animals to Improve
Global Food Security & Human Health
and XXVI Annual Convention of ISSAR,
G.B.P.U.A. & T., Pantnagar.
Guzeloglu, A., F. Michel and W.W. Thatcher. 2004.
Differential effects of interferon- on the
prostaglandin synthetic pathway in bovine
endometrial cells treated with phorbol ester. J.
Dairy Sci., 87: 2032-2041.
Kasimanickama, K., T.F. Duffielda, R.A. Fosterb,
C.J. Gartleya, K.E. Lesliea, J.S. Waltonc and
W.H. Johnsona. 2004. Endometrial cytology
and ultrasonography for the detection of
subclinical endometritis in postpartum dairy
cows. Theriogenology, 62: 9-23.
Kim, T.S., E.H. Kwon, S.U. Kim, H.S. Choi, H.J.
Lim and S.B. Park. 2010. Activation and
expression of urokinase-type plasminogen
activator are modulated by freezing/thawing
process through activation of redox signal
pathway in primary porcine endometrial
cells. Cryobiology, 60: 211-216.
Murakami, S., M. Shibaya, K. Takeuchi, D.J.
Skarzynski and K. Okudu. 2003. A passage
and storage system for isolated bovine
endometrial epithelial and stromal cells. J.
ACKNOWLEDGEMENTS
The project is funded by the National Fund
for Basic and Strategic Research in Agricultural
Sciences (NFBSRA) under the Indian Council
of Agricultural Research, New Delhi, India.
The authors thank the Director, IVRI, Izatnagar,
Bareilly, UP for necessary facilities to carry out
this research work.
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A.T. Grazul-Bilska and D.J. Skarzynski.
2007. Effects of storage and passage of bovine
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prostaglandin F2 production. J. Reprod
Dev., 53: 473-480.
Asselin, E., P. Drolet and M.A. Fortier. 1997.
Cellular mechanisms involved during
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in endometrial epithelial cells in vitro: Role of
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Specific properties of epithelial and stromal
cells from the endometrium of cows. J.
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Parent, J., C. Villenenve, A.P. Alexenko, A.D. Ealy
and M.A. Fortier. 2003. Influence of different
isoforms of recombinant trophoblastic
interferons on prostaglandin production
in cultured bovine endometrial cells. Biol.
Reprod., 68(3): 1035-1043.
Popov, Y.N. 1969. Diagnosis of occult endometritis
in cows (using white side test on cervical
mucus). Veterinariya Moscow, 4: 85-87.
Szostek, A.Z., M.J. Siemieniuch, A.M. Galvao,
K. Lukasik, D. Zieba, G.M. FerreiraDias and Skarzynski. 2012. Effects of cell
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282
Original Article
ADOPTION LEVEL OF BUFFALO FARMING PRACTICES IN

THE ARID ZONE OF RAJASTHAN
M.L. Meena and Dheeraj Singh
ABSTRACT
knowledge, non-availability of inputs, lack of time

and lack of veterinary services.
This study was conducted in Pali district.

This district was purposively selected for the study
as the first phase of the project was in operation
only in this district. The sample size was limited
to 200 participants and 100 non-participants
considering the time and other resources available
for the study. A sample of 10 participant farmers
were selected by adopting simple random sampling
technique from each of the selected 20 villages.
Likewise, 100 non-participant farmers were
selected from six villages of the non-project area
having similar socio-economic status. Thus, in all
300 farmers constituted the sample of the study. It
could be concluded that 47.50 and 42.50 percent
of the participant farmers were in the medium and
high levels of adoption of recommended practices,
respectively, whereas 54.00 and 45.00 percent of
the non-participant farmers in the control villages
were in the medium and low levels of adoption,
respectively. The extent of adoption of participant
farmers was strikingly more than that of the nonparticipant farmers and there existed a significant
difference between the two groups. In the case of the
non-participant farmers, the majority of them were
non-adopters of various recommended practices
except in practices such as period of grazing and
feeding of tree leaves in summer. The reasons given
by the majority of the participant farmers for nonadoption of recommended practices were lack of
Keywords: adoption, buffalo farming, technology,

Rajasthan
INTRODUCTION
The buffalo (Bubalus bubalis) holds the
greatest promise for food security and sustainable
development in the 21st century as these animals
form an integral part of the typical farming
system in India. In India, the buffalo has been the
backbone of the rural economy. It is the mainstay
in the production of butter and ghee. Not only
this, the buffalo is also considered more useful for
reasons of higher fat content in milk, its ability
to utilize agricultural byproducts more efficiently
and requirement for fewer kilo calories to produce
1 kg milk. The buffalo has been the friend of small
farmers, often their largest capital asset. It is a fit
companion for Indian farmers, who grow older with
their buffaloes developing a high degree of mutual
understanding. As with many of the live stock
spices maintained by rural families, there is a sociocultural linkage with buffaloes also. Livestock play
a pivotal role in the countrys rural economy. It
contributed nearly 16 percent of total income from
agriculture in 1970-71, which increased to over
25 percent in 2009-10. This will continue to be
Krishi Vigyan Kendra, CAZRI, Pali-Marwar, Rajasthan-306 401, India, *E-mail: moti2007m@yahoo.co.in
283
so in the coming period due to various economic

factors like increase in the population, urbanization
and per capita income growth. The role of livestock
is much pronounced in the arid zone of Rajasthan,
which is prone to drought and resultant crop failure.
At the time of crop failure, the livestock sector acts
as a cushion which the farmers can fall upon. As
per the 2007 Livestock Census, Rajasthan had 10.9
million cattle, 10.4 million buffalo, 10.0 million
buffalo and 16.8 million goats. The buffalo is one
of the major livestock species contributing to the
livelihood security of farmers in Rajasthan. It can
thrive in adverse climatic conditions using sparse
vegetation. The major buffalo breeds are Murrah,
Surti, Nili Ravi, Jafravadi and Mehsana. Buffaloes
are mainly reared by socially and economically
backward people of the society for whom they act
as a store of wealth of high liquidity, a resource
for meeting the household nutritional security
and a source of regular income. The farmers rear
buffalo mainly in an extensive management system
using traditional management practices relying
on common pastureland for grazing. Various
improved buffalo management technologies were
developed at the Central Arid Zone Research
Institute, Jodhpur to cater the needs of buffalo
farmers. These techno;ogies pertained to the broad
areas of breed management, nutrition, health care,
alleviation efforts.
In order to improve the socio-economic
status of these farmers by helping them cross
over the poverty line, a novel project, namely the
Farming Systems Research for Improved Buffalo
Production (FSRIBP). Central Arid Zone Research
Institute, aided by FSRIBP, was conceived and
launched in the arid zone of Rajasthan in the year
of 2007. A systematic evaluation on scientific lines
to understand the impact of the project in changing
the knowledge and behavioural aspects of target
farmers is essential because this would form a basis
for extending this project to the other target areas.
With this in mind, the present study was undertaken
with the following objectives:
1. To measures the extent of adoption of
recommended buffalo farming practices among
participant and non-participant farmers.
2. To ascertain the reasons for non-adoption
of recommended practices.
METERIALS AND METHODS

Raipur panchayat samiti of Pali district was
purposively selected for the study as the first phase
of the project was in operation only in this district.
The sample size was limited to 200 participants
and100 non-participants considering the time and
other resources available for the study. A sample of
10 participants farmers were selected by adopting
simple random sampling technique from each of the
selected 20 villages. Likewise, 100 non-participants
farmers were selected from two villages of nonproject area having similar socio-economic status.
Thus, in all 300 farmers constituted the sample of
the study (Sharma, 2005).
Rogers (1983) defined adoption as the
decision to make full use of an innovation as the
reproduction and product management. Adoption

of improved management practices is expected to
increase the income of farmers. The performance
and contribution of livestock in recent decades
towards agricultural production is phenomenal.
Livestock not only provide livelihood but also
offer employment opportunities to the poorer and
weaker sections of the society. Indian planners and
policy makers realized the need to recognize and
promote small scale livestock production, which
is thought to act as a key component of poverty
284
best course of action available. However, adoption

of improved management practices for buffaloes in
the field is low due to various reasons. Considering
the importance of buffaloes in the livelihood of
rural poor in marginalized environments such as
Rajasthan, it is imperative to enhance the adoption
of improved management technologies of buffalo
farming. The term adoption in this study means the
use of recommended practices by the respondents.
In consultation with the FSRIBP and Livestock
Development Project field functionaries, fifteen
skill oriented technologies/practices in buffalo
farming recommended through the programme were
identified for study of adoption. The recommended
practices were described to respondents and they
were requested to indicate their extent of adoption
during the previous years. The respondents who
had adopted the recommended practice were given
two scores and for those not adopted were given
zero score. The extent of adoption of recommended
practices for each respondent farmer was measured
by using an adoption quotient developed by
Sengupta (1967) and adopted by Swaminathan
(1986). Based on the adoption quotient, the
respondents were classified into three categories
using mean and standard deviation.
The findings on the presented and discussed

in terms of extent and practice wise adoption.
Extent of adoption of recommended practices
(overall adoption)
The data presented in Table 1 revealed that
about 42.50 and 47.50 percent of the participants
were high and medium adopters, respectively,
whereas only 01.00 and 54.00 percent of the nonparticipants were in these categories. The remaining
10.00 percent of the project farmers and 45.00
percent of the non-project farmers were in the low
level of adoption category. This indicates that the
extent of adoption of practices by beneficiaries was
higher in the target area than in the control area. The
reason is that the project has created a considerable
level of technological impact on the participant
farmers due to the utilization of services such as
veterinary and extension services rendered by the
project officials. Besides this, increased awareness
through mass media exposure and its utilization
and through contact with extension agents would
have certainly increased the level of adoption
along with knowledge gained regarding buffalo
husbandry practices. The non-availability of these
services and facilities in the control area would be
Table 1. Distribution of buffalo farmers according to their level of adoption behavior.
S. No.
1.
2.
3.
Level of adoption
Low -SD
Medium SD
High +SD
Overall
Adopters (N=200)
F
%
20
10.00
95
47.50
85
42.50
200
100
F = Frequency; % = Percentage
285
Non-adopters (N=100)
F
%
45
45.00
54
54.00
01
01.00
100
100
(N = 300)
Total (N=300)
F
%
65
21.66
149
49.67
86
28.67
120
100
the main reason for the considerable percentage

of low adoption. The findings are in line with
the findings of Mohan et al. (2005), Suresh et al.
(2008) and Meena et al. (2007). Thus, the buffalo
farmers who were selected as the beneficiaries of
the project had increased their level of adoption to
a moderately high level, indicating a considerable
amount of technological impact on them.
6 months of age and isolation of sick animals to

prevent the spread of diseases.
In the case of non-participant farmers,
the period of grazing and the feeding of tree
leaves such as neem (Azadirachta indica A Juss.),
khejari(Prosopis cinerria), babool(Acacia nilotica)
and jharberi pala (Ziziphus mauritiana) spices
were the practices adopted by 84.00 and 52.00
percent of farmers, while the majority of them were
non-adopters of almost all the remaining practices
in buffalo rearing. The reasons given were lack
of knowledge and non-availability of suitable
inputs. The main reason that the majority of the
participant farmers were in the adopter category
when compared to non-participants might be the
thrust of the project officials to improve buffalo
husbandry as a viable enterprise in the project area.
The findings are line with those of Kumaravel and
Krishnaraj (2005) and Meena (2005).
Practice wise adoption of recommended

practices
Fifteen practices in buffalo rearing as
recommended by Farming Systems Research
for Improved Buffalo Production (FSRIBP) were
considered for assessing the adoption by the
respondent farmers. The data generated in this
aspect were analyzed and are presented in Table 2.
It can be seen from the table that of the 15 practices
recommended by FSRIBP in buffalo rearing, the
practices namely selection and purchase of quality
buffalo (77.50 percent), provision of shelter for
animals (56.00 percent), feeding of colostrums to
new born calf (80.00 percent), period of grazing
(97.05 percent), feeding of crushed prosopis pods
and tamarind seeds (75.00 percent), feeding of
tree leaves in summer (83.00 percent), deworming
(73.00 percent), vaccination of buffalo (71.00
percent) and marketing of calf at 15 months of
Reasons for non-adoption of recommended

practices by participants farmers
The reasons for non-adoption of various
recommended practices were collected, tabulated
and are presented in Table 3. From the table it
can be observed that with regards to practice of
selection and purchase of quality animals, the
high cost of animals (67.70 percent) and the non-
age (67.05 percent) were adopted by the majority

of the participant farmers (Tajane et al., 2005).
The reason for the maximum level of adoption of
these technologies/ practices might be the farmers
conviction they had gained in these areas by way
of reduction in mortality and enhanced production
level. The adoption level seemed to be poor in
the areas of upgrading, ligation and disinfections
of naval cord, practice of weaning at 2 months of
age, ectoparasiticide application for eradication of
ticks and lice, castration of male buffalo calves at
availability of good breeds (32.30 percent) were

the reasons given by the farmers for non-adoption
of the recommended practices. The farmers stated
that they were unable to spend money on such
quality breed of buffalo.
With respect to provision of shelter for
animals, the participant farmers felt that for a few
animals the overhang in the house was enough
(22.70 percent) and some of them opined that
construction of separate shed for providing shelter
to the animals (77.30 percent) was a costly affair.
286
Table 2. Practice wise adoption of recommended technologies practices in buffalo.

Project area (N=200)
S. No.
Technologies
Adopter
Nonadopter
Non-project area (N=100)

Adopter
Non-adopter
No.
No.
No.
No.
Selection and purchase of

quality buffalo breed
155
77.5
45
22.5
08
8.00
92
92.00
Provision of shelter for

buffalo
112
56.0
88
44.0
40
40.00
60
60.00
3
4
Upgrading
Colostrums feeding
16
175
08.0
87.5
184
25
92.0
12.5
05
15
5.00
15.00
95
85
95.00
85.00
Ligation and disinfection of

naval
72
36.0
128
64.4
00
0.00
100
100
13
195
06.5
97.5
187
05
93.5
02.5
00
84
0.00
84.00
100
16
100
16.00
150
75.0
50
25.0
14
14.00
86
86.00
6
7
8
Practice of weaning
Period of grazing
Feeding of crushed
prosopis pods and tamarind
seeds
Feeding of tree leaves in

summer
166
83.0
34
17.0
52
52.00
48
48.00
10
Deworming
146
73.0
54
27.0
07
7.00
93
93.00
11
Ecto-parasiticide
application
54
27.0
146
73.3
03
3.00
97
97.00
12
13
14
Castration of lamb
Vaccination of buffalo
Isolation of sick buffalo
13
142
20
06.5
71.0
10.0
187
58
180
95.5
29.0
90.0
00
02
00
0.00
2.00
0.00
100
98
100
100
98.00
100
15
Marketing of ram cat 16

months of age
135
67.5
65
32.5
33
33.00
67
67.00
N=Number of respondents, % = Percent
287

Table 3. Reasons for non-adoption of recommended buffalo rearing practices (Farmers).
S.
No.
Recommended
practices
Selection and purchase
of quality animals
Provision of shelter for
animals
Non
adopter
Reasons for non-adoption
No.
(Percent)
30 ( 66.7)
15 (33.3)
20 (22.7)
68 (77.3)
18 (9.8)
166 (90.2)
Upgrading
184
Colostrum feeding
25
Ligation and disinfection

of naval cord
128
Practice of weaning
187
Period of grazing
Feeding of crushed
drosopis pods and
tamarind seeds
Feeding of tree leaves in
summer
05
a. High cost
b. Non-availability of breeds
a. Over hang in the house sufficient
b. Shed construction is costly
a. Non-availability of quality calves
b. lack of knowledge
a. Colostrum feeding will induce
diarrhea
b. Lack of time
a. Not necessary
b. Lack of knowledge
a. Lack of knowledge
b. lambs will feel lonely
50
50 (100.0)
b. Time consuming
b. Inadequacy of dewormers
a. Costly method
c. Time consuming
a. Lack of veterinary services
a. Buffalo go down in condition
b. Failure of vaccination
c. Lack of knowledge
a. Non-availability of isolation space
16 (47.1)
18 (52.9)
48 (88.9)
06 (11.1)
39 (26.7)
63 (43.2)
44 (30.2)
24 (12.8)
163 (87.2)
05 (08.6)
27 (46.6)
26 (44.8)
33 (18.3)
147 (81.7)
65 (100.0)
1
2
8
9
45
88
34
10
Deworming
54
11
Ecto-parsiticide
application
146
12
Castration of lamb
187
13
Vaccination of buffalo
58
14
Isolation of sick animals
180
15
Marketing of calf at 16
months of age
65
288
17 (68.0)
8 (32.0)
111 (86.7)
17 (13.3)
142 (75.9)
45 (24.1)
7 (100.0)
With regards to upgrading, lack of knowledge

about the scientific breeding practices (90.20
percent) and non-availability of quality calves
(09.80 percent) were the reasons mentioned by
most of the non-adopters. In the case of colostrums
feeding the farmers felt that feeding of colostrums
immediately after birth would induce diarrhea and
cause the animal to go down in condition (68.70
percent). The one third of the non-adopters of
this practice cited the non-availability of time as
the reason for non-adoption as they were engaged
in other activities. With regards to the practice of
ligation and disinfection of naval cord, the nonadopters felt that the mother itself would lick the
umbilical cord and it would heal automatically and
hence there was no need to cut and ligate the cord.
A few of the non-adopters said lack of knowledge
was their reason for not adopting this practice.
In the case of the practice of weaning,
lack of knowledge was cited by the majority of
the respondent farmers. Some of the farmers also
opined that weaned animals would feel lonely and
isolated. As far as period of grazing is concerned
very few gave lack of knowledge as the reason for
non-adoption of following the recommended time
of grazing. In the case of feeding of crushed prosopis
pods and tamarind seed, lack of knowledge was the
reason given by nearly half of the total participant
farmers. With regards to the feeding of tree leaves
in summer, some of the participant farmers stated
they lacked knowledge in the feeding of suitable
types of leaves, while the remainder felt that this
practice was a time consuming affair.
In the case of deworming, lack of
knowledge was given by the majority of the nonadopters (88.90 percent) while the remaining
11.10 percent of them stated that inadequacy of
dewormers with the local veterinarian and link
worker couples was their reason for non-adoption.
With regard to ectoparasiticide application, a large

proportion (43.20 percent) of respondents cited lack
of knowledge about the practice as their reason for
their non-adoption. The remaining 30.20 and 26.70
percent of non-adopters felt that the recommended
practice was time consuming and was a costly
method, respectively.
Lack of knowledge was the reason given by
the majority (87.20 percent) of non-adopters with
respect to the practice of castration of male calves,
followed by lack of veterinary services (12.80
percent) in their villages to perform this operation.
With respect to the vaccination of buffalo 46.60
percent of non-adopters gave bitter experience
of failure of vaccination leading to mortality of
buffalo as their major reason, followed by lack of
knowledge (44.80 percent) and the wrong notion
of the condition of animals would deteriorate if
vaccinated (8.60 percent). With regard to isolation
of sick animals, lack of knowledge (81.70 percent)
and non-availability of isolation space (18.30
percent) were the reasons cited for non-adoption.
In the case of marketing of male calves at 15
months of age, the non-adopters said that lack of
knowledge about organized marketing was their
reason for non-adoption.
CONCLUSION
About 47.50 and 42.50 percent of the
participant farmers were in the medium and high
levels of adoption of recommended practices,
respectively, whereas 54.00 and 45.00 percent
of non- participant farmers in the control
villages were in the medium and low levels of
adoption, respectively. The extent of adoption of
participant farmers was strikingly more than that
of the non-participant farmers and there existed a
289
Mohan, B., R.L. Sagar, K. Singh, P. Tripathi and

