Induction of parturition in water buffaloes (Bubalus bubalis )

Parturition was induced in ten buffaloes by combining treatments of dexamethasone and vetoestrol, so that they calved about one month before the expected term. Either dexamethasone alone (Group B) or dexamethasone and vetoestrol (Group C) was used. Another five animals served as controls (Group A). Calving was induced in four animals in group B and three animals in group C after two injections of the compounds four days apart. The average time from first injection to calving for these animals was 117.22 hr and 117.66 hr for groups B and C respectively. Induced calves weighed less at birth (P < 0.05) averaging 24.4 and 26.2 kg for groups B and C respectively, than controls (Group A; 30.20 kg). The body weight gain/week among calves was not significantly different (P > 0.05). The service period, number of services/conception and the milk yield of the buffaloes induced to calve was not significantly different (P > 0.05) from their previous records.     

Serum prolactin levels of non-cycling murrah buffaloes (Bubalus bubalis )

Prolactin levels were quantified by double antibody radio-immunoassay in the blood sampled daily for 24 days, from 9 non-cyclic Murrah buffaloes during hot months (ambient temp.41 to 43 degrees C). All buffaloes were kept in open loose housing. Three buffaloes were sprinkled with water for half an hour twice daily between 1200 and 1430 h to overcome partly the effect of heat during the period of investigation. Mean prolactin levels of 6 unsprinkled buffaloes ranged on different days from 249 to 739 ng/ml serum. The range of averages in case of 3 sprinkled animals was from 152 to 342 ng/ml serum. Buffaloes subjected to sprinkling had significantly (P / 0.01) lower prolactin levels than control buffalo.     

Physico-chemical characterization of growth hormone from water buffaloes (Bubalus bubalis)

A purified preparation of growth hormone from pituitaries of water buffaloes (Bubalus bubalis) has been extensively characterized with regard to physico-chemical properties. The molecular size of buffalo GH (buGH) by electrospray ionization mass spectroscopy (ES-MS) was found to be 21394.00+/-8.44Da and its stokes radius was determined as 2.3 nm. Size heterogeneity in buffalo GH was checked both by electrophoresis and molecular sieve chromatography using 125I-labelled buffalo GH. Similar size heterogeneity was found in standard preparations of ovine and bovine growth hormones. Isoelectric focussing and chromatofocussing indicated charge heterogeneity in buffalo GH preparation. Major charge isoforms having pI of 7.2, 7.7 and minor forms in the pI range of 5.7 to 7.0 were found. Lectin chromatography on Concanavalin A matrix showed that less than 1% of buffalo GH was glycosylated. Heterogeneity in NH2-terminal sequence was also observed, with alanine, phenylalanine and methionine as the NH2-terminal residues as checked by dansyl and DABITC methods. Estimation of tryptophan residue indicated that a single tryptophan residue was present. Ellman's method showed presence of two disulfide bridges per mole of buffalo GH. Intrinsic fluorescence spectrum of buffalo GH exhibited lambda emission maximum at 337 nm. UV-CD spectrum showed that almost 48% of the secondary structure of buGH was constituted by alpha-helicity. The T(M) of buGH as determined by differential scanning calorimetric (DSC) studies was found to be 63 degrees C.     

Physico-chemical and immunological characteristics of pituitary prolactin from water buffaloes (Bubalus bubalis)

