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Sample records for buffalo bubalus bubalus

  1. FATAL INTESTINAL COCCIDIOSIS IN A THREE-WEEK OLD BUFFALO CALF (BUBALUS BUBALUS)

    USDA-ARS?s Scientific Manuscript database

    The water buffalo (Bubalus bubalus) is important to the economy of several countries, especially in Asia and Brazil. Little is known regarding the impact of coccidiosis in buffaloes. Cattle and buffaloes are considered to have common species of Eimeria, but critical cross transmissions have not been...

  2. Congenital Malformations in River Buffalo (Bubalus bubalis).

    PubMed

    Albarella, Sara; Ciotola, Francesca; D'Anza, Emanuele; Coletta, Angelo; Zicarelli, Luigi; Peretti, Vincenzo

    2017-02-10

    The world buffalo population is about 168 million, and it is still growing, in India, China, Brazil, and Italy. In these countries, buffalo genetic breeding programs have been performed for many decades. The occurrence of congenital malformations has caused a slowing of the genetic progress and economic loss for the breeders, due to the death of animals, or damage to their reproductive ability or failing of milk production. Moreover, they cause animal welfare reduction because they can imply foetal dystocia and because the affected animals have a reduced fitness with little chances of survival. This review depicts, in the river buffalo (Bubalus bubalis) world population, the present status of the congenital malformations, due to genetic causes, to identify their frequency and distribution in order to develop genetic breeding plans able to improve the productive and reproductive performance, and avoid the spreading of detrimental gene variants. Congenital malformations most frequently reported in literature or signaled by breeders to the Department of Veterinary Medicine and Animal Production of the University Federico II (Naples, Italy) in river buffalo are: musculoskeletal defects (transverse hemimelia, arthrogryposis, umbilical hernia) and disorders of sexual development. In conclusion this review put in evidence that river buffalo have a great variety of malformations due to genetic causes, and TH and omphalocele are the most frequent and that several cases are still not reported, leading to an underestimation of the real weight of genetic diseases in this species.

  3. Folliculogenesis in buffalo (Bubalus bubalis): a review.

    PubMed

    Manik, R S; Palta, P; Singla, S K; Sharma, V

    2002-01-01

    The urgent need for improving the reproductive performance of buffalo necessitates a better understanding of the mechanisms controlling ovarian follicular growth and development. Attention needs to be focused on improving superovulation responses and conception rates, and reducing the variability in ovulation rate and embryo loss. Application of ultrasonic imaging has revealed that follicular turnover during an unstimulated oestrous cycle occurs in waves, with each wave involving synchronous development of a group of follicles, one dominant and several subordinate follicles. There is a predominance of two waves with the first wave beginning around Day 0 (day of ovulation) and the second wave around Day 9 or 10. Primary reasons for a lower superovulation response in buffalo compared with that in cattle is a lower number of primordial and antral follicles, a slower shift from small to large follicles during superovulation, a higher incidence of deep atresia and inability of several large follicles to ovulate, especially when superovulation is induced by equine chorionic gonadotrophin treatment. There is near complete lack of information in Bubalus bubalis on the factors controlling the selection of the dominant follicle, the period of functional dominance and the effects of environmental factors, such as climate and nutrition, on follicular dynamics.

  4. Congenital Malformations in River Buffalo (Bubalus bubalis)

    PubMed Central

    Albarella, Sara; Ciotola, Francesca; D’Anza, Emanuele; Coletta, Angelo; Zicarelli, Luigi; Peretti, Vincenzo

    2017-01-01

    Simple Summary Congenital malformations (due to genetic causes) represent a hidden danger for animal production, above all when genetic selection is undertaken for production improvements. These malformations are responsible for economic losses either because they reduce the productivity of the farm, or because their spread in the population would decrease the total productivity of that species/breed. River buffalo is a species of increasing interest all over the world for its production abilities, as proved by the buffalo genome project and the genetic selection plans that are currently performed in different countries. The aim of this review is to provide a general view of different models of congenital malformations in buffalo and their world distribution. This would be useful either for those who performed buffalo genetic selection or for researchers in genetic diseases, which would be an advantage to their studies with respect to the knowledge of gene mutations and interactions in this species. Abstract The world buffalo population is about 168 million, and it is still growing, in India, China, Brazil, and Italy. In these countries, buffalo genetic breeding programs have been performed for many decades. The occurrence of congenital malformations has caused a slowing of the genetic progress and economic loss for the breeders, due to the death of animals, or damage to their reproductive ability or failing of milk production. Moreover, they cause animal welfare reduction because they can imply foetal dystocia and because the affected animals have a reduced fitness with little chances of survival. This review depicts, in the river buffalo (Bubalus bubalis) world population, the present status of the congenital malformations, due to genetic causes, to identify their frequency and distribution in order to develop genetic breeding plans able to improve the productive and reproductive performance, and avoid the spreading of detrimental gene variants. Congenital

  5. Isolation of Arcobacter species in water buffaloes (Bubalus bubalis).

    PubMed

    Piva, Silvia; Serraino, Andrea; Florio, Daniela; Giacometti, Federica; Pasquali, Frederique; Manfreda, Gerardo; Zanoni, Renato Giulio

    2013-05-01

    This is the first report of Arcobacter spp. in rectal fecal samples from healthy water buffaloes (Bubalus bubalis) reared on a dairy farm. Arcobacter species were isolated after enrichment, and isolates were identified at species level by multiplex-polymerase chain reaction assay. Thirty samples were examined and Arcobacter spp. were isolated from 96.7% of water buffaloes tested: 38 Arcobacter spp. isolates were obtained, with A. cryaerophilus as the dominant species followed by A. butzleri and A. skirrowii. Nine animals (31%) were colonized by more than one Arcobacter species. The present study indicates that water buffaloes can harbor a variety of Arcobacter spp. and that healthy buffaloes may act as hosts. Water buffalo fecal shedding of Arcobacter spp. may be of significance to human health, considering the potential fecal contamination during harvesting of raw milk and slaughtering.

  6. Fatal onion (Allium cepa) toxicosis in water buffalo (Bubalus bubalis).

    PubMed

    Borelli, Vanessa; Lucioli, Joelma; Furlan, Fernando Henrique; Hoepers, Patrícia Giovana; Roveda, Juliano Fleck; Traverso, Sandra Davi; Gava, Aldo

    2009-05-01

    Toxicosis caused by the ingestion of onion (Allium cepa) by 5 water buffalo (Bubalus bubalis) occurred in the district of Caçador, Santa Catarina, Brazil. The water buffalo died after ingestion of a large quantity of onion that had been left in the pasture. Clinical signs started 8 days postingestion and were characterized by pale mucous membranes, lethargy, and dark urine. At necropsy, pieces of onions were found in the rumen of 1 animal. The carcass smelled strongly of onion, and the kidneys and urine were dark brown. Microscopic renal lesions included tubular degeneration and necrosis with deposits of eosinophilic material in the cytoplasm of renal tubular epithelial cells and tubular lumina. These changes were consistent with hemoglobinuric nephrosis. Centrilobular coagulation necrosis was observed in the liver accompanied by hemorrhage and macrophages containing brown cytoplasmic pigment. A diagnosis of hemolytic anemia caused by onion toxicosis was based on the epidemiological data, clinical signs, macroscopic changes, and histological lesions.

  7. A review of Neospora caninum in water buffalo (Bubalus bubalis).

    PubMed

    Reichel, Michael P; McAllister, Milton M; Nasir, Amar; Moore, Dadin P

    2015-09-15

    A number of countries in the world have reported infections with Neospora caninum in water buffalo (Bubalus bubalis), from Africa to Asia, Europe and South America and recently Australia. In general, clinical manifestations (such as abortion) seem rare, which has raised the prospect that buffalo may be inherently resistant to clinical effects of N. caninum infection. Worldwide, the seroprevalence of N. caninum infection (as a measure of exposure determined by the detection of antibody) in buffalo is high, at approximately 48%. This reported seroprevalence is three or four times higher than that reported from the world's cattle populations, which have collective seroprevalence rates of 16.1% for dairy cattle and 11.5% for beef cattle. However, there is a lack of standardisation in seroprevalence studies and some studies may well under-estimate the true level of infection. Epidemiologic evidence supports post-natal transmission, and in utero transmission has also been demonstrated. The causes for water buffalo to have markedly higher seroprevalence but apparently lower neosporosis abortion rates than cattle warrant further investigation. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Heterochromia iridis in water buffaloes (Bubalus bubalis).

    PubMed

    Misk, N.A.; Semieka, M.A.; Fathy, A.

    1998-01-01

    This study included 45 unaffected animals and 593 animals affected with heterochromia irides, and 85 enucleated eyeballs with heterochromia irides. The classification of heterochromia irides, morphology of normal and heterochromic irides, and the histology, ultrastructure, and scanning electron microscopy are presented. The incidence of heterochromia irides in water buffaloes was 7.62% affecting either one or both eyes. Both complete and partial heterochromia irides occurred. Complete heterochromia iridis is more frequent than the partial form in either bilateral or unilateral cases. The pupil has a dumb-bell-shape appearance. Granula iridica occurred at the upper (100%) and lower (30%) pupillary margins and originated from the posterior pigmented epithelium. In heterochromia irides, the melanocytes is absent in the anterior border and stromal layers, and iridal thickness appeared thinner than that of normal eyes.

  9. Production of wild buffalo (Bubalus arnee) embryos by interspecies somatic cell nuclear transfer using domestic buffalo (Bubalus bubalis) oocytes.

    PubMed

    Priya, D; Selokar, N L; Raja, A K; Saini, M; Sahare, A A; Nala, N; Palta, P; Chauhan, M S; Manik, R S; Singla, S K

    2014-04-01

    The objective of this study was to explore the possibility of producing wild buffalo embryos by interspecies somatic cell nuclear transfer (iSCNT) through handmade cloning using wild buffalo somatic cells and domestic buffalo (Bubalus bubalis) oocytes. Somatic cells derived from the ear skin of wild buffalo were found to express vimentin but not keratin and cytokeratin-18, indicating that they were of fibroblast origin. The population doubling time of skin fibroblasts from wild buffalo was significantly (p < 0.05) higher, and the cell proliferation rate was significantly (p < 0.05) lower compared with that of skin fibroblasts from domestic buffalo. Neither the cleavage (92.6 ± 2.0% vs 92.8 ± 2.0%) nor the blastocyst rate (42.4 ± 2.4% vs 38.7 ± 2.8%) was significantly different between the intraspecies cloned embryos produced using skin fibroblasts from domestic buffalo and interspecies cloned embryos produced using skin fibroblasts from wild buffalo. However, the total cell number (TCN) was significantly (p < 0.05) lower (192.0 ± 25.6 vs 345.7 ± 42.2), and the apoptotic index was significantly (p < 0.05) higher (15.1 ± 3.1 vs 8.0 ± 1.4) for interspecies than that for intraspecies cloned embryos. Following vitrification in open-pulled straws (OPS) and warming, although the cryosurvival rate of both types of cloned embryos, as indicated by their re-expansion rate, was not significantly different (34.8 ± 1.5% vs 47.8 ± 7.8), the apoptotic index was significantly (p < 0.05) higher for vitrified-warmed interspecies than that for corresponding intraspecies cloned embryos (48.9 ± 7.2 vs 23.9 ± 2.8). The global level of H3K18ac was significantly (p < 0.05) lower in interspecies cloned embryos than that in intraspecies cloned embryos. The expression level of HDAC1, DNMT3a and CASPASE3 was significantly (p < 0.05) higher, that of P53 was significantly (p < 0.05) lower in interspecies than in intraspecies embryos, whereas that of DNMT1 was similar between the two

  10. Evaluation of fasting metabolism of growing water buffalo (Bubalus, Bubalis).

    PubMed

    Qin, Guangsheng; Zou, Caixia; Pang, Chunying; Yang, Bingzhuan; Liang, Xianwei; Liu, Jianxin; Xia, Zhongsheng; Wen, Qiuyan; Yan, Tianhai

    2011-12-01

    The objectives of the present study were to evaluate fasting metabolism (FM) of water buffalo (Bubalus, Bubalis) at three stages of growth (12, 18 and 24 months) in Guangxi, China. Five female water buffalo were used for each age group and their live weight was on average 254, 326 and 338 kg, respectively. All animals were of average body condition, healthy and de-wormed before start of the study. Prior to a 6-day fasting period, buffalo were offered a mixed diet of forage and concentrates (70% and 30%, dry matter basis) on a restricted nutritional level (419 kJ/kg(0.75) of metabolizable energy, ME) for 15 days. Gas exchanges for each animal were determined for 3 days from day 4 of starvation, using open-circuit respiratory head hoods. Fasting body weight was 0.918 of live weight (P < 0.001, r(2) = 0.99). Both fasting heat production (FHP) and FM (MJ/day) increased significantly with increased age of animals (P < 0.05). Linear regression analysis indicated a positive relationship between fasting body weight (kg(0.75)) and FHP (MJ/day, P < 0.01, r(2) = 0.49) or FM (MJ/day P < 0.01, r(2) = 0.52) when using individual animal data across three groups. However, when expressed as kJ/kg(0.75) of fasting body weight, the differences in FHP or FM between three groups of animals were not significant. The present average FHP and FM (322 and 347 kJ/kg(0.75) of fasting body weight) were compatible to those published in the literature for water buffalo, beef and dairy cattle. The present FM data were also used to estimate net energy (NE(m)) and ME (ME(m)) requirements for maintenance for water buffalo. The results for these two parameters were similar to those for FHP and FM. There was no significant difference between three groups of buffalo in NE(m) or ME(m) when expressed as kJ/kg(0.75) of live weight. The present average NE(m) and ME(m) values (347 and 506 kJ/kg(0.75) of live weight) are close to those proposed by the Agricultural and Food Research Council adopted in UK for

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

    USDA-ARS?s Scientific Manuscript database

    Water buffaloes (Bubalus bubalis) are intermediate hosts for 4 species of Sarcocystis, i.e., S. fusiformis and S. buffalonis with cats as definitive hosts, S. levinei with dogs as definitive hosts, and S. dubeyi with an unknown definitive host, but thought to be zoonotic. Currently, the latter speci...

  12. Effects of co-stocking smallmouth buffalo, Ictiobus bubalus, with channel catfish, Ictalurus punctatus

    USDA-ARS?s Scientific Manuscript database

    Proliferative gill disease (PGD) in catfish is caused by the myxozoan Henneguya ictaluri. The complex life cycle requires Dero digitata as the oligochaete host. Efforts to control PGD by eradicating D. digitata have been unsuccessful. Smallmouth buffalo, Ictiobus bubalus, (SMB) are opportunistic bot...

  13. Redescription of Sarcocystis fusiformis sarcocysts from the water buffalo (Bubalus bubalis)

    USDA-ARS?s Scientific Manuscript database

    Five species of Sarcocystis have been reported from the water buffalo (Bubalus bubalis): Sarcocystis fusiformis and Sarcocystis buffalonis have macrocysts and cats act as definitive hosts; Sarcocystis levinei has microcysts and dogs act as definitive host; Sarcocystis dubeyi and S. sinensis have mic...

  14. Effects of co-stocking smallmouth buffalo, Ictiobus bubalus, with channel catfish, Ictalurus punctatus

    USDA-ARS?s Scientific Manuscript database

    Proliferative gill disease (PGD) in catfish is caused by the myxozoan Henneguya ictaluri. The complex life cycle requires Dero digitata as the oligochaete host. Efforts to control PGD by eradicating D. digitate have been unsuccessful. Smallmouth buffalo, Ictiobus bubalus, (SMB) are opportunistic bot...

  15. Cryopreservation of buffalo (Bubalus bubalis) semen in Bioxcell extender.

    PubMed

    Akhter, S; Ansari, M S; Rakha, B A; Andrabi, S M H; Iqbal, S; Ullah, N

    2010-10-01

    This study was designed to compare commercially available extender Bioxcell with tris-citric egg yolk extender for post thaw quality and in vivo fertility of buffalo semen. For comparison of post thaw semen quality: semen was collected from five adult Nili-Ravi buffalo (Bubalus bubalis) bulls of similar age group with artificial vagina (at 42 degrees C) for three weeks (replicates). Qualifying ejaculates having motility >60% from each buffalo bull were divided in two aliquots and diluted (at 37 degrees C having 50 x 10(6) spermatozoa/ml) in tris-citric egg yolk or Bioxcell extender. Diluted semen was cooled to 4 degrees C in 2 hours, equilibrated for 4 hours and filled in 0.5 ml straws. Semen straws were kept over liquid nitrogen vapors (5 cm) for 10 minutes. Straws were then plunged and stored in liquid nitrogen (-196 degrees C). After 24 hours of storage, semen straws were thawed at 37 degrees C for 30 seconds to assess sperm motility, viability, plasma membrane integrity, normal apical ridge, and abnormalities (head, mid piece, and tail). For comparison of in vivo fertility: semen from two buffalo bulls of known fertility was cryopreserved in tris-citric egg yolk and Bioxcell as described earlier, and used for inseminations under field conditions. Post-thaw percentage of sperm motility (45.3 +/- 1.1, 45.0 +/- 1.4), viability (66.2 +/- 1.1, 64.4 +/- 1.3) plasma membrane integrity (60.4 +/- 1.2, 59.2 +/- 1.4) and normal apical ridge (82.9 +/- 0.5, 80.7 +/- 0.5) did not differ (P > 0.05) in tris-citric egg yolk and Bioxcell extender, respectively. Similarly, sperm abnormalities of head (1.20 +/- 0.1, 1.20 +/- 0.1), mid piece (0.67 +/- 0.1, 0.87 +/- 0.1) and tail (11.7 +/- 0.2, 11.6 +/- 0.3) remained similar (P > 0.05) in tris-citric egg yolk and Bioxcell extender, respectively. In vivo fertility rates of buffalo semen cryopreserved in tris-citric egg yolk and Bioxcell also remained similar (44% vs. 47%). It is concluded that commercially available Bioxcell may be

  16. Factors affecting the quality of cryopreserved buffalo (Bubalus bubalis) bull spermatozoa.

    PubMed

    Andrabi, S M H

    2009-06-01

    Storage of buffalo (Bubalus bubalis) bull semen in the cryopreserved state is discussed in this article. Fertility rate in buffalo following artificial insemination with frozen-thawed semen is reviewed. To better understand the freezability of bubaline spermatozoa, the available data on biochemical components and the activity of specific enzymes of semen/spermatozoa are given. Moreover, the major factors that may influence the post-thaw viability and fertility of buffalo spermatozoa are examined in detail. In addition, suggestions for improvement in cryogenic procedures for buffalo spermatozoa are also given.

  17. Effects of smallmouth buffalo, Ictiobus bubalus biomass on water transparency, nutrients, and productivity in shallow experimental ponds

    USGS Publications Warehouse

    Goetz, Daniel B.; Kroger, Robert; Miranda, Leandro E.

    2014-01-01

    The smallmouth buffalo Ictiobus bubalus is a native benthivore to floodplain lakes in the Yazoo River Basin, USA. Based on evidence from other benthivorous fish studies we hypothesized high biomasses of I. bubalus contribute to poor water quality conditions. We tested this hypothesis in shallow (< 1.5 m) 0.05 ha earthen ponds at three stocking biomasses over a 10-week period during the summer of 2012. The most notable results from the permutational multivariate analysis of variance suggest I. bubalus at high and moderate biomasses significantly (p < 0.05) enhanced turbidity and suspended solid levels while decreasing Secchi depth. Our results suggest that effects of I. bubalus on water clarity may have considerable ecological implications in natural habitats such as shallow floodplain lakes.

  18. Seroprevalence of Toxoplasma gondii infection in water buffaloes (Bubalus bubalis) in Veracruz State, Mexico and its association with climatic factors

    USDA-ARS?s Scientific Manuscript database

    Background: Infection with Toxoplasma gondii in water buffaloes (Bubalus bubalis) is of epidemiological importance because of the risk for transmission to humans. We sought to determine the seroprevalence of T. gondii infection in 339 water buffaloes in Veracruz State, Mexico using the modified aggl...

