copy of BB25-21 - International Buffalo Information Centre

Volume 25 No.2

International Buffalo Information Center
Aims
(IBIC)
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
ISSN : 0125-6726
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.
Editor
S. Sophon
Publisher
International Buffalo Information Centre,
Main Library, Kasetsart University
Online availible:
http://ibic.lib.ku.ac.th/e-Bulletin
BUFFALO BULLEITN
IBIC, KASETSART UNIVERSITY, P.O. BOX 1084
BANGKOK 10903, THAILAND
URL : http://ibic.lib.ku.ac.th
E-mail : libibic@ku.ac.th
Tel : 66-2-9428616 ext. 344
Fax : 66-2-9406688

25
Buffalo Bulletin (June 2006) Vol.25 No.2
RUMINAL pH AS REGULATOR OF RUMEN METABOLISM IN BUFFALOES
The importance of the buffalo as a dairy
animal is due to its ability to produce milk efficiently,
effectively and economically. The process of
conversion of feed into milk is complicated and is
dependent upon the ruminal environment. The type
of diet affects the rumen environment by changing
its pH, causing various digestive disorders
influencing the metabolic activity: hypo-galactia and
agalactia. The present study was, therefore,
undertaken to investigate the effect of changes in
pH on rumen metabolism and production in lactating
Murrah buffaloes.
Materials and Methods
A total 144 clinical cases of buffaloes of third/
fourth lactation (12 months) suffering from primary
indigestion were studied for one year at the
Veterinary Clinics of the University. Similarly, a total
120 buffaloes (10 per month) at the same lactation
were randomly selected and maintained on normal
farm schedule at the Animal Farm of the University.
Rumen samples were collected from these animals
at monthly intervals. The pH strip was used
immediately to assess the pH of rumen liquor and
later the pH of strained rumen liquor (SRL) was
measured for confirmation. Total counts of rumen
protozoa and bacteria were done following the
method described by Moir (1951) and Gall et al.
(1945) and their classification were done as per the
method described by Pounden and Hibs (1948). The
total and individual volatile fatty acids were
analyzed by using gas chromatography (Bernard and
Bouchque, 1968). Ruminal biogenic amines were
estimated by HPLC (Kochler and Eitenmiller, 1975).
M.A. Akbar and Ramesh Kumari
Results and Discussion
For the past last thirteen years, studies were
carried out on 144 clinical cases and 120 healthy
buffaloes of third/fourth lactation every year. Out of
144 clinical cases of buffaloes, three cases (2.08%)
of bloat, six cases (4.17%) of acidic indigestion, 21
cases of impaction (14.58%) and 114 cases (79.16%)
of alkaline indigestion were observed.
The mean pH value of strained rumen
liquor (SRL) ranged from 6.99 to 7.3 in healthy
animals and 5.5 to 8.9 in digestive disorder cases.
On the basis of pH value, the clinical cases were
categorized into four kind of indigestion, viz., bloat
(pH 5.52 to 5.72), acidic indigestion (pH 5.73 to 6.99),
impaction (pH 7.00 to 7.49) and alkaline indigestion
(pH 7.5 to 8.9) as shown in the Table . The above
categorization on the basis of pH corroborates with
the reports of Bhaskar (1971), Gupta et al. (1976)
and Choudhary et al. (1981).
Most of the cases had alkaline indigestion
and had a history of feeding on too much sorghum,
pearl millet and berseem fodder. It was observed
that acidic indigestion occurred due to excessive
feeding on dry ration and grains. The consumption
of rotten wheat straw, pearl millet stover and paddy
straw caused impaction while the cases of bloat
were due to excessive intake of green fodders like
lucerne, mustard, etc.
On microscopic examination of SRL, the
predominant genera of protozoa found were
Entodenia followed by Isotricha and Dasytricha at
pH 5.5 to 8.9 in both healthy and anorexic
buffaloes. Other protozoan species like Entodinium
bursa, Diplodinium dentatum, Polyplastron
multivesiculatum were observed only in healthy
buffaloes. The bacteria Oscillospira guillermondii
College of Animal Sciences, CCS Haryana Agricultural University, Hisar-125 004, India

Buffalo Bulletin (June 2006) Vol.25 No.2
was also found in these animals during winter months,
and this is an index of positive nitrogen balance in
healthy buffaloes. Entodenia species like E. simplex,
E. bicarinatum, E. triacum and E. elongatum were
seen in healthy buffaloes at pH 6.99 to 7.30.
These species are very rarely found in buffaloes
suffering from primary indigestion because these are
very sensitive to pH changes in the rumen (Hungate,
1966).
Rumen bacteria like Gram negative short rods
in fours and multiples of four suggestive of window
pans (Hay flora, group II-c) remained prominent in
healthy as well as in buffaloes suffering from
indigestion. Gram positive cocci singly or in long
chains, Gram negative rods resembling coli form
organisms, grain flora group-II, large Gram positive
cocci in closely knit-pairs or tetrads, hay flora group
I and medium sized comparatively thin Gram
positive rods or grain flora group-I were present in
addition to miscellaneous organisms like quoin’s oval
in healthy buffaloes. In clinical cases, small Gram
negative rods (F-type grain flora group-II) in
alkaline pH 7.5 to 8.9 and the gram positive cocci
were predominant after d-type flora in impaction at
pH 7.0 to 7.4. In the cases of acidic (pH 5.5 to 6.9)
only a particular species, i.e. Gram positive cocci,
dominated because other species are very sensitive
and required a specific pH and the proper substrate
for their role and activity. These observations
corroborate with the findings of other workers
(Ryan, 1964; Hungate, 1966), which indicated that
the number of species was higher in the control group
due to favourable pH and proper substrate to
survive and thrives.
The total protozoan count ranged from 4.35
x 10 5 /ml to 7.26x10 5 /ml at normal pH from 6.99 to
7.3 in healthy animals. In clinical cases where pH
varied from 5.5 to 8.9 the total protozoan count
ranged from 2.35 to 4.12 x10 5 /ml, which was
significantly (P