Prolactin (PRL) was purified from freshly frozen pituitary glands of water buffaloes (Bubalus bubalis) by a combination of existing procedures of Ellis and Jiang and Wilhelmi involving serial extraction of different pituitary proteins. The partially purified preparation was further fractionated on DEAF-Sephadex followed by Sephadex G-100 chromatography. This was finally purified on HPLC. This preparation was found to be homogeneous by SDS-PAGE and HPLC and had a single N-terminus amino acid (Threonine). The molecular size was estimated to be 24K ± 0.5 by SDS-PAGE and 25K by GPC-HPLC. The buffalo PRL gave a dose dependent inhibition curve in a rat liver based radio receptor assay with a potency of 30–35 L U./mg and also in a partial homologous RIA using ¹²⁵I-buffalo PRL and rabbit anti-oPRL serum giving a potency of 30I.U./mg. Metabolic labelling studies using ³⁵SO₄ ^2– with buffalo pituitary minces showed the incorporation of radioactive sulfate into immunoprecipitable PRL-like material. Physico-chemical characterization of the site of the linkage between sulfate and PRL revealed the presence of Tyr-O-SO₄ in bu-PRL. A high affinity monoclonal antibody (MAB) with Ka of 10¹⁰ L/M, belonging to IgG₁ isotype, and capable of cross reacting with ovine and bovine PRL was generated. This MAB was conformation specific as reduced and carboxymethylated PRL did not react with it. A homologous RIA system using this MAB has been standardised.     

Bio-histopathological studies of endometrium in repeat breeding buffaloes (Bubalus bubalis)

A total of 24 uterine biopsies were successfully obtained from 40 repeat-breeding buffaloes. The endometrium of 15 buffaloes showed marked neutrophilic infiltration in the epithelium, stroma and uterine glands. In some cases, lymphocyte and plasma cell infiltration, cystic dilatation of the glands and periglandular fibrosis indicated chronicity of the endometritis. Most of the animals (85.0%) had uterine cultures positive for Streptococci, Staphylococci, Gram positive variable cocci, Gram negative bacilli, Anthracoids, and Gram positive bacilli with a predominance of Corynebacteria. With the treatment based on in vitro drug sensitivity tests, 57.1% repeat-breeding buffaloes conceived.     

Luteinizing hormone in blood plasma of post-partum buffaloes (Bubalus bubalis)

The changes in luteinizing hormone concentration were measured by heterologous double-antibody radioimmunoassay in blood plasma of 28 Murrah buffaloes. The LH concentration fluctuated between 0.22 to 0.78 ng/ml during first 21 days post-partum. The level increased significantly (P < 0.001) at estrus. The basal LH concentration of second and third week post-partum was inversely related to the first post-partum ovulation interval.     

Sarcocystis dubeyi (Huong and Uggla, 1999) infection in water buffaloes (Bubalus bubalis) from Egypt

Water buffaloes (Bubalus bubalis) are intermediate hosts for 4 species of Sarcocystis , i.e., Sarcocystis fusiformis and Sarcocystis buffalonis with cats as definitive hosts; Sarcocystis levinei with dogs as definitive hosts; and Sarcocystis dubeyi with an unknown definitive host but thought to be zoonotic. Currently, the latter species has been identified with certainty only from Vietnam. In the present study, sarcocysts of S. dubeyi are reported in 11 (30%) of 35 Egyptian water buffaloes from which the esophageal muscles were examined histologically. Sarcocysts were microscopic, measuring 180-250 × 70-110 μm in size. Ultrastructurally, the sarcocyst wall was 3.5-6.5 μm thick and had palisade-like villar protrusions which give it a striated appearance. The villar protrusions contained microtubules that were distributed along the whole villus. This is the first report of S. dubeyi from water buffaloes in Egypt.     

Ultrasonographic fetometry and determination of fetal sex in buffaloes (Bubalus bubalis)

The aim of the current study was to establish ultrasonic biometric threshold of different fetal parts in buffaloes and to evaluate the feasibility and accuracy of ultrasonic fetal sex determination. Serial ultrasonographic examinations were carried out on twelve pregnant buffalo-cows, during which fetal parts were measured, and fetal sex and presentation were determined. The obtained results revealed that embryo and amniotic vesicle (AV) were detected by the forth and fifth week of pregnancy, respectively. Organization was observed by the seventh week, while ossification was indicated between the eighth and 10th week. High correlations were found between different studied parameters and gestational age, where the highest correlation was found with the crown-rump length (CRL) and amniotic vesicle diameter (AVD) at the early-gestation; the biparietal diameter (BPD) at the mid-gestation; and the eyeball diameter (EBD) at the mid- and late-gestation. The results also revealed that the best window for fetal sexing was found between the 10th and 18th week of gestations, with an overall accuracy of 97.1%. The final polarity with all fetuses in anterior presentation was adopted by the 30th week. In conclusion, the overall data indicated the feasibility and value of ultrasonographic fetometry in buffaloes for evaluation of fetal development, estimation of gestational age and determination of fetal sex.     

