Clinical, hematological, and biochemical findings of uterine torsion in buffaloes (Bubalus bubalis)

The aim of this study was to investigate uterine torsion in buffaloes, examine factors influencing the outcome of the disease, and to characterize the related alterations in blood constituents. A total of 126 buffaloes with uterine torsion were examined for stage of gestation, duration, degree, site and direction of torsion, as well as the location of the pregnant horn. Methods of correction were documented along with dam and calf survival. Blood samples were obtained from 20 buffaloes with uterine torsion and 10 healthy buffaloes for hematological and biochemical comparisons. Results showed that uterine torsion in buffaloes occurred in multi- (81.7%) and primiparous (18.3%), during late pregnancy (58.4%) and at full term (41.6%), clockwise (96%) and counter- clockwise (4%), at post- (98.4%) and precervical (1.6%), and was of high (52.3%), moderate (31%) and mild (16.7%) degrees. Torsion was predominantly (P=0.01) on same direction of the pregnant horn. Fetal and maternal mortalities occurred in 78.6% and 23.8% of the cases, respectively. The stage of pregnancy, and degree and duration of uterine torsion were major risk factors for fetal mortality (P=0.0001), while the stage of pregnancy and fetal viability were important risk factors for maternal mortality (P<0.05). There were significant (P<0.05) increases in monocytes, albumin, aspartate aminotransferase, creatine phosphokinase, blood urea nitrogen, and phosphorus and decreases in mean corpuscular hemoglobin concentration and globulin in the affected buffaloes. Time of occurrence and duration of torsion affected some of these parameters. Uterine torsion appears to be a serious problem in buffaloes that has certain peculiarities including time of occurrence, site and direction of torsion, and the high mortality rates. Uterine torsion adversely affects liver and kidney functions. Multiparous might be at greater risk of uterine torsion. The stage of pregnancy, as well as degree and duration of uterine torsion are risk factors for fetal and maternal mortalities.     

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.     

Effect of PGF-2 alpha on serum progesterone levels in the swamp buffalo (Bubalus bubalis)

Induction of some oestrous phenomena was achieved. Treatment with 25 mg PGF-2 alpha cuased mucous discharge, within 48-72 h after injection, which lasted for 4-5 days. Rectal palpation indicated rapid regression of the CL, and in 9 treatments of 6 buffaloes serum progesterone levels declined from 1.76 +/- 0.01 (s.d.) ng/ml before treatment to less than 0.25 ng/ml within 24 h after injection. Concentrations increased at about Day 11 and reached a peak of 1.78 +/- 0.62 ng/ml on Day 18.50 +/- 2.45.     

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.     

Carbonic anhydrase and acid base balance in relation to thermal stress in buffaloes (Bubalus bubalis)

The blood samples from fifteen normal lactating buffaloes were taken from December 15th 1978 to 31st August, 1979. Depending upon the climatic conditions, the whole period of study was divided into four seasons. The mean values of carbonic anhydrase (moles CO₂/l/sec×10^?5) were 3.08±0.26, 4.94±0.44, 5.23±0.35, 6.44±0.32 in pregnant and 4.87±0.27, 4.53±0.41, 4.74±0.45, 6.36±0.40 in non-pregnant animals during winter, spring, hot and dry and hot and humid seasons. Mean values of pO₂ (mm Hg) were 31.26±1.41, 31.92±0.61, 35.90±0.59, 33.80 ±0.67 in pregnant and 31.89±0.44, 31.53±0.54, 35.52±0.69, 31.65±0.95 in non-pregnant buffaloes during winter, spring, hot and dry and hot and humid periods, respectively. There were highly significant (P< 0.01) differences between seasons with respect to pO₂, pCO₂, actual HCO₃ and heamoglobin. However, PCV changed significantly (P<0.01) with the physiological status of the animal. Different correlation of biochemical parameters with climatic elements were discussed. Thus, the shifts in the levels of carbonic anhydrase, HCO₃ and heamoglobin may prove to be a better tool/index for thermal stress in buffaloes.     