et al., loc. cit.).
The levels of ruminal biogenic amines, i.e.
histamine, tryptamine and tyramine, have positive
correlation with milk production (Singh et al., 1989).
Any kind of deviation in ruminal pH directly affects
the levels of biogenic amines and hence milk
production. The ruminal histamine level was found
to be more than double in bloat, i.e. at pH 5.00 to
5.50, and towards higher side in acidic indigestion,
where the pH was 5.60 to 6.90. High levels of
histamine in the above cases was responsible for
rumen atony and loss of appetite, thereby, reducing
metabolic activity and ultimately milk production.
The lower level of histamine in impaction and
alkaline indigestion, where the pH ranged from 7.5
to 8.9, was also observed to be associated with lower
milk production (Irwin et al., 1972; Gupta et al., loc.
cit.; Chaudhary et al., loc. cit.).
Ruminal tryptamine and tyramine levels
were found highest in alkaline indigestion followed
by impaction, acidic indigestion and bloat. Their
levels increased due to decarboxylation of tryptopha
and tyrosine for which optimum pH was observed
to be 7.00 to 8.90. These observations do not
correspond with the findings of Irwin et al. (loc. cit.;
1979). They reported higher values of these amines
at 10 per cent dietary protein level and the optimum
pH for decarboxylation was 5.5, while in the present
investigations it was observed to be alkaline. The
decarboxylation of tyrosine by mixed bacteria may
continue to occur when pH is in the neutral or in the
27
Buffalo Bulletin (June 2006) Vol.25 No.2
alkaline range. On the other hand, higher level of
tryptamine was probably due to its endogenous
release even at low protein concentration in the
rumen liquor. The higher level of both the amines
reduced the rumen motility, causing rumen atony
and various digestive disorders. This resulted in
reduced feed intake, low metabolic activity and
hence lower milk production through inhibition of
galactopoietic hormones and release of
catecholamine further impairing the action of
oxytocin towards release of milk.
Summary
Studies conducted on clinical cases of
lactating buffaloes suffering from primary
indigestion revealed that ruminal pH is one of the
most important factors. Any deviation in ruminal pH
causes different kinds of indigestion, which disrupt
the rumen environment resulting in reduced
metabolic activities and ultimately inhibiting milk
production.
Acknowledgement
Authors are thankful to Dr. Narendra Singh,
former Dean, College of Animal Sciences of this
University, for providing research facilities to carry
out this work under the National Fellow (ICAR)
project.

Buffalo Bulletin (June 2006) Vol.25 No.2
Table1 Microbial counts, VFA and biogenic amines found in rumen liquor of buffaloes suffering from
primary indigestion.
Attribute Digestive disorders
Healthy Bloat Acidic Impaction Alkaline
animals indigestion indigestion
pH 7.30±0.05 5.70±0.05 6.60±0.10 7.10±0.04 8.20±0.04
Total protozoan
counts (nx10 5 /ml) 5.59±0.39 1.50±0.04 4.18±0.20 4.59±0.06 3.14±0.07
Total bacterial
counts (nx10 9 /ml) 6.77±0.21 3.14±0.02 4.70±0.15 4.22±0.04 4.94±0.09
TVFA (m.eq./l) 110.09±6.73 50.02±0.76 51.30±1.033 53.08±0.68 54.63±0.26
Acetic acid (%) 70.42±2.02 54.69±0.94 55.23±1.46 56.62±0.68 54.63±0.26
Propionic acid (%) 14.06±0.73 30.13±0.32 28.51±0.22 24.76±0.34 22.62±0.12
Butyric acid (%) 10.93±0.29 7.38±0.09 8.09±0.06 9.61±0.06 10.14±0.03
Ruminal biogenic amines ( µg/ml)
Histamine 0.513±0.004 1.084±0.005 0.864±0.013 0.198±0.003 0.203±0.003
Tryptamine 0.298±0.004 0.285±0.001 0.351±0.001 0.460±0.008 0.453±0.003
Tyramine 0.268±0.004 0.197±0.001 0.211±0.002 0.277±0.006 0.357±0.002
28

REFERENCES
Ash, R.M. 1956. In: The Rumen and its Microbes.
Hungate R.E., ed. Academic Press, New York.
J. Physiol. 133: 75-78.
Bernard, G.C. and C.V. Bouchque. 1968. Rapid
method for the gas chromatographic
determination of volatile fatty acids in rumen
fluid. J. Agric. Food Chem. 16: 105-108.
Bhaskar, V.K. 1971.Studies on some of the
clinico-pathological aspects of indigestion
in buffaloes and cattle. M.V.Sc. Thesis,
Haryana Agricultural University, Hisar.
Choudhary, P.C., B. Prasad and S.K. Misra 1981.
Note on the use of rumen liquor in the
treatment of chronic alkaline indigestion in
cows. Indian J. Anim. Sci. 51: 356-360.
Dirksen, G. 1970. In: Physiology of Digestion and
Metabolism in the Ruminants. Phillipson, A.T.
ed. Oriels Press, London. p. 692.
Gall, L.S., W. Burroughs, P. Gerlaugh and B.H.
Edginton 1945. Special methods for rumen
bacterial studies in the field. J. Anim. Sci. 8:
433-436.
Gupta, G.C., B.P. Joshi and P. Rai. 1976. A note on
histamine concentration in rumen fluid and
blood in tropical bovine suffering from
indigestion. J. Anim. Sci. 46: 319-321.
Hungate, R.E. 1966. The Rumen and its Microbes.
Academic Press, New York.p.434.
Irwin, L.N., Jr. Mitchel, R.E. Tucker and G.T.
Shelling, 1972. Amine production by sheep with
glucose. J. Anim. Sci. 35: 267-271.
29
Buffalo Bulletin (June 2006) Vol.25 No.2
Irwin, L.N., G.E. Mitchell, R.E. Tucker and G.T.
Shelling. 1979. Histamine, tryptamine and
tyramine, and electrolytes during glucose
induced lactic acidosis. J. Anim. Sci.
48: 367-374.
Kochler, P.E. and R.R. Eitenimller. 1975.High
pressure liquid chromotatographic analysis of
tyramine, phenyl ethylamine and tryptamine in
sausage and chocolate. J. Food Sci.
43: 1245-1249.
Moir, R.J. 1951. The seasonal variation in the
ruminal micro organisms of grazing sheep.
Australian J. Agric. Res. 2: 322-331.
Pounden, W.D. and J.W. Hibs. 1948. The influence
of the ratio of grain to hay in the ration of dairy
calves in certain rumen micoorganisms.
J. Dairy Sci. 31: 1051-1057.
Ryan, R.K. 1964. In: The Rumen and its Microbes.
Ed. Hungate R.E., Academic Press, New York.
Am. J. Vet. Res., 25: 646-648.
Singh, N., J.P. Puri, O.P. Nangia and S.L. Garg.
1983. Early development of rumen function in
buffalo calves: Rumen microbes, metabolites
and cellulose digestion in vitro as a function of
age and diet. Indian J. Anim. Sci.
53: 933-938.
Singh, N., Kumari, R. and Akbar, M.A. 1989.
Effect of biogenic amines on milk production.
Indian J. Anim. Sci. 59: 1319-1323.