V.S. Vihan. 2007. Present status of goat
rearing under rural condition. Indian
Research Journal of Extension Education,
7(1): 43-45.
Rogers, E.M. 1983. Diffusion of Innovations. The
Free Press, New York: 132.
Sengupta, H. 2007. Adoption scale to measures
livestock production practices in arid zone
of Rajsthan (Unpublished), RAU, Bikaner,
Campus: SKN, COA, Jobner.
Sharma, N.K. 2006. Construction of adoption
scale about improved buffalo production
technology in Ago-climatic Zone IIIb of
Rajasthan. Rajasthan Journal of Extension
Education, 16(2): 23-26.
Suresh, A., D.C. Gupta and J.S. Mann. 2008.
Adoption of improved management
practices of buffalo in the semi arid region
of Rajasthan. Indian Journal of Extension
Education, 44(1&2): 100-105.
Swaminathan, M.S. 1986. Agricultural planningimproving the livestock in rural areas.
Indian Farmers Times, 6(11): 32.
Tajane, S.B., P.J. Rajkamal and Veeranna. 2005.
Work environment of the veterinary
surgeons. Indian Journal of Extension
Education, 41(1&2): 99-101.
significant difference between the two groups. The

recommended practices in buffalo rearing, namely
selection and purchase of quality buffalo, provision
of shelter for animals, feeding of colostrums for
new born calf, period of grazing of buffalo, feeding
of crushed prosopis pods and tamarind seeds,
feeding of tree leaves in summer, deworming,
vaccination and marketing of calf at 15 months of
age were adopted by majority of the participant
farmers. Most of the participant farmers had poor
adoption in areas such as upgrading of animals,
ligation and disinfection of naval cord, practice of
weaning, ectoparasiticide application, castration of
male calves and isolation of sick animals. In the
case of non-participant farmers, the majority of
them were non-adopters of various recommended
practices except practices such as period of grazing
and feeding of tree leaves in summer. The majority
of the participant farmers said that the reason for
their non-adoption of recommended practices were
lack of knowledge, non-availability of inputs, lack
of time and lack of veterinary services.
REFERENCES
Kumaravel, P. and R. Krishnaraj. 2005. Adoption
behaviour of livestock farmers. Indian
Journal of Extension Education, 41(1&2):
117-121.
Meena, M.L. 2005. Problems and prospects of
improved buffalo production in the flood
prone eastern plain zone of Rajasthan. Ph.D.
Thesis, RAU, Bikaner Campus: Jobner.
Meena, M.L., N.K. Sharma and A. Dudi. 2007.
Buffalo keepers adoption about improved
buffalo production practices. Indian
Research Journal of Extension Education,
7(2&3): 65-68.
290
Original Article
MILK PRODUCTION AND REPRODUCTION PERFORMANCE OF MURRAH BUFFALOES

OF TAMIL NADU, INDIA
A.K. Thiruvenkadan*, S. Panneerselvam, N. Murali, S. Selvam and V. Ramesh Saravanakumar
ABSTRACT
as service period, calving interval and dry period

were slightly higher than those observed in the
home tract of the Murrah buffalo and hence better
breeding management and introduction of genetic
evaluation programmes are needed for genetic
improvement of these traits.
The data pertaining to variable production

and reproduction traits of Murrah buffaloes (1980
lactation records of 698 Murrah buffaloes) were
collected from the Central Cattle Breeding Farm,
Alamadhi, Tamil Nadu, India. The overall leastsquares means ( SE) for peak yield, days to
attain peak yield, 305-day milk yield, lactation
length, lactation milk yield, service period, calving
interval and dry period were 8.87 0.05 kg, 53.4
0.8 days, 1804.9 14.7 kg, 297.8 1.9 days,
1855.6 16.1 kg, 225.0 5.5, 532.8 5.5 and
230.2 4.9 days, respectively. Period of calving
had a highly significant (P<0.01) effect on all the
traits studied except days to attain peak yield,
where it had only a significant (P<0.05) effect.
Season of calving had a significant (P<0.05) effect
on peak yield and lactation milk yield and a highly
significant (P<0.01) effect on days to attain peak
yield, 305-day milk yield, milk yield per day of
lactation and all the reproduction traits studied. The
lowest calving interval was observed in south-west
monsoon calvers, and they differed significantly
(P<0.05) from winter and summer calvers. Parity
had a highly significant effect (P<0.01) on all the
traits studied. Pairwise comparison revealed that
the lactation milk yield was lowest in first parity
and differed significantly (P<0.05) from other
parities. In general, the reproduction traits such
Keywords: coastal region, genetic parameters,

Murrah buffaloes, production, reproduction
INTRODUCTION
According to the 2003 livestock census,
India possesses 185.2 million cattle and 97.9
million buffaloes, which is about 13.7 percent
of the total cattle and 57.5 percent of the total
buffalo population of the world. The dairy industry
in India has made significant progress in the last
few decades. Today, India is the largest producer
of milk in the world. The milk production in the
country has risen to about 88.1 million tonnes in
2003-2004 from 17.0 million tonnes in 1950-51. At
present, Indias contribution to the total world milk
production is about 14.3 percent and the national
per capita milk availability is 231 g/day. In India,
although the proportion of buffaloes to cattle is 1:2,
the buffaloes contribute 56.86 percent of the total
milk obtained from cattle and buffaloes. Tamil Nadu
with 9.14 million cattle and 1.66 million buffaloes
Department of Animal Genetics and Breeding, Veterinary College and Research Institute, Orathanadu, Tamil
Nadu, 614 625, India, *E-mail: drthirusiva@gmail.com
291
produces an estimated quantity of 4.75 million

tonnes of milk (Report, 2006). The Murrah breed is
the finest genetic material of milk producing buffalo
not only in India but also probably in the world.
This breed has been used extensively throughout
the country to upgrade the non-descript buffalo
stock to improve milk production. The breeding
policy of Tamil Nadu state is to use Murrah or Surti
as the breeds of choice to improve non-descript
buffaloes. As a result of these measures, the Murrah
and graded Murrah population have been increased
over the years. Although the water buffalo in the
tropics out-produces other domestic animals,
commercial milk production is adversely affected
by a large number of detractive factors such as
late age at first calving, seasonality of oestrus, and
long calving interval and dry period. Therefore,
it is necessary to evaluate the relative importance
of various fixed environmental and physiological
effects in influencing the milk production and
reproduction traits in order to devise appropriate
feeding and other managemental practices. From
the literature reviewed, it was found that the bulk
of scientific information on buffaloes has come
from the analysis of records made available from
institutional and government farms in northern
India (Sethi and Khatkar, 1997; Dass and Sadana,
2000; Gogoi et al., 2002; Kundu et al., 2003a,b;
buffaloes in the coastal regions of Tamil Nadu to

suggest suitable managemental practices, selection
and breeding strategies for genetic improvement
of Murrah buffaloes under hot and humid climatic
conditions of India.

The study was based on the data pertaining
to the Murrah buffaloes born and bred at the Central
Cattle Breeding Farm, Alamadhi, Chennai, Tamil
Nadu, India from 1979 to 2006 (28 years). This
farm is located approximately at 13o N latitude and
80o E longitude at an altitude of about 20 metres
above mean sea level. The climate is generally hot,
humid and tropical in nature. The mean annual
maximum and minimum temperatures were 33oC
and 24.7oC, respectively, and the mean relative
humidity ranged between 69.2 and 76.2%. The
Murrah buffaloes were housed in permanent sheds
with open type ventilation and maintained under
stall-fed conditions. Roughage in the farm of green
fodder and paddy straw was provided. In addition,
concentrate mixture was provided to all age groups
as per the standard requirements. Cows were handmilked twice daily in the morning and evening.
Data on production and reproduction performance
of Murrah buffaloes (1980 lactation records form
698 Murrah cows) were extracted from History
and Pedigree Sheets. The traits studied were peak
yield, days to attain peak yield, 305-day milk yield,
lactation length, lactation milk yield, milk yield per
day of lactation, service period, calving interval,
dry period and number of services per conception.
Period and season were the fixed environmental
effects considered for all the traits studied. As the
calvings were less in a year, year-season analysis
was not done. To utilise all available data the entire
Yadav et al., 2007). Such information from the

southern peninsular region, especially under hot
and humid coastal regions of Tamil Nadu, is scanty.
The home tract of Murrah buffaloes is a hot and dry
climatic region in the north-western part of India.
Breeding these buffaloes in the southern peninsular
region of India, which is hot and humid, may affect
their performance and adaptability. Hence, the
present study has been made to understand the
performance and the influence of various nongenetic factors affecting economic traits of Murrah
292
to 2006), where the difference was not significant.

There was a steep increase in peak yield, 305-day
milk yield and lactation milk yield from the second
to the sixth periods and then a slight decline in
period 7 (2003 to 2006); the values observed in
period 2 differed significantly (P<0.05) from the
rest of the periods.
Season of calving had no effect on lactation
length; however, it had a significant (P<0.05) effect
on peak yield and lactation milk yield and a highly
significant (P<0.01) effect on days to attain peak
yield, 305-day milk yield and milk yield per day
of lactation. The peak yield of Murrah buffaloes
calving in the south-west monsoon season was
the highest and it differed significantly (P<0.05)
from that of north-east monsoon calvers. However,
there was no significant difference among the other
seasons. On the other hand, the lowest 305-day
milk yield, lactation milk yield and milk yield per
day of lactation were observed in cows calving
in the north-east monsoon season. Winter and
summer calvers had higher 305-day milk yield and
lactation milk yield than the monsoon calvers. The
yields observed among winter, summer and southwest monsoon calvers were not significant but they
differed significantly (P<0.05) from north-east
monsoon calvers.
Parity had a highly significant (P<0.01)
effect on all the traits studied. First lactation peak
yield was significantly (P<0.05) lower than the
rest. The yield increased from first to fourth parity
and from there on it started declining. Whereas,
the days to attain peak yield decreased from first
to fifth parity followed by a moderate increase in
sixth parity. The days to attain peak yield in first
lactation was significantly (P<0.05) higher (60.9
days) than the rest. The 305-day milk yield and
lactation milk yield increased up to third parity,
was maintained at fourth parity, and declined
duration was divided into seven periods each

with an interval of five years assuming that there
would not be any major managemental changes
within a period. Further each calendar year was
sub-divided into four seasons, viz. winter (January
and February), Summer (March to May), southwest monsoon (June to September) and north-east
monsoon (October to December). LSMLMW and
MIXMDL PC-2 VERSION computer programme
of Harvey (1990) was used to study the effect of
various non-genetic factors and the means were
compared using Duncans multiple range test. The
model used for analysis was Yijkl = + Pi + Sj + Ok +
eijkl, where, Yijkl = the lth observation in ith period, jth
season and kth parity, = overall mean when equal
subclass frequencies exist, Pi = effect of ith period
(i =1 to 7), Sj = effect of jth season (j =1 to 4), Ok
= effect of kth parity (k=1 to 6) and eijkl = random
errors NID (0, 2e).

Production Traits
Least-squares means (SE) of different
production traits are presented in Table 1. Period
of calving had a highly significant (P<0.01) effect
on the variation of peak yield, 305-day milk yield,
lactation length, lactation milk yield and milk yield
per day of lactation and had only a significant
(P< 0.05) effect on days to attain peak yield. The
highest peak yield, 305-day milk yield and lactation
milk yield were observed in period 6 (1999 to
2002) and they differed significantly from rest of
the periods, except with periods 5 and 7, where
the differences were not significant. The lowest
lactation length was observed in period 3 (1987 to
90) and this period differed significantly (P<0.05)
from the rest of the periods except period 7 (2003
293
294
48.3 0.9a
**
60.9 1.2b
54.0 1.3a
51.1 1.5a
50.6 1.7a
50.0 2.1a
53.5 1.9a
8.79 0.06a
**
7.73 0.08a
8.85 0.09b
9.37 0.10cd
9.54 0.12d
8.98 0.14b
8.74 0.13b
830
645
457
311
224
150
193
54.1 1.1b
9.03 0.07b
724
276
150
268
491
430
265
171
236
119
8.87 0.05
**
8.47 0.12b
7.96 0.09a
8.30 0.08b
9.16 0.11c
9.47 0.13ce
9.52 0.11e
9.18 0.16ce
*
8.81 0.10ab
8.84 0.14ab
1980
Days to attain
peak yield
(days)
53.4 0.8
*
50.7 1.7ad
54.9 1.3bc
52.0 1.3ab
52.2 1.6abc
57.2 1.9c
51.4 1.7ab
55.0 2.3bcd
**
53.0 1.5b
57.9 2.0b
**
1619.7 22.6a
1832.8 23.5c
1913.9 27.7d
1910.4 32.3d
1823.4 38.7c
1729.1 35.1b
1733.8 17.7a
1793.1 19.8b
1804.9 14.7
**
1670.2 32.6b
1584.2 23.6a
1632.0 23.4ab
1872.9 29.8c
1947.2 36.1cd
1974.1 31.1d
1953.7 43.2cd
**
1839.0 28.6b
1853.8 38.1b
305-day milk
yield (kg)
**
310.4 2.9c
301.1 3.1b
302.3 3.6bc
299.0 4.2bd
289.8 5.0abd
284.3 4.6a
297.4 2.3
298.8 2.6
301.8 3.7
293.3 5.0
297.8 1.9
**
295.6 4.2b
300.2 3.1b
285.1 3.0a
298.3 3.9b
307.2 4.7b
305.8 4.1b
292.5 5.6ab
Lactation
length (days)
**
1687.6 24.8a
1894.5 25.7cd
1967.3 30.4e
1966.5 35.5de
1853.7 42.4bc
1763.8 38.5ab
1801.6 19.4a
1849.6 21.8b
Lactation
milk yield
(kg)
1855.6 16.1
**
1706.2 35.8a
1629.9 25.8a
1658.5 25.6a
1925.2 32.7b
2030.3 39.6c
2055.6 34.1c
1983.2 47.4bc
*
1888.6 31.4b
1882.4 41.8b
**
5.38 0.05a
6.22 0.06b
6.45 0.07c
6.50 0.08c
6.33 0.09bc
6.09 0.09b
5.99 0.04a
6.14 0.05b
Mil yield
per day of
lactation (kg)
6.16 0.04
**
5.74 0.08b
5.38 0.06a
5.72 0.06b
6.43 0.07c
6.55 0.09c
6.66 0.08c
6.66 0.11c
**
6.19 0.07b
6.34 0.09b
n= Number of observations. * P<0.05, ** P<0.01.Means bearing same superscript within classes do not differ significantly (P>0.05).
Overall mean ()
Period of calving
P1 (1979-82)
P2 (1983-86)
P3 (1987-90)
P4 (1991-94)
P5 (1995-98)
P6 (1999-2002)
P7 (2003-2006)
Season of calving
Winter (Jan.-Feb.)
Summer (Mar.-May)
South-west monsoon
(Jun.- Sep.)
North-east monsoon
(Oct.- Dec.)
Parity
First
Second
Third
Fourth
Fifth
Sixth and above
Effect
Peak yield
(kg)
Table 1. Least-squares means ( SE) for different milk production traits of Murrah buffaloes
thereafter. Pairwise comparison revealed that the

305-day and lactation milk yields observed in first
parity differed significantly (P<0.05) from other
parities. Similar to days to attain peak yield, the
lactation length decreased with the advancement of
parity. There was an initial sharp reduction (3.09
percent) in lactation length in the second parity
followed by gradual decline later. The reduction in
lactation length between first and second parities
was statistically significant (P<0.05). The milk
yield per day of lactation increased linearly with
the advancement of parity up to 4th parity and then
declined; the values observed in first parity differed
significantly (P<0.05) from the rest of the parities.
The average peak yield obtained in the
present investigation was higher than the value
reported by a few earlier workers (Rao and Rao,
1994; Kundu et al., 2003b). However, Chhikara
et al. (1998) and Suresh et al. (2004) reported
higher values for Murrah buffaloes than those
observed in the present study. The days to reach
peak yield in the present study was substantially
higher than those reported for Murrah buffaloes
at different places in India (Kundu et al., 2003b;
Suresh et al., 2004). Lactation milk yield of cattle
and buffaloes up to 305-days of lactation is the
criterion most commonly used for the selection of
dairy animals and a study of the performance of this
trait is of paramount importance for carrying out
selection. The overall 305-day milk yield obtained
for Murrah buffaloes in this investigation was
comparable to the value reported by Ulaganathan et
al. (1983) and higher than the values reported by
other researchers (Kandasamy, 1987; Suresh et al.,
2004). The overall least-squares means of lactation
milk yield obtained was higher than those observed
by Ulaganathan et al. (1984) and Patnaik (1988) in
the same herd and lower than the present estimate
was also reported by Kandasamy (1987) and Suresh
et al. (2004). Differences in the estimates might be

due to sampling errors, genetic constitution of the
herds, agroclimatic variations and managemental
conditions. In general, the performance in terms of
the first lactation milk yield of Murrah buffaloes
at the Central Cattle Breeding Farm, Alamadhi
is quite similar (Sethi and Khatkar, 1997; Kumar
et al., 2002) to those herds in Haryana indicating
that there might not be any appreciable genotype x
environment interaction.
Reproduction Traits
Least-squares means (SE) of different
reproduction traits are set out in Table 2. Analyses
of variance revealed that the period of calving
influenced service period, calving interval, dry
period and number of services per conception in a
highly significant (P<0.01) manner although there
was no consistent trend over the periods. Period 3
(1987 to 90) had the lowest service period, calving
interval and number of services per conception,
and these differed significantly (P<0.05) from other
periods except period 1(1979 to 1982). Whereas, the
lowest and the highest dry periods were observed
in period 1 and period 5, respectively, and the
dry period observed in period 5 was significantly
(P<0.05) different from periods 1, 3 and 4.
Season of calving also influenced all the
reproduction traits in a highly significant manner
(P<0.01). Murrah buffalo heifers freshening in the
south-west monsoon had significantly (P<0.05)
shorter service period, calving interval, dry period
and number of services per conception than those
calving in other seasons. The highest service
period, calving interval and number of services per
conception were observed in winter season calvers.
The service period and calving interval observed in
south-west monsoon calvers differed significantly
(P<0.05) from other seasons except with north-east
295
Table 2. Least-squares means ( SE) for different reproduction traits of Murrah buffaloes.
Service period
(days)
Calving
interval (days)
Dry period
(days)
Number of
services per
conception
211
109
225.0 5.5
**
182.8 11.4af
241.3 8.1be
176.1 8.5a
218.9 10.4b
272.8 14.0cd
252.9 10.7cde
230.5 17.6bdef
**
246.8 10.3b
245.2 14.0b
532.8 5.5
**
488.8 11.4a
548.2 8.2be
481.9 8.5a
527.7 10.4b
580.8 14.0cd
559.7 10.7cde
542.4 17.7bd
**
554.4 10.3b
553.4 14.0b
230.2 4.9
**
181.6 10.2a
242.6 7.3bc
191.5 7.6a
228.6 9.3b
268.7 12.5c
253.3 9.6bc
245.1 15.8bc
**
248.0 9.2c
254.1 12.5c
2.31 0.05
**
1.92 0.11ad
2.45 0.08b
1.87 0.08ad
2.29 0.10b
2.82 0.13c
2.55 0.10bc
2.30 0.16bcd
**
2.56 0.10c
2.44 0.13bc
South-west monsoon
(Jun.- Sep.)
587
197.5 7.2a
505.5 7.2a
201.5 6.4a
2.02 0.07a
North-east monsoon
(Oct.- Dec.)
643
210.6 6.4a
517.9 6.5a
217.1 5.8b
2.24 0.06b
513
367
259
175
118
118
**
280.1 8.0c
237.1 8.3b
218.4 9.6ab
205.9 11.6a
187.4 13.7a
221.2 14.0ab
**
586.6 8.1c
544.9 8.3b
526.4 9.6ab
512.7 11.6a
495.5 13.7a
530.8 14.1ab
**
274.1 7.2c
240.3 7.4b
220.9 8.6ab
207.9 10.4a
201.2 12.2a
236.8 12.6ab
**
2.82 0.08c
2.40 0.08b
2.24 0.09ab
2.19 0.11ab
2.00 0.13a
2.24 0.13ab
Effect
Overall mean ()
Period of calving
P1 (1979-82)
P2 (1983-86)
P3 (1987-90)
P4 (1991-94)
P5 (1995-98)
P6 (1999-2002)
P7 (2003-2006)
Season of calving
Winter (Jan.-Feb.)
Summer (Mar.-May)
1550
Parity
First
Second
Third
Fourth
Fifth
Sixth and above
220
423
316
210
114
199
68
n= Number of observations.
* P<0.05, ** P<0.01. Means bearing same superscript within classes do not differ significantly (P>0.05).
296
monsoon calvers. On the other hand, the dry period

