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PubMed2005

Ovum pick-up in cycling and lactating postpartum swamp buffaloes (Bubalis bubalis).

The objective of this study was to evaluate the efficiency of Ovum Pick Up (OPU) in cycling (n = 5) and lactating, postpartum, swamp buffaloes (n = 6) with and without gonadotropin stimulation. The OPU was performed every two weeks in all groups of animals, for a total of six sessions. Thirty collections were performed in five cycling buffaloes and 36 collections in six lactating postpartum buffaloes. Buffaloes that received hormonal stimulation were given a total of 400 mg, follicle stimulating hormone (FSH), administered twice daily over 3 days in decreasing doses, together with 100 microg of GnRH, 24 h after the last FSH injection. Following a resting period of 1 month, the two groups of buffaloes, were subjected to the same OPU regimen, but without any hormonal treatment for an additional six OPU sessions. The number of aspirated follicles recorded from the hormonal stimulated, cycling animals and lactating, postpartum buffaloes was not significantly different, 7.2 +/- 3.7 and 9.0 +/- 3.2, respectively (p > 0.05). Recovered oocytes collected from the two groups of hormonally stimulated animals were also not statistically different: 3.7 +/- 2.7 in the cycling and 5.9 +/- 3.5 in the lactating postpartum group (p > 0.05). In the two groups of buffaloes not receiving hormonal stimulation, the number of aspirated follicles was not significantly different: 2.1 +/- 1.4 and 1.4 +/- 0.7 in cycling and lactating postpartum buffaloes respectively (p > 0.05). Recovered oocytes in the non-treated groups were also similar: 1.4 +/- 1.3 vs 0.7 +/- 0.8 in cycling and lactating buffaloes (p > 0.05). Among stimulated buffaloes, most aspirated follicles were small in size (< or =5 mm), whereas they were mostly medium and large sizes in the non-treated buffaloes. The oocyte recovery rate in both the groups, cycling and lactating postpartum, were 51.6% and 69.5% in stimulated groups and 55.0% and 53.1% in non-stimulated groups (p > 0.05). The majority of recovered oocytes were single- and multi-layered, and the number was greater in the cycling than in the lactating, postpartum buffaloes. The number and quality of recovered oocytes was similar in all groups of buffaloes whether they were received or did not receive hormonal stimulation. Moreover no difference was found in multi- and single-layered oocytes between cycling and lactating, postpartum buffaloes. In conclusion, OPU can be performed successfully in swamp buffalo in different reproductive status and FSH administration was shown to increase the number of aspirated oocytes in both cycling and lactating, postpartum buffaloes.

Animals↗
PubMed2001

Seroprevalence of Neospora caninum in female water buffaloes (Bubalus bubalis) from the southeastern region of Brazil.

Antibodies to Neospora caninum were assayed in sera of 222 female water buffaloes from Ribeira Valley of São Paulo State, Brazil, using an indirect fluorescent antibody test (IFAT) and Neospora agglutination test (NAT). IFAT antibodies were found in 64% of buffaloes with titers of 1:25 (42 buffaloes), 1:50 (53 buffaloes), 1:100 (31 buffaloes), 1:200 (10 buffaloes), 1:400 (3 buffaloes), or > or =1:800 (3 buffaloes). NAT antibodies were found in 53% of buffaloes; in titers of 1:40 in 52 buffaloes, 1:80 in 27 buffaloes, 1:160 in 21 buffaloes, and > or =1:320 in 17 buffaloes. Results indicate a high prevalence of N. caninum exposure in water buffaloes in Brazil and warrant an investigation of the role of N. caninum as an abortifacient in water buffaloes.

Animals↗
PubMed2005

Follicle turnover and pregnancy rates following oestrus synchronization protocols in Mediterranean Italian buffaloes (Bubalus bubalis).