  19. Mitochondrial DNA Variability of Domestic River Buffalo (Bubalus bubalis) Populations: Genetic Evidence for Domestication of River Buffalo in Indian Subcontinent.

    PubMed

    Nagarajan, Muniyandi; Nimisha, Koodali; Kumar, Satish

    2015-04-20

    River buffalo, Bubalus bubalis is a large bovine species frequently used livestock in southern Asia. It is believed that the river buffalo was domesticated from Bubalus arnee, the wild buffalo of mainland Asia, a few thousand years ago, probably during the period of Indus Valley civilization. However, the domestication history of the river buffalo has been the subject of debate for many decades mainly due to the lack of clear archeological evidence and the divisive conclusions of the genetic studies. Therefore, in order to understand the domestication history and genetic relationship among the various river buffalo populations, we analyzed 492-bp region of mitochondrial DNA control region sequences of 414 river buffalo sampled from India, Pakistan, Egypt, and Iran along with the available 403 swamp buffalo sequences. The phylogenetic analyses of our study along with the archaeological evidence suggest that the river buffalo was domesticated in an atypical manner involving continuous introgression of wild animals to the domestic stocks in Indian subcontinent prior to mature phase of Indus Valley civilization (2600-1900 BC). Specifically, our data exclude Mesopotamian region as the place of domestication of the river buffalo. © The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  20. Congenital ocular dermoid cyst in a river buffalo (Bubalus bubalis) calf.

    PubMed

    Sarrafzadeh-Rezaei, F; Farshid, A A; Saifzadeh, S

    2007-02-01

    Clinical and histopathological findings of a congenital ocular dermoid cyst, located at the lower eyelid of a river female buffalo (Bubalus bubalis) calf were presented. A soft, fluctuant, non-tender, hyperaemic cystic mass was detected overlaying the left eye. Fine needle aspirate revealed filamentous debris with no malignant cells. The cyst was treated surgically by orbital exenteration and subsequently subjected to histopathological examination. The histopathological study disclosed a conjunctival dermoid cyst. This report is novel, in that; such ocular cyst has not previously been described in river buffalo calves.

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

    PubMed

    Napolitano, F; Pacelli, C; Grasso, F; Braghieri, A; De Rosa, G

    2013-10-01

    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

  2. Isolation and characterization of bovine parainfluenza virus type 3 from water buffaloes (Bubalus bubalis) in Argentina

    PubMed Central

    2012-01-01

    Background Parainfluenza virus type 3 (PIV3) was isolated from dairy buffaloes (Bubalus bubalis) naturally affected with respiratory and reproductive clinical conditions. Results Examination of nasal and vaginal swabs collected from 12 diseased buffaloes led to the isolation of three paramyxovirus isolates from two animals. Antigenic, morphological and biological characteristics of these three isolates were essentially similar to those of members of the Paramyxoviridae family. Antigenic analysis by direct immunofluorescence and cross neutralization test placed these isolates together with bovine parainfluenza virus type 3 (BPIV3). Nucleotide and amino acid phylogenetic analysis of partial matrix gene sequences of the buffalo isolates and six field BPIV3 isolates from bovines in Argentina were studied. Buffalo isolates were similar to genotype B (BPIV3b) while the six BPIV3 isolates were similar to genotypes A (BPIV3a) and C (BPIV3c). Conclusions This is the first characterization of BPIV3 in water buffalo. According to the samples analyzed, in Argentina, the genotype B was found in buffalo and the genotypes A and C were found in cattle. PMID:22716217

  3. Traumatic Reticuloperitonitis in Water Buffalo (Bubalus bubalis): Clinical Findings and the Associated Inflammatory Response

    PubMed Central

    El-Ashker, Maged; Salama, Mohamed; El-Boshy, Mohamed

    2013-01-01

    The present study was carried out to describe the clinical picture of traumatic reticuloperitonitis (TRP) in water buffalo (Bubalus bubalis) and to evaluate the inflammatory and immunologic responses for this clinical condition. Twenty-two buffalo with acute local TRP were monitored in our study. Additionally, 10 clinically healthy buffalo were randomly selected and served as controls. Acute local TRP was initially diagnosed by clinical examination and confirmed by ultrasonographic (USG) examination and/or necropsy findings. Blood samples were collected from all examined buffalo to measure the respective levels of tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, IL-6, IL-10 and interferon gamma (INF)-γ, serum amyloid A (SAA), C-reactive protein (CRP), haptoglobin (Hp), fibrinogen (Fb), and serum sialic acid (SSA). It was found that TNF-α, IL-1β, IL-6, IL-10, SAA, CRP, Hp, Fb, and SSA were significantly higher in buffalo with TRP than the controls. Our findings suggest that the examined immunologic variables were helpful in documenting the inflammatory response in buffalo with TRP. However, their diagnostic usefulness only becomes apparent when considered in tandem with the clinical findings for any given animal, its anamnesis, and a subsequent USG assessment. Due to the frequent complications of TRP, more accurate indicators of its occurrence and severity would be useful. PMID:26464911

  4. Prevalence and etiology of subclinical mastitis among buffaloes (Bubalus bubalus) in Namakkal, India.

    PubMed

    Srinivasan, P; Jagadeswaran, D; Manoharan, R; Giri, T; Balasubramaniam, G A; Balachandran, P

    2013-12-01

    Milk samples from 206 apparently healthy buffaloes of marginal farmers maintained under the prevailing field conditions were screened for Subclinical Mastitis (SCM) to determine its prevalence and etiology by White Side Test (WST), California Mastitis Test (CMT), Somatic Cell Count (SCC) and Bacteriological examination. 26.20% of animals were positive for SCM in SCC and bacteriological examination. Prevalence in graded Murrah buffaloes were found to be higher compared to graded Surti and Non-descript breeds. The prevalence of SCM was highest in above 6th stage of lactation. Single quarter infection (51.85%) was more common compared to multiple quarter infection. Hind (83.34%) and left side (57.41%) quarters were more prone to SCM. The indirect tests such as WST and CMT were found to be closely agreement with SCC and bacteriological examination. Due to their efficacious, easy to perform and interpret, these tests can effectively be employed to detect SCM under field conditions. Somatic cell count of = 3,00,000/mL of milk was regarded as the upper limit for normal buffaloes. The SCC of = 3,00,000/mL of milk with positive bacterial growth were used to diagnose SCM in the present study. Staphylococcus sp. 25 (46.30%) was the most common pathogens isolated from SCM followed by Streptococcus sp. 11 (20.37%) and E. coli 06 (11.11%) of the 54 bacterial isolates. Monobacterial and mixed bacterial infections were observed in 47 (87.04%) and 7 (12.96%) cases respectively.

  5. Analysis of rumen methanogen diversity in water buffaloes (Bubalus bubalis) under three different diets.

    PubMed

    Franzolin, Raul; St-Pierre, Benoit; Northwood, Korinne; Wright, André-Denis G

    2012-07-01

    The water buffalo (Bubalus bubalis) is a prominent livestock species for the production of milk and meat in many countries. We investigated the diversity of rumen methanogens in Mediterranean water buffaloes maintained in Brazil under different diets: corn silage, grazing pasture, or sugar cane. A total of 467 clones were isolated from three methanogen 16S rRNA gene clone libraries that each represented a distinct feed type. The 467 clones were assigned to 19 species-level operational taxonomic units (OTUs). Four OTUs were represented in all three libraries, eight OTUs were library-specific, six OTUs were found in only the corn silage and pasture grazing libraries, and one OTU was shared only between pasture grazing and sugar cane libraries. We found that Methanobrevibacter-related sequences were the most abundant in the water buffaloes sampled for our analysis, in contrast to previously reported studies showing that Methanomicrobium mobile-like methanogens were the most abundant methanogens in water buffaloes of Murrah and Surti breeds sampled in India. Considering the worldwide distribution of water buffaloes and the likely wide variety of diets provided, our results combined with studies from other groups support that larger scope analyses of microbiomes for this livestock species would provide great insight into the contribution of geographical location, breed, and diet in determining the population structure of rumen microorganisms.

  6. Genome-wide profiling of sperm DNA methylation in relation to buffalo (Bubalus bubalis) bull fertility.

    PubMed

    Verma, Arpana; Rajput, Sandeep; De, Sachinandan; Kumar, Rakesh; Chakravarty, Atish Kumar; Datta, Tirtha Kumar

    2014-09-15

    The DNA methylation pattern in spermatozoa of buffalo bulls of different fertility status was investigated. Spermatozoa isolated DNA from two groups of buffalo bulls (n = 5), selected based on their artificial insemination-generated conception rate data followed by IVF efficiency, were studied for global methylation changes using a custom-designed 180 K buffalo (Bubalus bubalis) CpG island/promoter microarray. A total of 96 individual genes with another 55 genes covered under CpG islands were found differentially methylated in sperm of high-fertile and subfertile buffalo bulls. Important genes associated with biological processes, cellular components, and functions were identified to be differentially methylated in buffalo bulls with differential fertility status. The identified differentially methylated genes were found to be involved in germ cell development, spermatogenesis, capacitation, and embryonic development. The observations hint that methylation defects of sperm DNA may play a crucial role in determining the fertility of breeding bulls. This growing field of sperm epigenetics will be of great benefit in understanding the graded fertility conditions of breeding bulls in commercial livestock production system. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. DGAT1 and ABCG2 polymorphism in Indian cattle (Bos indicus) and buffalo (Bubalus bubalis) breeds.

    PubMed

    Tantia, Madhu S; Vijh, Ramesh K; Mishra, Bishnu P; Mishra, Bina; Kumar, S T Bharani; Sodhi, Monika

    2006-11-07

    Indian cattle (Bos indicus) and riverine buffalo (Bubalus bubalis) give a poor yield of milk but it has a high fat and protein percentage compared to taurine cattle. The identification of QTLs (Quantitative Trait Loci) on BTA14 and BTA6 and its subsequent fine mapping has led to identification of two non conservative mutations affecting milk production and composition. Our objective was to estimate the frequency of K232A (DGAT1--diacylglycerol-acyltransferase 1) and Y581S (ABCG2--ATP binding cassette sub family G member 2) polymorphisms in diverse cattle and buffalo breeds of India having large variation in terms of milk production. We screened the reported missense mutations in six cattle and five buffalo breeds. The DGAT1K and ABCG2Y alleles were found to be fixed in Indian cattle and buffalo breeds studied. This study provides an indirect evidence that all the Indian cattle and buffalo breeds have fixed alleles with respect to DGAT1 and ABCG2 genes reported to be responsible for higher milk fat yield, higher fat and protein percent.

  8. Effect of age, sex and physiological stages on hematological indices of Banni buffalo (Bubalus bubalis)

    PubMed Central

    Patel, Mehul D.; Lateef, Abdul; Das, Hemen; Patel, Ajay S.; Patel, Ajay G.; Joshi, Axay B.

    2016-01-01

    Aim: To determine the physiological baseline values for hematological indices of Banni buffalo (Bubalus bubalis) as well as to assess their alteration due to age, sex and physiological stages. Materials and Methods: A total of 42 clinically healthy Banni buffaloes were categorized into seven groups (n=6): Group I (male calves ≤1 year), Group II (bulls >1 year), Group III (female calves ≤1 year), Group IV (pregnant lactating buffaloes), Group V (non-pregnant lactating buffaloes), Group VI (pregnant dry buffaloes), and Group VII (non-pregnant dry buffaloes). Blood samples collected aseptically from all the experimental groups were analyzed employing automated hematology analyzer. The data obtained were statistically analyzed; the mean and standard deviations were calculated and set as the reference values. Results: The erythrocytic indices viz. total erythrocytes count (TEC), hemoglobin, and packed cell volume (PCV) were significantly higher in bulls as compared to that of male calves unlike mean corpuscular volume, mean corpuscular hemoglobin (MCH), and MCH concentration. The female calves had higher TEC and PCV than the adult buffaloes irrespective of sex. The total leukocyte count (TLC) and neutrophil counts in male calves were significantly lower than the bulls unlike the eosinophil, while monocyte and basophil remained unchanged with age. The TLC, differential leukocyte count and platelet count varied non-significantly among the adult female groups at different physiological stages. However, neutrophils were found to be apparently higher in lactating buffaloes. Conclusion: The present study would be helpful for physiological characterization of this unique buffalo breed of Gujarat. Further, data generated may be a tool for monitoring the health and prognosis as well as diagnosis of diseases. PMID:27051182

  9. Clinical and ultrasonographic observations of functional and mechanical intestinal obstruction in buffaloes (Bubalus bubalis)

    PubMed Central

    Khalphallah, Arafat; Aref, Nasr-Eldin M.; Elmeligy, Enas; El-Hawari, Sayed F.

    2016-01-01

    Aim: This study was designed for clinical and laboratory evaluation of intestinal obstruction (IO) in buffaloes (Bubalus bubalis) with special emphasis on the diagnostic value of ultrasonographic findings. Materials and Methods: A total number of 30 buffaloes were included in the study and divided into 2 groups: Healthy (n=10) and diseased group (n=20). Diseased buffaloes were admitted to the Veterinary Teaching Hospital at Assiut University, Egypt, with a history of anorexia, abdominal pain, various degrees of abdominal distention, and absence or presence of scanty mucoid faces. These animals were subjected to clinical and ultrasonographic as well as laboratory examinations. Results: Based on ultrasonographic findings, various forms of IO were diagnosed. Functional obstruction, paralytic ileus, was diagnosed in 17 cases (85%) while mechanical IO was diagnosed only in 3 cases (15%). Out of 17 cases of paralytic ileus, both proximal and distal ileuses were successfully imaged in 8 and 9 cases, respectively. Proximal ileus was imaged from the right dorsal flank region as a single dilated loop of diameter >6 cm, while distal ileus was imaged as multiple dilated loops of diameter <6 cm. Mechanical obstruction due to duodenal intussusception was visualized as two concentric rings with outer echogenic wall and hypoechoic lumen. All cases of IO showed leukocytosis, hypoproteinemia, and increased activity of alkaline phosphatase and aspartate aminotransferase. Conclusion: Ultrasonography proved to be an essential tool for diagnosis and differential diagnosis of various forms of IO in buffaloes. PMID:27284223

  10. Binucleate trophoblast giant cells in the water buffalo (Bubalus bubalis) placenta.

    PubMed

    Carvalho, A F; Klisch, K; Miglino, M A; Pereira, F T V; Bevilacqua, E

    2006-01-01

    The binucleate trophoblast giant cells (BNC) of the water buffalo, Bubalus bubalis, placenta were studied, with emphasis on the synthesis of BNC-specific proteins. Placentomal tissues of 27 water buffalos (2-10 months of pregnancy) were processed for light and electron microscopy. The frequency of BNCs was 20% of the trophoblastic cells in 2-3-month placentas and increased to 27% in the later stages. Ultrastructurally, binucleate cells displayed a prominent granular endoplasmic reticulum and Golgi apparatus, typical of cells involved with protein synthesis and exportation. The buffalo BNCs contained periodic acid-Schiff (PAS)-positive granules and reacted with antisera against bovine placental lactogen, prolactin-related protein-I, and pregnancy-associated glycoproteins. Lectin histochemistry with Dolichos biflorus agglutinin, Vicia villosa agglutinin, and Phaseolus vulgaris leucoagglutinin showed specific staining of BNCs. Different stages of BNC migration and fusion with uterine epithelial cells were observed. Trinucleate feto-maternal hybrid cells were the typical outcome of cell fusions. These cells underwent degeneration, with typical morphological features of apoptosis. The results revealed a strong homology between water buffalo and cattle BNCs concerning cell morphology, protein expression, glycosylation pattern, and characteristics of cell migration and fusion.

  11. Allelic diversity at MHC class II DQ loci in buffalo (Bubalus bubalis): evidence for duplication.

    PubMed

    Niranjan, Saket K; Deb, Sitangsu M; Kumar, Subodh; Mitra, Abhijit; Sharma, Arjava; Sakaram, Durgam; Naskar, Soumen; Sharma, Deepak; Sharma, Sita R

    2010-12-01

    The genetic diversity of MHC class II DQ genes was investigated in riverine buffalo (Bubalus bubalis) by PCR-RFLP and sequencing. Highly variable regions (exons 2-3) of DQ genes were amplified from 152 buffaloes and genotyped by PCR-RFLP. Alleles identified by differential restriction patterns were sequenced for the characterization. PCR-RFLP was a rapid method to discriminate between DQA1 and duplicated DQA2 genes in buffalo, however, the method appeared to be inadequate for determining the more complicated DQB genotypes. A total of 7 and 10 alleles were identified for DQA and DQB loci, respectively. Nucleotide as well as amino acid variations among DQ alleles particularly at peptide binding regions were high. Such variations were as expected higher in DQB than DQA alleles. The phylogenetic analysis for both genes revealed the grouping of alleles into two major sub-groups with higher genetic divergence. High divergence among DQ allelic families and the isolation of two diverse DQA and DQB sequences from individual samples indicated duplication of DQ loci was similar in buffalo to other ruminants.

  12. Detection of Bovine viral diarrhea virus from three water buffalo fetuses (Bubalus bubalis) in southern Italy.

    PubMed

    Martucciello, Alessandra; De Mia, Gian Mario; Giammarioli, Monica; De Donato, Immacolata; Iovane, Giuseppe; Galiero, Giorgio

    2009-01-01

    Bovine viral diarrhea virus (BVDV) is an important pathogen that primarily infects ruminants, leading to several clinical problems including abortion. BVDV-specific antibodies were reported in a wide range of hosts within domestic and wildlife animal populations, and serological studies also indicated BVDV infection in buffaloes. The purpose of this study was to analyze the presence of BVDV in 2 water buffalo (Bubalus bubalis) herds with a history of abortion. Virus isolation from aborted fetuses and from maternal buffy coat and the molecular characterization of the isolates confirmed the presence of BVDV in these animals. The sequence analysis based on the 5' UTR and N(pro) coding regions of the Pestivirus genome revealed that the isolates belong to subgenotype 1b of BVDV. The findings of this study also suggest a possible role of BVDV in causing congenital infection in water buffalo. Its presence in fetal tissues as well as in maternal blood raises questions about the possible development of clinical disease or its influence in abortions in water buffalo.

  13. Myxobolus ictiobus n. sp. and Myxobolus minutus n. sp. (Cnidaria: Myxobolidae) from the gills of the smallmouth buffalo Ictiobus bubalus Rafinesque (Cypriniformes: Castostomidae)

    USDA-ARS?s Scientific Manuscript database

    The smallmouth buffalo Ictiobus bubalus Rafinesque (Catostomidae) is native to North American waterways and occasionally grown in pond aquaculture. Species of Myxobolus Bütschli, 1882 have been reported from the gills, integument, and intestinal tract of buffalo fish, although there is ambiguity in ...

  14. Ameliorative effects of boron on serum profile in buffalo (Bubalus bubalis) fed high fluoride ration.

    PubMed

    Bharti, Vijay K; Gupta, Meenakshi; Lall, D

    2008-02-01

    An experiment was undertaken to evaluate the protective role of boron on the serum profile of buffalo calves fed a high fluoride ration. Twelve male Murrah buffalo (Bubalus bubalis) calves of 6-8 months age, divided into three groups of four calves in each, were fed basal diets and supplemented with sodium fluoride (NaF, 60 ppm) alone or in combination with borax (Na2B4O7.10H2O, 140 ppm) for 90 days. Boron (B) was added in the ration as borax to make @140 ppm boron (elemental B) on DM basis in treatment II. Dietary F caused a significant (p<0.05) depressing effect on serum Ca and Zn on day 90 which was improved with B supplementation. However, serum Fe and Cu did not show any significant change on F or F+B supplementation. The serum ALP and phosphorus level were increased significantly (p<0.05) on F feeding but declined significantly (p<0.05) when B was fed. The findings suggested beneficial effect of boron on serum minerals and ALP in buffalo calves fed high fluoride ration.