Embryo transfer in water buffalo (Bubalus bubalis)

A normal, live 35-kg water buffalo bull calf was born 300 days after it was nonsurgically collected as a 7-day blastocyst from a water buffalo donor and transferred nonsurgically to an unrelated water buffalo recipient. The development of estrus synchronization, superovulation and estrus detection methods in water buffalo are described.     

Genetic admixture and population structure analysis of Indian water buffaloes (Bubalus bubalis) using STR markers

Molecular Biology Reports - India has a vast riverine and swamp buffalo diversity adapted to various agro-ecological conditions. In the present study, genetic diversity data for 10 different...     

Serum FSH levels during estrus and a 4-week period following mating in Murrah buffaloes (Bubalus bubalis )

Circulating FSH levels of 13 Murrah buffaloes were investigated by a double-antibody radioimmunoassay using a homologous bovine system. The mean (+/- s.e.m.) peak FSH level on the day of estrus was if57.94 +/- 12.84 ng/ml of blood serum during hotter months (June-August) and 65.23+/-10.54 ng/ml during cooler months (October-December). These were much higher than the FSH concentrations recorded during the rest of the cycle post-mating. There were sizable fluctuations in FSH levels on different days of the estrous cycle within animals. However, the mean values did not exhibit much variation. The occurrence of the FSH peak and the LH peak coincided during the cycle.     

The behaviour and welfare of buffaloes (Bubalus bubalis) in modern dairy enterprises

This review deals with the behaviour of river buffaloes (Bubalus bubalis), in confinement and in extensive conditions, also focusing on the effects of different housing and rearing conditions on their welfare. The behavioural repertoire expressed by buffaloes in extensive and intensive conditions is similar to those displayed by other domestic ruminants. However, through natural selection, buffaloes have also acquired several morphological, physiological and behavioural (i.e. wallowing) adaptations to hot climatic conditions. Buffaloes kept in intensive conditions and having no access to pasture and water for wallowing extend their periods of idling and are less often involved in investigative activities. Confinement is also associated with a reduction of space; however, no specific studies have been carried out to determine the specific requirements of this species. Space restriction can adversely affect various aspects of buffalo welfare, such as health (increased levels of lesions and injuries), social behaviour (increased number of agonistic interactions) and heat dissipation. The buffaloes, originating from tropical areas, are well adapted to large variations in food availability and quality, and to dietetic unbalances. As to human animal relationship, it has been observed that the incidence of stepping and kicking behaviour of buffaloes in the milking parlour is positively correlated with the frequency of oxytocin injections, whereas the frequency of positive stockperson interactions with the animals such as talking quietly, petting and gentle touching are negatively correlated with the number of kicks during milking. Data from farms where both dairy cattle and buffaloes are present show that avoidance distance measured in the pen is lower in buffaloes than in cattle. This may be attributed to the fact that buffaloes are generally recognised to be curious animals. Finally, the effects of different farming practices on animal-related indicators are described. However, these measures should be integrated into a monitoring protocol, such as the Welfare Quality® scheme, to reliably assess buffalo welfare in the current intensive farming conditions.     