An ultrasound assessment of follicle turnover following two different protocols for synchronization of oestrus and ovulation, as well as an assessment of achieved synchronization between ovulation and AI and conception rates in nulliparous and pluriparous buffaloes were carried out during months of increasing day length. Nulliparous buffaloes (n = 30) were subjected only to Ovsynch protocol whereas pluriparous buffaloes (n = 31) were assigned to Ovsynch (n = 14) or to PRID-pregnant mare serum gonadotrophin (PMSG) (n = 17) protocol according to the presence of functional CL confirming cyclic and acyclic conditions. Ultrasound examination of ovarian follicular dynamics at critical days in the course of synchronization treatments was employed to monitor the fate of the largest available follicles at the beginning of treatments. Such available dominant follicle would persist throughout the protocol as ovulating follicle (no-follicle shift) or would regress giving way to a new follicle to become dominant and ovulate (follicle shift). Furthermore, ultrasound monitoring would determine the degree of synchronization of ovulation and final outcome represented by pregnancy rates. Pregnancy rate following Ovsynch protocol was 40% (12/30) and 42.8% (6/14) in nulliparous and pluriparous buffaloes respectively (p = 0.8575). Most ovulations were synchronized and recorded at AI and the following day in nulliparous (24/30; 80%) and pluriparous (12/14; 85.7%) buffaloes respectively (p = 1.000). A follicle shift was recorded in 14 of 30 (46.6%) and 11 of 14 (78.5%) in nulliparous and pluriparous buffaloes respectively (p = 0.0466). Among established pregnancies: eight derived from follicle shift (66.6%) and four from no-follicle shift (33.3%) in nulliparous buffaloes, p = 0.0729 whereas in pluriparous buffaloes five (83.3%) derived from follicle shift and one from no-follicle shift (16.6%), p = 0.6154. Collectively, from 18 pregnancies in nulliparous and pluriparous buffaloes following Ovsynch protocol, 13 derived from follicle shift (72.2%) and five from no-follicle shift (27.7%), p = 0.0860. Pregnancy rate in pluriparous buffaloes following PRID-PMSG protocol was 70.5% (12/17). The majority of ovulations were synchronized and recorded at first, second AI and following day (13/17; 76.4%). A follicle shift occurred in 15/17 buffaloes (88.2%) and among the 12 recorded pregnancies, 11 derived from follicle shift (91.6%), p = 0.5147. In conclusion, pregnancy rates following Ovsynch protocol were similar in nulliparous and pluriparous cyclic buffaloes. A progestagen treatment on acyclic buffaloes but still displaying some ovarian follicular dynamics, resulted in significantly higher pregnancy rate compared with Ovsynch (p = 0.0376). According to the time of scheduled AI, a high degree of synchronized ovulations were recorded following the implementation of both protocols.

Animals↗
PubMed2004

Ovarian follicular dynamics and hormonal profiles in heifer and mixed-parity Mediterranean Italian buffaloes (Bubalus bubalis) following an estrus synchronization protocol.

The primary objective was to elucidate ovarian follicular dynamics and hormonal profiles in nulliparous heifer (HE; n = 11 ) and mixed-parity (MP; n=10 ) Mediterranean Italian water buffaloes (Bubalus bubalis) following an estrus synchronization protocol. Both groups received a progesterone releasing intravaginal device (PRID) implant for 10 days; a luteolytic dose of synthetic prostaglandin was given 7 days after PRID insertion. Daily ultrasound monitoring and collection of blood to determine plasma concentrations estradiol and progesterone started 1 day after PRID removal and lasted for 55 and 65 days in HE and MP buffaloes, respectively. Data analysis was restricted to the first 5 days after PRID removal and to one estrus cycle following induced ovulation. The HE buffaloes were not inseminated and only one ovulated within 5 days after PRID removal; the remainder ovulated between 8 and 48 days after PRID removal (except one in which ovulation was never detected). All HP buffaloes were inseminated 72, 96 and 120 h after PRID removal; seven buffaloes ovulated within 5 days after PRID removal and two were pregnant. Mean diameter of the largest follicle was significantly smaller in HE than MP buffaloes the first 4 days after PRID removal. There was a parity by time interaction ( P=0.0047 ) for plasma progesterone concentrations; progesterone was higher in HE than MP buffaloes 1 day after PRID removal, but the converse was true 2 days after PRID removal. After induced ovulation, HE buffaloes exhibited a one-wave ( n=5; length of cycle, 8-12 days), two-wave ( n=4; range: 20-26 days) or three-wave cycle ( n=1; 25 days). In contrast, all non-pregnant MP buffaloes ( n=8 ) had a two-wave cycle (range: 19-25 days). For buffaloes with two-wave cycles, the growth rate and diameter of the largest follicle was significantly smaller in HE than MP buffaloes for both the first follicular wave (1.3mm versus 1.7 mm per day and 10.5 mm versus 13.3 mm, respectively) and the second follicular wave (1.0 mm versus 1.3 mm per day and 11.0 mm versus 13.8 mm). In conclusion, there were many significant morphological and endocrine differences between HE and MP buffaloes.