  15. Hand-made cloned buffalo (Bubalus bubalis) embryos: comparison of different media and culture systems.

    PubMed

    Shah, Riaz A; George, Aman; Singh, Manoj K; Kumar, Dharmendra; Chauhan, Manmohan S; Manik, Radhaysham; Palta, Prabhat; Singla, Suresh K

    2008-12-01

    Hand-made cloning (HMC) has proved to be an efficient alternative to the conventional micromanipulator-based technique in some domestic animal species. This study reports the development of an effective culture system for in vitro culture of zona-free cloned buffalo (Bubalus bubalis) embryos reconstructed using adult skin fibroblast cells as nucleus donor. Cleavage and blastocyst rates observed were 52 and 0% in modified Charles Rosenkrans 2 (mCR2), 61 and 4.6% in modified Synthetic Oviductal Fluid (mSOF), and 82 and 40.3% in Research Vitro Cleave (RVCL; Cook, Australia) medium, respectively. Similarly, higher blastocyst rates (24.5 +/- 4.1%) were observed when zona-free parthenotes were cultured in RVCL medium. Culturing zona-free cloned buffalo embryos on flat surfaces (FS) yielded significantly higher (p < 0.05) blastocyst rates than Well of the Wells (WOW) or microdrops (MD). Furthermore, development in WOW was found to be significantly better than MD culture. The quality of HMC blastocysts was examined using differential staining. This study establishes the application of zona-free nuclear transfer procedures for the production of hand-made cloned buffalo embryos and the development of efficient culture system and appropriate media requirements for enhancing their preimplantation development.

  16. Embryonic Development of Heart in Indian Buffalo (Bubalus bubalis)

    PubMed Central

    Gupta, Anuradha; Bansal, Neelam; Uppal, Varinder

    2014-01-01

    The present study was conducted on 35 buffalo foetuses from 0.9 cm CVRL (32 days) to 99.5 cm CVRL (298 days) to observe the morphogenesis and histogenesis of heart. The study revealed that, in 0.9 cm CVRL buffalo foetus, heart was unseptated and tubular which was clearly divided into common atrial chamber dorsally, primitive ventricles ventrally, primitive outflow tract with bulbous cordis region proximally, and aortic sac distally at 1.2 cm CVRL. Septum primum appeared at 1.9 cm CVRL whereas the truncal swellings and fold of interventricular septum appeared at 2.5 cm CVRL foetus. At 3.0 cm CVRL septum primum, endocardial cushions, septum secundum, and foramen ovale were observed. At 7.6 cm CVRL the endocardial cushions fused to form right and left atrioventricular openings and ventricular apex became pointed. Interventricular canal was obliterated and four-chambered heart was recognised along with atrioventricular valve, chordae tendineae, and papillary muscles in 8.7 cm CVRL (66 days) buffalo foetus. The endocardium as well as epicardium of the atria was thicker as compared to ventricle, whereas the myocardium of atria was thin as compared to ventricles in all the age groups. All the internal structures of heart were well differentiated from 50 cm CVRL onwards. The detailed structural components of buffalo heart during prenatal period have been discussed in the present paper. PMID:27355030

  17. Polymorphisms in the DGAT1 gene in buffaloes (Bubalus bubalis) in the Amazon.

    PubMed

    Silva, C S; Silva Filho, E; Matos, A S; Schierholt, A S; Costa, M R; Marques, L C; Costa, J S; Sales, R L; Figueiró, M R; Marques, J R F

    2016-09-16

    Water buffaloes (Bubalus bubalis) are quite well adapted to climatic conditions in the Amazon, and in this biome, they are noted for the considerable amount of meat and milk they produce and how hard they are able to work. Because of a lack of research dedicated to improving the rearing of buffaloes in the Amazon, the objective of this study was to genetically characterize the Murrah and Mediterranean breeds, as well as a mixed-breed population, based on polymorphisms in the diacylglycerol O-acyltransferase 1 gene (DGAT1), and associate the genotypes with milk production. By using the polymerase chain reaction-single-strand conformation polymorphism technique, the alleles A (0.79), B (0.20), and D (0.01) were found in the Murrah breed. In the Mediterranean and mixed-breed buffaloes, we found alleles A (0.69) and (0.77) and B (0.31) and (0.23), respectively. The Murrah breed had the genotypes AA (0.63), AB (0.29), BB (0.05), and AD (0.03), and the Mediterranean and mixed-breed buffaloes had the genotypes AA (0.44) and (0.61), AB (0.50) and (0.31), and BB (0.06) and (0.08), respectively. For the Murrah, Mediterranean, and mixed-breed buffaloes, respectively, the expected heterozygosity values were 0.34, 0.43, and 0.35, the inbreeding coefficients were 0.78, -0.15, and 0.17, and the Hardy-Weinberg probabilities were 0.70, 0.67, and 0.52. The genotypes evaluated did not have an effect on milk production; however, the single nucleotide polymorphisms can be used in studies on genetic variability.

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

    PubMed

    Presicce, Giorgio Antonio; Bella, Antonino; Terzano, Giuseppina Maria; De Santis, Giuseppe; Senatore, Elena Maria

    2005-03-15

    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

  19. Study of gastrointestinal parasites in water buffalo (Bubalus bubalis) reared under Mexican humid tropical conditions.

    PubMed

    Ojeda-Robertos, Nadia Florencia; Torres-Chablé, Oswaldo Margarito; Peralta-Torres, Jorge Alonso; Luna-Palomera, Carlos; Aguilar-Cabrales, Aguilar; Chay-Canul, Alfonso Juventino; González-Garduño, Roberto; Machain-Williams, Carlos; Cámara-Sarmiento, Ramón

    2017-03-01

    The objective was to determine the frequency of gastrointestinal parasites (GP) genera affecting water buffalo (Bubalus bubalis) reared under humid tropical conditions of Mexico. Three hundred eighty-three Munrah breed water buffalo were included, 251 adult females and 132 calves. Feces were directly collected form the rectum of the animals and processed by the McMaster technique. Coprocultures were made to identify the genera of the nematodes. The frequency of GP in B. bubalis was 42%, independently of their age, 60% of calves resulted parasitized. Age had a strong association with the presence of GP (Xi(2) = 77.4014, d.f. = 1, p = 0.001). The family Trichostrongylidae was found in both age groups. The genera identified were Strongyloides sp. (47.2%), Cooperia sp. (33.9%), and Haemonchus sp. (10.4%), as well as Eimeria sp., Moniezia sp., Trichuris sp., and Strongyloides sp. The highest parasite burden corresponded to the genus Strongyloides sp. with 1108.9 EPG. There is a need to carry out further studies in order to know the prevalence and incidence of nematode affecting to B. bubalis as an introduced animal species to Mexican tropics.

  20. Sarcocystis dehongensis n. sp. (Apicomplexa: Sarcocystidae) from water buffalo (Bubalus bubalis) in China.

    PubMed

    Chen, Xinwen; Wen, Tao; Hu, Junjie; Liu, Tingting; Esch, Gerald W; Liang, Yu; Li, Hongliang; Huang, Si

    2017-08-01

    Water buffalo (Bubalus bubalis) is the intermediate host for at least four species of Sarcocystis: S. fusiformis, S. buffalonis, S. levinei, and S. sinensis/S. dubeyi. Here, a new species, Sarcocystis dehongensis, is reported in 51 of 756 (6.7%) water buffaloes in China. By light microscopy, the cysts of S. dehongensis were macroscopic, up to 18.5 mm long and 95 μm in diameter; 4.9-11.9 μm villous protrusions extended beyond the sarcocyst wall. Using transmission electron microscopy, the sarcocyst wall had lancet- or leaf-like protrusions in longitudinal section, but the cross section showed that the protrusions appeared as mushroom-like in shape with a core of tightly packed microtubules, similar to "type 24." BLAST searches revealed that S. dehongensis shared the most similarities with the 18S rDNA sequence of S. hardangeri (92.4%) and mitochondrial cox1 gene sequence of S. ovalis (81.0%), whereas no sequences in GenBank were found to be significantly similar to the ITS-1 region of S. dehongensis. A phylogenetic analysis based on 18S rDNA and mitochondrial cox1 gene sequences suggested that S. dehongensis was closely related to Sarcocystis species from cervids that employ corvids as definitive hosts.

  1. First molecular characterisation of Cryptosporidium and Giardia from Bubalus bubalis (water buffalo) in Victoria, Australia.

    PubMed

    Abeywardena, Harshanie; Jex, Aaron R; von Samson-Himmelstjerna, Georg; Haydon, Shane R; Stevens, Melita A; Gasser, Robin B

    2013-12-01

    We conducted a molecular epidemiological survey of Cryptosporidium and Giardia from Bubalus bubalis (water buffalo) on two extensive farms (450 km apart) in Victoria, Australia. Faecal samples (n=476) were collected from different age groups of water buffalo at two time points (six months apart) and tested using a PCR-based mutation scanning-targeted sequencing-phylogenetic approach, employing markers within the small subunit of ribosomal RNA (designated pSSU) and triose phosphate isomerase (ptpi) genes. Based on pSSU data, Cryptosporidium parvum, Cryptosporidium bovis and Cryptosporidium genotypes 1, 2 (each 99% similar genetically to Cryptosporidium ryanae) and 3 (99% similar to Cryptosporidium suis) were detected in two (0.4%), one (0.2%), 38 (8.0%), 16 (3.4%) and one (0.2%) of the 476 samples tested, respectively. Using ptpi, Giardia duodenalis assemblages A and E were detected in totals of 56 (11.8%) and six (1.3%) of these samples, respectively. Cryptosporidium was detected on both farms, whereas Giardia was detected only on farm B, and both genera were detected in 1.5% of all samples tested. The study showed that water buffaloes on these farms excreted C. parvum and/or G. duodenalis assemblage A, which are consistent with those found in humans, inferring that these particular pathogens are of zoonotic significance. Future work should focus on investigating, in a temporal and spatial manner, the prevalence and intensity of such infections in water buffaloes in various geographical regions in Australia and in other countries.

  2. Seasonal variation in semen quality of swamp buffalo bulls (Bubalus bubalis) in Thailand.

    PubMed

    Koonjaenak, Seri; Chanatinart, Vichai; Aiumlamai, Suneerat; Pinyopumimintr, Tanu; Rodriguez-Martinez, Heriberto

    2007-01-01

    To test the hypothesis that season affects the semen quality of swamp buffalo (Bubalus bubalis) bulls used for artificial insemination (AI) under tropical conditions in Thailand, as it does in Bos taurus and Bos indicus. Clinical and andrological examinations, and monitoring of semen production and quality were carried out on five mature, healthy swamp buffalo AI bulls in Thailand from July 2004 to the end of June 2005. Sperm output, motility, morphology and plasma membrane integrity (PMI) were compared between three seasons of the year (rainy, i.e. July-October; winter, i.e. November-February; and summer, i.e. March-June) with distinct ambient temperature and humidity. All bulls were diagnosed as clinically healthy and with good libido throughout the study. Ejaculate volume, pH, sperm concentration, total sperm number and initial sperm motility did not differ between seasons, whereas PMI and the relative proportion of morphologically normal spermatozoa were highest in summer and lowest in winter (P<0.05). Buffalo age, week of collection and season influenced sperm morphology (P < 0.05-0.001). Among morphological abnormalities, only proportions of tail defects were affected by season, being highest in the rainy season and lowest in summer (P<0.001). In conclusion, climatic changes did not seem to largely affect semen sperm output or viability. Although the proportions of PMI and tail abnormalities were affected by season, they were always below what is considered unacceptable for AI bull sires. Seasonal changes did not appear to cause deleterious changes in sperm quality in swamp buffalo AI-sires in tropical Thailand.

  3. Comparison of two methods of seminal plasma removal on buffalo (Bubalus bubalis) sperm cryopreservation.

    PubMed

    Albuquerque, R S; Morais, R; Reis, A N; Miranda, M S; Dias, Ear; Monteiro, F M; Paz, Ccp; Nichi, M; Kawai, Gkv; Della'Aqua, Cpf; Papa, F O; Viana, R B; Gimenes, L U

    2017-10-01

    Cryopreservation causes damage to spermatozoa, and methods minimizing this damage are therefore needed. Although much discussed, seminal plasma removal has become an alternative to improve sperm quality and viability after freezing and has been applied to different species in attempt to obtain good results. The objective of this study was to evaluate semen quality in buffaloes submitted to two methods for seminal plasma removal (filtration and centrifugation). Semen samples were collected from seven Murrah buffalo bulls (Bubalus bubalis) once a week for 8 weeks. Each ejaculate was divided into three groups: control (presence of seminal plasma), centrifugation and filtration. Sperm kinetics was evaluated with the computer-assisted sperm analysis (CASA) system. Plasmalemma and acrosomal membrane integrity, mitochondrial membrane potential and reactive oxygen species (ROS) were measured by flow cytometry, and lipid peroxidation was evaluated by the thiobarbituric acid reactive substances (TBARS) assay. Seminal plasma removal did not improve sperm kinetics compared to the control group. Centrifugation increased the number of cells with damaged acrosomal membranes (0.77 ± 0.05) and filtration caused greater plasmalemma and acrosomal membrane damage (22.18 ± 1.07). No difference in the mitochondrial membrane potential was observed between groups. In contrast, ROS production was higher in the centrifugation group compared to the control and filtration groups, although no differences in TBARS formation were detected. In conclusion, seminal plasma removal did not improve the quality of thawed buffalo semen compared to control in terms of sperm kinetics, membrane integrity, mitochondrial membrane potential or lipid peroxidation. © 2017 Blackwell Verlag GmbH.

  4. Comparative pharmacokinetics of ceftiofur hydrochloride and ceftiofur sodium after administration to water buffalo (Bubalus bubalis).

    PubMed

    Nie, Haiying; Feng, Xin; Peng, Jianbo; Liang, Liu; Lu, Chunyan; Tiwari, Roshan V; Tang, Shusheng; He, Jiakang

    2016-06-01

    OBJECTIVE To evaluate pharmacokinetics and bioavailability after administration of ceftiofur hydrochloride and ceftiofur sodium to water buffalo (Bubalus bubalis). ANIMALS 5 healthy adult water buffalo (3 males and 2 nonlactating females). PROCEDURES All animals received a dose (2.2 mg/kg) of 3 ceftiofur products (2 commercially available suspensions of ceftiofur hydrochloride [CEF1 and CEF2, IM] and ceftiofur sodium [CEF3, IV]). Blood samples were collected for up to 196 hours. Concentrations of ceftiofur in plasma were determined by use of high-performance liquid chromatography, and pharmacokinetic parameters were calculated on the basis of noncompartmental methods. RESULTS Most of the pharmacokinetic parameters, except for bioavailability and the area under the concentration-time curve extrapolated to infinity, were significantly different between the 2 products administered IM. Mean ± SD bioavailability of CEF1 and CEF2 was 89.57 ± 32.84% and 86.28 ± 11.49%, respectively, which indicated good absorption of both products. In addition, there was a longer drug residence time for CEF1 than for CEF2. Data analysis for CEF1 revealed a flip-flop phenomenon. CONCLUSIONS AND CLINICAL RELEVANCE In this study, there was good absorption of CEF1, and CEF1 had a longer drug residence time in vivo than did CEF2. On the basis of pharmacokinetic parameters and the in vitro antimicrobial susceptibility, a dosage regimen of 2.2 mg/kg administered at 48- and 36-hour intervals for CEF1 and CEF2, respectively, could be an appropriate choice for the treatment of buffalo with infectious diseases.

  5. Follicular dynamics and oestrous detection in Thai postpartum swamp buffaloes (Bubalus bubalis).

    PubMed

    Yindee, M; Techakumphu, M; Lohachit, C; Sirivaidyapong, S; Na-Chiangmai, A; Rodriguez-Martinez, H; van der Weyden, G C; Colenbrander, B

    2011-02-01

    This study characterized follicular activity and oestrous behaviour from 5 to 9 days post-calving up to the 4th ovulation postpartum (pp) in 16 multiparous (range 2-7 parities) Thai swamp buffalo cows (Bubalus bubalis), aged 4-12 years and weighing from 432 to 676 kg. Ovarian follicular activity was examined by transrectal ultrasonography (TUS) every morning. Oestrous detection was performed twice daily by direct personal observation of behaviour and for presence of clear cervical mucus discharge and indirectly by video camera recording during 21 h/day. A follicular wave-like pattern was present before the 1st ovulation leading to short oestrous cycles. Growth rates and maximum diameters of the ovulatory follicles did not differ between the 1st and 4th ovulations. However, growth rate for non-ovulatory dominant follicles (DF) before the 1st ovulation was lower than for the ovulatory follicle (p<0.05). In addition, the diameter of all ovulatory follicles (14.3 ± 0.46 mm, n=39) was significantly larger (p < 0.01) than those of the preceding last but one non-ovulatory DF (10.8 ± 0.20 mm, n = 5), but similar to the last preceding non-ovulatory DF diameter (12.92 ± 0.96 mm, n = 14). Short oestrous cycles were most common between the 1st and 2nd ovulations (93.75%, 15/16 cows, 10.2 ± 0.38 days) decreasing in prevalence thereafter (50%, 3/6 buffaloes, 12.0 ± 1.53 days). Oestrous signs were relatively vague around the 1st ovulation pp to become more easily detectable thereafter. This study suggests that properly fed swamp buffaloes could be mated successfully within 2 months pp, at their 2nd spontaneous ovulation, provided oestrous detection is at least performed daily at 06:00-08:00 hour.

  6. Identity of Sarcocystis species of the water buffalo (Bubalus bubalis) and cattle (Bos taurus) and the suppression of Sarcocystis sinensis as a nomen nudum

    USDA-ARS?s Scientific Manuscript database

    There are uncertainties concerning the identity and host species specificity of Sarcocystis species of the water buffalo (Bubalus bubalis) and cattle (Bos taurus). Currently, in cattle three species are recognized with known endogenous stages, viz.: S. cruzi (with canine definitive host), S. hirsuta...

  7. Characterization of leukocyte subsets in buffalo (Bubalus bubalis) with cross-reactive monoclonal antibodies specific for bovine MHC class I and class II molecules and leukocyte differentiation molecules

    USDA-ARS?s Scientific Manuscript database

    Although buffalo (Bubalus bubalis) are a major component of the livestock industry worldwide, limited progress has been made in the study of the mechanisms regulating the immune response to pathogens and parasites affecting their health and productivity. This has been, in part, attributable to the l...