Hormonal dynamics and follicular turnover in prepuberal Mediterranean Italian buffaloes (Bubalus bubalis)

The aim of this study was the investigation of hormonal and ovarian follicular dynamics in prepuberal buffaloes (Bubalus bubalis) bred in Italy. Eleven 5-9-month old buffalo calves ranging in weight from 122 to 270kg, maintained under controlled nutritional and environmental conditions, underwent 50 days of ultrasonographic ovarian follicular monitoring in the months of October-December. Blood sampling for E(2) and FSH determination and ultrasonographic monitoring using a 7.5MHz linear probe and an ALOKA SSD-500 monitor were performed daily. No differences in any of the parameters under study were highlighted when calves were divided into two weight categories (<200 and >200kg) and thus data were pooled. In this study, values are reported as mean+/-S.D. A range of two-six regular follicular waves was reported among calves with an average of 4+/-1.1. Overall interval (days) between wave emergence was 9.9+/-2.8 and largest diameters (mm) of dominant and first subordinate follicles were 8.4+/-1.2 and 4.8+/-0.6, respectively (P<0.05). With the exception of one calf, some minor follicular waves (short waves or SWs; 1.6+/-1), lasting <10 days (6.1+/-1.2) were reported. They were monitored contemporaneously on the ovary contralateral (n=7) or ipsilateral (n=3) to the main follicular wave. Growth rate (mm per day) of dominant follicles (DF) was significantly faster than for corresponding subordinate follicles (SF) and follicles of SWs (1.08+/-0.2 versus 0.79+/-0.1 and 0.83+/-0.1, respectively, P<0.05). The static phase (days) lasted longer in DF compared to SF and SW (5.4+/-1.8 versus 2.4+/-1.2 and 2.6+/-1, respectively, P<0.05). The regressing phase (mm per day) was similar among DF, SF and SW (0.86+/-0.2, 0.94+/-0.2 and 0.84+/-0.1, respectively, P=0.09). Episodic spikes of E(2) and FSH were reported, corresponding to wave development throughout the course of investigation. In conclusion, the majority of buffalo calves displayed a typical pattern of regular follicular development in conjunction with a dynamic trend of ovarian and hypophyseal hormones. Some minor follicle turnover was reported with parallel main follicular waves.     

Reticulo-ruminal motility in cattle (Bos indicus) and water buffaloes (Bubalus bubalis) fed a low quality roughage diet

1. The effect of A and B sequence contractions of the reticulo-rumen on passage rate of digesta was compared in buffaloes and cattle fed low quality rhodes grass. 2. Both species ate the same amount per unit body weight but buffaloes spent 53% more time ruminating than cattle. 3. Buffaloes had fewer A and B sequence contractions each day and the rate of these contractions during eating, ruminating and at rest were slower. 4. A larger pool of fine feed particles in the rumen of buffaloes, generated by extra ruminating activity was associated with the 30% shorter mean residence time of particulate matter in the forestomach compared with cattle. 5. It is concluded that the difference in the number and frequency of contractions between the species was insufficient to affect passage rate of digesta from the stomach.     

Dominance of Methanomicrobium phylotype in methanogen population present in Murrah buffaloes (Bubalus bubalis)

Aims:  To study the diversity of rumen methanogens in Murrah buffaloes ( Bubalus bubalis ) from North India by using 16S rRNA gene libraries obtained from the pooled rumen content from four animals and using suitable software analysis.
Methods and Results:  Genomic DNA was isolated and PCR was set up by using specific primers. Amplified product was cloned into a suitable vector and the positive clones were selected on the basis of blue–white screening and sequenced. The resulting nucleotide sequences were arranged in the phylogenetic tree. A total of 108 clones were examined, revealing 17 different 16S rRNA gene sequences or phylotypes. Of the 17 phylotypes, 15 (102 of 108 clones) belonged to the genus Methanomicrobium , indicating that the genus Methanomicrobium is the most dominant component of methanogen populations in Murrah buffaloes ( Bubalus bubalis ) from North India. The largest group of clones (102 clones) was more than 98% similar to Methanomicrobium mobile . BLAST analysis of the rumen contents from individual animals also revealed 17 different phylotypes with a range of 3–10 phylotypes per animal.
Conclusion:  Methanomicrobium phylotype is the most dominant phylotype of methanogens present in Murrah buffaloes ( Bubalus bubalis ).
Significance and Impact of the Study:  Effective strategies can be made to inhibit the growth of Methanomicrobium phylotype to reduce the methane emission from rumen contents and thus help in preventing global warming.     