and number of services per conception observed
in south-west monsoon season calvers differed
significantly (P<0.05) from winter, summer and
north-east monsoon season calvers. The differences
of service period among parities were highly
significant (P<0.01). The service period decreased
with the order of lactation up to fifth parity and the
lowest value of 187.4 13.7 days was observed
at this parity. The reduction in service period in
the second and third parities was rather sharp and
further decline was gradual. In general, pairwise
comparison revealed that the mean service periods
of first and second parities differed significantly
(P<0.05) from other parities as well as between
them. Similarly the Murrah buffaloes calving for
the first time had the longest calving interval and
dry period, which declined thereafter. The calving
interval and dry period observed in the first parity
was significantly (P<0.05) different from second
and later parities. In addition, the mean calving
intervals and dry periods observed between the
second and fourth and fifth parities differed
significantly (P<0.05). The decline from third
to fifth parities was gradual and the differences
between means were not significant.
The mean service period, calving interval
and dry period of Murrah buffaloes observed were
in agreement with some research reports on Murrah
buffaloes (Kandasamy, 1987; Patnaik, 1988; Kundu
et al., 2003b). However, studies on these traits in
the Murrah breeding tract reported (Chhikara et al.,
1995a; Dass and Sadana, 2000; Banik and Tomer,
2003) much lower values than those observed in the
herd under study. The mean number of services per
conception observed (2.31 0.05) was also much
higher than that reported by Kumar et al. (2003).
However, Dutt and Yadav (1988) and Chhikara et
al. (1995b) found comparable estimates for Murrah
buffaloes maintained at the National Dairy Research

Institute, Karnal and the Buffalo Research Centre,
Hisar in India.
The main factor controlling variations in
the calving interval is the service period, which
in turn depends on post-partum oestrus days and
number of services per conception. In addition,
many other additional factors have been implicated
in lengthened calving intervals, such as embryonic
mortality, high milk production, seasonal and
environmental factors, age of cow and sire used for
service. The coefficient of variation obtained for
service period (67.3 percent) in the present study
indicates that the herd was more heterogeneous for
this trait. This strongly suggests better opportunities
for improvement through good breeding practices.
Hence, every effort should be made to reduce the
service period sufficiently to reduce the calving
interval.
The highly significant influence of period of
calving observed in the present study on different
production and reproduction traits was supported
by similar findings on Murrah buffaloes maintained
at different places in India (Kandasamy, 1987;
Sethi and Khatkar, 1997; Suresh et al., 2004;
Yadav et al., 2007). The difference in performance
of the animals among different periods might be
attributed to differences in management practices,
sires used for breeding, environmental conditions
such as ambient temperature, humidity, rainfall
etc., and variations in feed and fodder availability.
The significant to highly significant effect
of season of calving on different production traits
corroborated with the findings of the earlier workers
(Chhikara et al., 1998; Dass and Sadana, 2000) and
indicated that there was a pronounced seasonal
influence on the traits under study. The buffaloes
calving in the winter season had longer lactation
length and higher lactation milk yield than those
297
calving in the rainy seasons. This confirms the

findings in most of the earlier reports on Murrah
buffaloes (Rao and Rao, 1994; Dass and Sadana,
2000; Gogoi et al., 2002). The higher lactation
milk yield in the winter and summer seasons might
be due to the fact that the buffaloes calving in
those periods were found to have less gestational
stress due to longer service period and delayed
conception and also that during their descending
stage of lactation there was an abundant availability
of fodders, since it falls on monsoon seasons. The
lowest milk yield in monsoon calvers might be due
to the fact that they suffered from hot humid stress
and non-availability of quality fodder during the
major part of the lactation period.
The significant effect of season of calving on
service period and calving interval is in agreement
with findings reported in the literature for Murrah
buffaloes (Chhikara et al., 1995b; Kumar et al.,
2003; Suresh et al., 2004). It is generally observed
that buffalo cows are seasonally polyoestrus during
October to February and they breed regularly during
these periods. This might be one of the reasons for
shorter service period during monsoon seasons.
The higher number of services per
conception in winter calvers observed in this study
might be due to the fact that those animals calving
in winter exhibit post-partum heat in summer
load on conception rate should be implemented to

reduce number of services per conception. This in
turn will have a positive effect on service period
and calving interval.
The significant influence of parity on
different production and reproduction traits is
in accordance with the results obtained by other
researchers (Dass and Sadana, 2000; Kundu et
al., 2003b) on Murrah buffaloes. That the highest
305-day and lactation milk yields were obtained in
the third parity indicates that lactational maturity
is attained in the third lactation. These dindings
aresimilar to the reports of Ulaganathan et al.
(1983) and Kandasamy (1987). The significant
influence of calving sequence on service period
and calving interval and longer first calving interval
than the rest found in the present study concurred
with the other reports on Murrah buffaloes
(Kandasamy, 1987; Dass and Sadana, 2000; Kundu
et al., 2003a; Suresh et al., 2004). The reduction
in service period and calving interval over parities
might be due to differences in age of the animals
and periodic culling of buffalo cows with longer
calving intervals. The other plausible reason is that
following the irst two calvings, the physiological
rhythm may be maintained (i.e. reduced postpartum
oestrus days and better conception) which results in
shorter inter-calving periods in pluriparous buffalo
months and hence there would be reduction in

conception rate. Conception rate is related to
oestrous behaviour, time of oestrus detection and
insemination and site of semen deposition. Among
the different factors, accurate detection of oestrus
is of paramount importance in any reproductive
management programmes; however, this is
difficult in buffaloes during the summer season,
since most of them exhibit silent oestrus. Thus
accurate detection of oestrus and managemental
interventions to ameliorate the effects of heat
cows. Similarly the highest dry period in the first

parity and significant reduction in later parities
might be due to the reduction of calving interval.
The slight increase in reproduction traits from the
fifth to sixth and above parities might be due to the
lumping of all later records with the sixth parity.
CONCLUSIONS
The study revealed that non-genetic factors
298
such as period and season of calving had highly

significant effect on all the traits studied. In general,
the milk production performance of the farmbred
Murrah buffaloes at the Central Cattle Breeding
Farm, Alamadhi was comparable to that of animals
maintained in other government and institutional
herds in India. However, the comparatively lower
performance of Murrah buffaloes with respect
to fitness traits indicates less adaptability of the
breed to the hot and humid coastal region. Since
temporary environmental factors play a major role
on these fitness traits, better breeding management
is needed for improvement. In addition, multitrait evaluation with a combination of production
and reproduction traits may be carried out for
simultaneous improvement of production and
reproduction performances of Murrah buffaloes
in the herd. It is therefore imperative to emphasize
improvements in husbandry practices and
introduction of genetic evaluation programmes at
the same time.
Dairying, Foods and Home Sci., 14: 29-32.

Chhikara, S.K., N. Singh, S.S. Dhaka and R.S. Yadav.
1995b. Factors influencing reproduction
characters of Murrah buffaloes. Indian J.
Anim. Res., 29: 22-26.
Dass, G. and D.K. Sadana. 2000. Factors affecting
some economic traits in Murrah buffaloes.
Indian J. Anim. Res., 34:43-45.
Dutt, G. and M.C. Yadav. 1988. Factors affecting
number of services per conception and
calving interval in Murrah buffaloes. Asian
J. Dairy Res., 7: 168-170.
Gogoi, P.K., D. Das, R.N. Goswami, N. Nahardeka
and G.C. Das. 2002. Studies on age at
first calving in Murrah and Surti buffaloes
maintained in Assam. Indian Vet. J., 79: 854855.
Harvey, W.R. 1990. Users Guide for LSMLMW
and MIXMDL PC-2 Version. Mixed model
least-squares and maximum likelihood
computer program. Ohio State University,
Columbus, Ohio, USA.
Kandasamy, N. 1987. Genetic Analysis of Milk
Records in Farmbred Murrah Buffaloes
of Tamil Nadu. Ph.D. Thesis, Tamil Nadu
Agricultural University, Coimbatore, India.
Kumar, A., Z.S. Rana, S.S. Dhaka, D.S. Dalal,
S. Singh and R.K. Sethi. 2002. Test day
yield and lactation parameters of Murrah
buffaloes. In Proceedings of the 7th World
Congress on Genetics Applied to Livestock
Production, Montpellier, France.
Kumar, D., H. Singh, D. Kumar and C.V. Singh.
2003. Non-genetic factors affecting some
production and reproduction traits in Murrah
buffaloes. Indian J. Anim. Res., 37: 24-27.
Kundu, S., R.S. Pandey and S.K. Singh. 2003a. Nongenetic factors affecting some reproductive
traits in Murrah buffaloes. Indian J. Anim.
REFERENCES
Banik, S. and S.S. Tomar. 2003. Total determination
of calving interval by path analysis in
Murrah buffaloes. Indian J. Anim. Res., 37:
19-23.
Chhikara, S.K., N. Singh and S.S. Dhaka. 1998.
Effect of some non-genetic factors on peak
yield and days to attain peak yield in Murrah
buffaloes. In Proceedings of the 6th World
Congress on Genetics Applied to Livestock
Production, Armidale, Australia, 24: 481484.
Chhikara, S.K., N. Singh, S.S. Dhaka, R.S. Yadav.
1995a. Effect of certain factors on calving
interval of Murrah buffaloes. Journal
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Patnaik, A.K.C. 1988. Body weight and its effect
on first lactation yield in Murrah. M.V.Sc.
Thesis, Tamil Nadu Agricultural University,
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Sethi, R.K. and M.S. Khatkar. 1997. Genetic
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Suresh, R., D.K. Bidarkar, B. Ramesh Gupta,
B. Sudhakarrao and K. Sudhakar. 2004.
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of Murrah buffaloes. Indian J. Anim. Sci.,
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Sinha. 1984. Factors affecting first lactation
characteristics of Murrah buffaloes, p. 4.
In Proceedings of the Seminar on Buffalo
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Ulaganathan, V., K. Venkatarajulu and S.
Balachandran. 1983. Estimation of breeding
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300
Original Article
EFFECT OF GENETIC AND NON-GENETIC FACTORS ON MORPHOMETRIC TRAITS

OF BUFFALOES
Shashi Shankar 1, Dhirendra Kumar2 and K.G. Mandal3
ABSTRACT
order of lactation had the lowest magnitudes of all

the morphometric traits and significantly (P<0.05)
increased upto third parity indicating that the
skeletal maturity of the buffaloes might be attained
at the age of 3rd parity.
The present experiment was conducted

on 60 randomly selected dairy units consisting of
116 graded Murrah, 70 Diara type and 121 nondescript type buffalo cows utilizing the procedures
of stratified random sampling with proportional
allocation (Snedecor and Cochran, 1967) in and
around Patna (Bihar). The aim of this investigation
was to study the effect of genetic and nongenetic factors on various morphometric traits of
buffaloes under farmers management system. The
morphometric traits included in the study were
height at wither (haw, cm), body length (bl, cm)
and chest girth (cg, cm) as well as body weight of
the adult buffalo cows. The genetic factors were
the three different genetic groups of buffaloes
viz. graded Murrah, Diara and non-descript types
prevalent in Bihar (India), whereas the non-genetic
factors included in the study were location of
herd, farming system and sequence of lactation.
The farming system and the order of lactation had
significant (P<0.05) influence on HAW, BL and CG
whereas location of animals did not influence these
traits. The animals managed under mixed farming
system had significantly (P<0.05) higher estimates
of morphometric traits than those maintained in the
units involved dairying alone. The animals of first
Keywords: morphometric traits, buffaloes, genetic

factors, non-genetic factors
INTRODUCTION
Cattle and buffaloes are the main milk
producing animals in India. The buffalo forms the
backbone of Indias dairy industry and is rightly
considered as the bearer cheque of the rural
folk. Although being less than one third of the
total bovine population, the buffalo contributes
more than 50% of the total milk production in
the country. Although small buffalo herds are
distributed throughout the length and breadth of
the state, the area under South and North gangetic
plains of Bihar is densely populated with clusters
of buffaloes. The majority of them are of nondescript type, some are graded Murrah and the rest
of the population is phenotypically homogenous
in certain distinguishable characteristics. These
Mahatma Gandhi Veterinary College (M.G.V.C.), Bharatpur, Rajasthan 321 001, India
Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu (SKUAST-J), Jammu, India,
E-mail: dr.dhirendrakumar@yahoo.com
3
Department of Animal Genetics and Breeding, Bihar Veterinary College, Patna-14, India
1
2
301
to door survey method. Altogether 920 buffaloes

consisting of 331 graded Murrah, 221 Diara and
368 non-descript buffaloes were enumerated from
145 dairy units located in and around Patna.
Out of 145 enumerated units, only 120 units
were respondent units which provided relevant
information. Out of total 120 units, 50% i.e. 60
dairy units, consisting of 430 buffaloes of different
genetic groups, were randomly selected utilizing
the procedure of stratified random sampling with
proportional allocation (Snedecor and Chochran,
1967). Data were recorded from buffaloes of
defined genetic groups, which had completed at
least one calving interval. The morphometric traits
included in the study were height at wither (haw,
cm), body length (bl, cm) and chest girth (cg, cm)
as well as body weight of the adult buffalo cows.
The data were classified on the basis of genetic
group of the buffaloes, location of herd (Zone),
farming system and lactation order to study the
effect of various genetic and non-genetic factors
on the morphometric traits.
The experimental animals under study
were classified under three genetic groups: (i)
graded Murrah, (ii) Diara type (iii) non-descript
type.
Performance records of the buffaloes were
classified into four groups on the basis of sequence
of lactation: 1st parity, 2nd parity, 3rd parity, 4th
parity. The enumerated dairy units were grouped
according to the farming system adopted by the
farmers which were as follows:
i)
Mixed farming (animal husbandry
integrated with agriculture)
ii)
Only animal husbandry.
To quantify the variation due to various
genetic and non-genetic factors on different
economic traits, the data were subjected to least
squares analysis (Harvey, 1966) for which the
phenotypically homogenous buffaloes are almost

true to the breed and have acquired full adoption to
the socio-agro-climatic and ecological conditions
of Gangetic plains, particularly in the Tal and
Diara area of the rivers Ganga, Gandak and Sone
in the division of Patna, Bhagalpur and Magadh.
These true breeding buffalo populations are
particularly called as Diara by the farmers. It
is pertinent to mention here that the true breeding
buffalo population (about 1.3 million of the total
buffalo population of the state) locally known as
Diara/Deshila which contribute about 32% of the
State Milk Pool, is almost untouched, so far as
their breeding and development programmes are
concerned, and hence the present study has been
undertaken.

Buffaloes consisting of three genetic
groups namely graded Murrah, Diara and nondescript types maintained in private dairy units at
farmers door located in a radius of 15 km in and
around Patna were the experimental animals for the
present study. The whole area under study was
divided into three distinct zones, which were as
follows:
Zone I
North West Patna
Zone II
South West Patna
Zone III
East Patna
The primary survey was conducted in
private dairy units popularly known as Khatals
located in a radius of 15 km in and around Patna.
Those Khatals which consisted of at least two
or more buffaloes consisting of graded Murrah,
Diara or non-descript buffaloes either alone or in
combination were enumerated, utilizing a door
302
estimate of height of wither for Diara buffaloes

reported by Singh et al. (2006) was in consonance
with the result of the present study. Thus it may
be concluded that graded Murrah, Diara and nondescript buffaloes in and around Patna (Bihar) were
observed to be smaller and lighter than Murrah
but heavier than Mehsana, Surti, Bhadawari and
Nagpuri buffaloes.
As evident from Table 1, the average height
at wither was highest (130.4560.183 cm) in the
animals located in southwest Patna of the study
area followed by northwest (130.1800.177 cm.)
and east Patna (130.1090.198 cm.).
Farming system had a significant (p<0.01)
influence on height at wither. The animals reared
under a mixed farming system integrated with
agriculture had the higher estimate (130.5130.157
cm) for HAW than those managed in the units
exclusively involved in dairying by 0.53 cm (Table
1). The difference was statistically significant
(p<0.05). Significantly higher estimates of HAW
in all the age groups for animals maintained in
livestock research stations than those maintained
under farmers management systems as observed
by Singh (1995a) were in consonance with the
findings of the present study. Significant (p<0.05)
difference for HAW between animals managed
under different farming systems might be attributed
to the fact that animals managed under better
management systems may have better growth.
The average HAW was found to be
increased significantly (p<0.05) and steadily
from first (124.3910.197 cm) to third lactation
(133.9520.213 cm) by 9.561 cm followed by slight
decrease in fourth lactation. However, the difference
in HAW between third and fourth parity was not
statistically significant. The significant (p<0.01)
influence of parity on height at wither observed in
the present investigation was in accordance with
following mathematical model was utilized.

YiJKlm = + Gi + Zj + Fk + Pl + eijklm
where, Yijklm = The value of mth individual under
ith genetic group, Jth location, Kth farming system
and lth parity.
= The overall population mean.

Gi
= The effect of ith genetic group
(i = 1,2,3).
Zj
= The effect of jth location of herd
(j = 1,2,3).
Fk
= The effect of Kth farming system
(K = 1,2).
Pl
eijklm
= The effect of lth parity (l = 1,2,3,4).

= The random error associated with
individual which is randomly and
independently distributed with mean
zero and variance e2
The statistical significance of various fixed

effect was tested by the F test whereas DMRT, as
modified by Kramer (1957), was applied to carry
out the pair wise comparisons among least squares
means at the 0.05 and 0.01 levels of probability.