Buffalo Bulletin (June 2006) Vol.25 No.2
A SIGNIFICANT OBSERVATION ON THE STATUS OF CORTISOL IN NEONATAL BUFFALO
CALVES AND COW CALVES VIS-À -VIS THEIR DAMS
ABSTRACT
Cortisol levels were measured through ELISA
in 85 animals inclusive of periparturient dams and
their calves up to 91 days of life. The levels were
lower in the dams but in the early neonatal periods
the cortisol levels were significantly (P

RESULTS AND DISCUSSION
The values of serum cortisol in the periparturient
buffaloes and cows and their respective neonatal
calves have been presented in Table 2. The levels
of cortisol in the periparturient dams remained
almost identical. However, in the neonatal calves,
particularly within a month period, the cortisol levels
were significantly (p

Buffalo Bulletin (June 2006) Vol.25 No.2
Table 2. Status of serum cortisol of neonatal buffalo calves and cow calves along with their dams at different
periods.
Parameters Serum cortisol (µg/dl)
Dams Buffaloes Cows
Pre-partum 1.5±0.3 1.6±0.2
Post-partum 1.3±0.1 1.5±0.2
Calves Bc Cc
Pre-colostrum feeding 4.2±3.3* 4.5±0.8
6 h Post Colostrum feeding 7.2±2.8 7.8±1.4
1 st day 11.0±4.0* 8.4±2.0*
2 nd day 5.4±4.0 9.6±3.0
3 rd day 7.4±3.0 6.4±2.0
4 th day 8.0±4.0 7.0±3.0
5 th day 6.0±5.0 12.0±4.0
6 th day 9.0±6.0 10.0±4.0
14 th day 4.6±4.0 5.7±2.0
21 th day 4.0±0.4 8.0±4.0
28 th day 5.0±0.7 9.0±5.6
35 th day 3.0±2.0 6.0±3.0
42 th day 3.0±1.0 5.0±2.0
49 th day 3.0±1.0 4.0±1.0
56 th day 3.0±0.6 4.0±1.0
63 th day 5.0±1.0 3.0±1.3
70 th day 3.0±0.5 3.0±1.0
32

Contd.
*P

Buffalo Bulletin (June 2006) Vol.25 No.2
PROFILE OF SERUM IMMUNOGLOBULINS, GLUCOSE- 6-PHOSPHATE
DEHYDROGENASE AND L: N RATIO AS A MEASURE OF INTERNAL DEFENSE OF
NEONATAL BOVINE CALVES VS THEIR DAM : EFFECT OF NEEM OIL AS
IMMUNOMODULATOR
A.K. Jain, I. J. Sharma, M.A. Quadri, R.K. Tripathi and R.G.Agrawal
ABSTRACT
The internal defense of the animals is
accomplished through the combined effort of blood
cells and different immunoglobulins. The levels of
immunoglobulins were higher in sera samples of
periparturient buffaloes while reverse was true for
the levels in the colostrum. The immunoglobulin
content in the serum of buffalo calves after
colostrum feeding was lower that in sera of cow
calves. This clearly indicates poor absorption of
immunoglobulins from the gastrointestinal tract of
buffalo calves. The L:N ratio was also low in the
buffalo calves, whereas G-6PD was higher. Together
the low immunoglobulin and L: N ratio reduce the
immunocompetence of the buffalo calves.
Keyword: neonate, postpartum, immunology,
glucose-6-phosphate dehydrogenase, neem oil,
calves
INTRODUCTION
In bovines, the maternal transfer of
immunoglobulins through placenta is very little.
Maternal colostrum accumulates and provides the
immunoglobulins to the newborn calves, which give
passive immunity to the calves. Variation in the
immunoglobulin levels in neonates of different
genotypes was reported by Sangwan et al. (1987).
The death of the buffalo calves within 15 days was
reported by Sikka and Seth (1996) due to poor
Jawaharlal Nehru Krishi Vishwa Vidyalaya,Jabalpur,Madhya Pradesh 482001 (M.P.)India
34
absorption of immunoglobulin from the gastrointestinal
tract. Internal defense is the result of
a combined effort of blood cells and serum
immunoglobulins. Gupta et al. (1990) pointed out that
the foetuses of buffalo calves had negligible amounts
of immunoglobulins in their serum. Later on after
birth 70% of young calves had more than 15mg%
due to absorption of colostral immunoglobulins
through intestinal villi. The L:N ratio is important from
viewpoint of defense through antibody secretion.
The G-6PD is the first line of defense of the body
(Ganong, 1999). The interassociation of other
related cellular parameters and serum
immunoglobulins with high mortality of the neonatal
buffalo calves is not clear. In an attempt to work
out the possible cause of the heavy toll of neonatal
buffalo calves, this project was envisaged.
MATERIALS AND METHODS
The experiment was carried out from
December 1, 2004 to August 2005. The experimental
design and animals used are given in Table 1. The
immunoglobulin levels in the sera samples of the
calves and whey portion of colostrum of their
respective dams were estimated as per the method
of Selman et al. (1970) using the ZnSO 4 turbidity
method and the level of G-6-PD by the methods of
Kachmar and Moss (1976) in the red cell
haemolysate using ready-made kits. The