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.     

Effect of prostaglandin F(2alpha)- thamsalt and Estrumate (ICI 80996) on plasma progesterone levels in Indian water buffaloes (Bubalus bubalis )

Thirty six buffalo showing normal reproductive cyclas were given a single intramuscular injection of prostaglandin F(2alpha)- thamsalt (PGF(2alpha)) and Estrumate on day 9 or 15 of the estrous cycle in order to induce estrus. In PGF(2alpha) treated animals, plasma progesterone dropped from 2.99+/-0.29 to 0.10 +/- 0.02 ng/ml and from 5.53 +/- 0.53 to 0.09 +/- 0.03 ng/ml (mean +/- SEM) within 72 hours of treatment, on day 9 and 15, respectively. Similarly, the treatment with Estrumate on day 9 or 15 of the cycle reduced the level of proges--terone from 3.02 +/- 0.03 to 0.14 +/- 0.03 ng/ml and from 4.26 +/- 0.47 to 0.21 +/- 0.03 ng/ml, respectively, 72 hours aPter each treatment. It was inferred that both drugs induce estrus in buffalo without any observed residual effect.     

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.     

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.     

Serum testosterone concentrations in advanced pregnancy in water buffaloes (Bubalus bubalis)

Peripheral testosterone levels were measured by radioimmunoassay of 65 serum samples taken at various intervals from 11 buffaloes in advanced pregnancy. Average testosterone levels measured at day 210 of gestation (248 ± 39 pg/ml) remained similar up to the last week of gestation. No difference in testosterone levels was recorded between buffaloes carrying male and female fetuses. Pregnant buffaloes had significantly higher testosterone levels than non-pregnant buffaloes.     

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.     

An enzyme-linked immunosorbent assay (ELISA) to measure growth hormone level in serum and milk of buffaloes (Bubalus bubalis)

An indirect Sandwich ELISA to measure growth hormone level in serum and milk of buffaloes was developed. The assay was based on purified anti rbST IgG raised in rabbits and chicken and rabbit anti chicken IgG horseradish peroxidase. The assay was validated in terms of sensitivity, specificity, precision and recovery. Parallelism was demonstrated between the standard curve and serially diluted serum, milk and pituitary derived growth hormone. Sensitivity of the assay was 0.1 ng/ml. Recovery of exogenous bovine somatotropin from serum and milk ranged from 90 to 102% and 96 to 108% respectively. The intra and inter assay variations to measure growth hormone in serum and milk were 3.36 to 8.81% and 6.01 to 12.31% respectively. Statistical analysis for parallelism and cross-reactivity of rbST with serum of other species confirmed the reproducibility of the assay.     

Occurrence of flehmen in male buffaloes (Bubalus bubalis) with special reference to estrus

Pheromonal cues present in the urine and vaginal mucus during the estrus period elicit courtship behaviour in mammals including the characteristic flehmen behaviour. In the present study, the flehmen behaviour was assessed in male buffaloes by exposing them to heifers for three consecutive estrous cycles. Behavioural observations revealed that the bulls sniffed the female's external genitalia, responded to the chemical signal(s) and exhibited the flehmen behaviour similar to those reported in other ungulates. The flehmen behaviour was recorded daily during 15-min contact with females for three consecutive estrous cycles. Out of 180 observations during five days estrus periods (-3 to +1 days) which included high frequency of flehmen behaviour and out of 637 observations with or without flehmen behaviour during diestrus periods, the bull displayed 365 and 441 flehmen behaviour, respectively. Average numbers of all (2.03 +/- 0.66) and repeated flehmen (1.05 +/- 0.64) behaviour during estrus periods were significantly higher than those of diestrus periods of all (0.69 +/- 0.25) and repeated flehmen (0.11 +/- 0.10). The statistical significance was higher (P < 0.001) in repeated flehmen behaviour of estrus as compared to that in diestrus. In case of all flehmen behaviour, the statistical significance was higher (P < 0.01) in estrus when compared to that in diestrus. The occurrence of flehmen behaviour in male buffaloes clearly indicates that the specific pheromonal compound(s) present in the urine/vaginal mucus during estrus significantly influence the flehmen behaviour.     