  8. Saliva ferning, an unorthodox estrus detection method in water buffaloes (Bubalus bubalis).

    PubMed

    Ravinder, R; Kaipa, Onnureddy; Baddela, Vijay Simha; Singhal Sinha, Eshu; Singh, Prashant; Nayan, Varij; Velagala, Chandra Sekhar Naidu; Baithalu, Rubina Kumari; Onteru, Suneel Kumar; Singh, Dheer

    2016-09-15

    Estrus detection is a major problem in buffalo husbandry because of inconsistent expression of estrous signs at different seasons, and a high prevalence of the silent heat and postpartum anestrus in this species. Around 50% of the estrus events in buffaloes are currently undetected in the field conditions, resulting in a huge economic loss. Although the cervicovaginal fluid fern patterns confirm the estrus for a breeding decision, the fluid discharge is absent during the silent-heat condition. Therefore, the present study focused on the crystallization patterns of the saliva as an alternative method for estrus detection in buffaloes. Saliva is a body fluid available regularly, and its ferning ability before ovulation was established in women. In this study, eight female nonpregnant Murrah buffaloes (Bubalus bubalis) were considered during two experimental periods of 3 months each. One period was in summer with five animals, and another period was in rainy season with three animals. Estrus was determined by the estrus symptoms, ovarian ultrasonography, and salivary estradiol (E2) to progesterone (P4) ratio. A total of 450 saliva samples were collected from these animals on the daily basis. The salivary smear was prepared with 20 μL of the cell-free saliva on a clean glass slide, and its microscopic images were captured at a magnification of × 200. The images were used for fractal analysis as the salivary crystallization or fern patterns follow the fractal geometry. Saliva at estrus showed a typical symmetrical fern-like crystallization patterns with significantly (P < 0.05) lower fractal dimension values. Salivary estradiol levels and E2/P4 ratio were significantly (P < 0.05) higher at the estrus stage than those at the diestrus stage. An average period of an estrous cycle was 21.7 ± 2.7 days (n = 18 estrous cycles) in buffaloes on the basis of distinct salivary crystallization patterns. The proportion of estrus detection by the salivary fern patterns

  9. Evaluation of antifreeze protein III for cryopreservation of Nili-Ravi (Bubalus bubalis) buffalo bull sperm.

    PubMed

    Qadeer, S; Khan, M A; Ansari, M S; Rakha, B A; Ejaz, R; Husna, A U; Ashiq, M; Iqbal, R; Ullah, N; Akhter, S

    2014-07-01

    Lower fertility in buffaloes with frozen-thawed semen is attributed to sperm damage that is believed to be due to formation of ice crystals during freeze/thaw process. It was hypothesized that antifreeze proteins in the extender may improve the post thaw quality of buffalo bull sperm. For this purpose, two separate experiments were conducted to evaluate antifreeze proteins III (AFP III) at 0 (control), 0.1, 1 and 10 μg/mL (Experiment I) and 0 (control), 0.01, 0.1 and 1 μg/mL (Experiment II) for its effect on post thaw quality of buffalo bull semen. Semen was collected from three Nili-Ravi buffalo (Bubalus bubalis) bulls with artificial vagina (42 °C) for three weeks (replicate) per experiment. For each experiment, qualifying ejaculates (6 ejaculates/bull) were divided into four aliquots and diluted (at 37 °C having 50 × 10(6) sperm/mL) in tris-citric acid extender containing above mentioned concentrations of AFP III. Diluted semen was cooled to 4 °C in 2 h, equilibrated for 4 h, filled in 0.5 mL straws, kept over liquid nitrogen vapors for 10 min and plunged in the liquid nitrogen. After 24 h of storage, semen straws were thawed at 37 °C for 30 s to assess sperm progressive motility (SM), plasma membrane integrity (PMI), viability (live sperm with intact acrosome) and normal epical ridge (NAR). In experiment I, improvement (P<0.05) in percentage SM and sperm PMI was recorded in extender containing 0.1 μg/mL AFP III compared to control, the higher concentrations (1 μg/mL and 10 μg/mL) being inefficient. While evaluating the lower concentration (experiment II), 0.01 μg/mL of AFP III in the extender it was found to be ineffective to improve semen quality parameters, while 0.1 μg/mL AFP III in extender was found better in terms of progressive motility and plasma membrane integrity of buffalo bull semen compared to control. Sperm viability and NAR remained similar (P>0.05) in extenders containing different concentrations of AFP III and control in both of

  10. Transcriptional Dynamics of Homeobox C11 Gene in Water Buffalo Bubalus bubalis

    PubMed Central

    Rawal, Leena; Pathak, Deepali; Sehgal, Neeta

    2015-01-01

    The Hox complex contains 39 genes clustered into four groups involved in cell differentiation and development. We cloned full-length sequence of Hoxc11 gene from water buffalo Bubalus bubalis, assessed its copy number, localized the same onto the chromosome 5, and studied its evolutionary conservation across the species. Northern hybridization of Hoxc11 showed a 2.2 kb band in the tissues analyzed. Real-Time PCR showed highest expression of Hoxc11 gene in lung followed by spleen, spermatozoa, and testis. Six interacting partners of this gene showed higher expression in spleen, lung, testis, and spermatozoa. During the early stages of development, Hoxc11 and its interacting partners both showed lower expression, which then became prominent during the age of 1–3 years, regressed drastically thereafter, and remained so until the animal's life time (∼20 years). The high expression of Hoxc11 and its interacting partners in spermatozoa and testis during the onset of puberty suggests its likely role in the differentiation of gonads and subsequent reproductive activities. Additional work on Hoxc11 especially, in the context of respiratory, immunological, and in/fertility in other species, including humans would be useful for establishing its broader biological significance towards the enrichment of functional and comparative genomics. PMID:25760398

  11. Analysis of mitochondrial D-loop region casts new light on domestic water buffalo (Bubalus bubalis) phylogeny.

    PubMed

    Kierstein, Gerold; Vallinoto, Marcelo; Silva, Artur; Schneider, Maria Paula; Iannuzzi, Leopoldo; Brenig, Bertram

    2004-02-01

    The phylogeny of water buffaloes (Bubalus bubalis) is still a matter of discussion, especially if the two types of domestic water buffalo (swamp and river) derived from different domestication events or if they are products of human selection. To obtain more insight, we analyzed the entire mitochondrial D-loop region of 80 water buffaloes of four different breeds, i.e., 19 swamp buffaloes (Carabao) and 61 river buffaloes (Murrah, Jafarabadi, and Mediterranean), sampled in Brazil and Italy. We detected 36 mitochondrial haplotypes with 128 polymorphic sites. Pooled with published data of South-East Asian and Australian water buffaloes and based on comprehensive median-joining network and population demography analyses we show evidence that both river and swamp buffaloes decent from one domestication event, probably in the Indian subcontinent. However, the today swamp buffaloes have an unravelled mitochondrial history, which can be explained by introgression of wild water buffalo mtDNA into domestic stocks. We are also discussing indications for an independent domestication of buffaloes in China.

  12. Seroprevalence of Toxoplasma gondii infection in water buffaloes (Bubalus bubalis) in Veracruz State, Mexico and its association with climatic factors.

    PubMed

    Alvarado-Esquivel, Cosme; Romero-Salas, Dora; García-Vázquez, Zeferino; Cruz-Romero, Anabel; Peniche-Cardeña, Alvaro; Ibarra-Priego, Nelly; Aguilar-Domínguez, Mariel; Pérez-de-León, Adalberto A; Dubey, Jitender P

    2014-09-30

    Infection with Toxoplasma gondii in water buffaloes (Bubalus bubalis) is of epidemiological importance because of the risk for transmission to humans. We sought to determine the seroprevalence of T. gondii infection in 339 water buffaloes in Veracruz State, Mexico using the modified agglutination test (MAT, cut off 1:25). Seroprevalence association with general characteristics of buffaloes and their environment was also investigated. Antibodies to T. gondii were found in 165 (48.7%) of the 339 buffaloes with MAT titers of 1:25 in 104, 1:50 in 52, and 1:100 in 9. Bivariate analysis showed that seroprevalence of T. gondii infection was similar in buffaloes regardless of their general characteristics i.e., age, sex, and breed. In contrast, the seroprevalence in buffaloes varied significantly with environmental characteristics including altitude, mean annual temperature, and mean annual rainfall of the municipalities studied. Multivariate analysis showed that T. gondii seropositivity in buffaloes was associated with a mean annual rainfall between 1266-1650 mm (OR = 1.84; 95% CI: 1.15-2.94; P = 0.01). Results indicate that environmental characteristics may influence the seroprevalence of T. gondii infection in buffaloes. This is the first report on the seroprevalence of T. gondii infection in buffaloes in Mexico. Further research is needed to assess the risk for infection in humans associated with the ingestion of raw or undercooked meat from buffaloes infected with T. gondii.

  13. Plasma and milk kinetic of eprinomectin and moxidectin in lactating water buffaloes (Bubalus bubalis).

    PubMed

    Dupuy, Jacques; Sutra, Jean-François; Alvinerie, Michel; Rinaldi, Laura; Veneziano, Vincenzo; Mezzino, Laura; Pennacchio, Saverio; Cringoli, Giuseppe

    2008-11-07

    The pharmacokinetics and mammary excretion of moxidectin and eprinomectin were determined in water buffaloes (Bubalus bubalis) following topical administration of 0.5mgkg(-1). Following administration of moxidectin, plasma and milk concentrations of moxidectin increased to reach maximal concentrations (C(max)) of 5.46+/-3.50 and 23.76+/-16.63ngml(-1) at T(max) of 1.20+/-0.33 and 1.87+/-0.77 days in plasma and milk, respectively. The mean residence time (MRT) were similar for plasma and milk (5.27+/-0.45 and 5.87+/-0.80 days, respectively). The AUC value was 5-fold higher in milk (109.68+/-65.01ngdayml(-1)) than in plasma (23.66+/-12.26ngdayml(-1)). The ratio of AUC milk/plasma for moxidectin was 5.04+/-2.13. The moxidectin systemic availability (expressed as plasma AUC values) obtained in buffaloes was in the same range than those reported in cattle. The faster absorption and elimination processes of moxidectin were probably due to a lower storage in fat associated with the fact that animals were in lactation. Nevertheless, due to its high excretion in milk and its high detected maximum concentration in milk which is equivalent or higher to the Maximal Residue Level value (MRL) (40ngml(-1)), its use should be prohibited in lactating buffaloes. Concerning eprinomectin, the C(max) were of 2.74+/-0.89 and 3.40+/-1.68ngml(-1) at T(max) of 1.44+/-0.20 and 1.33+/-0.0.41 days in plasma and milk, respectively. The MRT and the AUC were similar for plasma (3.17+/-0.41 days and 11.43+/-4.01ngdayml(-1)) and milk (2.70+/-0.44 days and 8.49+/-3.33ngdayml(-1)). The ratio of AUC milk/plasma for eprinomectin was 0.76+/-0.16. The AUC value is 20 times lower than that reported in dairy cattle. The very low extent of mammary excretion and the milk levels reported lower than the MRL (20ngml(-1)) supports the permitted use of eprinomectin in lactating water buffaloes.

  14. Carcass Characteristics and Meat Quality of Swamp Buffaloes (Bubalus bubalis) Fattened at Different Feeding Intensities

    PubMed Central

    Lambertz, C.; Panprasert, P.; Holtz, W.; Moors, E.; Jaturasitha, S.; Wicke, M.; Gauly, M.

    2014-01-01

    Twenty-four male 1-year old swamp buffaloes (Bubalus bubalis) were randomly allocated to 4 groups. One group grazed on guinea grass (GG) and another on guinea grass and the legume Stylosanthes guianensis (GL). The other two groups were kept in pens and fed freshly cut guinea grass and concentrate at an amount of 1.5% (GC1.5) and 2.0% (GC2.0) of body weight, respectively. The effect of the different feeding intensities on carcass characteristics and meat quality were assessed. The mean body weight at slaughter was 398 (±16) kg. Average daily gain was higher in concentrate-supplemented groups (570 and 540 g/d in GC1.5 and GC2.0, respectively) when compared to GG (316 g/d) and GL (354 g/d) (p<0.01). Likewise, the warm carcass weight was higher in GC1.5 and GC2.0 compared to GG and GL. Dressing percentage was 48.1% and 49.5% in GC1.5 and GC2.0 in comparison to 42.9% and 44.8% observed in GG and GL, respectively. Meat of Longissimus throracis from GC1.5 and GC2.0 was redder in color (p<0.01), while water holding capacity (drip and thawing loss) was improved in pasture-fed groups (p<0.05). Protein and fat content of Longissimus thoracis was higher in animals supplemented with concentrate (p<0.01), as was cholesterol content (p<0.05), whereas PUFA:SFA ratio was higher and n-6/n-3 ratio lower (p<0.01) in pasture-fed buffaloes. Results of the present study showed that the supplementation of pasture with concentrate enhances the growth and carcass characteristics of swamp buffaloes expressed in superior dressing percentage, better muscling, and redder meat with a higher content of protein and fat, whereas animals grazing only on pasture had a more favorable fatty acid profile and water holding capacity. In conclusion, the supplementation of concentrate at a rate of about 1.5% of body weight is recommended to improve the performance and carcass quality of buffaloes. PMID:25049987

  15. 'Soya milk Tris-based phytoextender reduces apoptosis in cryopreserved buffalo (Bubalus bubalis) spermatozoa'.

    PubMed

    Mohan, R; Atreja, S K

    2014-10-01

    The objective of this study was to investigate the effect of newly developed soya milk Tris (SMT)-based phytoextender as an alternative to egg yolk Tris (EYT) extender used for cryopreservation of buffalo (Bubalus bubalis) spermatozoa on apoptosis. Fresh buffalo semen (control without dilution) was cryopreserved in conventional EYT (20% egg yolk v/v in Tris) and SMT (25% soya milk v/v in Tris) extender and used for the assessment of expression of apoptotic proteins. Proteins extracted from a total number of nine ejaculates from three individual buffalo bulls chosen at random were separated using SDS-PAGE followed by immunoblotting against caspase-8, caspase-9, caspase-3, poly(ADP-ribose)polymerase (PARP), cytochrome c and apoptosis inducing factor (AIF). In addition, fluorescence microscopy was used for the detection of mitochondrial membrane potential (JC-1 assay) and apoptotic cells (annexin V-FITC/PI assay). The results obtained clearly indicate the significant (p < 0.05) reduction in the expression of caspase-3 (27 kDa), caspase-8 (53 kDa), caspase-9 (50 kDa) precursor and cytochrome c (17 kDa) in semen cryopreserved in SMT extender in comparison with EYT extender. A non-significant (p > 0.05) reduction in expression of PARP-DNA-binding subunit (24 kDa) was observed in SMT extender. No expression of AIF was found in cryopreserved semen samples. A significant (p < 0.05) increase in the mean percentage of cells having high mitochondrial membrane potential and a non-significant (p > 0.05) decrease in late apoptotic cells (AN+/PI+) was observed in SMT extender when compared to EYT extender. The results demonstrated that cryopreservation of buffalo semen in SMT-based phytoextender can replace the traditional egg yolk extenders as it reduces the expression of apoptotic proteins maintaining high mitochondrial membrane potential and gives better protection to sperms in terms of its non-animal origin. © 2014 Blackwell Verlag GmbH.

  16. Carcass Characteristics and Meat Quality of Swamp Buffaloes (Bubalus bubalis) Fattened at Different Feeding Intensities.

    PubMed

    Lambertz, C; Panprasert, P; Holtz, W; Moors, E; Jaturasitha, S; Wicke, M; Gauly, M

    2014-04-01

    Twenty-four male 1-year old swamp buffaloes (Bubalus bubalis) were randomly allocated to 4 groups. One group grazed on guinea grass (GG) and another on guinea grass and the legume Stylosanthes guianensis (GL). The other two groups were kept in pens and fed freshly cut guinea grass and concentrate at an amount of 1.5% (GC1.5) and 2.0% (GC2.0) of body weight, respectively. The effect of the different feeding intensities on carcass characteristics and meat quality were assessed. The mean body weight at slaughter was 398 (±16) kg. Average daily gain was higher in concentrate-supplemented groups (570 and 540 g/d in GC1.5 and GC2.0, respectively) when compared to GG (316 g/d) and GL (354 g/d) (p<0.01). Likewise, the warm carcass weight was higher in GC1.5 and GC2.0 compared to GG and GL. Dressing percentage was 48.1% and 49.5% in GC1.5 and GC2.0 in comparison to 42.9% and 44.8% observed in GG and GL, respectively. Meat of Longissimus throracis from GC1.5 and GC2.0 was redder in color (p<0.01), while water holding capacity (drip and thawing loss) was improved in pasture-fed groups (p<0.05). Protein and fat content of Longissimus thoracis was higher in animals supplemented with concentrate (p<0.01), as was cholesterol content (p<0.05), whereas PUFA:SFA ratio was higher and n-6/n-3 ratio lower (p<0.01) in pasture-fed buffaloes. Results of the present study showed that the supplementation of pasture with concentrate enhances the growth and carcass characteristics of swamp buffaloes expressed in superior dressing percentage, better muscling, and redder meat with a higher content of protein and fat, whereas animals grazing only on pasture had a more favorable fatty acid profile and water holding capacity. In conclusion, the supplementation of concentrate at a rate of about 1.5% of body weight is recommended to improve the performance and carcass quality of buffaloes.

  17. Effect of Leptin on In Vitro Nuclear Maturation and Apoptosis of Buffalo (Bubalus bubalis) Oocyte

    PubMed Central

    Khaki1, Amir; Batavani, Rouzali; Najafi, Gholamreza; Tahmasbian, Hamid; Belbasi, Abolfazl; Mokarizadeh, Aram

    2014-01-01

    Background: Leptin, as a 16 kDa adipokine, is a pleiotropic cytokine-like hormone that primarily secreted from adipose tissue. It also involves in the regulation of energy homeostasis, neuroendocrine function, immunity, lipid and glucose homeostasis, fatty acid oxidation, angiogenesis, puberty and reproduction. The aim of this study was to investigate the effects of in vitro addition of leptin to in vitro maturation (IVM) medium on buffalo oocyte maturation and apoptosis. Materials and Methods: In this experimental study, Ovaries from apparently normal reproductive organs of slaughtered adult buffaloes (Bubalus bubalis) with unknown breeding history were collected from Urmia Abattoir, Urmia, Iran, and were transported immediately to the laboratory in a thermos flask containing sterile normal saline with added antibiotics. Oocytes were aspirated from 2-8 mm visible follicles of the ovaries using an 18-G needle attached to a 10 ml syringe. IVM medium included tissue culture medium-199 (TCM-199), 10% fetal bovine serum (FBS), 22 µg/ml sodium pyruvate, 0.5 IU/ml ovine follicle-stimulating hormone (oFSH), 0.5 IU/ml ovine luteinizing hormone (oLH), 1 μg/ml oestradiol, 50 μg/ml gentamycin, and leptin [0 (control), 10, 50, and 100 ng/ml]. The good quality buffalo oocytes (batches of 10 oocytes) were placed in a culture plate containing six 50 μl droplets of maturation medium, covered with sterilized mineral oil, and then incubated at 38.5˚C with 5% CO2 in air for 24 hours. The maturation of oocytes was evaluated under a stereomicroscope by detecting the first polar body extrusion of oocytes. FITC-Annexin V propidium iodide (PI) staining method was used to detect oocyte apoptosis. Results: From a total of 115 collected ovaries, 1100 oocytes were recovered among which 283 oocyte were suitable for IVM. In the groups of leptin treated with 0 (control), 10, 50 and 100 ng/ml, the percentage of oocytes maturation was 74.65, 83.81, 77.85, and 75.40%, while the percentage of

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

    PubMed

    Presicce, Giorgio Antonio; Parmeggiani, Albamaria; Senatore, Elena Maria; Stecco, Romana; Barile, Vittoria Lucia; De Mauro, Guillermo Javier; De Santis, Giuseppe; Maria Terzano, Giuseppina

    2003-08-01

    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

  19. Serum acute phase proteins in control and Theileria annulata infected water buffaloes (Bubalus bubalis).

    PubMed

    El-Deeb, Wael M; Iacob, Olimpia C

    2012-11-23

    This study was carried out to ascertain the changes in acute phase proteins (APPs) and pro-inflammatory cytokines in Theileria annulata infected water buffalo. Thirty infected water buffaloes and 20 parasitologically free were used. In the present study there was significant (P ≤ 0.05) increase in haptoglobin (Hp), serum amyloid A (SAA), ceruloplasmin, α1-acid glycoprotein (AGP) and fibrinogen levels (2.18 ± 0.29 g/l, 156.58 ± 3.48 mg/l, 31.23 ± 1.25mg/dl, 370.23 ± 33.21 mg/l and 16.17 ± 1.18 g/l, respectively) in T. annulata infected water buffaloes when compared to healthy ones (0.13 ± 0.01 g/l, 23.9 ± 0.56 mg/l, 21.23 ± 1.21 mg/dl, 240.53 ± 22.45 mg/l and 4.2 ± 0.1 6g/l, respectively). Moreover, there was significant (P ≤ 0.05) increase in the levels of TNF-α, IL-1α, IL-6, IL-12, IL-1β and IFN-γ (2.55 ± 0.12 ng/ml, 98.32 ± 4.21 pg/ml, 152.32 ± 5.62 pg/ml, 26.44 ± 1.43 ng/ml, 240.33 ± 20.45 pg/ml and 123.65 ± 5.67 pg/ml, respectively) in T. annulata infected water buffaloes when compared to healthy ones (0.42 ± 0.04 ng/ml, 55.32 ± 3.21 pg/ml, 88.23 ± 3.21 pg/ml, 7.45 ± 0.67 ng/ml, 98.33 ± 3.45 pg/ml and 34.76 ± 1.56 pg/ml, respectively). There was also significant decrease (P ≤ 0.05) in the Hb content, PCV%, RBCs and WBCs counts in the diseased water buffaloes compared to the control ones. Neutropenia, eosinopenia, lymphopenia, monocytopenia and thrombocytopenia were also recorded. The biochemical changes revealed significant (P ≤ 0.05) elevation in the levels of AST, ALT, ALP, LDL-c, VLDL-c, BHBA and NEFA, with significant (P ≤ 0.05) decrease in the levels of total proteins, albumin, globulins, cholesterol, triglyceride, glucose, G6PD, calcium and phosphorus in T. annulata infected water buffaloes when compared to healthy ones. It could be concluded that APPs and pro-inflammatory cytokines could be used as a valuable biomarkers in T. annulata infected water buffaloes (Bubalus bubalis). Copyright © 2012 Elsevier B.V. All

  20. Prevalence and distribution of Neospora caninum in water buffalo (Bubalus bubalis) and cattle in the Northern Territory of Australia.