Lymphocyte: Neutrophil (L:N) ratio was calculated
from their respective absolute figures recorded with
a Medonic Thor – 20 Auto cell counter. The
statistical analysis of date was done as per Snedecor
and Cochren (1989).
RESULTS AND DISCUSSION
The results of serum and colostral
immunoglobulin of dams, their L:N ratio and
G-6-PD have been presented in Table 2. The total
immunoglobulin was recorded to be always higher
in the periparturient buffalo sera and colostral
samples in comparison to the periparturient cows.
However, immediately after birth, before colostrum
feeding to the calves of both the species, the total
immunoglobulin content was almost nil, but it started
rising 6 h after colostral feeding and thereafter with
a slight ebb during the first week of the neonatal
period (Table 3). Administration of neem oil
enhanced the immunocompetence of the calves as
earlier reported by Katiyar et al. (1997). One fact
was evident from the observation that the levels of
serum immuno globulins in buffalo calves was
always lower than the cow calves. This might have
been the resultant of poor absorption from the G.I.
tract of the buffalo calves resulting into low levels
of immunoglobulins in serum and high mortality rate
of neonatal buffalo calves. Archana et al. (1996)
also reported that the higher environmental
temperature reduces the absorption from G.I. Tract.
The L:N ratio was quite high in periparturient
dams but fell down (Table 3) in the precolostral fed
calves. The ratio increased slowly to adult values
with age. The conspicuous observation was that the
L:N ratio of buffalo calves was significantly (P

Buffalo Bulletin (June 2006) Vol.25 No.2
Table 1. Place of procurement and number of the animals.
S. No. Experimentation Condition of Animals Location of Animals No. of animals used Calves died
1. Periparturient Buffaloes Reliable Dairy, Gwarighat, Jabalpur 19 -
Periparturient Cows LSF, Adhartal, COVS, Jabalpur 16 -
2. Control Buffalo calves Reliable Dairy, Gwarighat, Jabalpur 12 6
Cow calves LSF, Adhartal, COVS, Jabalpur 12 2
36
3. Treated with Buffalo calves Reliable Dairy, Gwarighat, Jabalpur 15 7
Neem oil
@ 10ml/Day
Cow calves LSF, Adhartal, COVS, Jabalpur 11 -

Table 2. Status of defensive parameters of periparturient buffaloes and cows
37
Buffalo Bulletin (June 2006) Vol.25 No.2
Stage Animal Total Immunoglobulin (mg/ml) G-6PD (U/g. Hb) L: N ratio
Colostrum Serum
Prepartum Buffaloes 127±11 79±15 1.57
Cows 92±16 17±3.3 1.76
Postpartum Buffaloes 254±33 202±11 77±12 1.84
Cows 206±32 149±28 22±6.3 4.97

Buffalo Bulletin (June 2006) Vol.25 No.2
Table 3. Status of defensive parameters of neonatal buffalo calves (Bc) and cow calves (Cc) along with
effect of Neem oil at different time intervals
Time intervals Neonatal Total immunoglobulin G6PD(U/gm Hb) L: NRatio
(mg/ml)
Calves Control Neemoil treat Control Neemoil treat. Control Neemoil treat.
Precolostral Bc 5.8±0.6 - 107±25 - 0.8±0.1* -
feeding
Cc 4.1±1.0 34±8 1.1±0.1
6 h post Bc 122±30* 123±10 124±21* 81±12 0.7±0.1* 0.8±0.1
colost. feed.
Cc 153±34 155±43 26±6.5 21±5.2 1.2±0.1 1.1±0.1
1 st Day Bc 65±9.3 78±20 117±15* 112±22 0.8±0.1* 0.8±0.1
Cc 96±25 95±17 23±6.4 33±6.0 1.2±0.1 1.2±0.1
2 nd Day Bc 91±13 86±27 198±17* 122±18 0.8±0.1* 0.8±0.1
Cc 95±19 94±14 24±5.0 38±10 1.4±0.1 1.4±0.1
3 rd Day Bc 98±18 110±25 150±18* 99±23 0.8±0.1* 0.9±0.1
Cc 103±21 105±7.6 23±5.0 21±5.4 1.5±0.1 1.5±0.1
4 th Day Bc 119±13 111±25 158±25* 109±17 0.8 0* 0.9±0.1
Cc 104±19 108±23 33±4.0 37±10 1.5±0.1 1.6±0.1
5 th Day Bc 123±22 101±20 166±17* 146±61 0.8±0.1* 0.9±0
Cc 134±29 139±16 44±9.2 61±8.1 1.6±0 1.7±0.1
6 th Day Bc 103±27 94±21 146±28* 90±25 0.9±0.1* 0.9±0
Cc 113±11 115±13 47±10 16±4.0 1.5±0.1 1.8±0.2
14 th Day Bc 101±22 114±17 56±18 121±24 0.9±0.1* 0.9±0
Cc 110±21 140±25 36±9.8 36±9.8 1.5±0.1 1.5±0.3
21 st Day Bc 109±10 99±31 65±21 82±23 1±0.1 1.0±0
Cc 115±26 92±16 38±4.4 35±8.5 1.5±00 2.0±0.1
38

39
Buffalo Bulletin (June 2006) Vol.25 No.2
Time intervals Neonatal Total immunoglobulin G6PD(U/gm Hb) L: NRatio
(mg/ml)
Calves Control Neemoil treat Control Neemoil treat. Control Neemoil treat.
28 th Day Bc 125±12 101±18 122±26* 69±24 1.0±00 0.9±0.1
Cc 104±11 124±23 32±12 37±10 1.6±0.1 2.3±0.2
35 th Day Bc 129±15 181±42 108±30* 38±8.9 0.8±0.2* 0.9±0.1
Cc 140±35 155±43 25±7.8 30±9.1 1.5±0.1 2.3±0.2
42 nd Day Bc 90±24* 159±32 33±6.6 67±16 0.8±0.1 0.9±0.1
Cc 148±60 123±15 22±10 44±12 1.4±0.1 2.2±0.2
49 th Day Bc 184±32 90±17 60±21 75±28 0.8±0.1* 0.9±0.1
Cc 142±32 171±17 26±3.8 28±6.7 1.5±0.1 2.3±0.1
56 th Day Bc 104±26 92±16 47±15 50±16 0.7±0.1* 0.8±0.1
Cc 139±16 187±39 17±3.8 28±5.8 1.5±0.1 2.3±0.1
63 rd Day Bc 123±22 91±11 123±22 91±11 0.7±0.1* 0.8±0.1
Cc 114±20 141±20 114±20 141±20 1.5±0.1 2.2±0.1
70 th Day Bc 95±13* 92±16 95±13 92±16 0.7±0.1* 0.7±0.1
Cc 174±32 146±29 174±32 146±29 1.4±0.1 2.4±0.1
77 th Day Bc 126±10 123±10 61±20 72±19 0.7±0.1 0.7±0.1
Cc 115±26 148±22 19±3.9 24±2.4 1.6±0.1 2.3±0.4
84 th Day Bc 115±15 108±12 265±14 53±11 0.6±0.1* 0.6±0.1
Cc 104±12 147±20 19±5.8 32±4.7 1.5±0.1 2.3±0.2
91 st Day Bc 107±25 142±19* 65±16 53±11 0.5±0.1* 0.6±0.1*
postpatum Cc 115±25 145±28 31±7.4 33±4.8 1.5±0.1 2.6±0.3
. *P