Administration, Intravaginal↗
PubMed2005

Embryonic mortality in buffaloes synchronized and mated by AI during the seasonal decline in reproductive function.

The aim was to determine the factors that contribute to embryonic mortality in buffaloes mated by AI during a period of increasing day length which corresponds to a natural decline in reproductive activity. Italian Mediterranean buffalo cows (n=243) showing regular estrous cycles were synchronized using the Ovsynch-TAI program and mated by AI at 16 and 40 h after the second injection of GnRH. Blood samples were collected on Days 10 and 20 after the first AI and assayed for progesterone (P4). Pregnancy diagnosis was undertaken on Days 26 and 40 after the first AI using rectal ultrasonography. Buffaloes with a conceptus on Day 26 but not on Day 40 were judged to have undergone embryonic mortality and for these animals uterine fluid was recovered by flushing and analysed for common infectious agents. Estrus synchronization was achieved in 86% of buffaloes and the pregnancy rate on Day 40 was 34%. Embryonic mortality between Days 26 and 40 occurred in 45% of buffaloes and was associated with the presence of significant infectious agents in only 10 buffaloes (8%). Concentrations of P4 on Day 10 after AI were higher (P<0.05) in buffaloes that established a pregnancy than in buffaloes that showed embryonic mortality that was not associated with infectious agents. Similarly, on Day 20 after AI P4 concentrations were higher (P<0.01) in pregnant buffaloes compared with non-pregnant buffaloes and buffaloes that had embryonic mortality. It is concluded that a reduced capacity for P4 secretion can explain around 50% of embryonic mortalities in buffaloes synchronised and mated by AI during a period of low reproductive activity and that other as yet unidentified factors also have a significant effect on embryonic survival.

Animals↗
PubMed1994

Genetic variants of ribosomal DNA and mitochondrial DNA between swamp and river buffaloes.

To clarify the genetic relationship between Swamp and River buffaloes, the restriction fragment length polymorphisms (RFLPs) of nuclear genomic ribosomal DNA (rDNA) and cytoplasmic mitochondrial DNA (mtDNA) were analysed. Blood or liver samples from 73 Swamp and three River buffaloes were collected in East and South-east Asian countries. DNA samples from cattle, goats and sheep were used for comparisons. The analysis of rDNA allowed water buffaloes, cattle, goats and sheep to be characterized by four distinct repeat-types. However, swamp and river buffaloes showed the same repeat-type. Divergence of water buffalo and cattle is considered to have occurred approximately four to six million years ago. The RFLPs for mtDNA divided water buffaloes into three haplotypes, swamp-1, swamp-2 and river types. Swamp-1 accounted for 91% of all swamp buffaloes while swamp-2 was observed only in water buffaloes from Thailand (9%). All river buffaloes were of the same haplotype. No differences were observed between swamp and river buffaloes at the rDNA level. In contrast, a few distinct differences between them were found at the mtDNA level. Therefore, mtDNA polymorphisms provide an adequate means for classifying water buffaloes into either swamp or river buffaloes.

Animals↗
PubMed2001

The epidemiology of tuberculosis in free-ranging African buffalo (Syncerus caffer) in the Kruger National Park, South Africa.