The average height at wither of graded
Murrah was estimated to be 132.1130.187 cm.,
which differed significantly (p<0.01) from Diara
and non-descript buffaloes by 2.295 cm and 3.299
cm, respectively. The mean values obtained in
the present investigation for all the three genetic
groups viz. graded Murrah, Diara and non-descript
types, were lower than those reported elsewhere
for Murrah (Jawarkar and johar, 1975; Sreedharan,
1976 and Manik and Iqbalnath, 1981).The average
303
the findings of many workers (Jawarkar and Johar,

1975; Saini and Gill, 1986; Jogi and Patel, 1990;
Singh et al., 1995.
a significant (p<0.01) influence on body length.

The animals managed in the units integrated with
agriculture farming had significantly (p<0.05)
higher mean body length (137.0030.197 cm) in
comparison to those involved in the dairying alone
by 1.151 cm (Table 1). Differences in body length
were also observed by Singh et al. (1995a) in all
the age groups of Mehsana buffaloes managed in
LRS herd from those managed in milk union sheds
under farmers management systems which supports
the finding of the present study.
The lactation order had highly significant
(p<0.01) effect on body length. The lowest average
body length pooled overall the three genetic groups
viz. graded Murrah, Diara and non-descript types,
was estimated to be 130.9470.247 cm in the first
parity and then increased significantly (p<0.05)
over the lactations. The highest average body
length (140.0070.268 cm) was observed in third
parity, which was significantly (p<0.05) longer than
the average body length observed during first and
second parity by 4.279 and 9.06 cm, respectively.
The average body lengths during third and fourth
order of lactations were also significantly (p<0.05)
longer than in second parity by 4.781 and 4.505
cm, respectively. However, though the body length
in fourth parity was little bit shorter than in third
parity, the two did not differ significantly.
Body length
Overall least squares mean for body
length pooled over three different genetic groups
of buffaloes viz. Diara, graded Murrah and nondescript types included in this study, was reckoned
to be 136.4780.138 cm (Table 1).
Genetic group had a highly significant
(p<0.01) effect on body length. As revealed from
Table 1, the graded Murrah had the longest body
length (138.5980.235 cm) followed by Diara
(135.5070.232 cm), and non-descript had the
shortest body length (135.3280.231 cm). The
average body length of graded Murrah differed
significantly (p<0.05) from Diara and non-descript
types. The average estimates for body length in all
the three genetic groups obtained in the present
study were lower than the body length of Murrah
buffaloes as reported elsewhere by Jawarkar and
Johar (1975), Sreedharan (1976) and Manik and
Iqbalnath (1981). The significant (p<0.05) effect
of genetic group on body length reported by Sinha
(2006) and Singh et al. (2006) was in agreement
with the findings of the present study.
As evident from Table 1, the average body
length was found to be highest (136.7960.230 cm)
in the animals located in southwest Patna of the
study area followed by northwest (136.3840.22
cm) and lowest in east Patna (136.2530.249 cm).
The least squares analysis of variance revealed
non-significant effect of zone on body length. The
non-significant effect of location on body length as
observed in the findings of the present study is in
agreement with the results reported by Singh et al.
(1995) in Mehsana buffaloes.
The farming system was found to have
Significant influence of parity on body

length as observed in the present study has also
been reported in Murrah buffaloes by Jawarkar
and Johar (1975), Saini and Gill (1986), Jogi and
Patel (1990) and in Mehsana buffaloes by Singh et
al. (1995) and Singh et al. (2000), who stated that
body length in buffaloes increased significantly up
to third parity.
Chest girth
The overall least squares mean for chest
304
girth pooled over the three different genetic groups

of buffaloes, viz. Diara, graded Murrah and nondescript, included in this study was estimated to be
195.6920.384 cm (Table 1).
Genetic group had highly significant
(p<0.01) influence on chest girth.. As evident
from Table 1 the graded Murrah had the largest
chest girth (199.1530.653 cm) followed by nondescript (196.6400.6421 cm) and Diara buffaloes
(191.2820.646 cm). The chest girth in Diara
buffaloes was found to be significantly (p<0.05)
lower than the graded Murrah and non-descript
buffaloes by 7.871 and 5.358 cm, respectively.
The findings of the present study were
not in agreement the findings of Jawarkar and
Johar (1975), Sreedharan (1976) and Manik and
Iqbalnath (1981) who reported higher estimates of
chest girth in Murrah buffaloes.
As evident from Table 1, the highest average
chest girth was found to be 196.9530.641 cm in
the animals located in southwest Patna of the study
area followed by northwest Patna (195.2570.619
cm) and lowest in east Patna (194.8660.693 cm).
The least squares analysis of variance revealed nonsignificant effect of zone on chest girth. The nonsignificant effect of location on chest girth reported
by Singh et al. (1995a) in Mehsana buffaloes is in
agreement with the results of the present study.
The farming system was found to have
highly significant (p<0.01) influence on chest
girth. The animals managed in the units integrated
with agriculture farming had significantly (p<0.05)
higher average chest girth (197.5840.549 cm) in
comparison to those involved in dairying alone
by 3.785 cm (Table 1). Similar findings have also
been reported by Singh et al. (1995) who observed
significant difference in heart girth among all the
age groups of Mehsana buffaloes between LRS
herd and milk union sheds where animals were
under the farmers management system.

The order of lactation had highly significant
(p<0.01) influence on chest girth in buffaloes in and
around Patna. The average chest girth pooled over
all the three genetic groups viz. graded Murrah,
Diara and non-descript buffaloes was found to be
the lowest (186.4670.688 cm) at first parity.
The average estimates of chest girth in
second, third and fourth parities were observed to
increase significantly (p<0.05) over the average
chest girth at first parity by 7.85, 15.362 and
13.687 cm, respectively. A significant effect of
parity on chest girth is also reported by Jawarkar
and Johar1975), Saini and Gill (1986), Jogi and
Patel (1990) in Murrah and (Singh et al. (1995) and
Singh et al. (2000) in Mehsana buffaloes.
SUMMARY
Graded Murrah had significantly (P<0.05)
higher estimates of all the morphometric traits
(HAW, BL and CG) than the Diara and non-descript
types, whereas, Diara buffaloes had superiority
over non-descript types for height at wither and
chest girth. The average estimates of height at
wither, body length and chest girth of graded
Murrah were 132.113+0.187, 138.598+0.235 and
199.153+0.0653 cm respectively. Therefore, the
size of Diara buffaloes was in between Graded
Murrah and Non-descript types. The farming
system and the order of lactation had significant
(P<0.05) influence on HAW, BL and CG whereas
location of animals did not influence these traits.
The animals managed under mixed farming system
had significantly (P<0.05) higher estimates of
morphometric traits, than those maintained in the
units involved dairying alone.
305
Table 1. Least squares meansSE and CV% of morphometric traits of buffaloes in and around Patna.
Particulars
Overall Mean ()
Morphometric traits (cm)

HAW (MeanS.E.)
B.L. (MeanS.E.)
C.G. (MeanS.E.)
130.2480.110
136.4780.138
195.6920.384
(1.47)
(1.77)
(3.43)
132.113a0.187
138.598a0.235
199.153a0.653
(1.41)
129.818b0.185
(1.69)
135.507b0.232
(3.27)
191.282b0.646
(1.42)
128.814c0.184
(1.71)
135.328b0.231
(3.41)
196.640c0.642
(1.42)
(1.70)
(3.34)
130.1800.177
136.3840.222
195.2570.619
(1.44)
130.4560.183
(1.72)
136.7960.230
(3.35)
196.9530.641
(1.44)
130.1090.198
(1.73)
136.2530.249
(3.35)
194.8660.693
(1.44)
(1.72)
(3.35)
129.983a 0.149
135.952b0.187
193.800a0.520
(1.45)
130.513b0.157
(1.73)
137.003a0.197
197.584b0.549
(1.46)
(1.74)
124.391a 0.197
130.947a0.247
186.467a0.688
(1.49)
128.907b 0.188
(1.77
135.226b0.236
(3.48)
194.317b0.657
(1.42)
133.952c 0.213
(1.70)
140.007c0.268
(3.31)
201.829c0.745
(1.38)
133.743c0.273
(1.66)
139.731c0.343
(3.31)
200.154c0.955
(1.38)
(1.66)
(3.23)
Factors
Genetic group
Graded Murrah
Diara
Non-descript
Location
1. Northwest Patna
2. Southwest Patna
3. East Patna
Farming System
1. Animal Husbandry
alone
2. Mixed Farming
Lactation order
1st
2nd
3rd
4th
Means with different superscripts (column-wise) differed significantly (P<0.05)

Values in parentheses are CV%
306
CONCLUSION
different physical characteristics in Murrah

type heifers and dry buffaloes. Indian J.
Anim. Prod. Mgmt., 3(4): 193-199.
Singh, D.V., R.K. Pundir, G. Sahana, A.S. Dave
and A.E. Nivsarkar. 2000. Characterization
of Mehsana buffaloes. NBAGR Research
Bulletin, 8: 20-27.
Singh, D.V., V.N. Tripathi and A.S. Dave. 1995.
The animals of first lactation had the lowest

magnitudes of all the morphometric traits, and these
magnitudes significantly (P<0.05) increased upto
third parity indicating that the skeletal maturity
of the buffaloes might be attained at the age of 3rd
parity. On the basis of the findings of the present
study it can be concluded that the Diara buffaloes
were significantly different from Graded Murrah
and non-descript types in respect to morphometric
traits;therefore, work of a similar type may be
repeated in the entire Tal and Diara areas of the
river Ganges, Gandak and Sone pertaining to Bihar
to identify and enumerate the number of Diara
buffaloes so that a suitable breeding plan can be
chalked out for improvement of Diara buffaloes
and to improve the livelihood of the dairy farmers
in the state.
Studies on morphological traits in Mehsana

buffalo cows. Indian J. Dairy Sci., 48(1):
33-38.
Singh, D.V., V.N. Tripathi and A.S. Dave. 1995a.
Body measurements in female Mehsana
buffaloes. Indian J. Dairy Sci., 72: 12821285.
Singh, S.R., K.G. Mandal, P.K. Singh, R. Kumar
and R.N. Singh. 2006. Phenotypic parameter
of true breeding buffalo population in Tal
and Diara areas in Bihar. In Proceedings of
National Symposium on Conservation and
Improvement of Animal Genetic Resources
under Low Input System : Challenges and
Strategies, NBAGR, Karnal, Haryana,
India.
Sinha, R.K. 2006. Characterization of buffalo
genetic resources in Tal and Diara areas in
and around Barh (Patna). M.V. Sc. Thesis,
RAU, Pusa (Samastipur), Bihar, India.
Snedecor, G.W. and W.G. Cochran. 1967. Statistical
Methods, 6th ed. Iowa State Univ. Press,
Ames, USA.
Sreedharan, S. 1976. Inheritance of body size
measures in relation to production
efficiency in buffaloes. Ph.D. Thesis, Punjab
University.
REFERENCES
Harvey, W.R. 1966. Least Squares Analysis of Data
with Unequal Subclass Number. United
States Department of Agriculture (USDA).
Jawarkar, K.V. and K.S. Johar. 1975. A study on
some of the body measurements on Murrah
buffaloes. Indian J. Dairy Sci., 28(1): 5456.
Jogi, R.V.K. and D. Patel, U.G. 1990. Various body
measurements and their correlation with
milk yield and fat percentage. II Studies on
Surti buffaloes. Buffalo Bull., 9(2): 35-38.
Manik, R.S. and Iqbalnath. 1981. Relationship
of certain body measurements with milk
production in Murrah buffaloes. Indian J.
Dairy Sci., 34(1): 118-119.
Saini, A.L. and R.S. Gill. 1987. Relationship among
307
Original Article
OBSERVATIONS ON BUFFALO SARCOPTIC MANGE IN JAMMU, INDIA
T. Nazir1, R. Katoch1, R. Godara1,*, Anish Yadav1 and Vijay Pandey2
ABSTRACT
INTRODUCTION
Observations on sarcoptic mite infestation

in 1440 buffaloes belonging to 72 unorganised
dairy farms of Jammu revealed a 3.33% prevalence
rate. Prevalence was significantly (p0.05-0.01)
higher during the winter (5.55%) as compared
with the summer (2.91%) and the rainy (1.33%)
seasons. The disease was recorded in 3.75% and
2.5% of buffaloes up to 1 year and above 1 year of
age, respectively. Females showed a significantly
(p<0.05) higher prevalence rate (4.07%) than
males (1.96%). A strong negative correlation
(r=-0.725) was observed between the prevalence
rate and temperature whereas relative humidity had
little or no association (r=0.2). The initial lesions
were observed on the face and the upper part of
neck as papules and exudative crusts, amd then
the infestation spread quickly causing generalized
mange. In severe cases, animal had emaciated body
with no sign of fat deposits and loss of normal skin
colour and texture. Infested buffaloes showed a
significant (p<0.05) association with inappetance.
Mite infestation, particularly with the

obligate, burrowing mite Sarcoptes scabiei, is an
extremely pruritic, contagious and debilitating
disease of buffaloes (Naresh et al., 2005; Chhabra
and Pathak, 2011). It is recognized as one of the
most serious diseases of dairy animals in the
Indian subcontinent (Gill et al., 1989). The disease
spreads rapidly in dairy and breeding herds due to
overcrowding, malnutrition and poor managemental
conditions. Though economic losses must be of a
very high magnitude due to hide damage, decreased
milk and meat production, morbidity and mortality,
no accurate figures are available. The morbidity rate
varies from 1.5 to 82%, and reaching up to 100% in
severely affected herds, with heavy losses of young
animals (Gill et al., 1989; Patel et al., 2002a).
Despite this, no epidemiological study vis--vis
the incidence of sarcoptic mange in buffaloes was
undertaken previously in the state of Jammu and
Kashmir of India. The present work was therefore
conducted to determine the prevalence and clinical
signs associated with sarcoptic mite infestation
in buffaloes (Bubalus bubalis) and the effect of
climatic factors on the prevalence of sarcoptic
Keywords:
buffaloes,
prevalence, Jammu
Sarcoptes
scabiei,
Division of Veterinary Parasitology, Faculty of Veterinary Sciences and Animal Husbandry, Sher-e-Kashmir
University of Agricultural Sciences and Technology, R.S. Pura-181 102, Jammu, India, *E-mail: godara1972@
gmail.com
2
Department of Biochemistry, Veterinary College Mathura, Pandit Deen Dayal Upadhaya Pashu Chikitsa
Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan, Mathura, India
1
308
RESULTS
mange at Jammu, India.
Observations on the prevalence of sarcoptic

mange
During the year a total of 1440 buffaloes
belonging to 72 unorganised dairy farms of
Jammu were examined. Out of that total, 3.33% of
buffaloes were found to be infested with S. scabiei
var. bubalis (Table 1). The highest prevalence was
found in the month of February (7.5%), followed
by March (6.66%) and December (5.0%). The
lowest prevalence (0.83%) was found in the month
of May (Figure 1). None of the animals examined
were found positive during the months of June and
July. Significant association (p0.05-0.01) was
found between the prevalence rate and the seasons
and the highest prevalence rate (5.55%) was
found in the winter season (Table 1). There was
a strong negative correlation (r=-0.725, r2=0.525)

Study area, animals and techniques
The study was carried out in 1440
buffaloes belonging to 72 unorganised dairy farms
of Jammu of Jammu and Kashmir state, located
in north-western region of India, between 74o 50
east longitude and 30o 40 north latitude. The area
lies about 332 m above sea level and experiences
a subtropical humid climate. The area experiences
four seasons; summer (March to June), rainy (July
to September), post-rainy (October-November)
and winter (December to February).
Skin scrapings of buffaloes with visible
dermatitic lesions were collected using the
method given by Tikaram and Ruprah (1986).
These samples were brought to the laboratory and
examined under stereomicroscope using standard
techniques described elsewhere (Soulsby, 1982).
The data were recorded according to the age (up to
1 year and above 1 year) and sex (male/ female).
between the prevalence rate and the mean monthly

temperature (Figure 2) and a weak positive
correlation (r=0.2, r2=0.04) was found between
the prevalence rate and mean monthly relative
humidity (Figure 3). Sarcoptic mange infestation
was observed in 3.75% and 2.5% buffaloes up to
1 year and above 1 year of age, respectively (Table
1). However, the infestation was not significant.
The females had a significantly (p<0.05) higher
prevalence rate (4.07%) than the males (1.96%)
(Table 1).
Statistical analysis
Differences in the prevalence of mite
between seasons, age groups and sexes were
tested for significance by the chi-square test. The
relationship between the monthly variation of
the infection index and the one of the climatic
parameters: temperature and relative humidity were
determined by Pearson correlation analysis using
SPSS 16.0. A p-value of 0.05 was considered
significant.
Clinical observations
The initial lesions were often localized
accompanied by alopecia and were found on the
body parts which had thin hair and were prone to
scratches/ injuries such as the face and the upper
part of the neck. Small papules with intense itching
were evidenced as a result of early inflammatory
reactions to the mites invasion and injury to the
309
Figure 1. The prevalence (%) of sarcoptic mange in buffaloes from January 2010 to December 2010.
Figure 2. Scatter graph showing a strong negative correlation (r=-0.725, r2=0.525) between the prevalence
rate (%) of sarcoptic mange and mean monthly temperature (oC).
310
Figure 3. Scatter graph showing a weak positive correlation (r=0.2, r2=0.04) between the prevalence rate (%)
of sarcoptic mange and mean monthly relative humidity (%).
Figure 4. The prevalence (%) of sarcoptic mange on buffaloes in respect of site locations.
311
360
480
360
240
1440
Winter
Summer
Rainy
Postrainy
Total
48
14
20
No.
infested
3.33
3.33ab
1.66b
2.91b
5.55a
Percent
Male
Female
508
88
117
159
144
10
1.96p
2.27
1.88
3.47
932
152
243
321
216
38
11
15
examined infested percent examined infested
Figures with different superscripts differ significantly (p 0.05-0.01).
No.
examined
Season
Up to 1 year
4.07q
3.94
2.46
3.42
6.94
960
160
240
320
240
36
10
14
percent examined infested
Table 1. The prevalence of Sarcoptes scabiei on buffaloes in respect of season, sex and age.
3.75
3.75
2.5
3.12
5.83
percent
Above 1 year
480
80
120
160
120
12
2.5
2.5
2.5
examined infested percent
312
The occurrence of sarcoptic mange is

usually considered to be seasonal and is often
restricted to a few months of the year (during winter
and early summer months) (Tikaram and Ruprah,
1986; El-Khodery et al., 2010). Probably, with
increase in the mean temperature during summer
months, the mite numbers decrease rapidly due to
less active feeding and decreased oviposition by
the mites (Soulsby, 1982). Further the maximum
survival of both sarcoptic and psoroptic mites
has been recorded at 20-27oC rather than at 3139oC (Maske and Ruprah, 1981). During the hot
summer months, the disease is recognized as
subacute (dormant or quiescent) or chronic with
focal lesions mainly in the groin and submaxillary
regions. During this quiescent phase a spontaneous
cure has occurred. Since the more severe lesions
may subside, itching and pruritus are relieved and
new hair growth starts. These quiescent lesions are
potential sources of clinical infection and the risk
of a sudden outbreak of mange within a herd with
the change of season poses a serious threat.
The young animals showed a higher
infestation rate of S. scabiei than the adults. This
could be attributed to soft and tender skin, dense
hair coat, huddling tendency and relatively low level
of immunity in young animals (Patel et al., 2003a).
However, Cernea and Cernea (2006) concluded
that the mange-mite infestation develops in adult
animals irrespective of sex with higher prevalence
of psoroptic mange at 5-13 years of age. Although
male and female animals are kept under similar
managemental conditions, a significantly (p<0.05)
higher infestation rate in females than the males
suggests that differences in susceptibility between
sexes may exist.
In mite infestation, host reactions such as
cutaneous inflammation and intense pruritus are
associated with response to the mites burrowing
skin, which leads to rubbing of the affected parts