    PubMed

    Neverauskas, Claudia E; Nasir, Amar; Reichel, Michael P

    2015-10-01

    The seroprevalence of Neospora caninum infection in water buffalo (Bubalus bubalis) and domestic cattle in the Northern Territory (NT) of Australia has never been determined. A total of 480 serum samples from water buffalo and 192 serum samples from cattle, collected by the NT Government from 1993 through to 2001, at 18 different survey sites throughout the Northern Territory were tested by commercial ELISA for anti-N. caninum antibodies. The water buffalo samples demonstrated a seroprevalence of 88.3% (95% CI ± 2.9%), while 31.8% (±6.1%) of the cattle sera tested positive for N. caninum antibodies. Individual buffalo from the same herd, sampled over years, showed considerable fluctuations in S/P ratios. Overall, seropositivity was consistent across buffalo herds, and showed a slight decline over the years. The study presents evidence for the first time that N. caninum infection in water buffalo in the Northern Territory is a highly endemic and that infection rates are higher than those for cattle. This is important for an understanding of any potential sylvatic life cycle of N. caninum in Northern Australia. This survey also tests cattle from that territory for the first time for evidence of N. caninum infection and makes an important contribution to the understanding of disease management issues for the beef industry in the region. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  1. Molecular analyses detect natural coinfection of water buffaloes (Bubalus bubalis) with bovine viral diarrhea viruses (BVDV) in serologically negative animals.

    PubMed

    Craig, María I; König, Guido A; Benitez, Daniel F; Draghi, María G

    2015-01-01

    Infection of water buffaloes (Bubalus bubalis) with bovine viral diarrhea viruses (BVDV) has been confirmed in several studies by serological and molecular techniques. In order to determine the presence of persistently infected animals and circulating species and subtypes of BVDV we conducted this study on a buffalo herd, whose habitat was shared with bovine cattle (Bossp.). Our serological results showed a high level of positivity for BVDV-1 and BVDV-2 within the buffalo herd. The molecular analyses of blood samples in serologically negative animals revealed the presence of viral nucleic acid, confirming the existence of persistent infection in the buffaloes. Cloning and sequencing of the 5' UTR of some of these samples revealed the presence of naturally mix-infected buffaloes with at least two different subtypes (1a and 1b), and also with both BVDV species (BVDV-1 and BVDV-2). Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  2. Biological and genetic analysis of a bovine-like coronavirus isolated from water buffalo (Bubalus bubalis) calves.

    PubMed

    Decaro, Nicola; Martella, Vito; Elia, Gabriella; Campolo, Marco; Mari, Viviana; Desario, Costantina; Lucente, Maria Stella; Lorusso, Alessio; Greco, Grazia; Corrente, Marialaura; Tempesta, Maria; Buonavoglia, Canio

    2008-01-05

    We describe the isolation, biological and genetic characterization of a host-range variant of bovine coronavirus (BCoV) detected in water buffalo (Bubalus bubalis). By conventional and real-time RT-PCR assays, the virus was demonstrated in the intestinal contents of two 20-day-old buffalo calves dead of a severe form of enteritis and in the feces of additional 17 buffalo calves with diarrhea. Virus isolation, hemagglutination and receptor-destroying enzyme activity showed that the buffalo coronavirus (BuCoV) is closely related to BCoV but possesses some different biological properties. Sequence and phylogenetic analyses of the 3' end (9.6 kb) of the BuCoV RNA revealed a genomic organization typical of group 2 coronaviruses. Moreover, the genetic distance between BuCoV and BCoV was proven to be the same or even higher than the distance between other ruminant coronaviruses and BCoV. In conclusion, our data support the existence of a host-range variant of BCoV associated with enteritis in buffaloes.

  3. Effect of pen size on behavioral, endocrine, and immune responses of water buffalo (Bubalus bubalis) calves.

    PubMed

    Grasso, F; Napolitano, F; De Rosa, G; Quarantelli, T; Serpe, L; Bordi, A

    1999-08-01

    Female water buffalo (Bubalus bubalis) calves (n = 28) aged 7 to 10 d were divided into four groups of seven animals each to examine the effects of space allowance (Group A: 2.6 indoor m2 + 2.0 outdoor m2/calf; Group B: 2.6 indoor m2/calf; Group C: 1.5 indoor m2/calf; Group D: 1.0 indoor m2/calf) on behavioral, endocrine, and immune variables for a period of 60 d. Animals were offered 7 L/d of a commercial acidified milk substitute. The calves averaged 45.9 kg initially and 92.4 kg finally. The behavior observations were conducted 7 d after grouping and fortnightly thereafter. At wk 4 and 8, the phytohemagglutinin (PHA) skin test was performed to induce aspecific delayed hypersensitivity. At wk. 1 and 3, calves were injected i.m. with keyhole limpet hemocyanin. Antibody titers were determined at weekly intervals for 7 wk. Calves in pens with greater space allowance (Groups A and B) were less active than Groups C and D (P<.001). The latter groups were also observed feeding more often at wk 7 (P<.01). Calves provided with an outdoor paddock spent less time standing than Groups C and D (P<.01), and lay with a greater number of outstretched legs (P<.001). Groups C and D showed a lower reaction to PHA in both skin tests than did Groups A and B (P<.001 and P<.05, respectively). Group A showed an antibody response consistently higher than groups B, C, and D (P<.01, P<.05, and P<.05, respectively). At the end of the experimental period, the calves were subjected to an isolation test lasting 10 min. Group D showed a longer duration of movement with respect to Groups A and B (P<.01); animals from Group C walked more than did Group A (P<.05). Cortisol concentration evaluated 0, 10, 45, 90, 150, and 225 min after separation from the group was higher in Groups C and D than in Groups A and B (P<.01). For all animals, the highest cortisol level was observed immediately after the isolation test (P<.001). Space restriction resulted in evidence of stress in the animals as shown by

  4. BRCA-1 Gene Expression and Comparative Proteomic Profile of Primordial Follicles from Young and Adult Buffalo (Bubalus bubalis) Ovaries.

    PubMed

    Govindaraj, Vijayakumar; Krishnagiri, Harshini; Chauhan, Manmohan Singh; Rao, A J

    2017-04-03

    In our previous study, we demonstrated that the repair efficiency of DNA double-strand breaks declines with increasing age in rat primordial follicles. In the present study, we extended our studies to buffalo (Bubalus bubalis) wherein we studied the expression of BRCA-1 related DNA repair genes in primordial follicles of young (12 months-22 months) and adult (72-96 months) buffaloes. The relative expression of selected genes, as determined by RT-PCR, revealed a significant (p < 0.05) decrease in mRNA levels of BRCA1, MRE11, RAD51, ATM, and H2AX in adult primordial follicles as compared to the young. Western blot analysis revealed a significant (p < 0.05) decrease in the expression of phosphorylated protein levels of BRCA1 and H2AX in adult buffalo primordial follicles. The protein expression profile of young and adult buffalo primordial follicles revealed differential expression of proteins involved in mitochondrial function, cell survival and cell metabolism. Similar to reports from aging rodent and human primordial follicles, our findings support the fact that impairment of DNA repair may be an universal mechanism involved in oocyte aging.

  5. Characterization of immune cell infiltration in the placentome of water buffaloes (Bubalus bubalis) infected with neospora caninum during pregnancy.

    PubMed

    Cantón, G J; Konrad, J L; Moore, D P; Caspe, S G; Palarea-Albaladejo, J; Campero, C M; Chianini, F

    2014-05-01

    Neospora caninum infection in cattle stimulates host immune responses, which may be responsible for placental damage leading to abortion. Susceptibility of water buffaloes (Bubalus bubalis) to neosporosis is not well understood, although vertical transmission and fetal death have been documented. The aim of this study was to characterize the immune response in the placentome of water buffalo following experimental infection in early gestation with the Nc-1 strain of N. caninum. Placentomes were examined by immunohistochemistry using antibodies specific for T-cell subsets, natural killer cells and CD79(αcy) cells. Placental inflammation was characterized by the infiltration of CD3(+) and CD4(+) T cells and T cells expressing the γδ T-cell receptor. The distribution of these cellular subsets in buffalo placentomes was similar to that previously described in cattle infected with N. caninum in early gestation, but the lesions were milder, which may explain the lower number of abortions observed in this species after infection. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Genetic variation of foot-and-mouth disease virus isolates recovered from persistently infected water buffalo (Bubalus bubalis).

    PubMed

    Barros, José Júnior F; Malirat, Viviana; Rebello, Moacyr A; Costa, Eliane V; Bergmann, Ingrid E

    2007-02-25

    Genetic variation of foot-and-mouth disease virus (FMDV) isolates, serotype O, recovered serially over a 1-year period from persistently infected buffalos was assessed. The persistent state was established experimentally with plaque-purified FMDV, strain O(1)Campos, in five buffalos (Bubalus bubalis). Viral isolates collected from esophageal-pharyngeal (EP) fluids for up to 71 weeks after infection were analyzed at different times by nucleotide sequencing and T(1) RNase oligonucleotide fingerprinting to assess variability in the VP1-coding region and in the complete genome, respectively. Genetic variation increased, although irregularly, with time after infection. The highest values observed for the VP1-coding region and for the whole genome were 2.5% and 1.8%, respectively. High rates of fixation of mutations were observed using both methodologies, reaching values of 0.65 substitutions per nucleotide per year (s/nt/y) and 0.44s/nt/y for nucleotide sequencing and oligonucleotide fingerprinting, respectively, when selected samples recovered at close time periods were analyzed. The data herein indicate that complex mixtures of genotypes may arise during FMDV type O persistent infection in water buffalos, which can act as viral reservoirs and also represent a potential source of viral variants. These results fit within the quasi-species dynamics described for FMDV, in which viral populations are constituted by related, non-identical genomes that evolve independently from each other, and may predominate at a given time.

  7. Local Immune Responses of the Chinese Water Buffalo, Bubalus bubalis, against Schistosoma japonicum Larvae: Crucial Insights for Vaccine Design

    PubMed Central

    McWilliam, Hamish E. G.; Piedrafita, David; Li, Yuesheng; Zheng, Mao; He, Yongkang; Yu, Xinling; McManus, Donald P.; Meeusen, Els N. T.

    2013-01-01

    Asian schistosomiasis is a zoonotic parasitic disease infecting up to a million people and threatening tens of millions more. Control of this disease is hindered by the animal reservoirs of the parasite, in particular the water buffalo (Bubalus bubalis), which is responsible for significant levels of human transmission. A transmission-blocking vaccine administered to buffaloes is a realistic option which would aid in the control of schistosomiasis. This will however require a better understanding of the immunobiology of schistosomiasis in naturally exposed buffaloes, particularly the immune response to migrating schistosome larvae, which are the likely targets of an anti-schistosome vaccine. To address this need we investigated the immune response at the major sites of larval migration, the skin and the lungs, in previously exposed and re-challenged water buffaloes. In the skin, a strong allergic-type inflammatory response occurred, characterised by leukocyte and eosinophil infiltration including the formation of granulocytic abscesses. Additionally at the local skin site, interleukin-5 transcript levels were elevated, while interleukin-10 levels decreased. In the skin-draining lymph node (LN) a predominant type-2 profile was seen in stimulated cells, while in contrast a type-1 profile was detected in the lung draining LN, and these responses occurred consecutively, reflecting the timing of parasite migration. The intense type-2 immune response at the site of cercarial penetration is significantly different to that seen in naive and permissive animal models such as mice, and suggests a possible mechanism for immunity. Preliminary data also suggest a reduced and delayed immune response occurred in buffaloes given high cercarial challenge doses compared with moderate infections, particularly in the skin. This study offers a deeper understanding into the immunobiology of schistosomiasis in a natural host, which may aid in the future design of more effective vaccines

  8. Generation of induced pluripotent stem cells from buffalo (Bubalus bubalis) fetal fibroblasts with buffalo defined factors.

    PubMed

    Deng, Yanfei; Liu, Qingyou; Luo, Chan; Chen, Shibei; Li, Xiangping; Wang, Caizhu; Liu, Zhenzhen; Lei, Xiaocan; Zhang, Huina; Sun, Hongliang; Lu, Fenghua; Jiang, Jianrong; Shi, Deshun

    2012-09-01

    Ectopically, expression of defined factors could reprogram mammalian somatic cells into induced pluripotent stem cells (iPSCs), which initiates a new strategy to obtain pluripotent stem cell lines. Attempts have been made to generate buffalo pluripotent stem cells by culturing primary germ cells or inner cell mass, but the efficiency is extremely low. Here, we report a successful method to reprogram buffalo fetal fibroblasts (BFFs) into pluripotent stem cells [buffalo induced pluripotent stem cell (biPSCs)] by transduction of buffalo defined factors (Oct4, Sox2, Klf4, and c-Myc) using retroviral vectors. The established biPSCs displayed typical morphological characteristics of pluripotent stem cells, normal karyotype, positive staining of alkaline phosphatase, and expressed pluripotent markers including Oct4, Sox2, Nanog, Lin28, E-Cadherin, SSEA-1, SSEA-4, TRA-1-81, STAT3, and FOXD3. They could form embryoid bodies (EBs) in vitro and teratomas after injecting into the nude BALB/C mice, and 3 germ layers were identified in the EBs and teratomas. Methylation assay revealed that the promoters of Oct4 and Nanog were hypomethylated in biPSCs compared with BFFs and pre-biPSCs, while the promoters of Sox2 and E-Cadherin were hypomethylated in both BFFs and biPSCs. Further, inhibiting p53 expression by coexpression of SV40 large T antigen and buffalo defined factors in BFFs or treating BFFs with p53 inhibitor pifithrin-a (PFT) could increase the efficiency of biPSCs generation up to 3-fold, and nuclear transfer embryos reconstructed with biPSCs could develop to blastocysts. These results indicate that BFFs can be reprogrammed into biPSCs by buffalo defined factors, and the generation efficiency of biPSCs can be increased by inhibition of p53 expression. These efforts will provide a feasible approach for investigating buffalo stem cell signal pathways, establishing buffalo stem cell lines, and producing genetic modification buffaloes in the future.

  9. Effect of bypass fat supplementation on productive performance and blood biochemical profile in lactating Murrah (Bubalus bubalis) buffaloes.

    PubMed

    Ranjan, Amit; Sahoo, Biswanath; Singh, Vijay Kumar; Srivastava, Susant; Singh, Suresh Pratap; Pattanaik, Ashok Kumar

    2012-10-01

    The study investigated the effect of dietary supplementation of bypass fat on productive performance and blood biochemical profile of lactating Murrah buffaloes (Bubalus bubalis). Fifteen multiparous buffaloes (2-4 lactation) of early to mid lactation were divided in three homogenous groups T(1) (control), T(2), and T(3) of five each. The animals in T(1) were fed with a basal diet consisting of a concentrate mixture, green sorghum, and wheat straw as per requirements, while the animals in group T(2) and T(3) were fed with same ration supplemented with 0.7 % (100 g/day) and 1.4 % (200 g/day) bypass fat (on dry matter intake (DMI) basis), respectively. The feed intake, milk yield, and milk composition were not influenced by supplemental bypass fat. However, fat-corrected milk (6.5 %) yield was higher (P < 0.05) in T(3) (14.21) than that of T(1) (9.83) and similar with T(2) (11.05). Feed efficiency (milk yield/kg DMI) was higher (P < 0.05) in group T(3) (0.51) than that of T(1) (0.38) and T(2) (0.41) indicating that buffaloes fed with bypass fat which is 1.4 % (200 g/day) of the diet were economically more efficient. The serum cholesterol level was higher (P < 0.01) in bypass fat-supplemented group (T(2) and T(3)) of animals. Serum high density lipoprotein (HDL) cholesterol (good cholesterol) level was more (P < 0.05) than LDL cholesterol (bad cholesterol) level with higher dose of bypass fat in T(3) than T(2). It was concluded that bypass fat supplementation with 1.4 % of the diet (200 g/day) increased the fat-corrected milk production and feed efficiency along with serum HDL cholesterol level in lactating Murrah buffaloes.

  10. Molecular cloning, sequencing and structural studies of granulocyte-macrophage colony-stimulating factor (GM-CSF) from Indian water buffalo (Bubalus bubalis).

    PubMed

    Sugumar, Thennarasu; Pugalenthi, Ganesan; Harishankar, Murugesan; Dhinakar Raj, G

    2014-02-01

    Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that is essential for growth and development of progenitors of granulocytes and monocytes/macrophages. In this study, we report molecular cloning, sequencing and characterization of GM-CSF from Indian water buffalo, Bubalus bubalis. In addition, we performed sequence and structural analysis for buffalo GM-CSF. Buffalo GM-CSF has been compared with 17 mammalian GM-CSFs using multiple sequence alignment and phylogenetic tree. Three-dimensional model for buffalo GM-CSF and human receptor complex was built using homology modelling to study cross-reactivity between two species. Detailed analysis was performed to study GM-CSF interface and various interactions at the interface. © 2013 John Wiley & Sons Ltd.

  11. DIGE based proteome analysis of mammary gland tissue in water buffalo (Bubalus bubalis): lactating vis-a-vis heifer.

    PubMed

    Jena, Manoj K; Janjanam, Jagadeesh; Naru, Jasmine; Kumar, Saravanan; Kumar, Sudarshan; Singh, Surender; Mohapatra, Sushil K; Kola, Srujana; Anand, Vijay; Jaswal, Shalini; Verma, Arvind K; Malakar, Dhruba; Dang, Ajay K; Kaushik, Jai K; Reddy, Vanga S; Mohanty, Ashok K

    2015-04-24

    Mammary gland is an exocrine and sebaceous gland made up of branching network of ducts that end in alveoli. Milk is synthesized in the alveoli and secreted into alveolar lumen. Mammary gland represents an ideal system for the study of organogenesis that undergoes successive cycles of pregnancy, lactation and involution. To gain insights on the molecular events that take place in pubertal and lactating mammary gland, we have identified 43 differentially expressed proteins in mammary tissue of heifer (non-lactating representing a virgin mammary gland), and lactating buffaloes (Bubalus bubalis) by 2D-difference gel electrophoresis (2D-DIGE) and mass spectrometry. Twenty one proteins were upregulated during lactation whereas 8 proteins were upregulated in heifer mammary gland significantly (p<0.05). Bioinformatics analyses of the identified proteins showed that a majority of the proteins are involved in metabolic processes. The differentially expressed proteins were validated by real-time PCR and Western blotting. We observed differential expressions of certain new proteins including EEF1D, HSPA5, HSPD1 and PRDX6 during lactation which have not been reported before. The differentially expressed proteins were mapped to available biological pathways and networks involved in lactation. This study signifies the importance of some proteins which are preferentially expressed during lactation and in heifer mammary gland. This work is important because we have generated information in water buffalo (B. bubalis) for the first time which is the major milk producing animal in Indian Subcontinent. Out of a present production of 133milliontons of milk produced in India, contribution of buffalo milk is around 54%. Its physiology is somewhat different from the lactating cows. Buffalo milk composition varies from cow milk in terms of higher fat and total solid content, which confers an advantage in preparation of specialized cheese, curd and other dairy products. Being a major milk

  12. Effect of reactive oxygen species on capacitation and associated protein tyrosine phosphorylation in buffalo (Bubalus bubalis) spermatozoa.