The presence of bovine tuberculosis (Mycobacterium bovis) in the Kruger National Park (KNP) was determined for the first time in 1990. It was diagnosed in an African buffalo (Syncerus caffer) bull, which was found recumbent and in an emaciated and moribund state near the south-western boundary fence. This prompted an investigation into the bovine tuberculosis (BTB) status of the KNP, with emphasis on its epidemiological determinants and risk factors. This report documents the findings of surveys that were conducted from 1990 to 1996. It was found that BTB had entered the KNP ecosystem relatively recently (+/- 1960), and has found favourable circumstances for survival and propagation in a fully susceptible and immunologically naive buffalo population. Indications are that it entered the KNP from across the southern river boundary, where the presence of infected domestic cattle herds had been documented. From there the infection spread through the southern buffalo population and is currently spreading in a northward direction. It was estimated that this northward spread took place at a rate of about 6 km per year; the prospect being that, if this rate of spread is maintained, the entire KNP may be affected in less than 30 years from now. Spillover from buffalo had already occurred in species such as chacma baboon (Papio ursinus), lion (Panthera leo), cheetah (Acinonyx jubatus), kudu (Tragelaphus strepsiceros) and leopard (Panthera pardus). Although there is no indication yet that these species act as maintenance hosts, the possibility is raised that these, or an as yet overlooked species, might assume such a role in future. In the KNP, BTB manifests itself as a chronic and predominantly subclinical disease in buffalo. It may take years for clinical signs to develop, and then only at a terminal stage, when emaciation is a constant feature. It is suspected that the time from infection to death is variable and dependent on the animal's immune response, which can be weakened by such factors as stress, old age or droughts. It was found that, in the interim, buffalo have a normal reproductive life. On necropsy, buffalo show almost exclusively lung and upper respiratory tract involvement, pointing to an aerogenous mode of transmission. Histologically, little sign of encapsulation of lesions was detected, which suggests that they are exceptionally susceptible to BTB and that most lesions are open and infectious and progressive, leading ultimately to death of the individual. Evidence also indicates that BTB is progressive within the herd context (92% being the highest prevalence rate thus far determined in a buffalo herd) as well as progressive within the KNP buffalo population (the implication being that virtually all buffalo herds in the KNP will eventually be infected). Preliminary data suggest a positive correlation between disease prevalence and mortality, with potential mortality reaching up to 10% in buffalo herds having BTB prevalence rates of 50 % and higher. Only the future will tell what the effect of the disease on the population dynamics of buffalo will be.

Animals↗
PubMed2002

Comparison of two Ovsynch protocols (GnRH versus LH) for fixed timed insemination in buffalo (Bubalus bubalis).

We evaluated the efficiency of replacing GnRH with LH in the ovulation synchronization protocol in buffaloes. Buffaloes received GnRH on Day 0, (Buserelin; Conceptal, 20 microg), PGF2alpha (Luprostiol; Prosolvin, 15 mg) on Day 7 and GnRH (Buserelin; Conceptal, 10 microg; Group 1) or porcine LH (LH; Lutropin-V, 12.5 mg; Group 2) on Day 9. In Experiment 1, we studied the follicular dynamics of 30 buffaloes (Group 1, n = 15 and Group 2, n = 15). We performed ultrasonography every 12 h from Days 0 to 2, then on Day 7 and then every 6 h from the time of GnRH or LH treatment (Day 9) until the time of ovulation. All females not ovulating by 48 h after the second GnRH or LH injection were considered as nonresponders. In Experiment 2, we evaluated 305 buffaloes (Group 1, n = 154; Group 2, n = 151), using the same two treatments studied in Experiment 1. We also recorded and evaluated aspects like parity, lactational status, the presence of mucus, and uterine tone at the time of artificial insemination (Al). In Experiment 1, ovulation rate after the first GnRH was 86.6% (26/30). Ovulation rates were 93.3% (14/15; Group 1) after the second dose of GnRH and 93.3% (14/15) after LH (Group 2). Ovulation occurred 36.4+/-10.4 h after the first GnRH. The interval for treatment to ovulation was 26.5+/-9.6 h for buffaloes treated with GnRH (Group 1) and 24.4+/-7.9 h for buffaloes treated with LH (Group 2); the time of ovulation did not differ statistically between the two groups (GnRH versus LH; P > 0.05). In Experiment 2, conception rates of the animals AI in the field were 56.5% (Group 1) and 64.2% (Group 2), respectively (P = 0.08). The response to the treatment with LH was not different to the treatment with GnRH; however, multiparous buffaloes had higher conception rates than the primiparous buffaloes in both groups (P > 0.05). Buffaloes with mucus at the time of AI in Group 2 had higher conception rates than the buffaloes that had mucus in Group 1 (P < 0.05). Uterine tone and lactational status did not influence conception rates (P > 0.05). In summary, the results showed that both treatments resulted in synchronization of ovulation and acceptable conception rates. Therefore, the exogenous injection of LH can substitute the GnRH injections in the Ovsynch program in buffaloes.