with manger or the walls. The infestation spread
quickly from these localized lesions causing
generalized mange. Although the skin lesions were
found all over the body, the neck (47.5%) and the
face (39.6%) were the most affected parts (Figure
4). However, some animals had the lesions on two
sites together (Figure 4). The lesions appeared as
exudative crusts, scales, haemorrhagic and nonhaemorrhagic fissures. A decrease in feed intake
and a rapid fall in milk yield in lactating buffaloes
were prominent features of the infestation. In severe
cases, excess keratinization and thickening of skin
were observed, which resulted in corrugations
or folds often spread over with a fine, chalk-like
covering of scurf. Severely infested buffaloes had
emaciated bodies with no sign of fat deposits. The
characteristic signs of the infestation were loss of
normal skin colour and texture.
The infested buffaloes showed a significant
(p<0.05) association with inappetance. Out of 48
infested buffaloes, 29 (60.41%) showed inappetance
while 19 (39.58%) had normal appetite.
DISCUSSION
Sarcoptic mange is one of the few diseases
that seriously threaten the well-being of the water
buffalo (Bubalus bubalis). A higher incidence rate
of 27.6% of sarcoptic mange in buffaloes has been
reported from organised dairy farms of Haryana
state of India (Tikaram and Ruprah, 1986). ElKhodery et al. (2010) observed Psoroptes and
Chorioptes spp. infestation in 16.66% buffaloes
of smallholder farms in the Nile Delta region of
Egypt. As against the above observations, there is
an incidence rate of 3.33% currently recorded in
Jammu.
313
and feeding activities, which usually occur three

weeks after the initial infestation (Colebrook and
Wall, 2004; Rahbari et al., 2009). The reaction time
is longer accompanied by severe itching during
early stages of infestation when the mite numbers
are low. Subsequently, the reaction time becoming
much shorter after repeated exposures (Gary
and Durden, 2009). The induction of cutaneous
hypersensitivity in response to mite faecal antigens
may cause proliferation of mast cells and resultant
increase in chymase and tryptase activities (Noviana
et al., 2004) as well as decrease in anti-oxidant
enzyme activities and trace minerals suggesting
its association with compromise in anti-oxidant
defense and resulting in oxidative stress (Dimri et
al., 2008); these phenomena are supposed to play
an important role in development of pathogenicity.
Itching may develop as a result of histamine that is
released from the destroyed cells and that is a potent
pruritogen (Greaves and Wall, 1996). The rubbing
and scratching of the affected areas in an attempt to
alleviate the itchiness, result in the shedding of mites
in the surroundings for subsequent transmission to
other animals. Excessive rubbing and scratching
leads to excoriation and haemorrhage on the skin
surface and even death, particularly in animals
below one year of age when the lesions are large
(Patel et al., 2002a).
The mite infestation is often accompanied
by inappetance, and as a result, loss of weight gain
may occur (Rahbari et al., 2009). El-Khodery et al.
(2010) reported a significant (p<0.01) inappetance
in buffaloes infested with Psoroptes spp. mites.
Inappetance in mite infestation may develop as a
result of alteration in hepatic function and structure
and decrease in digestibility of nutrients (Dimri
and Sharma, 2004b). Damage of hepatic tissues in
cattle calves infested with Psoroptes ovis has also
been reported (Fisher and Crookshank, 1982).
On the basis of the results of the study, it may

be concluded that the season and sex are important
and significant predisposing factors for sarcoptic
mange in buffaloes. The application of suitable
acaricides would help to reduce the infestation and
to sustain the optimal growth and productivity of
buffaloes and minimise the perpetuating financial
losses to resource-poor communities in the region.
REFERENCES
Cernea, C. and M. Cernea. 2006. Establishing
the causal factors in the development of
psoroptic mange in buffalo from S.C.D.C.B.
Sercaia, Brasov District. Lucrari Stiintifice,
40: 349-354.
Chhabra, M.B. and K.L.M. Pathak. 2011. Sarcoptic
mange in domestic animals and human
scabies in India. J. Vet. Parasitol., 25: 1-10.
Colebrook, E. and R. Wall. 2004. Ectoparasites of
livestock in Europe and the Mediterranean
region. Vet. Parasitol., 120: 251-274.
Dimri, U. and M.C. Sharma. 2004b. Effects of
sarcoptic mange and its control with oil of
Cedrus deodara, Pongamia glabra, Jatropha
curcas and benzyl benzoate, both with and
without ascorbic acid on growing sheep:
assessment of weight gain, liver function,
nutrient digestibility, wool production and
meat quality. J. Vet. Med. A, 51: 79-84.
Dimri, U., S.C. Sharma, D. Swarup, R. Ranjan and
M. Kataria. 2008. Alterations in hepatic lipid
peroxidase and antioxidant profile in Indian
water buffaloes suffering from sarcoptic
mange. Res. Vet. Sci., 85: 101-105.
El-Khodery, S.A., S.A. Osman, M. Ishii and M.H.
Al-Gaabary. 2010. Risk factors of infestation
by Psoroptes spp. mites in buffalo (Bubalus
314
Rahbari, S., S. Nabian and A.R. Bahonar. 2009.

Some observations on sheep sarcoptic
mange in Tehran province, Iran. Trop. Anim.
Health Pro., 41: 397-401.
Soulsby, E.J.L. 1982. Helminths, Arthropods and
ELBS and Bailliere Tindall, London, UK. p.
1-809.
Tikaram, S.M. and N.S. Ruprah. 1986. Incidence
of sarcoptic mange in buffaloes in India.
Trop. Anim. Health Prod., 18: 86-90.
bubalis) at smallholder farms in the Nile

Delta region, Egypt. Trop. Anim. Health
Pro., 42: 275-281.
Fisher, W.F. and H.R. Crookshank. 1982. Effects
of Psoroptes ovis (Acarina: Psoroptidae) on
certain biochemical constituents of cattle
serum. Vet. Parasitol., 11: 241-251.
Gary, R.M. and Durden. 2009. Medical and
Veterinary Entomology, 2nd ed. Elsevier
Publication, Amsterdam. p. 469-470.
Gill, B.S., J. Singh, B.S. Gill, A. Singh, S.S.
Khehra, A. Rai and O. Hussain. 1989.
Efficacy of ivermectin against mange
and gastrointestinal nematodes of buffalo
(Bubalus bubalis). Vet. Parasitol., 31: 141147.
Greaves, N.W. and P.D. Wall. 1996. Pathophysiology
of itching. Lancet, 348: 938-940.
Maske, D.K. and N.S. Ruprah. 1981. Note on
in vitro survival of psoroptic mange in
buffaloes at different temperature and
relative humidities. Indian J. Anim. Sci., 51:
563-564.
Naresh, R., D. Swaroop, M.C. Sharma and R.
Ranjan. 2005. Clinical management of
sarcoptic mange in Indian buffalo calves
with the botanical ointment. Vet. Rec., 156:
684-685.
Noviana, D., D. Harjanti, Y. Otsuka and Y. Horri.
2004. Proliferation of protease-enriched
mast cells in sarcoptic skin lesions of racoon
dogs. J. Comp. Pathol., 131: 28-37.
Patel, J.S., P.R. Patel and H.H. Panchasara. 2002a.
Economic losses due to sarcoptic mange in
buffalo calves. Vet. Pract., 3: 186-189.
Patel, J.S., P.R. Patel, H.H. Panchasara and K.G.
Brahmaxtri. 2003a. Epizootiology of
sarcoptic mange in buffalo calves. Indian
Vet. J., 80: 972-974.
315
Original Article
BUFFALO MILK MARKETING BY THE GUJJARS (TRIBALS) IN THE JAMMU AND

KASHMIR STATE IN INDIA
Tarunvir Singh1 and Sudhakar Dwivedi2
is actually marketed. Marketed surplus may be

less, equal or more than the marketable surplus
depending upon the external factors operating on
the market economy.
It is often argued that marketed surplus of a
commodity is a function of the level of production
and consumption of the commodity. Eventually
large producers would market more both in
absolute terms as well as in relative terms. But in
the case of milk, being a perishable commodity,
sometimes there is distress sale, particularly among
small and marginal farmers, who will have a cash
crunch unless adequate market infrastructure is
provided, and it is often seen that the majority of
large producers sell proportionately less milk than
what they had produced. Thus, farms, unless they
happen to be specializing in dairy farming, do not
enter the market in a big way. Because of higher
home consumption of milk by large producers, what
is just non-marketing level for them is sufficiently
a specialized market level for small or marginal
farmers or landless labourers (Singh, 1978). The
retention capacity of the producer depends upon
his financial position, the credit facilities available
to him and the price level in the market. The proper
incentives and adequate market infrastructure for
the excess production of milk will not only help in
increasing milk production, but also increase the
marketed surplus. The marketed surplus is mainly
governed by production, domestic consumption,
ABSTRACT
The present investigation is an attempt to
study the marketing of buffalo milk produced by a
well-known pastoral community-the Gujjars of the
Jammu and Kashmir state. The study was undertaken
in the Jammu region. About 200 households were
studied in different districts of the Jammu region.
Marketed surplus of milk and factors affecting
it, marketing cost, margin and price spread were
studied. The Cobb-Douglas production function
was used to study the relationships between
marketed surplus of milk and factors affecting it,
and it was found that milk production and the price
of milk had a positive and significant impact on
the marketed surplus of milk. The producers share
in the consumers price was maximum in the first
channel (Producer- Consumer).
Keywords: marketed surplus, marketing margin,
marketing cost, price spread
INTRODUCTION
The excess of output over sectoral retention
refers to what is termed as marketable surplus. This
is distinct from marketed surplus. which represents
only that portion of the marketable surplus which
Institute of Economic Growth, New Delhi, India, E-mail: vir.tarun@gmail.com

Division of Agricultural Economics and Statistics, Sher-e-Kashmir University of Agricultural Sciences and
Technology- Jammu, 180009, India
1
2
316

based on the classification into three groups on
the basis of number of milch buffaloes (cube root
frequency method) namely small farms (up to 5),
medium farms (6-10) and large farms (10 & above).
Primary data on marketing of milk were collected
by the personal interview method with the help of
a structured and pre-tested schedule.
The quantity of milk available for sale
depends upon a number of factors like production,
price demand, etc. Milk being a perishable cannot
be stored for future date for sale and therefore,
depend upon immediate demand. Although a
large number of studies have been carried out to
develop marketed surplus functions in agricultural
commodities, little empirical evidence is available
on the marketed surplus of milk. Therefore, in this
section the empirical nature of relationship between
the marketed surplus of milk and its determinants
was examined. The explanatory variables
considered to develop the marketed surplus function
were total milk production, family size, education
level of the head of the family1, number of milch
animals and price of milk. The education status
(literate or illiterate) of the head of the household
was considered as a variable affecting the marketed
surplus of milk. A critical examination of zero order
correlation matrix among explanatory variables
revealed the absence of multicollinearity problem.
The estimated marketed surplus function for milk
in respect to Jammu, Udhampur, Rajouri, Poonch
and overall are reported in Table 1.
The following model was employed to find
the marketed surplus for different households in the
present study.
MS = f (X1, X2, X3, X4, X5)
MS = Marketed surplus of milk per
household per day in kg
X1 = Total milk production per household
in kg
price and market infrastructure.

In order to provide the minimum
nutritional requirements of milk to the majority
of population, the marketed surplus has to be
increased substantially. The disposal of milk in a
milk producing household consist of liquid milk
consumption and conversion of milk into various
milk products at the producer household, and
the balance is available for sale of milk, which
constitutes as marketed surplus.
The present investigation was an attempt
to study the marketing aspects of dairying of one of
the well-known pastoral communities-the Gujjars
of the Jammu and Kashmir state in India. Though
information on sociological and anthropological
aspects of these pastoral communities has been
generated in various studies elsewhere, socioeconomic studies are scanty. No concerted efforts
had heretofore been made to study the economics of
dairying among the Gujjars, who are well known for
rearing buffaloes only, particularly with reference
to various important economic parameters like
marketing of milk. With a view to identifying the
factors responsible for marketed surplus of milk,
marketed surplus functions were fitted separately
for different types and categories of milk producers
in the study area.

Four districts of the Jammu region were
selected purposively and from each district two
blocks were also selected purposively. One village
was selected purposively from each block:that in
which the population of the Gujjars involved in
rearing of buffaloes was highest. From each block
25 Gujjar dairy farmers were selected randomly
to constitute a total sample size of 200 farmers,
317
in Jammu, Udhampur, Rajouri, Poonch districts

and the overall Jammu region with R2 values at
X2 = Family size of household

X3 = Education level of head of household
X4 = Average price of milk
X5 = Number of milch animals per
household
0.956, 0.925, 0.996, 0.917 and 0.969 respectively,

was statistically significant meaning that about 95
percent, 92 percent, 99 percent, 92 percent and 96
percent, respectively, of the total variation in milk
production in the respective districts was explained
by the independent or explanatory variables under
consideration.
The functional analysis revealed that milk
production was found significant at the 1 percent
level of probability in Jammu, Udhampur, Rajouri
and Poonch districts and the overall Jammu region.
The value of regression coefficients for milk
production were 1.101, 0.971, 0.924, 1.068 and
0.979, respectively. The regression coefficient for
price of milk was significant and positive in Jammu
district at the 5 percent level of significance with
the value at 0.193, whereas in the overall Jammu
region price of milk was found statistically
significant at the 5 percent level of significance
with values at 0.138. The values for family size in
Rajouri (0.016) and Poonch (0.015) districts, for
education status in Jammu (0.002), Rajouri (0.001)
Poonch (0.001) and overall (0.083) and for number
of milch animals in Udhampur (0.039), Rajouri
(0.083) and overall (0.059) were, however, positive
but not significant.
The results of marketed surplus function
(Table 1) showed that the values of regression
coefficients of milk production had a positive
and significant impact on the marketed surplus of
milk in all the four districts and the overall Jammu
region indicating thereby that one percent increase
in the use of this inputs after keeping the use of all
other inputs constant, the marketed surplus could
increase by 1.101 percent in the case of Jammu
district, 0.971 percent in the case of Udhampur
district, 0.924 percent in the case of Rajouri,
The scale prepared by Trivedi and Pareek

(1963) to measure the education level of rural
families was used, the details of which are given
below:
Education
Weightage
Illiterate
0
Can read only
1
Can read and write
2
Primary
3
Middle
4
High School
5
Intermediate and above
6
1
To work out the marketing cost and price

spread during the marketing of milk, the lot method
was used. A specific lot or consignment was selected
and chased through the marketing system until it
reached the ultimate consumer. The cost and margin
involved at each stage was assessed (Acharya and
Aggarwal, 1998). This method is appropriate for
such perishable commodities as fruits, vegetables
and milk because the lag between the time the
commodity enters the marketing system and time
of its final consumption is very small.

Marketed surplus function for buffalo milk
A perusal of Table 1 indicates that the
production function for marketed surplus of milk
(variables included in the regression model explain
more than 91 percent of the total variation for milk)
318
1.068 percent in the case of Poonch district and

in the case of overall Jammu region, the increase
in marketed surplus of milk was 0.979 percent.
This clearly implies that as the milk production
increases, there will be more marketed surplus of
milk. The quantity of marketed surplus of milk also
increases with the increase in the price of milk in
the case of Jammu district and the overall Jammu
region, where the price of milk shows a positive
and significant impact on the marketed surplus of
the milk. In the other districts; the sign for the price
of milk was however positive but did not show
any significant impact. The other variables like
family size did not show any significant impact on
the marketed surplus. However, their signs were
negative in some cases, which clearly indicated that
as the family size increases, the marketed surplus
of milk will decrease. The effect of educational
status of the head of the household did not have
any significant influence on the marketed surplus
as most of the heads of household were illiterate.
All the variables included in the regression model
explained more than 90 percent of variation. These
findings are supported by the findings of Dhaka et
al. (1998) and Das (2004).
to be 289, 363, 469 and 335 gm in small, medium,

large and overall farms, in Jammu Region.
This shows that in Jammu region overall
production of milk was 34.85 litre per household
per day, in which large farms had the more
production per day as compared to other groups,
which was due to the large number of milking
animal kept by large farms in Jammu district.
These findings are supported by the findings of
Prabhakaran and Ramasamy (1984) and Kumar
(2006). The marketed surplus of milk per day per
household was found to be 33.29 litres. The overall
marketed surplus measured in terms of ratio to total
milk production was 95.53 percent for the overall
Jammu region, which was highest in large farms.
With the increase of herd size, marketed surplus of
milk also increased, which was observed in all the
districts of the overall Jammu region. More milk
was retained by small farms and medium farms
than large farms due to big family size. The per
capita consumption of milk in Jammu region was
found to 335 gm per day.
Marketing cost and price spread in buffalo milk
marketing
The following channels were usually
found for the distribution of milk from producer to
consumer in the Jammu region.
Producer - Consumer,
Producer - Vendor - Consumer
Producer - Vendor - Processor (sweet shop)
In the first channel, either consumers
themselves go to take milk from the producers
houses or producers themselves go to the houses of
consumers to sell the milk. In the second channel, a
vendor collects milk from the producer and supplies
it to the consumers. In the third channel, the vendor
collects the milk from the producers and supplies it
to the processor (sweet shop) for making different
Production, consumption and marketed surplus

of buffalo milk in Jammu region
Production as well as marketed surplus was
highest in large farms followed by medium farms
and small farms (Table 2). On average, the daily
milk production per house hold in different groups
was 21.44, 37.13, 85.21 and 34.85 litres for small,
medium, large and overall farms, respectively, of
which 20.25, 35.34, 82.35 and 33.29 litres was sold,
representing marketed surpluses of 94.45, 95.18,
96.64 and 95.52 percent on small, medium, large
and overall farm sample households in the Jammu
region. The per capita milk consumption was found
319
Table 1. Marketed surplus function for milk for the Jammu region.
Regression Coefficients
Milk
production
(X1)
Family
size
(X2)
Education
status
(X3)
Price of
milk
(X4)
No. of
milch
animals
(X5)
-0.845
1.101**
(0.049)
-0.025
(0.021)
0.002
(0.001)
0.193*
(0.091)
-0.073
(0.051)
0.956
Udhampur
-0.007
0.971**
(0.037)
-0.013
(0.014)
0.001
(0.001)
0.008
(0.044)
0.039
(0.040)
0.925
Rajouri
0.006
0.924**
(0.058)
0.016
(0.015)
0.001
(0.001)
0.023
(0.070)
0.082
(0.061)
0.996
Poonch
-0.675
1.068**
(0.075)
0.015
(0.010)
0.001
(0.001)
0.142
(0.089)
-0.057
(0.082)
0.917
Overall
-3.702
0.979**
(0.014)
-0.029
(0.039)
-0.083
(0.070)
0.138*
(0.081)
-0.059
(0.120)
0.969
Districts
Constant
Term
Jammu
R2
Note : Figure in parentheses indicate the standard error of regression coefficients.