Biometry of frozen-thawed sperm from eight breeds of Indian buffaloes (Bubalus bubalis)

Sperm morphometry, in combination with other objective traits, can be useful for developing a fertility index. The objective of the present study was to measure various biometric end points of frozen-thawed sperm from eight breeds of Indian buffaloes (Murrah, Surti, Tarai, Mehsana, Jaffrabadi, Bhadawari, Pandharpuri and Nili-Ravi). The sperm head of Pandharpuri buffaloes had the greatest length (10.21 microm), width (6.05 microm), area (52.31 microm(2)) and perimeter (31.86 microm). The ratio of sperm width to length was also greatest (0.61) in Pandharpuri as well as in two other breeds, viz. Nili-Ravi and Jaffrabadi. Murrah had the smallest sperm head width (4.75 microm), area (41.65 microm(2)) and perimeter (29.17 microm), but its sperm tail was longest (57.02 microm), along with that of Jaffrabadi buffaloes (56.96 microm). Based on mean values of sperm tail length, mid piece length and its width the eight buffalo breeds were categorized into three, four and five groups, respectively. Multivariate analysis and clustering put six breeds (Surti, Tarai, Mehsana, Jaffrabadi, Bhadawari and Nili-Ravi) in one cluster, whereas Murrah and Pandharpuri appeared as separate entities.     

Pregnancy in suboestrus buffaloes (Bubalus bubalis) after treatment with Prostaglandin F(2) alpha

Seventy six apparently anoestrous buffaloes, with a palpable corpus luteum at day 43 to 547 post partum, were treated intramuscularly with Prostaglandin, using either 25 mg single dose (Group I), 25 mg double dose 11 days apart (Group II), or 50 mg single dose (Group III). Animals exhibiting oestrus, without any treatment, during the period of experimentation served as control (Group IV). The overall conception rate for the treated animals was 28.8, 51,5, and 69.7 percents after one, two, and three inseminations, respectively. The conception rate, at induced as well as subsequent oestrus, was comparable to control animals. The intergroup differences among treated animals were not significant. The conception rate was greater when the induced heat was more intensely expressed. The subsequent mean oestrus cycle length was similar to controls when all of the treated animals were considered together.     

Attenuation of luteolytic response following fish meal supplementation in dairy buffaloes (Bubalus bubalis)

Luteolysis of corpus luteum, due to un-inhibited PGF(2α) secretion, has been reported to be a cause of early embryonic mortality in dairy animals. The objective of this study was to determine the effects of fish meal (FM) supplementation on the uterine secretion of PGF(2α) and hence establish its supplementation as an antiluteolytic strategy in dairy buffaloes. Five cycling Murrah buffaloes were supplemented with 250g FM daily for 55 days in addition to their routine feed and seven buffaloes were kept as non-supplemented control. After 30 days of FM supplementation, the oestrus was synchronized in all the buffaloes using Ovsynch protocol. On day 15 of synchronized cycle, animals were challenged with oxytocin (OT; 100IU) intravenously and blood samples were collected at 15min interval, 1h before to 4h after OT challenge. The PGF(2α) response was measured as the venous concentration of 13,14-dihydro-15-keto PGF(2α) (PGFM). The mean hourly concentration of PGFM in FM supplemented buffaloes was lower than in the control buffaloes at all the occasions. During peak response (1h post-OT challenge), PGFM concentration was significantly lower (P<0.05) in FM supplemented buffaloes than in the control (197.4±41.7pg/ml versus 326.3±33.5pg/ml, respectively). Also the percent rise in PGFM after OT-challenge in FM supplemented buffaloes was less than the control (11.73% versus 22.47%). The dietary supplementation did not affect the size of corpus luteum (CL) and plasma progesterone concentration. Plasma glucose and total protein concentrations remained within the normal physiological limits during FM supplementation. The present study indicated that supplementing FM decreased the concentrations of PGF(2α) without alterations in the size of CL and plasma progesterone concentrations in dairy buffaloes.