    PubMed

    Roy, S C; Atreja, S K

    2008-08-01

    In the present study, the effect of two particular reactive oxygen species (ROS), superoxide anion (O(2)(-)) and hydrogen peroxide (H(2)O(2)) on buffalo (Bubalus bubalis) sperm capacitation and associated protein tyrosine phosphorylation was studied. Ejaculated buffalo spermatozoa were suspended in sp-TALP medium at 50 x 10(6)/mL and incubated at 38.5 degrees C for 6h with or without heparin (10(g/mL; a positive control), or xanthine (X; 0.5mM)-xanthine oxidase (XO; 0.05 U/mL)-catalase (C; 2100 U/mL) system that generates O(2)(-) or NADPH (5mM) that stimulates the endogenous O(2)(-) production or H(2)O(2) (50 microM). The specific effect of O(2)(-), H(2)O(2) and NADPH on buffalo sperm capacitation and protein tyrosine phosphorylation was assessed by the addition of superoxide dismutase (SOD), catalase and diphenylene iodonium (DPI), respectively, to the incubation medium. Each of X+XO+C system, NADPH and H(2)O(2) induced a significantly higher percentage (P<0.05) of capacitation in buffalo spermatozoa compared to control. However, DPI inhibited this NADPH-induced capacitation and protein tyrosine phosphorylation and suggested for existence of an oxidase in buffalo spermatozoa. Using immunoblotting technique, at least seven tyrosine-phosphorylated proteins (20, 32, 38, 45, 49, 78 and 95 kDa) were detected in capacitated buffalo spermatozoa. Out of these, the tyrosine phosphorylation of p95 was induced extensively by both O(2)(-) as well as exogenous source of H(2)O(2) and using specific activators and inhibitors of signaling pathways, it was found this induction was regulated through a cAMP-dependent PKA pathway. Further, immunofluorescent localization study revealed that these ROS-induced tyrosine-phosphorylated proteins are mostly distributed in the midpiece and principal piece regions of the flagellum of capacitated spermatozoa and suggested for increased molecular activity in flagellum during capacitation. Thus, the study revealed that both O(2)(-) and H(2)O(2

  13. Production of recombinant buffalo (Bubalus bubalis) and goat (Capra hircus) growth hormones from genetically modified E. coli strains.

    PubMed

    Mukhopadhyay, Utpal Kumar; Sahni, Girish

    2002-08-28

    The growth hormone cDNAs of Indian reverine buffalo (Bubalus bubalis) and beetal goat (Capra hircus) were cloned in Escherichia coli through RT-PCR technique. Nucleotide sequencing revealed several silent mutations in both cDNAs and only one amino acid change in the case of goat when compared to reported bovine (Bos taurus) sequence. The high level expression of both the polypeptide hormones was achieved in E. coli (> or =30% of soluble intracellular proteins) through the construction of two-cistronic gene expression system. The solubilisation of recombinant growth hormones from inclusion bodies and subsequent oxidation to correctly folded monomeric form was also carried out. A combination of reverse-phase HPLC and non-reducing SDS-PAGE was successfully applied to distinguish between reduced and oxidised forms of growth hormones. A moderate yield ( approximately 40% of starting material, with potential for upscaling), two-step purification process comprising of hydrophobic interaction and ion-exchange chromatographies was developed. The process eliminates the need for costly, laborious and time-consuming steps of ultrafiltration and dialysis, as reported earlier for the purification of many recombinant animal growth hormones. The biophysical, biochemical and functional analyses of purified refolded polypeptides showed that the hormones produced in this study were identical to natural pituitary bovine growth hormone.

  14. Prevalence of antibodies against Bubaline herpesvirus (BuHV-1) among Mediterranean water buffalo (Bubalus bubalis) with implications in buffalo trade.

    PubMed

    Caruso, C; Prato, R; Ingravalle, F; Vecchio, D; Sciarra, A; Ternavasio, M; Ceccarelli, L; Martucciello, A; Galiero, G; De Carlo, E; Masoero, L

    2016-12-01

    Both Bovine herpesvirus (BoHV-1) and Bubaline herpesvirus (BuHV-1) have been reported to cross the species barrier. Antibody seroconversion in glycoprotein E (gE) blocking ELISA during BuHV-1 infection has been documented. Recent diagnostic efforts have focused on the development and application of discriminatory tests to distinguish between infections with BoHV-1 and BuHV-1. To evaluate the impact and distribution of these two infections in water buffalo farms in two regions (Piedmont (n = 3) and Campania (n = 10), Italy) where infectious bovine rhinotracheitis control programs have been implemented. Sampling was carried out on 13 buffalo farms comprising 1089 animals using specific gE-indirect ELISA's test able to discriminate among BoHV-1 and BuHV-1 infections. 59.0% of animals reacted positive to ELISA (irrespective of whether BoHV-1 or BuHV-1 antigen was used) and 86.4% of these were reactive to BuHV-1 only, whereas 11.8% showed absorbance values for both antigens and were classified as inconclusive. There was a statistically significant age-related difference in BuHV-1 infection rates but not in overall individual (47% vs. 58%) or herd prevalence (100% vs. 90%) of infection between the two regions. The low percentage of sera reactive to BoHV-1 (1.8%, 12/643) indicates that BuHV-1 may be the main circulating alphaherpesvirus infection in Mediterranean water buffalo in the two study areas. Since Bubalus bubalis is included in Directive 64/432/EEC on animal health problems affecting intra-community trade in bovine animals, diagnostic testing with nonspecific ELISA for BoHV-1 infection in buffalo may yield false-positive reactions. This scenario could lead to economic losses and hamper buffalo trade and movement, particularly for reproduction purposes.

  15. A diagnostic protocol to identify water buffaloes (Bubalus bubalis) vaccinated with Brucella abortus strain RB51 vaccine.

    PubMed

    Tittarelli, Manuela; Atzeni, Marcello; Calistri, Paolo; Di Giannatale, Elisabetta; Ferri, Nicola; Marchi, Enrico; Martucciello, Alessandra; De Massis, Fabrizio

    2015-01-01

    The use of live vaccine strain RB51 for vaccination of domestic water buffaloes (Bubalus bubalis) at risk of infection with Brucella abortus is permitted notwithstanding the plans for the eradication and only under strict veterinary control. The antibodies induced by RB51 vaccination are not detectable using conventional diagnostic techniques; therefore, it is necessary to have a specific diagnostic tool able to discriminate vaccinated from unvaccinated animals. The combination of a complement fixation test (CFT) with specific RB51 antigen (RB51-CFT) and a brucellin skin test has been demonstrated to be a reliable diagnostic system to identify single cattle (Bos taurus) vaccinated with RB51. So far, no data are available in the international scientific literature regarding the use of this test association in water buffalo. For this reason the suitability of this test combination has been evaluated in a water buffalo herd. One hundred twenty-seven animals farmed in a herd of Salerno province (Campania, Southern Italy), in the context of a presumptive unauthorized use of RB51 vaccine were chosen for this study. All tested animals resulted negative to Rose Bengal test (RBT) and complement fixation test (CFT) used for the detection of specific antibodies against Brucella field strains. Seventy-one animals (56%) developed RB51 antigen-specific CFT (RB51-CFT) antibodies against RB51 vaccine in a first sampling, while 104 animals (82%) gave positive result to a second serum sampling conducted 11 days after the intradermal inoculation of the RB51 brucellin. One hundred and seven animals (84%) showed a positive reaction to the RB51-CFT in at least 1 sampling, while 111 animals (87%) resulted positive to the RB51 brucellin skin test. Thus, analysing the results of the 3 testing in parallel, 119 animals (94%) were positive to at least 1 of the performed tests. The results suggest that the use in parallel of the RB51 brucellin skin test with RB51-CFT may represent a reliable

  16. Trehalose improves semen antioxidant enzymes activity, post-thaw quality, and fertility in Nili Ravi buffaloes (Bubalus bubalis).

    PubMed

    Iqbal, Sajid; Andrabi, Syed Murtaza Hassan; Riaz, Amjad; Durrani, Aneela Zameer; Ahmad, Nasim

    2016-03-15

    Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profile during cryopreservation (after dilution, before freezing, and after thawing), (2) in vitro quality (after thawing), and (3) in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Semen samples (n = 20) from four buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of trehalose (0.0, 15, 30, 45, and 60 mM) and frozen in French straws. At post dilution, profile of sperm catalase (U/mL) was higher (P < 0.05) in extenders containing 15, 30, and 45 mM of trehalose as compared to control. Although profiles of superoxide dismutase (U/mL) and total glutathione (μM) were higher (P < 0.05) in extenders containing 15 and 30 mM of trehalose as compared to control. At prefreezing, sperm catalase, superoxide dismutase, and total glutathione profiles were higher (P < 0.05) in all the treatment groups as compared to control. At post thawing, the profiles of catalase and total glutathione were higher (P < 0.05) in extender containing 30-mM trehalose as compared to other treatment groups and control. Whereas, profile of superoxide dismutase was higher (P < 0.05) in extenders containing 30, 45, and 60 mM of trehalose as compared to control and 15mM group. Post thaw total sperm motility (%) was higher (P < 0.05) in extender containing 30-mM trehalose as compared to control and 15 and 60-mM groups. Although sperm progressive motility (%), rapid velocity (%), average path velocity (μm/s), straight line velocity (μm/s), curvilinear velocity (μm/s), plasma membrane (structural and functional, %), acrosome (%), and DNA (%) integrity were higher (P < 0.05) in extender containing 30 mM trehalose as compared to other treatment groups and control. The fertility rates (61% vs. 43%) were higher (P < 0.05) in buffaloes inseminated with semen doses cryopreserved in extender containing 30 mM of trehalose than the control. It is

  17. Serological survey of Neospora caninum and Toxoplasma gondii in cattle (Bos indicus) and water buffaloes (Bubalus bubalis) in ten provinces of Brazil.

    PubMed

    da Silva, Jenevaldo Barbosa; Nicolino, Rafael Romero; Fagundes, Gisele Maria; Dos Anjos Bomjardim, Henrique; Dos Santos Belo Reis, Alessandra; da Silva Lima, Danillo Henrique; Oliveira, Carlos Magno Chaves; Barbosa, José Diomedes; da Fonseca, Adivaldo Henrique

    2017-06-01

    The objective of this study was to determine the prevalence of antibodies to Neospora caninum and Toxoplasma gondii among 500 cattle (Bos indicus) and 500 buffaloes (Bubalus bubalis) using the indirect fluorescent antibody test (IFAT) technique. Blood samples from were collected from water buffalo and cattle in 10 municipalities in the northern region of Brazil. The frequency of cattle and water buffaloes seropositive for Neospora caninum in Pará state, Brazil, was 55% and 44%, respectively, and the frequency of cattle and water buffaloes seropositive for Toxoplasma gondii was 52% and 39%, respectively. Seropositivity for both N. caninum and T. gondii was detected in 10.6% of the cattle samples and 14.8% of the buffalo samples. The frequency of cattle positive for N. caninum and T. gondii was significantly (p<0.05) higher than that of buffalo in two and three provinces, respectively. Buffaloes had a lower seroprevalence for N. caninum or T. gondii in all of the provinces studied. These results suggest that both species, when exposed to the same risks for N. caninum and T. gondii infection, have a high serological prevalence. Cattle showed a higher probability of being seropositive when exposed to the same risks for N. caninum and T. gondii. Our study, which included an extensive number of blood samples, provides important epidemiological information pertinent to buffalo production in tropical countries that can be used as a basis for disease-management practices in Latin America. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Glutathione-supplemented tris-citric acid extender improves the post-thaw quality and in vivo fertility of buffalo (Bubalus bubalis) bull spermatozoa.

    PubMed

    Ansari, Muhammad S; Rakha, Bushra A; Andrabi, Syed M H; Ullah, Nemat; Iqbal, Razia; Holt, William V; Akhter, Shamim

    2012-11-01

    In this study we evaluated the effects of semen extender supplementation with different concentrations of glutathione (GSH) on buffalo (Bubalus bubalis) bull sperm motility, plasma membrane integrity, viability and DNA integrity as well as in vivo fertility. Semen from three Nili-Ravi buffalo bulls was collected, and qualified semen ejaculates (n=18) were split into five aliquots for dilution (37°C; 50×10(6)spermatozoaml(-1)) with experimental tris-citric acid extender containing 0, 0.5, 1.0, 1.5 or 2.0 mM GSH. Extended semen was cooled to 4°C, equilibrated and filled in French straws. The straws were kept on liquid nitrogen vapors (5 cm above the LN(2) level) for 10 min and plunged in liquid nitrogen for storage. Sperm motility (%), plasma membrane integrity (%), viability (%) and DNA integrity (%) were assessed at 0, 2 and 4h post-thawing (37°C). Extender supplementation with GSH (0.5, 1.0, 1.5 and 2.0 mM) increased sperm motility, plasma membrane integrity and viability in a dose dependent manner. Sperm DNA integrity was higher (p<0.05) in all experimental extenders containing GSH when compared to the control extender (0 mM GSH). The in vivo fertility rate of cryopreserved buffalo bull (n=2) spermatozoa was higher (p<0.05) in extender containing 2.0 mM GSH compared to that of control. In summary, tris-citric acid extender supplemented with glutathione improved the freezability of buffalo bull spermatozoa in a dose dependant manner. Moreover, the addition of 2.0 mM GSH to the extender enhanced the in vivo fertility of buffalo (Bubalus bubalis) bull spermatozoa. Copyright © 2012. Published by Elsevier Urban & Partner Sp. z o.o.

  19. Organization and differential expression of the GACA/GATA tagged somatic and spermatozoal transcriptomes in Buffalo Bubalus bubalis

    PubMed Central

    Srivastava, Jyoti; Premi, Sanjay; Kumar, Sudhir; Ali, Sher

    2008-01-01

    Background Simple sequence repeats (SSRs) of GACA/GATA have been implicated with differentiation of sex-chromosomes and speciation. However, the organization of these repeats within genomes and transcriptomes, even in the best characterized organisms including human, remains unclear. The main objective of this study was to explore the buffalo transcriptome for its association with GACA/GATA repeats, and study the structural organization and differential expression of the GACA/GATA repeat tagged transcripts. Moreover, the distribution of GACA and GATA repeats in the prokaryotic and eukaryotic genomes was studied to highlight their significance in genome evolution. Results We explored several genomes and transcriptomes, and observed total absence of these repeats in the prokaryotes, with their gradual accumulation in higher eukaryotes. Further, employing novel microsatellite associated sequence amplification (MASA) approach using varying length oligos based on GACA and GATA repeats; we identified and characterized 44 types of known and novel mRNA transcripts tagged with these repeats from different somatic tissues, gonads and spermatozoa of water buffalo Bubalus bubalis. GACA was found to be associated with higher number of transcripts compared to that with GATA. Exclusive presence of several GACA-tagged transcripts in a tissue or spermatozoa, and absence of the GATA-tagged ones in lung/heart highlights their tissue-specific significance. Of all the GACA/GATA tagged transcripts, ~30% demonstrated inter-tissue and/or tissue-spermatozoal sequence polymorphisms. Significantly, ~60% of the GACA-tagged and all the GATA-tagged transcripts showed highest or unique expression in the testis and/or spermatozoa. Moreover, ~75% GACA-tagged and all the GATA-tagged transcripts were found to be conserved across the species. Conclusion Present study is a pioneer attempt exploring GACA/GATA tagged transcriptome in any mammalian species highlighting their tissue, stage and species

  20. Alginate encapsulation preserves the quality and fertilizing ability of Mediterranean Italian water buffalo (Bubalus bubalis) and Holstein Friesian (Bos taurus) spermatozoa after cryopreservation.

    PubMed

    Perteghella, Sara; Gaviraghi, Alessandro; Cenadelli, Silvia; Bornaghi, Valeria; Galli, Andrea; Crivelli, Barbara; Vigani, Barbara; Vigo, Daniele; Chlapanidas, Theodora; Faustini, Massimo; Torre, Maria Luisa

    2017-03-30

    The use of artificial insemination (AI) in buffalo (Bubalus bubalis) is limited by poor ovarian activity during the hot season, seasonal qualitative patterns in semen, low resistance of sperm cells in the female tract, difficulties in estrus detection, and variable estrus duration. Although AI procedures are commonly used in bovine, use of AI has been limited in buffalo. In the zootechnical field, different studies have been conducted to develop techniques for improvement of fertilizing ability of buffalo spermatozoa after AI. In this study, for the first time, the use of alginate encapsulation and cryopreservation of buffalo spermatozoa is described, and the same procedure was performed with Holstein Friesian (Bos taurus) semen. Results obtained from in vitro analyses indicate that the encapsulation process does not have detrimental effects (compared to controls) on quality parameters (membrane integrity, progressive motility, path average velocity) in either species. Similarly, there were no detrimental effects after cryopreservation in either species. The fertilizing potential of encapsulated and cryopreserved semen was evaluated after AI in 25 buffalo and 113 bovine females. Pregnancy rates were not affected in either species. The results of this study show proof of concept for the use of frozen semen controlled-release devices in buffalo.

  1. Alginate encapsulation preserves the quality and fertilizing ability of Mediterranean Italian water buffalo (Bubalus bubalis) and Holstein Friesian (Bos taurus) spermatozoa after cryopreservation

    PubMed Central

    Perteghella, Sara; Gaviraghi, Alessandro; Cenadelli, Silvia; Bornaghi, Valeria; Galli, Andrea; Crivelli, Barbara; Vigani, Barbara; Vigo, Daniele; Faustini, Massimo; Torre, Maria Luisa

    2017-01-01

    The use of artificial insemination (AI) in buffalo (Bubalus bubalis) is limited by poor ovarian activity during the hot season, seasonal qualitative patterns in semen, low resistance of sperm cells in the female tract, difficulties in estrus detection, and variable estrus duration. Although AI procedures are commonly used in bovine, use of AI has been limited in buffalo. In the zootechnical field, different studies have been conducted to develop techniques for improvement of fertilizing ability of buffalo spermatozoa after AI. In this study, for the first time, the use of alginate encapsulation and cryopreservation of buffalo spermatozoa is described, and the same procedure was performed with Holstein Friesian (Bos taurus) semen. Results obtained from in vitro analyses indicate that the encapsulation process does not have detrimental effects (compared to controls) on quality parameters (membrane integrity, progressive motility, path average velocity) in either species. Similarly, there were no detrimental effects after cryopreservation in either species. The fertilizing potential of encapsulated and cryopreserved semen was evaluated after AI in 25 buffalo and 113 bovine females. Pregnancy rates were not affected in either species. The results of this study show proof of concept for the use of frozen semen controlled-release devices in buffalo. PMID:27456772

  2. Ultrasonographic and endocrine aspects of follicle deviation, and acquisition of ovulatory capacity in buffalo (Bubalus bubalis) heifers.