* Significant at 5 percent level.
** Significant at 1 percent level.
Table 2. Production, consumption and marketed surplus of milk in the Jammu region (litres Household/
day).
Farm size
groups
Small
Medium
Large
Overall
Avg. no.
of milch
animal
Milk
production
Milk surplus
marketed
Milk Marketed
as % to milk
production
Per capita
consumption
(litres/day)
4.27
8.26
20.27
7.65
21.44
37.13
85.21
34.85
20.25
35.34
82.35
33.29
94.44
95.18
96.64
95.53
0.289
0.363
0.469
0.335
320
milk products.
rupee was maximum in first marketing channel due

to the absence of any marketing intermediatory.
Maximum marketing cost was found in third channel
whereas minimum marketing cost was found in
the case of the second channel. The marketing
cost in the first marketing channel in the overall
Jammu region was high due to high transportation
cost incurred by the producers in Udhampur and
Poonch districts. The maximum marketing margin
was found in the third channel.
The marketed surplus function for
milk for the Jammu region indicated that milk
production and the price of milk had a positive and
significant impact on the marketed surplus of milk.
This clearly implies that as the milk production
increases, there will be more marketed surplus of
milk and the quantity of marketed surplus will also
increase with the increase in the price of milk. The
production as well as marketed surplus in absolute
terms was highest in large farms, followed by
medium farms and small farms.. The per capita
consumption was found to be higher in large
farms. In estimating the cost of the marketing
of milk in the overall Jammu region, it was found
that the producers share in the consumers price
was maximum in the first channel followed by the
second and the third. Marketing cost and marketing
margin was maximum in the third channel.
Marketing cost, marketing margin and price

spread in marketing of buffalo milk in Jammu
region
Table 3 showing the marketing of milk
in Jammu region revealed that producers share
in the consumer rupee was 93.14 percent, 88.78
percent and 86.17 percent in the first (ProducerConsumer), second (Producer -Vendor - Consumer)
and third (Producer - Vendor - Processor) marketing
channels, respectively, whereas marketing cost
was 6.86 percent in the case of the first channel,
6.12 percent in the case of the second channel
and 7.45 percent in the case of the third channel,
respectively. The marketing margin was found nil
in the first channel, while it was 5.10 percent and
6.38 percent for the second and the third channels,
respectively.
It was observed in marketing cost and
price spread of the overall Jammu region that 93.14
percent of consumer rupee was received by the
milk producer in first marketing channel (ProducerConsumer), which was the maximum share of the
consumer rupee, while in third marketing channel
(Producer-Vendor-Processor),
the
minimum
producers share in consumer rupee was found
(86.17%). The producers share in the consumer
Table 3. Marketing cost, marketing margin and price spread of milk in the Jammu region (in percentage).
Marketing Channels
Particulars
Producers share
Marketing cost
Marketing margin
Consumers price
Producer to
consumer
Producer to vender
to consumer
Producer to vender to
processor(sweet shop)
93.14
6.86
0.00
100.00
88.78
6.12
5.10
100.00
86.17
7.45
6.38
100.00
321
REFERENCES
Acharya, S.S. and N.L. Agarwal. 1999. Agricultural
Marketing in India. Oxford and IBH
Publishing Co. Pvt. Ltd., New Delhi. p.
401.
Dhaka, J.P., D.K. Jain, V.K. Kesavan and L. Singh.
1998. A study of production and marketed
surplus functions for milk in India. IndoSwiss Project, National Dairy Research
Institute, Karnal.
Das, S. 2004. Economic efficiency of milk
production and marketed surplus in rural
area of Burdwan district (W.B.). M.Sc.
Thesis, NDRI, Karnal, India.
Kumar, R. 2006. Economic analysis of dairy
farming among nomadic and resettled Van
Gujjars in Uttaranchal: A comparative
study. Ph.D. Thesis, NDRI, Karnal, India.
Prabhakaran, R. and C. Ramasamy. 1984. Influence
of certain selected factors on marketed
surplus of milk. Cheiron, 13(2): 106-108.
Singh, K. 1978. Who feeds the milk market? Indian
Dairyman, 30(4): 277-280.
322
Original Article
PREVALENCE OF TICKS IN BUFFALOES IN THE UPPER SINDH PAKISTAN
Majid Hussain Soomro1, Shahida Parveen Soomro2, Muhammad Bachal Bhutto3, Zeeshan Akbar4,
Muhammad Yaqoob4 and Abdullah G. Arijo3
ABSTRACT
and tactile control of ticks in local breeds of dairy

animals in the Sindh province.
Tick infestation is still a major economic

dilemma for the dairy owners in Pakistan. The
current study reports the prevalence and bionomics
of tick in the areas of upper Sindh, Pakistan. The
study was carried out to identify and to quantify
variation in the prevalence of bovine tick
infestation with respect to host (age and species)
and area studied. Random sampling was used and
1600 samples of Kundi buffaloes from the different
areas were selected from extensive management
systems. Prevalence of bovine tick infestation
did not differ significantly (OR = 0.876; p>0.05)
in Kundi (179/800; 24.75%) and Nili-R av i
buffaloes (172/800; 22.3%). Hyalomma was the
major tick species (10.2%; 163/1600), followed by
Rhipicephalus (5.6%; 99/1600). The prevalence of
ticks in calves (< 1 year) was significantly (p < 0.05)
higher compare to the adult animals (1-2 years and
> 2 year animals). However, the prevalence of tick
infestation was not associated (p > 0.05) with the
location of the district. Moreover, the results of the
prevalence of the ticks in the studied area provide
the better understanding for evolving the strategic
Keywords: buffalo, tick, prevalence, Hyalomma,

Rhipicephalus
INTRODUCTION
Livestock is the key feature of the rural
based economy of Pakistan like other tropical and
sub-tropical countries. Cattle and buffaloes are
the major animals which contribute satisfying
the ever increasing demand for milk and meat in
the country but the major contribution is fulfilled by
buffaloes. Indigenous cattle breeds (Sahiwal, Red
Sindhi etc.) also shared but these breeds are now
being replaced by crossbred cattle owing to high
milk production. Although these breeds are resistant
to harsh climatic conditions and resistant to some
tropical diseases, farmer breed preference is utmost.
These crossbred cattle as compared to buffaloes are
greatly influenced by diseases, nutrition and climate
changes but parasitism is thought to be a major
cause of lower productivity (Sajid et al., 2007).
Parasitism also results in mortality, lowered
Department of Pathobiology, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan 60000,
Pakistan, E-mail: majidsoomro@hotmail.com
2
Department of Plant Protection, Sindh Agriculture University, Tandojam, 70060, Pakistan,
3
Department of Veterinary Parasitology, Faculty Animal Husbandry and Veterinary Sciences, Sindh Agriculture
University, Tandojam 70060, Pakistan
4
Faculty of Veterinary and Animal Sciences, Pir Mehr Ali Arid Agriculture University, Rawalpindi 46000,
Pakistan
323
general health condition, retarded growth, lower

output of work, and decrease in the production
of milk and meat. Moreover, financial losses are
also significantly associated with nagging irritation
and depreciation of the value of skins and hides
(Chaudhry et al., 2011).
Pakistan being a tropical and sub-tropical
region country, optimal climatic conditions
are very conducive for raising the buffaloes
and these conditions are also favorable for the
growth and development of many tick species
(Durrani and Shakoori, 2009). So under these
conditions crossbred cows are considered
unfit for survivability and adaptability. Poor
managemental practices further enhance the severity
of the problem (Irshad et al., 2010). Under these poor
conditions the buffalo is the only animal that survives
with optimal production. The buffalo is also prone
to tick infestation but its adaptability in tropical and
sub-tropical conditions is excellent. Previous findings
of tick prevalence report that exotic breeds suffer
more as compared to local breeds of cattle and
buffaloes in the climatic conditions of Pakistan..
With regard to the tick prevalence in buffaloes
there is scanty information available concerning
Pakistan (Manan et al., 2007; Sajid et al., 2008,
2009) and there is no report is available on tick
prevalence from the area of Sindh. Therefore, the
present cross- sectional study was focused on
prevalence and identification of genera of ticks
infesting the buffaloes in the upper areas of Sindh
province, Pakistan.
and eastern part consists of the Nara Desert. The

study area lies in the sub-tropical region between
273148N and 684423E. District Khaipur
has eight sub-divisions (Talukas): Khairpur
city, Kingri, Gambat, Sobhodero, Kotdeji, Nara,
Mirwah and Faizgunj. This cross-sectional
epidemiological survey was conducted at
different locations of the district to analyze the
variation in tick infestation during 2008 and 2009.
Statistical analyses
Statistical analyses were carried out by
using statistical package for social science (SPSS)
version 11.5. Multiple logistic regression tests were
performed to determine the association between
Sampling of animals
Random sampling was used to select a
total of 1600 buffaloes from the study area of
Khairpur District. The samples were collected
from small dairy herds having no history of use
of acaricide for tick control. About 107 farms and
1600 animals were selected for sampling of ticks.
Animals were categorized on the basis of b r e e d
(Kundi vs. Nili-Ravi buffaloes) and age (< 1 year
vs. 1-2 years vs. > 2 years).
Sampling and morphological identification of
ticks
Ticks were collected from body parts
of the infested animals and careful sampling
was made using forceps to avoid de-capitulation
(Solusby, 2006). After collection ticks were kept
in capped bottles properly labeled containing
70% ethyl alcohol for preservation. Morphological
identification of the ticks was made in a laboratory
at the Department of Veterinary Parasitology, Sindh
Agriculture University Tandojam (Soulsby, 1982;
Walker et al., 2007).
The study area included the district of

Khairpur Mirs of the upper Sindh (Pakistan). It
is situated south of the Indus River in the Sindh
324
of Sindh Pakistan.
The results of host susceptibility for tick
infestation findings represent that both breeds
(Nili-Ravi vs. Kundi) were prone to the infestation
at equal rates. These findings of prevalence were
not different from the previous reports in different
parts of the country (Manan et al., 2007; Sajid
et al., 2008). But the findings of the Sajid et al.
(2009) in the lower Punjab area do not correspond
with this current report. This difference might be
due to dry and hot environment of Sindh province
which reduces the activity of ticks. Moreover,
the local breeds of cattle (Sahiwal, Red Sindhi)
are resistant due to loose skin coat, and similarly,
their thick skin and wallowing habits prevent the
buffaloes (Nili-ravi, Kundi) from tick infestation
(Sajid et al., 2009). For the further investigation a
longitudinal study needs to be done to understand
the frequency of tick infestation during different
seasons in the Sindh province climate.
Data of host age determined that younger
stock were at more tick infestation risk than the
adult animals. These findings agrae with the report
of Swai et al. (2005) who found that calves are
more susceptible than adults. But reports that age
has no significant association with tick infestation
of Sajid et al. (2009) and Manan et al. (2007) do
not agree with our findings . The increased rate of
infestation in calves than adult might be due to
hairy coat of the calves, less attention of the farmer
and unhygienic conditions in places where calves
are kept that favor the ticks growth, development
and reproduction. Moreover, the thin and easily
penetrable skin of calves facilitates the easy
approach to the circulation for the nourishment of
ticks.
This study of the prevalence of ticks
suggests that there is of a need of a longitudinal study
to understand comprehensively all epidemiological
the prevalence of ticks and animal breeds, ages,

study area and tick species. Odds ratio measured
the possible association tick prevalence between
the factors studied. The significance level was set
at P value less than 0.05.
RESULTS
The results of the present study demonstrate
that the overall prevalence of tick infestation in both
breeds was 23% . However, the prevalence of ticks
did not differ (OR=0.876; p>0.05) significantly
and both breeds were at almost equal risk of tick
infestation (Table 1). The study of prevalence of tick
species in bovine infestation found that Hyalomma
(10.5%; 163/1600) was most abundant, followed
by Rhipicephalus (5.6%; 90/1600), Boophilus
(5.5%; 88/1600) and Ablyomma (2.3%; 36/1600)
(Table 1).
In district Khairpur, the current data
showed that the tick prevalence (p>0.05) was found
to be similar in all Talukas and all were under the
risk of tick infestation equally. The results of host
age prevalence showed that a significantly higher
prevalence of tick infestation was observed in
calves (p < 0.05) than in adult hosts and that calves
were more prone to tick infestation as compared to
adults (Table 1).
DISCUSSION
The issue of tick infestation and prevalence
was reported from all over the world in connection
with different parts of the Pakistan. So keeping
this in view the present study was designed to
investigate the different aspects of prevalence and
identification of ticks in buffaloes from upper areas
325
Table 1. Prevalence of ticks in buffaloes in upper area of Sindh Pakistan.

Parameter
Variable
Levels
Prevalence
(%)
p-value
Odd
ratios
Lower
95% CI
Upper
95% CI
Host
species
Bovine
Buffaloes
198/800
(24.75)
0.263
0.876
0.696
1.104
Cow
179/800
(22.38)
< 1 year
1-2 years
>2 years
City
Kingri
Gambat
Sobhodero
Kotdeji
Nara
Mirwah
Faizganj
182/518 (35.1)
116/436 (18.0)
79/336 (18.1)
44/200 (22.0)
43/200 (21.5)
44/200 (22.0)
63/200 (31.5)
53/200 (26.5)
43/200 (21.5)
50/200 (25.0)
37/200 (18.5)
0.425
0
0.73
0.78
0.78
0.11
0.36
0.78
0.46
-
2.448
0.999
1.206
1.171
1.171
2.073
1.588
1.171
1.468
-
1.808
0.729
0.738
0.715
0.715
1.303
0.987
0.715
0.909
-
3.315
1.37
1.971
1.917
1.917
3.298
2.555
1.917
2.371
-
Amblyomma
36/1600 (2.3)
0.042
0.625
0.284
1.377
Boophilus
88/1600 (5.50)
0.691
1.13
0.601
2.122
Hyalomma
163/1600
(10.2)
0.269
0.156
0.463
Rhipicephalus
90/1600 (5.6)
Age
Buffalo
Origen
Khairpur
Parasite
Tick
Species
Value less than P<0.05 shows significant difference.
326
infestation in two districts of lower Punjab,

Pakistan. Preventive Veterinary Medicine,
92: 386-391.
Sajid, M.S., Z. Iqbal, M.N. Khan and G.
Muhammad. 2008. Point prevalence of
hard ticks infesting domestic ruminants
of lower Punjab, Pakistan. International
Journal of Agricultural and Biological
Science, 10: 349-351.
Sajid, M.S., Z. Iqbal, M.N. Khan, G. Muhammad
and M.U. Iqbal. 2007. Effect of Hyalomma
ticks (Acari: Ixodidae) on milk production
of dairy buffaloes (Bos bubalus bubalis) of
Punjab (Pakistan). Italian Journal of Animal
Sciences, 6: 939-941.
Soulsby E.J.L. 1982. Helminthes, Arthropods and
Bailliere tindall, London, UK.
Soulsby, E.J.L. 2006. Helminthes, Arthropods and
Elsevier New Delhi, India
Walker, A.R., A. Bouattour, J.L. Camicas, A.
Estrada-Pena, I.G. Horak, A.A. Latif, R.G.
Pegram and P.M. Preston. 2007. Ticks of
Domestic Animals in Africa, Bioscience
Reports, Edinburgh, UK.
aspects of such as frequency, distribution and risk

factors associated with tick infestation in the Sindh
province of Pakistan, in order to design preventive
medicine programs.
ACKNOWLEDGEMENT
This part of study was conducted with
cooperation t h e Pak-USAID research project
Building molecular capacity for preventing tick
transmitted diseases in Pakistan.
REFERENCES
Chaudhry, Z.I., A. Saiddain, N. Sabir, N.A. Malik,
S. Azeem and A. Sajid. 2006. Prevalence of
pathological conditions causing skin damage
and consequently reducing its market value
in domestic ruminants of Punjab, Pakistan.
Veterinary Science Development, 1(4): 1923.
Durrani, A.Z and A.R. Shakoori. 2009. Study on
Ecological Growth Conditions of Cattle
Hyalomma Ticks in Punjab, Pakistan. Iran.
J. Parasitol., 4(1): 19-25.
Irshad, N., M. Qayyum, M. Hussain and M.Q.
Khan. 2010. Prevalence of tick infestation
and theileriosis in sheep and goats. Pak. Vet.
J., 30(3): 178-180.
Manan, A., Z. Khan, B. Ahmed and Abdullah. 2007.
Prevalence and identification of Ixodid tick
genera in Frontier Region Peshawar. Journal
of Agricultural and Biological Science, 2:
21-25.
Sajid, M.S., Z. Iqbal, M.N. Khan, G. Muhammad
and M.K. Khan. 2009. Prevalence and
associated risk factors for bovine tick
327
Original Article
COMPARISON OF BLOOD ACID BASE GAS PARAMETERS IN VENOUS AND ARTERIAL

BLOOD OF HEALTHY BUFFALOES
S.A. Hussain* and S.K. Uppal
of information for early phase of the disease and

they influence the development of clinical signs
and therapeutic effectiveness (Brobst 1975). Some
authors (Brobst, 1975; Singh et al., 1981; Naito
and Murakami, 1982) think that with the exception
of the respiratory components (pCO2 and pO2)
ABSTRACT
The present study was carried out on ten
clinically healthy adult buffaloes of 4 to 6 years of
age. The parameters of acid-base balance (pH, pCO2,
pO2, HCO3, SBC, ABE, SBE, SAT and O2ct) were
measured simultaneously in venous and arterial
blood. The results from this study revealed that the
mean, minimum and maximum values of pH, pO2,
ABE, SAT and O2ct were lower in the venous blood
and the SAT, other components are not different

in venous and arterial blood. Others think that the
venous blood is unsuitable for acid bas gas analysis
(Bergman and Coleman, 1983; Gemelli, 1981). The
reference values for every parameter help in correct
interpretation of the results. The reference values
in calves are available in the literature. The limited
overall research of acid-base balance in buffaloes
prompted us for this research, whose purpose was
to describe the blood acid-base gas information for
the venous and arterial blood in buffaloes.
than in the arterial blood while the values of pCO2

and HCO3 were higher in venous blood than in the
arterial blood. Wider differences were observed in
the values of the respiratory parameters of venous
and arterial blood. The differences found in the
parameters of acid-base balance between both the
kinds of blood revealed the restricted informative
significance of the venous blood. This calls for the
use of arterial blood when it is necessary to assess
the acid-base balance.
MATEERIALS AND METHODS

The study was carried out on ten clinical
healthy buffaloes of age between 4 to 6 years. The
selected buffaloes had no history of disease for the
current lactation and were clinically healthy at the
time of sampling. Blood samples were collected
in heparinised (1:1000) disposable syringes and
were examined by apparatus ABL- 77 v1.41,
produced by Radiometer- Denmark. Venous blood
samples were collected from the jugular vein and
Keywords: acid-base balance, arterial blood,

buffalo, venous blood
INTRODUCTION
The changes in blood acid-base gas status
are the early manifestation of many diseases in
humans as well as animals. So, they are source
Department of Veterinary Medicine, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana,
Punjab 141 004 India, *E-mail: draashiqhussain@gmail.com
328
0.66 mmol/L higher than those in arterial blood.