Animals↗
PubMed2001

Safety and efficacy of an oil-adjuvant vaccine against haemorrhagic septicaemia in buffalo calves: cross-protection between the serotypes B:2,5 and E:2,5.

The safety, efficacy and duration of immunity of an improved oil-adjuvant vaccine against haemorrhagic septicaemia, containing inactivated cells of Pasteurella multocida serotype B:2,5, were tested in young buffalo calves in Pakistan. For safety testing, five buffalo calves were vaccinated intramuscularly with twice the normal dose, and six weeks later with a normal dose. Except for a transient rise in rectal temperature at six hours after the vaccinations, no systemic reactions were observed. The buffaloes remained in good condition and had a normal appetite. No local reactions were observed at the injection site. For efficacy testing two trials were carried out. In the first, buffalo calves were vaccinated intramuscularly either with two doses two-and-a-half months apart, or with a single dose, or left unvaccinated. They were challenged subcutaneously with virulent P multocida after eight, 13 or 15 months. After challenge at eight months the four buffaloes given two doses and the buffalo given one dose were protected, whereas the control animal developed the typical signs of the disease. After the challenges at 13 and 15 months, the vaccinated animals were still protected whereas the control animals died. In the second trial, buffalo calves were vaccinated intramuscularly either with two doses two months apart, or with a single dose at two months or left unvaccinated. The buffaloes were challenged after eight or 14 months. After challenge at eight months the four control animals died, whereas three of the four buffaloes given a single dose were protected. After challenge at 14 months, the three control animals died, whereas four of the five buffaloes given two doses and both the buffaloes given a single dose were protected. To test for cross-protection against the heterologous serotypes E:2,5 and B:3,4, groups of mice were vaccinated once or left unvaccinated. Four weeks later, the vaccinated and control groups were challenged with a dilution series of the different challenge cultures. The vaccine appeared to induce protection against challenge with different strains of serotypes B:2,5 and E:2,5 but not against strains of serotype B:3,4.

Adjuvants, Immunologic↗
PubMed2005

Effect of suckling on basal and GnRH-induced LH release in post-partum dairy buffaloes.

Suckling, a common practice in smallholder dairy-farming systems in the developing world, delays the onset of post-partum ovarian activity in dairy buffalo. The present study was designed to assess the effect of suckling on pituitary function in lactating buffaloes 25-35 days post-partum. Six suckled and nine non-suckled buffaloes were challenged intravenously with a bolus injection of GnRH (20microg buserelin acetate; Receptal). Heparinized venous blood samples were collected at 15min intervals for 2h before and up to 4h after GnRH for luteinizing hormone (LH) estimation. Pretreatment basal LH concentrations were similar in the suckled (0.6+/-0.2ng/ml) and the non-suckled (0.5+/-0.1ng/ml) buffaloes. All but one suckled buffaloes released a LH surge, starting 15-60min post-GnRH treatment, which lasted for 180-225min. While one suckled buffalo did not respond to GnRH, the LH response in the remaining suckled buffaloes was significantly less than in the non-suckled buffaloes in terms of peak LH concentrations (14.3+/-2.7ng/ml versus 26.2+/-4.3ng/ml) and area under the LH curve (1575.6+/-197.4mm(2) versus 2108.9+/-323.9mm(2)). The LH response was least in suckled buffaloes challenged with GnRH while in the luteal phase of an oestrus cycle and with plasma progesterone concentration >1ng/ml. In conclusion, suckling suppressed pituitary responsiveness to exogenous GnRH challenge in post-partum buffaloes.

Animals↗
PubMed2004

The intravenous glucose tolerance test in water buffalo.