Buffalo Bulletin (June 2006) Vol.25 No.2
HAEMATO-BIOCHEMICAL PROFILES OF BUFFALO IN ANTHELMINTICS TREATMENT
AGAINST FASCIOLA GIGANTICA INFECTION
ABSTRACT
Haemato-biochemical profiles were studied on
Fasciola gigantica infected buffalo in Triclabendazole,
Rafoxanide and Oxyclozanide treatment
Haemetological analysis showed reduced
erythrocytic count, haemoglobin percent and PCV
level and eosinophilia in all groups before treatment.
The values were improved 15 days after treatment.
But the statistically significant improvement was seen
in eosinophil percent in Rafoxanide and Oxyclozanide
treated animals while Triclabendazole treated
animals showed low level of eosinophilia.
Biochemical analysis revealed hypoproteinemia,
hypoalbuminemia, hypoglobulinemia, increased
levels of total serum bilirubin, AST, ALT and ALP
before treatment. After treatment, total serum
bilirubin and AST values were decreased significantly
in all treated groups while a significant level of total
protein and globulin were found only in Oxyclozanide
treated animals.
INTRODUCTION
Fasciolosis is a serious economic disease of
livestock with worldwide distribution. Of the three
species (F. hepatica, F. giantica and F. jacksoni)
F. giantica is regarded as one of the most important
parasitic infection of ruminants in countries with
tropical climates (Boray, 1985; Harrison et al., 1996
and Tantrawatpan et al., 2003). It is a major
constraint for livestock industries due to high
mortality and morbidity (Sharma et al., 1989),
decrease in weight gain (Bhatia et al., 1989 and
S. Pal and C.K. Dasgupta
Department of Veterinary Parasitology, West Bengal University of Animal and Fishery Sciences, Belgachia,
Kolkata, West Bengal 700 037, India
40
Mandal, 1997), fertility, milk yield and wool
production and liver condemnation (Swarup et al.,
1987). Prevalences of F. giantica in fection in some
ruminants range up to 80-100% in many countries
like West Africa (Schillhorn Van Veen, 1980), India
(Sharma et al., 1989) and Pakistan (Kendall, 1954)
etc. Among the different definitive hosts, buffalo is
the susceptible.
Effective strategies for the control of fasciolosis
are mainly based on the proper diagnosis and the
use of very effective drugs. But, resistance of the
liver fluke to flukicides is becoming a great problem
to farmers (Mitchell et al., 1998; Estuningsih, et al.
1990 and Sanyal, 1996). It was therefore, decided
to undertake this study to see the haematobiochemical
changes in buffalo infected with F.
gigantica in leading flukicides therapy.
MATERIALS AND METHODS
Sixteen adult buffaloes (Murrah breed) of
either sex naturally infected with Fasciola gigantica
at Mayakole, Krishnagar, Nadia, West Bengal were
used for this study. They were divided into four groups
of four in each. Animals of Group-I were treated
with Triclabendazole (Fasinex, Trend Pharma
Private Ltd.) at 12 mg/kg body weight orally.
Group-II was treated with Rafoxanide (Rafoxin
2000, Concept Pharmaceuticals Ltd.) at 7.5 mg/kg
body weight and Group-III was treated with
Oxyclozanide (Distodin, Pfizer) at 10 mg/kg body
weight orally while Group-IV was left as an untreated
infected control. Efficacy of the chemotherapeutic

agents was determined on the basis of pre and 15 th
day post-treatment haematological and biochemical
values.
Haematological profiles:
The blood samples collected in EDTA
(sodium salt of ethylenediamine tetraacetic acid)
vials were processed immediately as per the method
describe by Jain (1986) to note the parameters viz.,
total erythrocyte count (10 6 /cumm), total leukocyte
count (10 3 /cumm), haemoglobin (gm/dl),
differential leukocyte count (%) and PCV (%).
Biochemical profiles:
The sera samples collected in the sterilized
plastic vials were preserved at 20 C till used for
biochemical analysis. The samples were subjected
to liver function tests viz., Serum Aspartate
Aminotransferase(AST), Serum Alanine
Aminotransferase (ALT), Alkaline Phosphatase
(ALP), total protein, albumin, globulin and serum
bilirubin as per the spectrophotometric methods
supplied by ‘SPAN’ diagnostic kits.
RESULTS AND DISCUSSION
Haematological profiles in bubaline fasciolosis :
The blood samples of the 16 buffaloes
confirmed infected with F. gigantica were subjected
to haematological analysis. The values of total
erythrocyte count, haemoglobin and PCV before the
treatment were below the mean reference values
and increased following treatment in all treated groups
(Table 1). But a significant difference (P