A difference was also found for the ABE, whose
average value in the venous blood was 1.03 mmol/L
lower than that in the arterial blood.
The analysis of the results for the average,
minimal and maximal values of the acid-base
balance of the venous and arterialized capillary
blood revealed that these values of the pH, pO2,
ABE, SAT and O2ct in the venous blood were
lower, and for pCO2 and HCO3 were higher than
those of the arterial blood. Also the acquired results
between the maximal and minimal values of pH,
HCO3 and ABE of the venous and arterial blood
were narrower, and the ones for pCO2, pO2, and
arterial blood samples from auricular artery. The

blood acid-base parameters determined were pH,
partial pressure of carbon dioxide (pCO2), partial
pressure of oxygen (pO2), actual bicarbonate
(HCO3), standard bicarbonate (SBC), actual base
excess (ABE), standard base excess (SBE), oxygen
saturation (SAT) and oxygen capacity (O2ct). The
students t-test was used to analyze the significance
of difference between parameters of venous and
arterial blood. The significance level was kept at
p0.05 and p0.01.
SAT are substantially wider. Van Sluijs et al. (1983)

and Waizenhofer and Mulling (1978) discussed
similar differences between the indexes of the
venous and arterialized capillary blood of dogs and
calves. According to these authors the values of the
indexes of acid-base balance in the arterial blood
are closer to the arterialized capillary blood rather
than the venous blood.
The results of the present study for the venous
blood of buffaloes were different that the ones
of Nangia and Sukhija (1981) and Sobti et al.
(1995) who found higher indexes for pH and lower
for pCO2 and HCO3. The values of pH, pO2 and
HCO3 of the arterial capillary blood found in this
study were closer to those of Singh et al. (1981)
who used arterial blood, rather than Nangia and
Sukhija (1981) and Sobti et al. (1995) who used
venous blood. Apart from the bigger differences
of the established values for the respiratory
indexes (pCO2 and pO2) and SAT between arterial
and venous blood, the values of the metabolic
component (HCO3 and ABE) of the acid-base
balance and the blood pH of both the blood types
overlapped to a certain degree. For example, the
average and minimal values of HCO3 in the venous
The mean, minimum and maximum values

of acid base balance are shown in Table 1. The
average (7.3860.023) and the maximal (7.43)
indexes of pH in the venous blood were over taken
with the average (7.4630.015) and maximal (7.52)
pH in the arterial blood. The mean pH value in the
venous blood was 0.077 lower than that of the
arterial blood. The difference between the minimal
pH indexes of the two kinds of blood was 0.07 and
between the maximal pH indexes it was 0.09.
The pCO2 in the venous blood was 7.33
mmHg higher than that in the arterial blood.
The biggest differences were found between the
maximal indexes of the venous and arterial blood
(11 mmHg), the lowest were between their minimal
indexes (4 mmHg). In comparison to pCO2, pO2 in
the venous was 53.6% to 72.6% lower as compared
to that in the arterial blood. Similar substantial
differential rate was found for SAT and O2ct. The
average, minimal and maximal indexes for SAT
and O2ct in the venous blood were lower than those
of the arterial blood.
The average values of HCO3 and SBC in the
venous blood were respectively 2.17 mmol/L and
329
Table 1. Mean, minimum and maximum values of acid base gas parameters in venous and arterial blood
of healthy buffaloes.
pH
pCO2 (mmHg)
pO2 (mmHg)
HCO3 (mmol/L)
SBC (mmol/L)
ABE (mmol/L)
SBE (mmol/L)
SAT (%)
O2ct (%)
Venous blood
MeanS.E
Minimum
7.3860.023
7.35
41.332.27
35
49.003.26
37
26.201.43
23.1
26.031.89
22.4
1.100.52*
-1.9
2.130.52*
-1.7
80.174.23**
71.7
12.371.23**
10.2
Maximum
7.43
46
69
30
28.8
3.4
5.7
95.2
14.8
Arterial blood
MeanS.E Minimum Maximum
7.4630.015
7.42
7.52
34.01.52
31
35
86.672.64
79
95
24.030.66
22.9
25.2
25.371.06
23.8
27.4
2.130.54
-0.7
5.4
0.630.48
-0.9
2.4
97.133.83
96.2
97.6
15.501.32
10.8
21.6
*Difference significant at p0.05; **Difference significant at p0.01.
blood overlapped with the maximum and minimum

values of the arterial blood and the found borders
for ABE of the venous blood covered most of the
values of arterial blood. Thus, it was inferred that
the venous blood of buffaloes could be used for
assessing pH and HCO3 only. For the respiratory
of the organism. Minirva Medica, 72(34):

2289-2292.
Nangia, O.P. and S.S. Sukhija. 1981. A comparative
study of acid base components in venous
and capillary blood. Indian J. Exp. Biol.,
19(5): 475-476.
Naito, Y. and D. Murakami. 1982. Blood gas
acid base values in the coccygeal artery of
Holstein-Friesian cows. Jpn. J. Vet. Sci.,
44(5): 777-780.
Singh. J., K. Mirakhur, B. Prasad and R.N.
Kohli. 1981. Acid base status and gases in
cerebrospinal fluid of healthy calves and
buffalo calves. Zbl. Vet. Med. A, 28(2): 131134.
Van Sluijs, F.J., H.W. de Vries, J.J. De Bruijne and
W.E. van den Brom. 1983. Capillary and
venous blood compared with arterial blood
in measurement of acid base and blood gas
status of dog. Am. J. Vet. Res., 44(3): 459462.
Sobti, V., S. Singh and B. Prasad. 1995. Effect of
components (pCO2 and pO2) and SAT the venous

blood did not give a clear idea.
The overall assessment of the venous blood
parameters showed that it was not suitable for the
evaluation of acid base balance. Because of this,
arterial blood is recommended for assessment of
acid base balance.
REFERENCES
Bergman, N. and J. Coleman. 1983. Choice of blood
for acid-base balance. Lancet, 2: 1135.
Brobst, D. 1975. Evaluation of clinical disorders of
acid-base balance. J. Am. Vet. Med. Assoc.,
166(4): 359-364.
Gemelli, A. 1981. Problems of acid-base equilibrium
330
storage on acid base status of buffalo bubalis

blood. Indian J. Vet. Surg., 6(1): 34-40.
Waizenhofer, H. and M. Mulling. 1978. Behavior
of pHact, PO2 and PCO2 in venous, capillary
and arterial blood of newborn calves. Berl
Munch Tierarztl., 91(9): 173-176.
331
Original Article
PREVALENCE OF SALMONELLA AND ESCHERICHIA COLI ASSOCIATED WITH

DIARRHEA IN BUFFALO AND COW CALVES
M. Anwarullah*, J.A. Khan, M.S. Khan, K. Ashraf and M. Avais
ABSTRACT
economically important agents (Acha et al., 2004;

House et al., 1978). Salmonella typhimurium is
a major cause of calf morbidity and mortality
in the United States and in Europe (Tsolis et al.,
1999; Rice et at., 1997). Khan et al. (2009) found
the prevalence of calf diarrhea due to Salmonella
was up to 16.3%. Calf diarrhea is a clinical sign
of disease that may have many causes but E. coli
has been considered as the primary bacterial cause
in calves (Yamamoto and Nakazawa, 1997). The
morbidity and mortality rates associated with many
outbreaks of gastrointestinal diseases caused by
Escherichia coli have highlighted the threat that
these organisms pose to public health (Paton et
al., 1998). The prevalence of enterohemorrhagic
Eschrechia Coli O157:H7 in the feces of dairy
calves and feedlot cattle is low. Eschrechia coli
have been isolated from 0.3 to 2.2% of fecal
samples collected from healthy calves or cattle in
the United States, the United Kingdom, Germany,
and Spain (Blanco et al., 1993).
Calf diarrhea caused by Salmonella and

Escherichia coli was the particular focus in the
current study. A total 150 calves up to 6 months of
age suffering from diarrhea were examined. Fresh
fecal and blood samples were collected. Fecal
samples were cultured on McConkey and on S-S
agar for isolation of bacteria. Bacterial colonies
were identified by using an API 20 E Kit. Overall
prevalence of Salmonella and E. coli was 33.3%.
Prevalence of Salmonella and E. coli was 18.6%
and 14.6%, respectively. Occurrences of diarrheal
disease due to various risk factors like age, sex
were recorded for determining the effect of these
risk factors.
Keywords: buffalo, Bubalus bubalis, cow, Bos
taurus, Escherichia coli, Salmonella, diarrhea
INTRODUCTION
Major causes of mortality of neonatal

calves were conditions like diarrhea and pneumonia
(Khan et al., 2009, Shimizu and Nagatoma 1978).
The neonatal calf mortality in the first month of age
is more than 80% of the total mortality in calves
(Jenny et al., 1981). Salmonella and Escherichia
coli are known to be the most common and
Selection of site: The research was

conducted at different private livestock farms and
civil veterinary hospitals of district Jhelum. Total
duration of the study was of three months.
Animals: A total number of 150 cow and
Department of Clinical Medicine and Surgery, Department of Parasitology, Faculty of Veterinary Science,
University of Veterinary and Animal Sciences Lahore-54000 Pakistan, *E-mail: jawaria.khan@uvas.edu.pk
332
buffalo calves under six months of age, suffering

from diarrhea were studied for determination of
Salmonella and E. coli.
Collection of samples: A fecal sample
was collected directly from the rectum of each of
the calves under six months of age. Then the fecal
material was shifted to sterilized plastic containers
and again those containers were placed in zip bags
to prevent entry of foreign particles. Fecal material
was kept in contact with ice to avoid further changes
in fecal material.
Blood: Blood samples from the same
individuals were collected from the jugular vein
after applying disinfectants and immediately shifted
into anticoagulant coated vaccutainer tubes.
Bacterial culture: One gram of fecal
material was dissolved in 10 ml of autoclaved
normal saline and cultured on MacConkey agar
with the help of safety buds. Then it was incubated
in an incubator for 24 h.The next day the colorless
colonies were again cultured on S-S agar by
dissolving those colorless colonies first in 5 ml
autoclaved normal saline and then culturing on S-S
agar with safety buds.
The API 20E commercial kit was used for
the identification of E. coli and Salmonella (Popovic
et al., 2004; API 20E Kit Biomerieux, France).
Blood parameters: Following blood
parameters were compared according to Benjamin
(1979): (a) Total leukocyte count: Total leukocyte
counts were performed with the help of leukocyte
diluting pipette. (b) Differential leukocytic
count: The Giemsa stain blood smear method,
was preferred for differential leukocyte count.
(c) hemoglobin estimation: Sahlis method was
used to determine the Hb level. (d) Erythrocyte
sedimentation rate: Erythrocyte sedimentation
rate was determined with a Wintrobe hematocrit
tube. The upper level of sedimenting erythrocytes
sas read in millimeters on the left scale after the

time interval of 8-24 h for cow and buffalo calves.
Seasonal effect: The effect of season, environment,
temperature, rainfall and humidity was observed.
Risk factors: The effects of age, sex, breed, diet,
managemental practices and vaccination were also
studied.
Statistical analysis: The data collected were
tabulated and analyzed statistically by chi-square
test (Steel et al., 1997).
RESULTS
Of the total of 150 cow and buffalo calves
under six months of age studied with the signs
of diarrhea in district Jehlum, the total number
of E.coli positive samples was 22, and 28 were
Salmonella positive. The prevalence of E. coli was
14.6%, and that of Salmonella was 18.6% The data
were analyzed by chi-square test and value of P
was greater than 0.05, which means no significant
difference was found. The positive samples of
Salmonella and E. coli in 1-30 day, 31-90 day, 91180 day age groups were 3, 11, 14 for Salmonella
and 2, 6, 14 for E. coli. Thus the prevalence of
Salmonella and E. coli in 1-30 days, 31-90 days,
and 91-180 were 2%, 7.33%, 9.33% for Salmonella
and 1.33%, 4%, 9.33% for E. coli, respectively.
The value of P showed was less than the
level of significance so there was no difference for
the values of different age groups. The prevalence
of positive bacterial samples in the age groups of
1-30 days 31-90 days 91-180 days was noted 14%,
23% and 63% for E. coli and 11%, 29% and 60%
for Salmonella, respectively. The total number of
positive samples in male calves was 18 and the
prevalence was 12%, and in the case of female
calves, the total number of positive samples was
333
32 and the prevalence was 21%.

Out of the total number of 150 diarrheic
calves, the number of positive samples for cow
calves was 31 and that for buffalo was 19. The
prevalence of Salmonella and E. coli in cow and
buffalo calves was 21% and 12%, respectively. The
P value for the test was greater than 0.05 and data
showed non-significant. Total numbers of positive
samples of Salmonella and E. coli in cow calves
was 0, 17, 14 for 1-30 days, 31-90 days, 91-180
days and for Salmonella were 6, 0, 13 for 1-30
days, 31-90 days, and 91-180 days. The prevalence
of bacteria (Salmonella and E. coli) was 0, 11.33%,
9.33% for cow calves and 4%, 0, 8.67% for buffalo
calves for the different age groups.
The prevalence in males in the 1-30 day
age group was 0, that in those in the 31-90 day
group was 5.33%, and that in those in the 91-180
daygroup was 7.33%. Similarly in females in the
1-30 daygroup, the prevalence was 4%; in the 3190 daygroup, it was 5.33% , and in the 91-180
daygroup, it was 11.33%. The prevalence of E. coli
in male calves was 0 %, 6%, 4.66% for the 1-30
day, 31-90 day, and 91-180 day age groups, and that
for Salmonella was 0%, 0% and 3.33% for the 1-30
day, 31-90 day, 91-180 day groups. The prevalence
of E. coli in female calves was 2%, 2%, 3% for
the 1-30 day, 31-90 day, and 91-180 day, and that
for Salmonella was 1%, 3.330% and 5.33% for the
1-30 day, 31-90 day, 91-180 daygroups as shown in
Figure 1.
Blood parameters varied in the 1-30 day,
31-90 day, 91-180 day age groups as shown in
Table 1. Total leukocyte count values in these
groups were 9%, 19%, 13% respectively. Similarly,
hemoglobin values were 14%, 13%, 14%
erythrocyte sedimentation rates were 11%, 8%,
19%, neutronphil values were 19%, 35%, 26%,
eosinophil values were 6%, 11%, 19%, basophil
values were 13% 9%, 19%, lymphocyte values

were 37%, 38%, 59%, monocyte values were 16%,
8%, 10% for the three age groups, respectively. The
effect of season was observed for three months (July,
August, and September). The prevalence rate was
high in August and least in July. Rainfall increased
the number of bacterial positive samples.
DISCUSSION
The prevalence of diarrhea due to
bacteria and E. coli in overall calves was 33.3%
in the present study. Although that is very high, a
similar prevalence was also seen in Mozambique.
In Mozambique, Baule et al. (1995) reported an
overall prevalence of diarrhoeic calves as high
as 36% but this percentage includes values of
prevalences of diarrhoea from other farms not
included in the present study. This might be due to
an outbreak in a particular area in those days or due
to outbreaks among different farms as sampling was
done from certain private livestock farms as well.
Among bacteria, enterotoxigenic Escherichia coli
and Salmonella are known to be the most common
and economically most important agents that cause
diarrhea in young calves (House et al., 1978).
The values according to different age
groups showed that total leukocytes were greater in
age group of 31-90 days, hemoglobin in the younger
group of 1-30 days, erythrocyte sedimentation was
greater in the 91-180 day age group, neutrophils
were higher in the 31-90 day group, eosinophils,
basophils and lymphocytes were greater in the
oldest age group (91-180 days) and monocytes
were highest in the youngest age group. In the
current study, the average of the total leukocyte
count value increased in the cow as compared
to the buffalo calves. Similarly, the average of
334
Figure 1. Prevalence of Salmonella and E. coli in calves in different age groups.
Table 1. Average values of blood parameters in cow calves according to different age groups.
Age
1-30 days
31-90 days
91-180 days
TLC
9%
19%
13%
Hb
14%
13%
14%
ESR
11%
8%
19%
N
19%
35
26%
335
E
6%
11%
19%
B
13%
9%
19%
L
37%
38%
59%
M
16%
8%
10%
Escherichia coli 0157 in cull dairy cows on

farm and at slaughter. J. Food Protect., 60:
386-387.
Shimizu, T. and H. Nagatoma. 1978. Current status
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Andreas. 1999. Contribution of Salmonella
typhimurium Virulence Factors to Diarrheal
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Yamamoto, T. and M. Nakazawa. 1997. Detection
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Escherichia coli heat- stable enterotoxin
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isolated from piglets and calves with
diarrhea. J. Clin. Microbiol., 35: 223-227.
hemoglobin was higher in the cow calves as

compared to the buffalo calves, but the averages
of erythrocyte sedimentation rate and neutrohils
were higher in the buffalo calves than in the cow
calves. Again. as far as basophiles, eosinophils and
monocytes are concerned, the average values were
greater in the cow calves than in the buffalo calves,
but lymphocytes values were higher in buffalo
calves than in cow calves.
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Blanco, J., E.A. Gonzalez, S. Garcia, M. Blanco,
B. Regueiro and I. Bernardez. 1988.
Production of toxins by Escherichia coli
strains isolated from calves with diarrhoea
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Maqbool, M. Salman and Z.U. Rehman.
2009. Epidemiology of major bacterial and
viral causes of diarrheoa in buffalo calves
in three districts of the Punjab province of
Pakistan. J. Zool., Suppl. Ser. (9): 187193.
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and Diagnosis of Shiga Toxin-Producing
Escherichia coli Infections. Clinical
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D.E. Herriott and L.V. Carpenter. 1997.
336
Original Article
DISTRIBUTION OF SALMONELLA SPECIES IN BUFFALOES IN

SOME MIDDLE GOVERNORATES OF IRAQ
Afaf Abdulrahman Yousif* and Ali, D.M. Al-Hashimi
ABSTRACT
The antibiotic susceptibility pattern

of Salmonella against 15 antimicrobial drugs
revealed that all isolates were resistant (100%)
to
chloramphenicol,
suphamethoxazole,
erythromycin, cloxacillin, and tetracycline,
and all isolates were sensitive to amicacin and
trimethoprimwhile neomycine, gentamycine,
cefixime,
ciprofloxacin,
kanamycin
and
streptomycin gave intermediate results.
This study was planned to isolate and

type Salmonella from buffalo in three middle
governorates in Iraq. One hundred fifty milk samples
were collected from 150 lactating buffaloes in the
field, and in the slaughter house (150) fecal samples
were collected at slaughter time and (900) samples
were collected post slaughtering:150 samples from
each organ: gall bladder, mesenteric lymph nodes,
spleen, liver, small intestine and cecum.
Diagnostic study depended upon the
morphological and cultural properties of the isolates
on some selective media like Brilliant Green , XLD,
SS agar and Hicrome rajhans medium, modified
(Salmonella agar modified) were used in addition
to different biochemical tests, API-20E and KB
003 Hi25^TM Enterobacteriaceae Identification
Kit, latex test and serotyping of isolates.
Twenty-two isolates were obtained from the
fecal samples and organs; these isolates belonged
to three serotypes (Salmonella anatum (68.18%),
S.muenchen (18.18%) and S. enteritidis (13.64%),
while no isolate was obtained from milk samples.
In animals from slaughter houses, the
percentage of infection varied in examined organs
and feces: in bile duct and liver, the percentage was
3.33% each; in mesenteric lymph nodes, 2.67%;
in spleen and cecum, 2% each; and in feces,
1.33%.
Keywords: Salmonella, buffaloes, serotyping of