OBJECTIVE: The response to intravenous glucose loading in the buffalo using the intravenous glucose tolerance test (IGTT) was investigated to provide a reference for intravenous glucose injection in buffaloes. METHOD: Twelve healthy, fasted, male swamp buffaloes were divided into three groups. Group I: six buffaloes were given 50% glucose at a dosage of 1 g/kg body weight via the jugular vein. Group II: three buffaloes received normal saline. Group III: three buffaloes were not injected. Blood samples were taken from the opposite vein at 60 and 10 min pre-injection (pre60 and pre10), and at 1, 5, 10, 30, 60, 120, 180, 240, 300, 360 and 420 min post-glucose injection (PGI). Plasma glucose was analyzed by the oxidase method. Insulin and glucagon were soon determined with a human radioimmunoassay kit. The insulin (pmol/l)/glucose (mmol/l) ratios (IGR) were also calculated for each sampling time. RESULTS: Mean plasma glucose, insulin and glucagon concentrations of buffaloes in groups II and III were similar at all the sampling times (p > 0.05) and the curves of the IGR for group II and group III were flat throughout. Group I Buffaloes showed an immediate 20 times increase in the mean plasma glucose concentration PGI, over the pre60 and pre10. The peak plasma insulin concentration occurred at 30 min PGI. The mean plasma glucose and insulin concentrations remained above pre-administration levels until 420 min PGI (p < 0.05). However, the mean plasma glucagon concentrations were different only at 1 and 5 min PGI sampling times. The curve of the IGR for group I showed an initial decrease at 1 min PGI, and fluctuated from 10.18 to 25.55 for the remainder of the sampling period. The correlation analysis showed that the mean plasma glucose concentration was positively correlated with insulin level (r = 0.73, p < 0.005), and significantly negatively correlated with mean plasma glucagon (r = -0.58, p < 0.05). The mean plasma insulin level did not show significant correlation with the glucagon (r = 0.06, p > 0.05). CONCLUSION: The hyperglycemia, high insulin, and protracted glucose and insulin curves, the initial decrease in the insulin/glucose ratio indicates that there was an unexpected glucose tolerance to acute intravenous glucose loading in water buffalo compared with other ruminants. The possibly suggested intravenous glucose load in buffaloes is about 5.09-8.28 mmol/l.

Animals↗
PubMed2005

Postpartum ovarian follicular dynamics in primiparous and pluriparous Mediterranean Italian buffaloes (Bubalus bubalis).

The objective of this study was to monitor ovarian function in postpartum primiparous and pluriparous Mediterranean Italian buffaloes (Bubalus bubalis) during months of increasing daylength. Ovarian ultrasound monitoring was carried out for a total of 60 days from calving in 10 primiparous and 10 pluriparous buffaloes. Progesterone was determined from calving until a week after first postpartum ovulation. The study was undertaken during months of increasing day length. Time required for complete postpartum uterine involution was 31 +/- 1.0 and 33 +/- 1.3 days in primiparous and pluriparous buffaloes respectively (P = 0.1). The first postpartum ovulation was recorded on 4 primiparous and 8 pluriparous buffaloes (P = 0.16). Time for first postpartum ovulation to occur was 25.5 +/- 6.9 and 15.5 +/- 1.3 days in primiparous and pluriparous buffaloes, respectively (P = 0.07). Overall, 8 of the 12 first postpartum ovulations (66.6%) occurred in the ovary contra-lateral to the one bearing the gravidic CL, one out of 4 in primiparous and 3 out of 8 in pluriparous buffaloes (P = 1.0). Following a first postpartum ovulation, 3 primiparous and 4 pluriparous buffaloes displayed a complete wave of follicular development leading to a new ovulation. Ovulation following parturition was not recorded in 6 primiparous and two pluriparous buffaloes for the 60 days of ultrasound monitoring. Growth rate (mm/d) and largest size (mm) of first postpartum ovulating follicle was 0.95 +/- 0.18 and 1.07 +/- 0.07 (P = 0.4), and 13.5 +/- 0.8 and 14.1 +/- 0.4 (P = 0.4) in primiparous and pluriparous buffaloes, respectively. Following calving, the total number of available antral follicles (> or =2 mm) declined gradually towards the end of the study period. Follicles greater or equal to 3 mm in diameter on the contrary showed a prominent increase in the first 2 weeks from calving. The number of follicles greater or equal to 3 mm in diameter was significantly higher in the ovary contra-lateral to the one bearing the gravidic CL. A balance in the number of such follicles was reached toward the end of the first month. In conclusion, although some follicular activity was recorded in the ovaries of all buffaloes, true postpartum resumption of cyclicity in the months of increasing daylight hours was delayed in the majority of animals.

Animals↗