Buffalo Bulletin (June 2006) Vol.25 No.2
Table 1 Haematological profiles of F. gigantica infected buffalo before (BT) and after treatment (AT):
Anima1 Drug TEC(X10 6 /ul) Hb(g/dl) PCV(%)
BT AT BT AT BT AT
1 Group-I 4.14 4.50 8.45 8.89 23 26
2 Triclabendazole 5.85 5.70 10.18 10.83 30 30
3 @ 12 mg/kg B.wt. 5.53 5.62 9.84 10.03 29 30
4 5.76 6.10 10.21 10.81 30 31
Avg.±S.E. 5.32±0.39 5.48±0.34 9.67±0.41 10.41±0.45 28.00±1.68 29.25±1.26 ab
5 Group-II 5.02 5.28 10.06 10.74 25 26
6 Rafoxanide @ 5.58 5.47 10.06 10.44 26 27
7 7.5mg/kg b. wt. 4.61 5.03 9.18 9.48 20 24
8 5.61 6.08 10.47 10.63 27 31
Avg. ±S.E. 5.21±0.24 5.47±0.22 9.94±0.27 10.32±0.28 24.50±1.55 27.00±1.47 b
9 Group-III 5.96 6.15 11.37 11.53 30 32
10 Oxyclozanide @ 5.34 5.86 10.88 11.03 26 29
11 10Mg/kg b. wt. 5.12 5.87 9.33 10.02 24 30
12 5.67 6.08 11.20 11.47 27 31
Avg. ±S.E. 5.52±0.18 5.99±0.07 10.70±0.46 11.01±0.34 26.75±1.25 30.50±0.65 a
13 Group-IV 5.60 5.52 10.12 10.28 28 27
14 Untreated 5.41 5.46 10.33 10.26 27 27
15 Control 5.78 5.62 11.07 10.69 28 26
16 5.27 5.30 9.93 10.13 25 24
Avg. ±S.E 5.52±0.11 5.48±0.06 10.36±0.24 10.34±0.12 27.00±0.71 26.00±0.71 bc
Means under the same superscripts do not differ significantly
42

Table 3 Biochemical profiles of F. gigantica infected buffaloes before (BT) and after treatment (AT).
Anima1 Drug TLC Neutrophil Eosinophil Basophil Lymphocyte Monocyte
(X103ul) (%) (%) (%) (%) (%)
BT AT BT AT BT AT BT AT BT AT BT AT
43
Buffalo Bulletin (June 2006) Vol.25 No.2
1 Group I 10.85 11.95 32.00 37.00 19.00 12.00 1.00 1.00 45.00 48.00 3.00 2.00
2 Triclaben 9.25 8.70 32.00 34.00 9.00 7.00 2.00 1.00 58.00 54.00 3.00 4.00
3 dazole @ 10.05 10.79 36.00 35.00 11.00 7.00 - 1.00 55.00 54.00 4.00 3.00
4 12 mg/kgB. wt 10.22 11.32 32 36.00 11.00 10.00 1.00 1.00 53.00 48.00 3.00 5.00
Avg. ±S.E. 10.09 10.69 33.00 35.50 12.50 9.00 1.00 1.00 52.75 51.00 3.25 3.50
±0.32 ±0.70 ±1.00 ±0.64 ±2.21 ±1.22b ±0.41 ±0.00 ±2.78 ±1.73 ±0.25 ±0.6
5 Group II 8.90 8.95 33.00 38.00 14.00 6.00 2.00 1.00 47.00 52.00 4.00 3.00
6 Rafoxa 9.95 11.55 26.00 34.00 13.00 4.00 - 1.00 58.00 57.00 3.00 4.00
7 nide@ 10.05 11.40 30.00 30.00 17.00 6.00 - 1.00 47.00 58.00 6.00 5.0
8 7.5 mg/kg b. wt.12.95 12.85 35.00 36.00 13.00 6.00 1.00 1.00 44.00 54.00 7.00 4.00
Avg. ±S.E. 10.46 11.19 31.00 34.50 14.25 5.50 0.75 1.00 49.00 55.25 5.00 4.00
±0.86 ±0.81 ±1.95 ±1.70 ±0.94 ±0.50a ±0.48 ±0.00 ±3.08 ±1.37 ±0.91 ±0.40
9 Group III 8.15 9.65 34.00 33.00 10.00 5.00 - 1.00 53.00 58.00 3.00 3.00
10 Oxyclo 9.90 10.56 36.00 36.00 12.00 6.00 - - 50.00 55.00 2.00 3.00
11 zanide @ 8.70 10.05 30.00 35.00 18.00 7.00 1.00 1.00 47.00 53.00 4.00 4.00
12 10 mg/Kg b. wt 10.12 10.88 32.00 33.00 11.00 6.00 1.00 1.00 52.00 58.00 4.00 2.00
Avg. ±S.E. 9.22 10.29 33.00 34.25 12.75 6.00 0.50 0.75 50.50 56.00 3.25 3.00
±0.47 ±0.27 ±1.29 ±0.75 ±1.79 ±0.40a ±0.29 ±0.25 ±1.32 ±1.22 ±0.48 ±0.41
13 Group IV 9.35 9.41 36.00 35.00 12.00 11.00 1.00 1.00 47.00 50.00 4.00 3.00
14 Untreated 8.80 8.65 29.00 30.00 10.00 8.00 - 1.00 58.00 57.00 3.00 4.00
15 Control 8.44 8.40 27.00 29.00 11.00 8.00 1.00 - 57.00 59.00 4.00 4.00
16 10.07 10.21 34.00 34.00 11.00 14.00 - - 52.00 48.00 3.00 4.00
Avg. ±S.E. 9.17 9.17 31.50 32.00 11.00 10.25 0.50 0.50 53.50 53.50 3.50 3.75
±0.35 ±0.40 ±2.10 ±1.47 ±0.41 ±1.44b ±0.29 ±0.29 ±2.53 ±2.66 ±0.29 ±0.25
Means under the same superscripts do not differ significantly