Prognosis prediction of uterine torsion mechanical treatment (rolling) after estimation of calcium and creatinine level in the serum of buffaloes (bubalus bubalis)

The present study was carried out to investigate the relationship between creatinine and calcium concentration in buffalo serum in cases of uterine torsion before rolling, and 1 h and 24 h after calving. The degree, duration and site of uterine torsion, as well as fetus viability, time needed for cervical dilation, and the occurrence of uterine rupture were recorded. A total of 150 pregnant buffaloes suffering from colic and anorexia were brought to our clinic and clinically examined. Of these, 35 buffaloes were diagnosed with uterine torsion (different durations, directions and degrees) and the viability of the foeti and any evidence of uterine rupture were determined using ultrasonography and the serum levels of creatinine and calcium were estimated using calorimetric method. The animals were also examined for incidence of uterine rupture after rolling and calving. The calcium level significantly (P < 0.05) decreased with increasing duration and severity of uterine torsion, however, it was higher in cases where a live fetus was delivered compared with a dead one. Conversely, the creatinine level significantly (P < 0.05) increased with increasing duration and severity of uterine torsion but was lower in cases that delivered a live fetus compared with a dead one. The calcium and creatinine levels returned to approximately normal concentration within 24 h after calving. In conclusion, calcium and creatinine serum concentration have a correlation with duration and severity of uterine torsion. Animals with low levels of calcium (below 8.44 mg/dL) and high levels of creatinine (above 2.25 mg/dL) did not usually respond to rolling or suffer from uterine rupture during calving. The calcium and creatinine levels can be used as indicators for the prognosis of mechanical treatment of uterine torsion in buffaloes.     

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.     

Effect of Bromocryptine on hormones and milk secretion in Murrah buffaloes ( Bubalus bubalis)

An experiment was carried out on 10 advance pregnant Murrah buffaloes to determine the role of hormones in milk secretion around parturition. Experimental animals were administered with a single injection of bromocryptine, @ 100 μg/kg BW, for 5 days before expected calving, whereas control group buffaloes were injected with placebo injections. Blood samples collected before parturition (-5,-4,-3,-2,-1 days), on day of parturition (day-0) and on day 1, 2, 3, 4, 5, 10 and 15 post partum were analyzed for growth hormone (GH), insulin like growth factor-I (IGF-I) and prolactin (PRL) by radioimmunassay methods. Milk samples were collected daily for 5 days and on day 10 and 15 after parturition. Milk fat, protein, lactose, citric acid, non-esterified fatty acids (NEFAs) and somatic cell counts (SCCs) were determined in milk samples. Bromocryptine treatment significantly (P < 0.01) decreased pre partum PRL and increased GH levels (P < 0.01) on day of parturition in experimental buffaloes without influencing plasma IGF-I level. Milk yield was significantly lower (P < 0.01) in experimental than in control group. Further, effect of bromocryptine on milk yield was only for a week. Milk yield increased (P < 0.01) gradually and was similar to control group on day 15 post partum. Bromocryptine treatment significantly increased milk SCC (P < 0.01) and protein content (P < 0.01) but there was no effect of treatment on fat, lactose, citric acid, glucose, milk and plasma NEFA concentration. It was concluded that prepartum suppression of PRL by bromocryptine impairs milk secretion temporarily in ensuing lactation. The significant rise in GH level before parturition and on day of parturition suggests a role of it in milk secretion of buffaloes.     

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.     

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.     

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.