    PubMed

    Gimenes, L U; Carvalho, N A T; Sá Filho, M F; Vannucci, F S; Torres-Júnior, J R S; Ayres, H; Ferreira, R M; Trinca, L A; Sartorelli, E S; Barros, C M; Beltran, M P; Nogueira, G P; Mapletoft, R J; Baruselli, P S

    2011-02-01

    The objectives of this study were to determine the interval from ovulation to deviation and the diameter of the dominant (DF) and largest subordinate (SF) follicles at deviation in buffalo (Bubalus bubalis) heifers. Two methods of evaluation (observed vs. calculated) were used. FSH and LH profiles encompassing follicle deviation (Experiment 1), and the follicular diameter when the DF acquired ovulatory capacity (Experiment 2) were also determined. The time of deviation and the diameter of the DF and the largest SF at deviation did not differ between observed and calculated methods. Overall, follicle deviation occurred 2.6 ± 0.2d (mean ± SEM) after ovulation, and the diameters of the DF and SF at deviation were 7.2 ± 0.2 and 6.4 ± 0.2mm, respectively. No changes in plasma levels of FSH or LH were observed (P=0.32 and P=0.96, respectively). Experiment 2 was conducted in two phases according to the diameter of the DF during the first wave of follicular development at the time of LH challenge (25mg of pLH). In the first phase, follicles ranging from 5.0 to 6.0mm (n=7), 6.1 to 7.0mm (n=11), or 7.1 to 8.0mm (n=9) were used, and in the second phase, follicles ranging from 7.0 to 8.4mm (n=10), 8.5 to 10.0mm (n=10), or 10.1 to 12.0mm (n=9) of diameter were used. After the pLH treatment, the DF was monitored by ultrasonography every 12h for 48h. No ovulations occurred in heifers in the first phase. However, in the second phase, an effect of follicular diameter was observed on ovulation rate [7.0-8.4mm (0.0%, 0/10), 8.5-10.0mm (50.0%, 5/10), and 10.0-12.0mm (55.6%, 5/9)]. In summary, follicle deviation occurred 2.6d after ovulation in buffalo (B. bubalis) heifers, when the diameters of the DF and SF were 7.2 and 6.4mm, respectively. No significant changes in plasma concentrations of FSH or LH were detected. Finally, the acquisition of ovulatory capacity occurred when the DF reached 8.5mm in diameter.

  3. Identity of Sarcocystis species of the water buffalo (Bubalus bubalis) and cattle (Bos taurus) and the suppression of Sarcocystis sinensis as a nomen nudum.

    PubMed

    Dubey, J P; Fayer, R; Rosenthal, B M; Calero-Bernal, R; Uggla, A

    2014-09-15

    There are uncertainties concerning the identity and host species specificity of Sarcocystis species of the water buffalo (Bubalus bubalis) and cattle (Bos taurus). Currently, in cattle three species are recognized with known endogenous stages, viz.: S. cruzi (with canine definitive host), S. hirsuta (feline definitive host), and S. hominis (primate definitive host). Recently, a fourth Sarcocystis species with an unknown life cycle has been reported from cattle. In the water buffalo, four species of Sarcocystis have been described: S. fusiformis (feline definitive host), S. buffalonis (feline definitive host), S. levinei (canine definitive host), and S. dubeyi (definitive host unknown but not cat or dog). Besides, there are studies of Sarcocystis infections in buffalo and cattle from China with results that are difficult to interpret and validate. For example, some of the studies report transmission of Sarcocystis species between cattle and buffalo, but steps to preclude exogenous exposures were not reported. A species of the water buffalo, 'S. sinensis', was proposed at a Chinese national conference in 1990, and published as an abstract without figures and with no archived type specimens for verification. The International Code of Zoological Nomenclature Articles 9 and 10 state that "abstracts of articles, papers, posters, text of lectures, and similar material when issued primarily to participants at meetings, symposia, colloquia or congress does not constitute published work"; therefore, S. sinensis is a nomen nudum. Published by Elsevier B.V.

  4. Design and validation of a 90K SNP genotyping assay for the water buffalo (Bubalus bubalis).

    PubMed

    Iamartino, Daniela; Nicolazzi, Ezequiel L; Van Tassell, Curtis P; Reecy, James M; Fritz-Waters, Eric R; Koltes, James E; Biffani, Stefano; Sonstegard, Tad S; Schroeder, Steven G; Ajmone-Marsan, Paolo; Negrini, Riccardo; Pasquariello, Rolando; Ramelli, Paola; Coletta, Angelo; Garcia, José F; Ali, Ahmad; Ramunno, Luigi; Cosenza, Gianfranco; de Oliveira, Denise A A; Drummond, Marcela G; Bastianetto, Eduardo; Davassi, Alessandro; Pirani, Ali; Brew, Fiona; Williams, John L

    2017-01-01

    The availability of the bovine genome sequence and SNP panels has improved various genomic analyses, from exploring genetic diversity to aiding genetic selection. However, few of the SNP on the bovine chips are polymorphic in buffalo, therefore a panel of single nucleotide DNA markers exclusive for buffalo was necessary for molecular genetic analyses and to develop genomic selection approaches for water buffalo. The creation of a 90K SNP panel for river buffalo and testing in a genome wide association study for milk production is described here. The genomes of 73 buffaloes of 4 different breeds were sequenced and aligned against the bovine genome, which facilitated the identification of 22 million of sequence variants among the buffalo genomes. Based on frequencies of variants within and among buffalo breeds, and their distribution across the genome, inferred from the bovine genome sequence, 90,000 putative single nucleotide polymorphisms were selected to create an Axiom® Buffalo Genotyping Array 90K. This 90K "SNP-Chip" was tested in several river buffalo populations and found to have ∼70% high quality and polymorphic SNPs. Of the 90K SNPs about 24K were also found to be polymorphic in swamp buffalo. The SNP chip was used to investigate the structure of buffalo populations, and could distinguish buffalo from different farms. A Genome Wide Association Study identified genomic regions on 5 chromosomes putatively involved in milk production. The 90K buffalo SNP chip described here is suitable for the analysis of the genomes of river buffalo breeds, and could be used for genetic diversity studies and potentially as a starting point for genome-assisted selection programmes. This SNP Chip could also be used to analyse swamp buffalo, but many loci are not informative and creation of a revised SNP set specific for swamp buffalo would be advised.

  5. The complete coding region sequence of river buffalo (Bubalus bubalis) SRY gene.

    PubMed

    Parma, Pietro; Feligini, Maria; Greppi, Gianfranco; Enne, Giuseppe

    2004-02-01

    The Y-linked SRY gene is responsible for testis determination in mammals. Mutations in this gene can lead to XY Gonadal Dysgenesis, an abnormal sexual phenotype described in humans, cattle, horses and river buffalo. We report here the complete river buffalo SRY sequence in order to enable the genetic diagnosis of this disease. The SRY sequence was also used to confirm the evolutionary divergence time between cattle and river buffalo 10 million years ago.

  6. G-Protein-Coupled Chemokine Receptor Gene in Lumpy Skin Disease Virus Isolates from Cattle and Water Buffalo (Bubalus bubalis) in Egypt.

    PubMed

    El-Tholoth, M; El-Kenawy, A A

    2016-12-01

    Lumpy skin disease virus (LSDV), sheep poxvirus (SPV) and goat poxvirus (GPV) are the most serious poxviruses of ruminants. In this study, we analysed the G-protein-coupled chemokine receptor (GPCR) genes of LSDV isolates from cattle and water buffalo (Bubalus bubalis) in Egypt during the summer of 2011. Multiple alignments of the nucleotide sequences revealed that the water buffalo LSDV isolate differed from the cattle isolate at four nucleotide positions, and both isolates had nine nucleotide mutations from the reference strain, Egyptian tissue culture-adapted cattle LSDV/Ismailyia88 strain. Compared with the GPCR sequences of SPV and GPV strains, a 21 nucleotide insertion and a 12 nucleotide deletion were identified in the GPCR genes of our used isolates and other LSDVs. The amino acid sequences of GPCR genes of our isolates contained the unique signature of LSDV (A11 , T12 , T34 , S99 and P199 ). Phylogenetic analyses showed that the GPCR genes of cattle and water buffalo LSDVs were closest genetically, indicating a potential transmission of cattle LSDV to water buffalo. © 2015 Blackwell Verlag GmbH.

  7. Some qualitative and chromatic aspects of thawed buffalo (Bubalus bubalis) meat.

    PubMed

    Tateo, A; De Palo, P; Quaglia, N C; Centoducati, P

    2007-06-01

    After thawing, the meat of beef calves (Italian Frisian breed) and buffalo calves (Mediterranean breed) slaughtered at 4, 8 and 12 months of age was examined. Both the pH and the thawing loss confirmed that the meat of buffalo calves is more suitable for preservation by freezing. With increased age and time of exposure to air the lightness of the non-renewed surface was reduced. The lightness of the fresh cut surface remained stable in the various thawing phases though it was less in the older animals. The a(∗) index increased with animal age but decreased during the 4 days post-thawing. The fresh cut surface of buffalo meat from calves slaughtered at 4 and 8 months was not darker than beef slaughtered at the same age. On the contrary at 12 months of age, the buffalo meat had a lower redness index than beef and a higher haematin concentration.

  8. Somatic cell cloning in Buffalo (Bubalus bubalis): effects of interspecies cytoplasmic recipients and activation procedures.

    PubMed

    Kitiyanant, Y; Saikhun, J; Chaisalee, B; White, K L; Pavasuthipaisit, K

    2001-01-01

    Successful nuclear transfer (NT) of somatic cell nuclei from various mammalian species to enucleated bovine oocytes provides a universal cytoplast for NT in endangered or extinct species. Buffalo fetal fibroblasts were isolated from a day 40 fetus and were synchronized in presumptive G(0) by serum deprivation. Buffalo and bovine oocytes from abattoir ovaries were matured in vitro and enucleated at 22 h. In the first experiment, we compared the ability of buffalo and bovine oocyte cytoplasm to support in vitro development of NT embryos produced by buffalo fetal fibroblasts as donor nuclei. There were no significant differences (p > 0.05) between the NT embryos derived from buffalo and bovine oocytes, in fusion (74% versus 71%) and cleavage (77% versus 75%) rates, respectively. No significant differences were also observed in blastocyst development (39% versus 33%) and the mean cell numbers of day 7 cloned blastocysts (88.5 +/- 25.7 versus 51.7 +/- 5.4). In the second experiment, we evaluated the effects of activation with calcium ionophore A23187 on development of NT embryos after electrical fusion. A significantly higher (p < 0.05) percentage of blastocyst development was observed in the NT embryos activated by calcium ionophore and 6-DMAP when compared with 6-DMAP alone (33% versus 17%). The results indicate that the somatic nuclei from buffalo can be reprogrammed after transfer to enucleated bovine oocytes, resulting in the production of cloned buffalo blastocysts similar to those transferred into buffalo oocytes. Calcium ionophore used in conjunction with 6-DMAP effectively induces NT embryo development.

  9. Molecular cloning and expression analysis of the STAT1 gene in the water buffalo (Bubalus bubalis).

    PubMed

    Deng, Tingxian; Pang, Chunying; Zhu, Peng; Liao, Biyun; Zhang, Ming; Yang, Bingzhuang; Liang, Xianwei

    2015-01-01

    Signal transducer and activator of transcription 1 (STAT1) is a critical component of the transcription factor complex in the interferon (IFN) signaling pathways. Of the seven STAT isoforms, STAT1 is a key mediator of type I and type III IFN signaling, but limited information is available for the STAT genes in the water buffalo. Here, we amplified and identified the complete coding sequence (CDS) of the buffalo STAT1 gene by using reverse transcription polymerase chain reaction (RT-PCR). Sequence analysis indicated that the buffalo STAT1 gene length size was 3437 bp, containing an open reading frame (ORF) of 2244 bp that encoded 747 amino acids for the first time. The buffalo STAT1 CDS showed 99, 98, 89, 93, 86, 85, and 87% identity with that of Bos taurus, Ovis aries, Homo sapiens, Sus scrofa, Rattus norvegicus, Mus musculus, and Capra hircus. The phylogenetic analyses revealed that the nearest relationship existed between the water buffalo and B. taurus. The STAT1 gene was ubiquitously expressed in 11 buffalo tissues by real-time PCR, whereas STAT1 was expressed at higher levels in the lymph. The STAT1 gene contained five targeted microRNA sequences compared with the B. taurus by the miRBase software that provide a fundamental for identifying the STAT1 gene function.

  10. Demonstration of alternative and classical complement pathway activity in colostrum from buffalo (Bubalus bubalis).

    PubMed

    Matheswaran, K; Dhinakar Raj, G; Nachimuthu, K

    2003-09-01

    Buffalo colostrum caused lysis of unsensitized red blood cells (RBC) from sheep, goats, rabbits and chickens. RBC from cattle and buffalo were resistant to lysis. That lysis was due to the presence of natural antibodies to these RBC was ruled out since there was no reduction in haemolytic titres even after adsorption with the respective RBC. The addition of EGTA to the diluent had no effect on the haemolytic activity. These findings indicate the presence of alternative complement pathway (ACP) activity in buffalo colostrum. The haemolytic activity of buffalo complement for unsensitized rabbit RBC was reduced to very low levels by heating at 50 degrees C for 45 min. Treatment with zymosan also inhibited the haemolytic activity, while inulin had no effect. The maximum activity of ACP occurred in the presence of 4 mmol/L Mg(2+) in the diluent. The range of ACP activities in colostrum from buffaloes varied from 4.06 to 8.48 CH50 units/ml. Using a standard system for titrating the classical complement pathway and rabbit red blood cells sensitized with goat haemolysin, the range of complement activity in buffalo colostrum was 4.81-6.77 CH50/ml.

  11. Molecular characterization of oxytocin receptor gene in water buffalo (Bubalus bubalis).

    PubMed

    Arunmozhi, N; Singh, S K; Sarath, T; Agarwal, S K; Doiphode, A; Shankar, U

    2014-10-01

    Buffaloes are known for their productivity as compared to average yielding cows due to higher fat percentage, better feed conversion ability and disease resistance. On the other hand, the reproductive performances of buffaloes are often considered as poor owing to late sexual maturity, weak/silent oestrus, repeat breeder and prolonged intercalving interval. The study of cascade of events during oestrus and oestrous cycle can be useful for the improvement of reproductive efficiency of buffaloes. More precisely, the hormonal changes initiated at the molecular level within the animal determine the reproductive nature of the species. Nucleotide/protein sequence analysis serves as a vital tool in analysing the binding of the hormones for their effect or functions. In this study, we have reported cloning and characterization of the complete coding (cDNA) sequence of oxytocin receptor gene (OXTR) in buffaloes. Buffalo OXTR gene contains an uninterrupted ORF of 1176 nucleotides corresponding to an inferred polypeptide length of 391 amino acids (aa). The molecular weight of the deduced aa sequence was found to be 43 kDa with an isoelectric point of 9.253 and 16.328 charge at pH 7.0. The deduced protein sequence consists of 38 strongly basic (+) (K,R), 22 strongly acidic (-) (D,E), 186 hydrophobic (A, I, L, F, W, V) and 95 Polar (N, C, Q, S, T, Y) aa. Results indicated that aspartate (D) at aa position 85 and D, R and C at aa positions 136, 137 and 138, respectively, are conserved in buffaloes. The buffalo OXTR gene shared a per cent similarity ranging from 84.7 to 98.1 and 88.5 to 97.7 at nucleotide and deduced aa sequence levels, respectively, with that of other species. Phylogram constructed on the basis of either nucleotide or deduced aa sequences of buffalo OXTR gene showed that buffalo, cattle and sheep have diverged from human and swine and formed a separate clad. The buffalo sequence has shown maximum similarity and closeness with cattle followed by sheep both at

  12. Cryogenic changes in proteases and antiprotease activities of buffalo (Bubalus bubalis) and cattle (Bos taurus) semen.

    PubMed

    Gurupriya, Vijayasaraswathy S; Divyashree, Bannur C; Roy, Sudhir C

    2014-02-01

    The postthaw motility and fertility of buffalo and cattle semen is reduced when they are cryopreserved for artificial insemination. In the present study, an attempt was made to characterize the cryogenic changes in proteases and antiprotease activities (APA) of buffalo and cattle semen because these proteolysis regulators have been reported to be associated with sperm motility and fertility. Buffalo sperm demonstrated at least two major proteases of 45 and 42 kDa and three minor proteases of 95, 52, and 33 kDa. Similarly, cattle sperm demonstrated three major proteases of 62, 45, and 42 kDa and two minor proteases of 85 and 78 kDa. Buffalo seminal plasma demonstrated at least three major proteases of 78, 68, and 62 kDa and one minor protease of 98 kDa and cattle seminal plasma demonstrated one major protease of 68 kDa and two minor proteases of 78 and 75 kDa. Except for the 45 kDa protease, most of the previously mentioned proteases were found to be metalloproteinases. Compared with fresh sperm, cryopreserved buffalo and cattle sperm demonstrated a major protease band of 52/49 kDa and the activity of this protease reduced progressively with the duration of cryopreservation. On the contrary, compared with the fresh seminal plasma, cryopreserved buffalo and cattle semen extenders displayed the presence of a new protease band of 45 kDa and demonstrated that this protease activity was leaked from buffalo and cattle cryopreserved spermatozoa. Buffalo and cattle seminal plasmas displayed at least two major APA of 86 and 26 kDa. Compared with buffalo, cattle seminal plasma demonstrated significantly greater APA. Thus, the present study demonstrated the presence of an array of proteases and APA in buffalo and cattle semen and the activities of which changed during cryopreservation. The leakage of the specific protease activity and changes in the proteases and APA might be attributed to reduced motility and fertility of cryopreserved semen in these species. Copyright

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

    PubMed

    Presicce, G A; Senatore, E M; De Santis, G; Bella, A

    2005-10-01

    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

  14. Establishment and Characterization of a Buffalo (Bubalus bubalis) Mammary Epithelial Cell Line

    PubMed Central

    Anand, Vijay; Dogra, Nilambra; Singh, Surender; Kumar, Sudarshan N.; Jena, Manoj K.; Malakar, Dhruba; Dang, Ajay K.; Mishra, Bishnu P.; Mukhopadhyay, Tapas K.; Kaushik, Jai K.; Mohanty, Ashok K.

    2012-01-01

    Background The objective of this study was to establish the buffalo mammary epithelial cell line (BuMEC) and characterize its mammary specific functions. Methodology Buffalo mammary tissue collected from the slaughter house was processed enzymatically to obtain a heterogenous population of cells containing both epithelial and fibroblasts cells. Epithelial cells were purified by selective trypsinization and were grown in a plastic substratum. The purified mammary epithelial cells (MECs) after several passages were characterized for mammary specific functions by immunocytochemistry, RT-PCR and western blot. Principal Findings The established buffalo mammary epithelial cell line (BuMEC) exhibited epithelial cell characteristics by immunostaining positively with cytokeratin 18 and negatively with vimentin. The BuMEC maintained the characteristics of its functional differentiation by expression of β-casein, κ-casein, butyrophilin and lactoferrin. BuMEC had normal growth properties and maintained diploid chromosome number (2n = 50) before and after cryopreservation. A spontaneously immortalized buffalo mammary epithelial cell line was established after 20 passages and was continuously subcultured for more than 60 passages without senescence. Conclusions We have established a buffalo mammary epithelial cell line that can be used as a model system for studying mammary gland functions. PMID:22792341

  15. Molecular detection of bovine immunodeficiency virus in water buffaloes (Bubalus bubalis) from the Amazon region, Brazil.

    PubMed

    Albernaz, Tatiane Teles; Leite, Rômulo Cerqueira; Reis, Jenner Karlison Pimenta; de Sousa Rodrigues, Ana Paula; da Cunha Kassar, Telissa; Resende, Claudia Fideles; de Oliveira, Cairo Henrique Sousa; Silva, Rafaela das Mercês; Salvarani, Felipe Masiero; Barbosa, José Diomedes

    2015-12-01

    Bovine immunodeficiency is a chronic progressive disease caused by a lentivirus that affects cattle and buffaloes. Although the infection has been described in cattle in some countries, including in Brazil, there are only two reports of infection in buffaloes: one in Pakistan and one in Cambodia. The aim of the present study was to survey the occurrence of bovine immunodeficiency virus (BIV) in water buffaloes from the Amazon region, Pará state, Brazil. BIV proviral DNA was surveyed in 607 whole blood samples of water buffaloes from 10 farms located in the state of Pará using semi-nested polymerase chain reaction (PCR) (PCR-SN) to amplify the pol region of the viral genome. Of the 607 samples tested, 27 (4.4 %) were positive for BIV proviral DNA. The amplified fragments were confirmed by sequence analysis after cloning and nucleotide sequencing. The sequence obtained had 99 % similarity to the reference strain (R-29). The present study provides important epidemiological data because BIV was detected for the first time in water buffaloes in Brazil. Further, the results suggest the possibility of the virus being a risk factor for herd health because it may be a potential causal agent of chronic disease and, also may be associated to other infectious diseases.