Buffalo Bulletin (June 2006) Vol.25 No.2
Table 3 Biochemical profiles of F. gigantica infected buffaloes before (BT) and after treatment (AT).
Animal Drug TotalProtein Albumin Globulin A/G Total serum
No. (g/dl) (g/dl) (g/dl) Bilirubin (mg/dl)
BT AT BT AT BT AT BT AT BT AT
1 Group I 6.70 6.98 2.54 2.89 4.16 4.09 0.61 0.71 0.38 0.18
2 (Triclaben 6.94 7.16 2.78 3.03 4.16 4.13 0.67 0.73 0.32 0.18
3 dazole 7.16 7.62 3.30 3.61 3.86 4.01 0.85 0.90 0.27 0.19
4 Treated) 6.52 6.81 3.07 3.35 3.45 3.46 0.89 0.97 0.21 0.10
Avg. ±S.E. 6.83 7.14 2.92 3.22 3.90 3.92 0.76 0.83 0.29 0.16
±0.13 ±0.17b ±0.16 ±0.16 ±0.16 ±0.15b ±0.06 ±0.06 ±0.22 ±0.22b 5 Group II 6.21 6.87 2.43 3.18 3.78 3.69 0.64 0.86 0.20 0.06
6 (Rafoxa 6.93 7.22 2.50 2.62 4.43 4.60 0.56 0.57 0.35 0.02
7 nide 6.72 7.06 2.92 3.11 3.80 3.95 0.77 0.79 0.29 0.01
8 treated) 7.20 7.65 3.17 3.49 4.03 4.16 0.79 0.84 0.12 0.05
Avg. ±S.E. 6.77 7.20 2.76 3.10 4.01 4.10 0.69 0.76 0.24 0.04
±0.20 ±0.16b ±0.17 ±0.18 ±0.15 ±0.19ab ±0.05 ±0.06 ±0.07 ±0.21a 9 Group III 6.85 8.55 2.96 3.92 3.89 4.63 0.76 0.85 0.32 0.05
10 (Oxyclo 7.73 9.83 2.63 3.50 5.10 5.33 0.52 0.66 0.23 0.01
11 zanide 7.40 8.95 2.65 3.77 4.75 5.18 0.56 0.73 0.23 0.03
12 treated) 6.36 6.77 2.29 2.91 4.07 3.86 0.56 0.75 0.30 0.06
Avg. ±S.E. 7.09 8.53 2.63 3.53 4.45 4.75 0.6 0.75 0.27 0.04
±0.25 ±0.50a ±0.13 ±0.22 ±0.23 ±0.33a ±0.05 ±0.03 ±0.28 ±0.33a 13 Group IV 6.87 6.48 3.01 3.07 3.3.86 3.41 0.78 0.90 0.34 0.28
14 (Untreated 6.60 6.71 2.82 2.73 3.78 3.98 0.75 0.69 0.23 0.26
15 Control) 6.41 6.28 3.43 3.41 2.98 2.87 1.15 1.19 0.27 0.29
16 7.43 7.12 3.64 3.60 3.79 3.52 0.96 1.02 0.31 0.35
Avg. ±S.E. 6.83 6.65 3.23 3.20 3.60 3.45 0.91 0.95 0.29 0.30
±0.17 ±0.19b ±0.17 ±0.19 ±0.16 ±0.24b ±0.07 ±0.11 ±0.24 ±0.16c 44
Means under the same superscripts do not differ significantly

45
Buffalo Bulletin (June 2006) Vol.25 No.2
Table 4 Biochemical profiles of F. gigantica infected buffaloes before (BT) and after treatment (AT).
Animal Drug AST(SGOT) ALT(SGPT) ALP
No. (U/ml) (U/ml) (K.A. Unit)
BT AT BT AT BT AT
1 Group-I 56 54 32 30 33.70 28.16
2 (Triclaben 47 45 30 29 17.86 15.52
3 dazole 52 49 29 26 20.06 17.01
4 treated) 50 47 21 22 27.14 18.55
Avg. ±S.E. 51.25 48.75 28.00 25.75 24.69 19.81
±1.89 ±1.93 a ±2.42 ±2.02 ±3.59 ±1.65
5 Group-II 53 46 26 22 17.40 16.26
6 (Rafo 51 45 22 24 16.41 15.17
7 xanide 54 48 28 25 30.05 20.34
8 treated) 46 42 38 30 52.40 37.87
Avg. ±S.E. 51.00 45.25 28.50 25.25 29.06 22.41
±1.78 ±1.25 a ±3.40 ±1.70 ±8.37 ±5.27
9 Group-III 46 44 29 26 20.73 14.61
10 (Oxyclo 53 46 23 21 30.47 23.28
11 zanide 64 53 34 31 25.05 19.94
12 treated) 51 47 36 31 18.29 12.74
Avg. ±S.E. 53.50 47.50 30.50 27.25 23.63 17.64
±3.80 ±1.94 a ±2.90 ±2.39 ±2.67 ±2.42
13 Group-IV 54 54 31 26 32.25 31.66
14 (Untreated 54 55 25 28 17.14 21.41
15 Control) 51 52 28 27 20.41 20.80
16 57 56 30 29 34.05 27.73
Avg. ±S.E. 54.00 54.25 28.50 27.50 25.96 25.40
±1.22 ±0.85 b ±1.32 ±0.65 ±4.21 ±2.60
Means under the same superscripts do not differ significantly

Buffalo Bulletin (June 2006) Vol.25 No.2
REFERENCES
Bhatia, B.B., D.S. Upadhaya and P.D. Juyal. 1989.
Epidemiology of Fasciola gigantica in
buffaloes, goat and sheep in Tarai region of
Uttar Pradesh. J. Vet. Parasitol, 3: 25-29.
Bharti, P. and K.D. Prasad. 2001. Biochemical
profiles of cattle and buffalo infected with
Paramphistomum spp and Fasciola
gigantica. J. Vet. Parasitol., 15(2): 149-151.
Boray, J.C. 1985. Flukes of domestic animals. In
Gaafar, S.M., W.E. Howard and R.E.Marsh
(eds.). Parasites, Pests and Predators.
Elsevier, New York, pp. 179-218.
Estuningsih, S.E., P. Stevenson, Beriajaya and M.R.
Knox. 1990. Triclabendazole in the treatment
of Fasciola gigantica infection in swamp buffalo
(Bubalus bubalis). Australian Vet. J., 67:
234-235.
Harrison, L.J.S., J.A. Hammond and M.M.H.
Sewell. 1996. Studies on helminthosis at the
centre for Tropical Veterinary Medicine
(CTVM). Trop. Anim. Hlth Prod., 28: 23-39.
Jain, N.C. 1986. Schalm Veterinary Haematology.
4 th ed. Lea and Febiger, Philadelphia.
Kendall, S.B. 1954. Fascioliasis in Pakistan. Ann.
Trop. Med. Parasitol., 48: 307-313.
Kumar, M., K.M.L. Pathak and S.P. Pachauri. 1982.
Clinicopathological studies on naturally
occurring bovine fascioliasis in India. British
Vet. J., 138: 241-246.
Maiti, S.K., V.N. Rao, S.L. Ali, G. Dutta and A.
Mishra. 1999. Haematobiochemical and
therapeutic aspects of biliary amphistomiasis
in buffalo. Indian J. Vet. Medicine, 19:
49-51.
Mandal, S. 1997. Evaluation of Fasciola gigantica
antigens for immunodiagnosis of fasciolosis
in ruminants. M.V.Sc. Thesis, I.V.R.I.,
Izatnagar, India.
46
Martinez-Moreno, A., V.M. Jimenez, M.S. Martinez-
Cruz, F.J. Martinez, C. Becerra and
S.Harnandez. 1999. Triclabendazole treatment
in experimental goat fasciolosis: anthelmintic
efficacy and influence in antibodody response
and pathophysiology of the disease. Vet.
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Mitchell, G.B., L.Maris and M.A. Bonniwell. 1998.
Triclabendazole resistant liver flukes in Scottish
sheep. Vet. Record, 143: 399.
Moreno, A., F.J. Martinez, V. Jimenez, M.S. Cruz,
F.J. Martinez-Moreno, B.C. Martinez and S.
Hernandez. 1997. Triclabendazole treatment in
experimental goat fascioliasis: Anthelmintic
efficacy and influence in antibody response and
pathophysiology of the disease. Vet. Parasitol.,
68: 57-67.
Sanyal, P.K. 1996. Kinetic disposition and critical
efficacy of triclabendazole against
experimental bovine and bubaline fasciolosis.
J. Vet. Parasitol., 10(2): 147-152.
Schillhorn van Veen, T.W. 1980. Fascioliasis
(Fasciola gigantica) in West Africa: a review.
Vet. Bulletin, 50: 529-533.
Sharma, R.L., D.N. Dhar and O.K. Raina. 1989.
Studies on the prevalence and laboratory
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Swarup, D., S.P. Pachauri, B. Sharma and S.K.
Bandhopadhyay. 1987 Serodiagnosis of
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S. Wongkham, P.M. Intapan and K.
Nakashima. 2003. Characterization of
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363-372.