Salmonella, Iraq
INTRODUCTION
Buffalo raising is a major contributor to
the agriculture and livestock industry in many
Asian countries through the production of good
quality meat and farmyard manure (Singh, 2010).
The buffalo is also recognized as the world second
most important milk producing species (Bhatti et
al., 2009). Dairy buffaloes are also called the black
gold of South Asia, where 95% of the worlds
buffalo milk is produced (Javaid et al., 2009)
Therefore, the pathogens either causing disease in
buffaloes and their progeny or transmitted through
their production are important because they affect
milk production and overall livestock production.
Salmonellosis is associated with medium to
College of Veterinary Medicine, Department of Internal Medicine, University of Baghdad, Iraq, *E-mail:
afaf_a.rahman@yahoo.com
337
Brilliant Green agar, Xylose-Lysine Deoxycholate

agar, Salmonella-Shigella agar and Hicrome
rajhans modified Salmonella agar and incubated
aerobically at 37oC for 24 h. Salmonella suspected
colonies were identified by Gram staining, in
addition to different biochemical tests: motility,
triple sugar iron agar, indole, methyl red and citrate
utilization tests, API-20E and the use of KB 003
Hi25^TM-Enterobacteriaceae Identification Kit,
which is a standardized colorimetric identification
system utilizing thirteen conventional biochemical
tests and eleven carbohydrate utilization tests
based on the principle of pH change and substrate
utilization (Thangamaalr et al., 2009).
Then, the isolates were grouped by the
Wellcolex color latex test for Salmonella (Rohner
et al., 1992). Description of kit contents (Figure
1).
1. Latex Reagent 1: one drop of gray-brown
suspension of polystyrene latex particles in buffer
containing 0.05% Bronidox preservation. The
latex particles are coated with rabbit antibody with
the following specificity:
Red latex Salmonella group B; Blue latex
S. group C; Green latex; S. group D.
2. Latex Reagent 2: one drop of gray-brown
suspension of polystyrene latex particles in buffer
containing 0.05% Bronidox preservation. The
latex particles are coated with rabbit antibody with
the following specificity:
Red latex Vi; Blue latex S. group E and G;
Green latex S. group A.
3. Red positive control : Killed bacterial suspension
of organisms with Salmonella group B and Vi
antigens containing 0.05% Bridox and 0.5%
formalin as preservative.
4. Blue positive control : Killed bacterial suspension
of organisms with Salmonella group C and E
antigens containing 0.05% Bridox and 0.5%
severe morbidity and even mortality in farm animals,

representing a major economic productivity loss
in the food and animal industries (Malkawi et al.,
2004). Salmonella has been widely reported in
buffaloes (Hassanain et al., 2010; Khan et al., 2009;
Ribeiro, 2000; Abdulwahid and Raheem 1981), and
infected animals may shed the organism in their
feces without showing any clinical signs of disease
(Fardsanei et al., 2010). Therefore, buffaloes may
carry this organism undetected into an abattoir at
the time of slaughter.
This study aimed at determining the
distribution of Salmonella in different organs, feces,
and milk samples of buffaloes.in three governorates
in Iraq and at determining the sensitivity of the
obtained isolates to different antimicrobials.

The study was performed on 300 buffaloes
distributed as follows: 150 dairy buffaloes in the
field and 150 buffaloes in slaughter houses.
Information about animals concerning age,
sex, and any signs of diarrhea were recorded. One
hundred fifty milk samples were collected from
lactating buffaloes in the field , and in the slaughter
house 150 fecal samples were collected at slaughter
time and (900) samples postslaughtering samples
were collected, distributed as follows: 150 samples
of gall bladder, 150 samples of mesenteric lymph
nodes, 150 samples of spleen, 150 samples of liver,
150 samples of small intestine and 150 samples of
cecum.
The isolation and biochemical identification
of Salmonella was carried out according to standard
laboratory methods (Quinn et al., 2004). Each
sample was transferred into tetrathionate broth
for Salmonella enrichment before streaking onto
338
Figure 1. Wellcolex color latex test for Salmonella.
***Results of KB003 Hi25
Enterobacteriaceae Identification :
Malonate
utilazation
Lactose
Figure 3. Showing results to KB003 Hi25 Enterobacteriaceae.

339
Figure 4. Salmonella serogroup : E (blue agglutination- pink background with Reagent 2).
Figure 5. Salmonella serogroup D : (green agglutination-pink background) with reagent 1.
Figure 6. Salmonella serogroup G (blue agglutination-pink background) with reagent 1.
340
black center, on Brilliant green agar the colonies

appeared gray reddish/pink slightly convex and on
Hicrome rajhans modified medium the colonies
appeared as pink in color.
The results of biochemical tests by
Api and
KB003 Hi25
Enterobacteriaceae
Identification system showed that this bacterium
was positive for: Lysine utilization, Ornithine
utilization, Nitrate reduction, H2 S production,
Citrate utilization, Methyl red, Arabinose, Xylose,
Rhamnose, Melibiose, Glucose; and negative for:
ONPG,Urease ,phenylalanine Deamination, Voges
Proskauers, Indole, Malonate utilization, Esculin
hydrolysis, Adonitol, Cellobiose, Saccharose,
Raffinose, Trehalose, Lactose, Oxidase (Figure 3).
The results of serogrouping of Salmonella
isolate according to serogrouping Wellcolex Color
Salmonella showed that and 4 isolates gave blue
agglutination-pink background with reagent 2 [it
belong to group E, (Figure 4)]; 3 isolates gave
[green agglutination- pink background with reagent
1 [it belongs to serogroup D, (Figure 5)] and 15
isolates gave blue agglutination-pink background
with reagent 1 [it belong to group G, (Figure
6)]. Table 1 showed of Salmonella grouping and
serotyping.
formalin as preservative.
5. Green positive control: Killed bacterial
suspension of organisms with Salmonella group
A and D antigens containing 0.05% Bridox and
0.5% formalin as preservative.
Finally, serotypings of Salmonella isolates
were confirmed in the Central Public Health
Laboratories by using specific antisera.
Antibiotic susceptibility tests for Salmonella
isolates were performed according to the Kirby
Bauer method (Bauer et al., 1996). Mueller Hinton
agar was used as growth medium for standard disc
diffusion test and growth was spread on plates
with the help of a sterilized cotton swab to form
a smooth bacterial lawn. The discs were placed on
to the agar surface using sterile forceps. Each disc
was gently pressed with the point of sterile forceps
to ensure complete contact with the agar surface.
Plates were incubated overnight at 37C.
Characterization of strains as sensitive or resistant
was based on the size of the inhibition zone around
the disc compared with the interpretation standards
provided by the manufacturers. The antimicrobial
drugs used were ampicillin, bacitracin,
chloramphenicol, erythromycin, gentamycin,
kanamycin, novobiocin, penicillin, spectinomycin,
streptomycin, tetracycline and trimethoprim.
Percentage of infection with Salmonella in

buffaloes
Microbiological examination of samples
that had been collected from slaughter houses
(organs and feces ) revealed that seven out of 150
were positive for Salmonella. The milk samples of
buffalo in the field revealed negative results so that
the statistic analysis showed significant differences
at p>0.05 between animals at the slaughter house
and animals in the field.
RESULTS
Isolation and identification of Salmonella spp.
Isolation and identification of Salmonella
were confirmed in different media as showed
in Figure 2. The cultural characteristics showed
different colonies, On Salmonella Shigella agar ,the
colonies appear as small pale, rounded with black
center, on Xylose-Lysine Deoxycholate agar, they
showed slightly transparent zone of reddish and
Percentage of infection with Salmonella species

341
in buffaloes in the slaughter house

The results of serotyping of Salmonella spp.
from (1050) samples at the slaughter house were
that22 isolates of three different serotypes were
recognized according to the Central Public Health
Laboratories. These serotypes were Salmonella
anatum (68.18%), Salmonella muenchen (18.18%)
and Salmonella enteritidis (13.64%). The statistical
analysis showed significant differences at p>0.05
between Salmonella anatum and the others .
age groups of buffaloes was found as shown in

Figure 7.
Clinical signs
Out of 41 animals, three animals showed
different clinical signs: diarrhea (1), respiratory
signs (1) and more than one symptom (1) were
affected with Salmonella, while four animals out
of 109 examined that gave positive results for
Salmonella did not show clinical signs (Table 2). In
the field, all animals with acute or chronic mastitis
and without clinical signs gave negative results for
Salmonella infection.
Distribution of Salmonella spp according to

month of year
Figure 8 shows the distribution of Salmonella
spp. according to month of year. No isolates
appeared in November and December, one affected
animal (1 isolate) was found in January, but in
February, three affected animals (12 isolates) were
found, two affected animals (6 isolates) were found
in March, and one affected animals (3 isolates) was
found in April.
Percentage of Salmonella isolated from different

samples
Salmonella was isolated from different
samples with different percentages except samples
of small intestine and milk. The highest percentages
appeared in the gall bladder (3.33%) and the liver
(3.33%) followed by mesenteric lymph nodes
(2.67%). Statistical analysis showed significant
difference at (p<0.05) (Table 3).
Results of antimicrobial sensitivity tests

The antibiotic susceptibility pattern
of Salmonella against the antimicrobial disc
revealed that all isolates were resistant (100%)
to
chloramphenicol,
suphamethoxazole,
erythromycin, cloxacillin, and tetracycline.
Aall isolates were sensitive to amicacin and
trimethoprim.
Distribution of Salmonella isolated according to

age
According to age of buffaloes at the
slaughter house, the results shows that Salmonella
was highest (6.67%) in the 1.5 - 3 year age group
and lower (3.23%) in animals more than 3 years
old. Statistically no significant difference between
DISCUSSION
Distribution of Salmonella species according to

sex
Thirteen isolates from 97 samples were
recorded in males and nine isolates from 53were
recorded in females. Statistically, there was no
significant difference between males and females
at p 0.05.
The results of the present study showed that

Salmonella were isolated at a percentage of 2.33%
buffaloes, and this is in a agreement with the study
of Hassanain 2008 which recorded a percentage
2.16% of Salmonella infection in buffaloes in
342
Egypt, and with a study of Al-Nakshabandy

(2001) in buffaloes, which found that the overall
ratio of infection with Salmonella in Mosul city in
Iraq was 2.23%). while Sen et al. (1988) recorded
the low percentage 1.60% of Salmonella from
rectal swabs of buffalo in Bangladesh. Phillips et
al. (2008) isolated Salmonella from 1.1% of ground
buffalo beef samples.
Other studies have also shown different
results, Hassanain et al. (2010) found a high
percentage (11.11%) of Salmonella in buffaloes
in Eygpt. Also Khan et al. (2009) found higher
percentage (16.3%) of Salmonella during study of
diarrhea in buffalo calves in Pakistan .
Boonmar et al.(2008) isolated Salmonella
from fecal samples of buffaloes in Japan at a
percentage of 8%. Sharma et al. (1989) found
16.53% of meat samples were positive to Salmonella
in Indian buffaloes.
No significant difference between
Salmonella infection in the three governorates may
explained by their being in the same geographical
region and having the same climate. Also. there
was easy transmission of animals between them.
No study is available concerning the distribution
of Salmonella in buffaloes at slaughterhouses in all
governorates in Iraq.
These results resemble those of other
studies. In Pakistan, Ali et al.(2008) recorded
negative results for Salmonella species in 200
mastitis quarters in buffaloes. Also, negative results
for Salmonella in Iraqi buffaloes were recorded by
Abdul Razak.(1982).
Our study is compatible with many studies
which showed that Salmonella was not isolated
from milk samples, such as Coroian et al. (2010) in
Romanian buffaloes ; Khan et al. (2009) in Pakistan
; Ali et al . (2008) in Pakistan, and Moroni et al.
(2006) in northern Italy.
In contrast, a study in Pakistan by Iqbal

et al. (2004) showed isolation of Salmonella at a
percentage of 2.41% in milk samples of lactating
dairy buffaloes .
The results of present study were in
agreement with those of Gunasegaran, 2011 who
found that Salmonella was resistant to tetracycline
and chloramphenicol and sensitive to kanamycin.
Also, Singh, et al. (2010) recorded that 46 strains of
Salmonella enteric.were resistant to streptomycin
and kanamycin but disagreed about resistance to
gentamicin and amoxycillin.
The present study showed that the
Salmonella anatum and Salmonella enteritidis were
resistant to seven antimicrobials and Salmonella
meunchen was resistant to eight antimicrobials ,
while a study by Boonmar et al. (2008) recorded
that the buffalo isolates were susceptible to most
of antimicrobials (tetracycline, streptomycin,
ampicillin,
sulfamethoxazole-trimethoprim,
chloramphenicol, amoxicillin-clavulanic acid and
nalidixic acid) and all isolates showed sensitivity
to cefotaxime, norfloxacin and ciprofloxacin.
In the present study, some animals showed
different clinical signs of Salmonellosis, and these
signs were similar to those recorded by Santana et
al.(2008) in their study on experimental infection
with Salmonella dublin in buffalo calves, which
showed the main clinical signs, i.e., diarrhea, fever,
respiratory signs and dehydrationwhile other animals
without clinical signs but gave positive results for
Salmonella infection. This revealed a number of
carrier animals without clinical signs, confirming
Nabbut and Al-nakhlil (1982), Galland et al.(2000),
and Radke et al. (2002) who found that animals
which had recovered from Salmonellosis may
continue shedding of Salmonella microorganisms
from 2-12 weeks post infection, whereas shedding
of Salmonella microorganisms from animals from
343
Figure 7. Distribution of Salmonella isolated according to age.
Figure 8. The distribution of Salmonella spp. according to month.
344
Table 1. Results of Salmonella grouping and serotyping.

Total No. of Salmonella
isolates
22
No. of
isolates
15
3
4
No. isolate
Serotyping at center of Salmonella
Group E
Group D
Group G
Salmonella anatum (68.18%)

Salmonella enteritidis (13.64%)
Salmonella muenchen (18.18%)
Statistical results showed significant difference at p < o.o5.
Table 2. Distribution of affected animals according to clinical signs before slaughtering.

Clinical signs
With clinical signs
No. of animals
41
Without clinical signs
109
No. of affected animals with Salmonella

3 animals (diarrhea(1), respiratory signs (1) and
more than one sings (1)
4 animals
345

Table 3. Percentage of Salmonella isolated from different samples.
Collection of
samples
No. of examined No. of positive No. of Salmonella

sample
sample
spp.
4 S. anatum,
Gall bladder
150
5
1 S. munchen
3 S. anatum
Mesenteric lymph node
150
4
1 S. munchen
2 S. anatum
Spleen
150
3
1 S. munchen
3 S. anatum
Organ and feces
Liver
150
4
1 S. munchen
at slaughter
1 S. enteritidis
house
2 S. anatum
Cecum
150
3
1 S. munchen
Small intestine
150
0
0
1 S. anatum
Feces
150
2
1 S. enteritidis
Total at slaughter house
1050
22
22
Milk
150
0
Field
Total
1200
22
Sample
Table 4. Antibiotic susceptibility pattern of Salmonella.
No.
Antibiotics used
Numbers of Salmonella isolates

R
Chloramphenicol
22
100
Neomycin
9.1
15
68.2
22.7
Suphamethoxazol
22
100
4
5
6
Gentamicin
Cefixime
Erythromycin
21
18
22
95.5
81.9
100
0
4
0
0
18.2
0
1
0
0
4.5
0
0
Ciprofloxacin
4.5
21
95.5
Kanamycin
4.5
11
50
10
45.5
Amoxicillin
21
95.5
4.5
10
Streptomycin
4.5
21
95.5
11
Tetracyclin
22
100
12
13
14
15
Amicacin
Trimethoprim
Cloxacillin
Nitrofurantoin
0
0
22
1
0
0
100
4.5
0
0
0
2
0
0
0
9.1
22
22
0
18
100
100
0
81.8
346
which clinical signs had disappeared may be found

with chronic infection of Salmonellosis. Carrier
animals may also be clinical cases if resistance is
lowered by environmental stress or inter current
infection.
In the present study, data demonstrated
that 22 Salmonella isolates were isolated from
1050 samples, constituting about 2.09%., and that
three different species were found. This results is
consistent with Singh et al. (2010) who recorded
different species in buffaloes (S. anatum, 13; S.
weltevreden, 13; S. rostock, 6; S. typhimurium, 5;
S. gallinarum, 5; S. stockholm, 1; S. dublin, 1; and
S. orion, 2) and with two studies: Hassanain et al.
(2008) and Hassanain (2010) who found two species
in buffaloes (S. enteritidis and S. typhimurium).
In India, Sharma et al. (1989) recorded
different species in buffaloes: S. anatum, S.
bareilly, S. stanley, S. Weltevreden, S. newport, S.
saintpaul, S. typhimurium, S. agona, S. chester,
and S. senftenberq, while Boonmar et al. (2008)
in Japan found two species only: S. derby and S.
javiana.
The results of the present study resemble
a study in Pakistan by Ali et al. (2008) which
recorded no infection for Salmonella in 200 mastitic
quarters in buffaloes, and also a study by Abdul
Razak (1982) in Iraqi buffaloes which showed no
infection in mastitic milk in buffaloes. Also, many
previous studies reported similar findings.
In the present study, the data demonstrated
a higher percentage of Salmonella isolates from
the gall bladder and the liver, whereas all of the
Salmonella serotypes (S. anatum, S.enteritidis,
S.muenchen) were isolated from the gall bladder
and the liver at a percentage of 40% from the
total of Salmonella isolates in this study followed
by isolates from the mesenteric lymph nodes, the
spleen and the cecum.
Our results also showed that the higher

percentage of Salmonella isolated from organs
as compared with feces. The reason for this
might be by that the body does not shed
Salmonella microorganisms continuously in feces:
particularly, carrier animals with Salmonella
shed microorganisms intermittently. This is in
agreement with the study of Molla et al. (2002)
which reported a high percentage of Salmonella
isolates (4.2%) in organs (mesenteric lymph node)
and a low percentage in feces (1.9%).These results
resemble those of Poernomo et al. (1986) who
found that the percentage of Salmonella isolates
from fecal samples (1.7%) was lower compared
with the percentage of Salmonella isolates from
organs (3.12%).
Our study revealed a difference in
the distribution of Salmonella in organs and
feces. This indicates that an animal may be a
carrier of Salmonella in its organs or actively
excretSalmonella in its feces This is in agreement
with a study of Singh et al. (2010) who mentioned
that if the animal was infected with Salmonella
organisms, it may become a clinical case or an
active or latent carrier . In the active carrier cases,
the organisms are localized in the intestine and gall
bladder, from whence they are excreted with the
feces, contaminating the environment and posing
to a threat to susceptible hosts. In latent carrier
cases, the organisms are localized in the lymph
nodes, liver, spleen and tonsils, but the organisms
are not excreted with the feces.
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International Buffalo Information Center (IBIC)

Dr. Rafat Al Jassim

School of Agriculture and Food Sciences, Faculty of

Assistant Journal Manager

International Buffalo Information Center, Office of the

International Buffalo Information Center, Office of the