RESEARCH ABSTRACTS
ANATOMY
N. Singh, K. S. Roy. Punjab Agricultural
University, Ludhiana, Punjab 141 004,
India.Histochemical study on the
mammary gland of Indian buffalo (Bubalus
bubalis). Indian Journal of Animal Sciences
(2006) 76: 43-45.
The distribution of neutral and acid
mucopolysaccharides, basic protein and lipids in the
mammary glands of lactating and non-lactating
buffaloes was studied using histochemistry. It was
shown that the neutral mucopolysaccharides and
basic protein were mainly found in the alveoli of the
mammary glands of lactating buffaloes, while acid
mucopolysaccharides were mainly found in the
stromal mammary tissue of non-lactating animals.
Sudanophilic lipids were also present in the alveolar
and duct system epithelium of the mammary glands
of lactating buffaloes. The corpora amylacea was
positive for neutral mucopolysaccharides and basic
protein in the mammary gland of non-lactating
buffaloes.
47
Buffalo Bulletin (June 2006) Vol.25 No.2
PHYSIOLOGY
Sheikh, P. A., F. D. Merry, D. G. McGrath.
Department of Biology, Pennsylvania State
University, University Park, PA 16802, USA.
Water buffalo and cattle ranching in the
Lower Amazon Basin: comparisons and
conflicts. Agricultural Systems (2006) 87
(3):313-330.
From 1975 to 2000, the water buffalo
population in the Brazilian Amazon increased at
nearly 13% per year, making it one of the fastest
growing herds in the world. On the floodplains of
the Amazon River buffalo are managed in a similar
manner to cattle, but often earn superior production
figures. This production advantage, however, is
tempered by the role buffalo play in conflicts
between landowners; buffalo are prone to altering
the floodplain environment and interfering with
production activities such as fishing and farming. In
this research, we show that buffalo are kept on the
floodplains 24% longer than cattle throughout the
year, and 37% longer than cattle during periods when
landowner conflicts are most likely to occur. We also
show that buffalo productivity is greater than cattle
in this system, which gives an opportunity to design
management regimes for buffalo that may increase
production costs, but that will lower the
environmental and social problems that involve
buffalo. Specifically , when the waters begin to rise,
buffalo should be removed from the floodplains at
the same time as cattle. Although this will not lessen
the damage done while the buffalo is on the
floodplain, it will place buffalo on the floodplain
during the dry season when the erosion potential is
at its lowest, and reduce the time that buffalo may
interfere with other production activities such as
fishing. The additional production costs incurred by
early removal will not dissipate the production
advantage over cattle. Without specific management
that addresses the socio-economic and environmental
problems caused by buffalo, the continued high
growth rate for the buffalo population on the
Amazon floodplains may not be sustainable and
conflicts will become commonplace.

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Manuscript. In 12 point Times or Times New Roman. Type on one
side of A4 paper. Use one inch margins. Number all pages. Send an
original manuscript and 1 photocopy.
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containing the manuscript in Microsoft Word.
Mail manuscript to:
By post: International Buffalo Information Center
Office of University Library
Kasetsart University,
50 Pahonyothin Road, Chatuchak,
Bangkok 10900, Thailand
Tel. 66-2-942-8616
By e-mail: libibic@ku.ac.th

Buffalo Bulletin (June 2006) Vol.25 No.2
CONTENTS
Ruminal pH as regulator of rumen metabolism in buffaloes
M.A. Akbar and Ramesh Kumari................................................................................. 25
A significant observation on the status of cortisol in neonatal buffalo calves and cow
calves VIS-À -VIS their dams
I.J.Sharma, A.K. Jain,and R.K. Tripathi...................................................................... 30
Profile of serum immunoglobulins, glucose-6-phosphate dehydrogenase and L: N
ratio as a measure of internal defense of neonatal bovine calves vs their dam: effect
of neem oil as immunodrlator
A.K. Jain, I. J. Sharma, M.A. Quadri, R.K. Tripathi and R.G.Agrawal............................ 34
Haemato-biochemical profiles of buffalo in anthelmintics treatment against Fasciolo
gigantica infection
H.S. Raghuvanshi, S.Nema, S.K Jain and B.C Sarkhe................................................ 40
RESEARCH ABSTRACT........................................................................................... 47
BUFFALO BULLETIN
IBIC, KASETSART UNIVERSITY, P.O. BOX 1084
BANGKOK 10903, THAILAND
URL : http://ibic.lib.ku.ac.th
E-mail : libibic@ku.ac.th
Tel : 66-2-5795017
Fax : 66-2-5611369
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