  16. High nucleotide and amino acid sequence similarities in tumour necrosis factor-alpha amongst Indian buffalo (Bubalus bubalis), Indian cattle (Bos indicus) and other ruminants.

    PubMed

    Gupta, P K; Bind, R B; Walunj, S S; Saini, M

    2004-08-01

    Tumour necrosis factor-alpha (TNF-alpha) mRNA from Indian water buffalo (Bubalus bubalis) and Indian cattle (Bos indicus) was reverse transcribed and amplified using reverse transcriptase-polymerase chain reaction (RT-PCR). The nucleotide sequences of cDNAs were determined after cloning into pGEM-T-Easy vector (Promega, Madison, WI) and compared with reported nucleotide sequences of TNF-alpha cDNA from other species. The nucleotide sequences of TNF-alpha from Indian cattle revealed significantly high similarities at nucleotide (99.2%) and amino acid (100%) levels with those of cattle (Bos taurus; Zebu). The sequences from buffalo had 98.4% nucleotide and 99.1% amino acid similarities with Indian cattle, indicating functional cross-reactivity. One amino acid deletion at position 63 and one substitution (A-->P) at position 64 were observed in buffalo compared with Indian cattle. The amino acid deletion at position 63 was predicted due to differences in pre-mRNA splicing.

  17. Pheno- and genotyping of Brucella abortus biovar 5 isolated from a water buffalo (Bubalus bubalis) fetus: First case reported in the Americas.

    PubMed

    Martínez, Diana; Thompson, Carolina; Draghi, Graciela; Canavesio, Vilma; Jacobo, Roberto; Zimmer, Patricia; Elena, Sebastián; Nicola, Ana M; de Echaide, Susana Torioni

    2014-09-17

    An isolate of Brucella spp. from an aborted water buffalo (Bubalus bubalis) fetus was characterized based on its pheno- and genotype. The phenotype was defined by carbon dioxide requirement, hydrogen sulfide production, sensitivity to thionin and basic fuchsin and agglutination with Brucella A and M monospecific antisera. The genotype was based on the amplification of the following genes: bcsp31, omp2ab, and eri and the species-specific localization of the insertion sequence IS711 in the Brucella chromosome via B. abortus-B. melitensis-B. ovis-B. suis (AMOS)-PCR. Unexpectedly, the isolate showed a phenotype different from B. abortus bv 1, the most prevalent strain in cattle in Argentina, and from vaccine strain 19, currently used in bovines and water buffaloes. Genotyping supported the phenotypic results, as the analysis of the omp2ab gene sequence showed an identical pattern to either B. abortus bv 5 or B. melitensis. Finally, the AMOS PCR generated a 1700-bp fragment from the isolate, different than those amplified from B. abortus bv 1 (498bp) and B. melitensis (731bp), confirming the presence of B. abortus bv 5. The OIE/FAO Reference Laboratory for Brucellosis confirmed this typing. This is the first report of B. abortus bv 5 from a water buffalo in the Americas.

  18. Detection of pseudocowpox virus in water buffalo (Bubalus bubalis) with vesicular disease in the state of São Paulo, Brazil, in 2016.

    PubMed

    Laguardia-Nascimento, Mateus; de Oliveira, Ana Paula Ferreira; Fernandes, Fernanda Rodas Pires; Rivetti, Anselmo Vasconcelos; Camargos, Marcelo Fernandes; Fonseca Júnior, Antônio Augusto

    2017-12-01

    Parapoxviruses are zoonotic viruses that infect cattle, goats and sheep; there have also been reports of infections in camels, domestic cats and seals. The objective of this report was to describe a case of vesicular disease caused by pseudocowpox virus (PCPV) in water buffalo (Bubalus bubalis) in Brazil. Sixty buffalo less than 6 months old exhibited ulcers and widespread peeling of the tongue epithelium. There were no cases of vesicular disease in pigs or horses on the same property. Samples were analysed by PCR and sequencing. Phylogenetic analysis in MEGA 7.01 was reconstructed using major envelope protein (B2L) by the Tamura three-parameter nucleotide substitution model and the maximum likelihood and neighbor joining models, both with 1000 bootstrap replicates. The genetic distance between the groups was analysed in MEGA using the maximum composite likelihood model. The rate variation among sites was modeled using gamma distribution. The presence of PCPV in the buffalo herd could be demonstrated in epithelium and serum. The minimum genetic distance between the isolated PCPV strain (262-2016) and orf virus and bovine papular stomatitis virus was 6.7% and 18.4%, respectively. The maximum genetic distance calculated was 4.6% when compared with a PCPV detected in a camel. Conclusions/Clinical Importance: The peculiar position of the isolated strain in the phylogenetic trees does not necessarily indicate a different kind of PCPV that infects buffalo. More samples from cattle and buffalo in Brazil must be sequenced and compared to verify if PCPV from buffalo are genetically different from samples derived from cattle.

  19. Clinical, haematological and therapeutic studies on tropical theileriosis in water buffaloes (Bubalus bubalis) in Egypt.

    PubMed

    Osman, Salama A; Al-Gaabary, Magdy H

    2007-05-31

    Thirty buffaloes naturally infected with Theileria annulata and 10 parasitologically free controls were used to determine the potential clinical, haematological and therapeutic impact of tropical theileriosis in Egypt. The clinical signs in the infected buffaloes were pyrexia (40.5-41.5 degrees C), enlargement of superficial lymph nodes, slight nasal and ocular discharges, salivation, anaemia and respiratory distress. Eye lesions also were recorded. There was a significant decrease in erythrocyte counts and haemoglobin content and a significant decrease in total leucocyte counts in infected buffaloes compared to controls. Early treatment with buparvaquone was 100% effective in eliminating the protozoan parasites from the blood and lymph nodes and led to an improvement in the clinical state whereas treatment in the later stages of the disease whilst eliminating the parasites failed to improve the clinical condition of the animal.

  20. Soya-lecithin in extender improves the freezability and fertility of buffalo (Bubalus bubalis) bull spermatozoa.

    PubMed

    Akhter, S; Ansari, M S; Andrabi, S M H; Rakha, B A; Ullah, N; Khalid, M

    2012-10-01

    Egg yolk is routinely used as a cryoprotectant in semen extenders. However, it may contain cryoprotective antagonists, and there are hygienic risks associated with its use. Proteins of plant origin, like soya-lecithin, lack these hazards. The aim of this study was to use soya-lecithin as a cryoprotectant in extender and to investigate its effects on in vitro quality and in vivo fertility of buffalo semen. Semen from three buffalo bulls was frozen in tris-citric extender containing 5.0%, 10% or 15% soya-lecithin or 20% egg yolk. Sperm motility, plasma membrane integrity and viability were assessed post-dilution, pre-freezing and post-thaw. In Post-dilution and pre-freezing, the values for motility, plasma membrane integrity and viability remained higher (p ≤ 0.05) in extenders containing 10% soya-lecithin and control compared with extender containing 5% and 15% soya-lecithin. However, motility, plasma membrane integrity and viability were higher (p < 0.05) in extender containing 10% soya-lecithin compared with control and extenders containing 5% and 15% soya-lecithin. Semen from two buffalo bulls was frozen in tris-citric extender containing either 10% soya-lecithin or 20% egg yolk. Higher (p < 0.05) fertility rate was recorded in buffaloes inseminated with semen containing 10% soya-lecithin (56%) compared with 20% egg yolk (41.5%). The results suggest that 10% soya-lecithin in extender improves the freezability and fertility of buffalo bull spermatozoa and can be used as an alternate to egg yolk in cryopreservation of buffalo semen. © 2011 Blackwell Verlag GmbH.

  1. Effect of straw size and thawing time on quality of cryopreserved buffalo (Bubalus bubalis) semen.

    PubMed

    Ansari, Muhammad S; Rakha, Bushra A; Andrabi, Syed M H; Akhter, Shamim

    2011-03-01

    This study was designed to compare the effect of straw size (0.25 vs. 0.5 ml) and thawing time (30 vs. 60 sec) on the quality of cryopreserved buffalo bull semen. Sperm motility, plasma membrane integrity and viability were higher (p ≤ 0.05) in 0.25 ml than 0.5 ml straw, thawed at 37°C either for 30 or 60 sec. In conclusion, cryopreservation of buffalo semen in 0.25 ml straw resulted in a higher post-thaw quality.

  2. Seasonal variations of Italian Mediterranean Buffalo (Bubalus bubalis) Mozzarella cheese quality.

    PubMed

    Ranucci, David; Garofalo, Angela; Urbani, Eleonora; Rea, Stefano; Loschi, Anna Rita; Stocchi, Roberta; Miraglia, Dino; Branciari, Raffaella

    2016-11-01

    Water buffalo Mozzarella cheese is more appreciated in the summer, but milk production is higher during wintertime, as water buffalo are seasonally polyoestrous animals. The aim of the study reported in this Research Communication was to evaluate the effect of the reversing of the calving period on Mozzarella cheese composition and quality traits. Ten batches of Mozzarella cheeses produced during the summer and winter periods were analysed for pH, colour, chemical composition. Seasonal differences were observed for cheese yield (26·66% in winter and 25·61% in summer), moisture content (66·54% in winter and 61·18% in summer) colour and consumer evaluation.

  3. Power of exclusion of 19 microsatellite markers for parentage testing in river buffalo (Bubalus bubalis).

    PubMed

    Kathiravan, P; Kataria, R S; Mishra, B P

    2012-08-01

    In the present study, 19 microsatellite markers were assessed for their power of exclusion to test parentage in river buffalo. Microsatellite genotypes of 216 unrelated buffaloes belonging to five different breeds were utilized for the study. The probabilities of exclusion were calculated for three hypothetical situations viz. paternity testing (PE1), one parental genotype unavailable (PE2) and exclusion of both parents i.e. substituted offspring (PE3). The mean probability of exclusion across 19 investigated markers in buffalo was 0.578 (PE1), 0.405 (PE2) and 0.764 (PE3) respectively. The probability of exclusion for paternity (PE1) ranged between 0.297 and 0.814 across different markers. The exclusion probability for the cases one parent unavailable (PE2) and substituted offspring (PE3) varied from 0.143 to 0.688 and 0.465 to 0.946 respectively. Polymorphism information content and expected heterozygosity were found to have significantly high correlation with probability of exclusion of microsatellite markers. The cumulative PE1 of nine marker loci was estimated to be 0.9999 while in case of absence of one of the parental genotypes, a minimum of 11 markers were required to achieve a cumulative PE2 of 0.999. In conclusion, the present study proposes two multiplex sets with four and five markers respectively for routine parentage testing in buffalo and an additional set of four markers for doubtful cases of paternity.

  4. Comparison of three different extenders on Murrah buffaloes (Bubalus bubalis) semen freezability.

    PubMed

    Zorzetto, M F; Martin, I; Sancler-Silva, Y F R; Zoca, S; Freitas-Dell'Aqua, C P; Papa, F O; Ramos, A A; Nunes, J F; Salgueiro, C C M; Oba, E

    2017-05-10

    The use of frozen semen for artificial insemination is the main approach utilised for the genetic improvement of most domesticated species. The advantages include lower transportation costs, continuous availability of semen, fewer occurrences of sexually transmitted diseases and the incorporation of desirable genes in a relatively short amount of time. Nevertheless, the use of frozen semen in buffalo herds remains limited due to the loss of sperm quality when buffalo semen is frozen. So, the goal of this study was to evaluate the pre- and post-cryopreservation quality of buffalo semen diluted in three distinct freezing media: Tris-egg yolk, Botu-bov® (BB) and ACP-111®. Thirty-two ejaculates from four bulls were analysed in terms of kinetics, morphology and sperm viability by epifluorescence microscope. Thawed samples were also evaluated for capacitation-like damage, DNA fragmentation and plasma and acrosomal membrane integrity using flow cytometry. The Tris-egg yolk and BB® extenders yielded better results than the ACP-111® extender for kinetics parameter (total motility, progressive motility and percentage of rapid cells). However, semen samples were similar for parameters evaluated by flow cytometry. Taken together, the data indicate that in comparison with Tris-egg yolk and BB extender, ACP-111® can also be used as an extender for buffalo semen cryopreservation. © 2017 Blackwell Verlag GmbH.

  5. Gas chromatographic-mass spectrometric analysis of chemical volatiles in buffalo (Bubalus bubalis) urine.

    PubMed

    Barman, Purabi; Yadav, M C; Kumar, H; Meur, S K; Ghosh, S K

    2013-10-01

    Isolation of active fraction and characterization of chemosignals from urine have been attempted in several mammalian species in the recent years. The objective of this study was to identify the urinary volatiles across various reproductive stages of buffalo cow, namely, estrus, diestrus, and pregnancy, and in bull, by chemical extraction followed by gas chromatography-linked mass spectrometry (GC-MS). Urine samples were collected from six buffalo cows at two different phases of estrous cycle, namely, estrus and diestrus. Besides, urinary samples were collected from five pregnant buffalo cows (60-75 days after artificial insemination (AI)) and six adult bulls. Thin-layer chromatography was performed as a preliminary test for qualitative comparison of different compounds extracted by organic solvents. Identification of the urinary compounds was carried out in a gas chromatograph (Perkin Elmer, Autosystem XL) linked to a mass spectrometer (Turbomass). The results of GC-MS analysis indicated the presence of 21 compounds with varying molecular weights and retention time, which were further categorized as diestrus-specific, pregnancy-specific, and bull-specific urinary compounds. No compound, however, could be identified as estrus-specific. We concluded that qualitative differences do exist in estrus, diestrus, and pregnant buffalo cow urine and in bull urine, as evidenced by GC-MS. Copyright © 2013 Elsevier Inc. All rights reserved.

  6. Collagen-IV supported embryoid bodies formation and differentiation from buffalo (Bubalus bubalis) embryonic stem cells

    SciTech Connect

    Taru Sharma, G.; Dubey, Pawan K.; Verma, Om Prakash; Pratheesh, M.D.; Nath, Amar; Sai Kumar, G.

    2012-08-03

    Graphical abstract: EBs formation, characterization and expression of germinal layers marker genes of in vivo developed teratoma using four different types of extracellular matrices. Highlights: Black-Right-Pointing-Pointer Collagen-IV matrix is found cytocompatible for EBs formation and differentiation. Black-Right-Pointing-Pointer Established 3D microenvironment for ES cells development and differentiation into three germ layers. Black-Right-Pointing-Pointer Collagen-IV may be useful as promising candidate for ES cells based therapeutic applications. -- Abstract: Embryoid bodies (EBs) are used as in vitro model to study early extraembryonic tissue formation and differentiation. In this study, a novel method using three dimensional extracellular matrices for in vitro generation of EBs from buffalo embryonic stem (ES) cells and its differentiation potential by teratoma formation was successfully established. In vitro derived inner cell masses (ICMs) of hatched buffalo blastocyst were cultured on buffalo fetal fibroblast feeder layer for primary cell colony formation. For generation of EBs, pluripotent ES cells were seeded onto four different types of extracellular matrices viz; collagen-IV, laminin, fibronectin and matrigel using undifferentiating ES cell culture medium. After 5 days of culture, ESCs gradually grew into aggregates and formed simple EBs having circular structures. Twenty-six days later, they formed cystic EBs over collagen matrix with higher EBs formation and greater proliferation rate as compared to other extracellular matrices. Studies involving histological observations, fluorescence microscopy and RT-PCR analysis of the in vivo developed teratoma revealed that presence of all the three germ layer derivatives viz. ectoderm (NCAM), mesoderm (Flk-1) and endoderm (AFP). In conclusion, the method described here demonstrates a simple and cost-effective way of generating EBs from buffalo ES cells. Collagen-IV matrix was found cytocompatible as it

  7. Fertility response in postpartum anoestrus buffaloes (Bubalus bubalis) using modified Ovsynch based timed insemination protocols

    PubMed Central

    Gupta, K. K.; Shukla, S. N.; Inwati, P.; Shrivastava, O. P.

    2015-01-01

    Background and Aim: Suboestrus constitutes the largest factor responsible for poor reproductive efficiency in buffaloes. Therefore, oestrus synchronization (OS) and fixed-time artificial insemination (AI) is considered an alternative approach to enhance reproductive efficiency in buffaloes. Thus, the present study was carried out to study the efficacy of modified Ovsynch protocol with fixed time insemination in post-partum anoestrus buffaloes. Materials and Methods: Total 50 post-partum anoestrus dairy buffaloes were selected and randomly divided into 5 Groups, each comprising ten animals (n=10). Animals of Group I received buserelin acetate 10 μg(GnRH) at day 0 and 9, and prostaglandin F2α (PGF2α) at day 7; in Group II similar to Group I except double dose of Gn RH (20 μg) at day 0; in Group III, similar to Group I plus additional administration of insulin on day 0, 1 and 2; in Group IV, similar to Group II plus additional administration of insulin on day 0,1 and 2; in Group V similar to Group I except replacement of first Gn RH with insulin on day 0,1 and 2. Animal of all groups were inseminated at fixed time using frozen semen at 60 h and 72 h after PGF2α administration and confirmed for pregnancy at day 60 post-insemination. Results: The results revealed satisfactory and comparable synchronization of oestrus (60-80%) and conception rate (20-40%) among the various treatment groups in the present study. The better synchronization was observed in modified protocols. However, non-significantly higher conception rate was recorded in modified Ovsynch protocol (40% each in Group II-V) as compared to 20% in Group-I (p>0.05). Conclusion: In this study comparatively higher OS and conception following insulin modified Ovsynch based timed AI protocols in post-partum anoestrus buffaloes was found satisfactory and comparable. PMID:27047090

  8. Efficiency of antifreeze glycoproteins for cryopreservation of Nili-Ravi (Bubalus bubalis) buffalo bull sperm.

    PubMed

    Qadeer, S; Khan, M A; Ansari, M S; Rakha, B A; Ejaz, R; Iqbal, R; Younis, M; Ullah, N; DeVries, Arthur L; Akhter, S

    2015-06-01

    Experiments were conducted to evaluate the effect of Antarctic fish antifreeze glycoproteins, (AFGP) size 1-5 (34-10.5 kDa) and 7-8 (3.2 and 2.4 kDa) in extender on buffalo bull sperm at cooling (4 °C) and at post thawing. Semen was collected from three Nili-Ravi buffalo bulls with artificial vagina for 3 weeks. Qualifying ejaculates from each buffalo bull were diluted (at 37 °C having 50×10(6) sperm/mL) in tris-citric acid extender containing AFGP at 0 (control), 0.1, 1 and 10 μg/mL. An aliquot of diluted semen was evaluated for sperm progressive motility and plasma membrane integrity, while the remaining fraction was cooled to 4 °C in 2 h. Further, an aliquot of cooled semen was evaluated for the previously described variables and the remaining fraction was cryopreserved (-196 °C). After 24 h of storage, straws were thawed at 37 °C for 30 s to assess post-thaw sperm quality. Inclusion of AFGP in the extender did not affect (P>0.05) sperm progressive motility and plasma membrane integrity of buffalo bull sperm at cooling stage (4 °C). However, at post thawing, improvement (P<0.05) in sperm progressive motility and plasma membrane integrity was recorded in extender containing AFGP 1-5 and AFGP 7-8 at 1 μg/mL compared to the control. Percentage of live sperm with an intact acrosome remained similar (P>0.05) in extenders containing different amounts of AFGP and control. In conclusion, supplementation of 1 μg/ml of AFGP in extender improved the motility and plasma membrane integrity of Nili-Ravi buffalo sperm after thawing. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Expression and molecular cloning of interferon stimulated genes in buffalo (Bubalus bubalis).

    PubMed

    Thakur, Nipuna; Singh, Girjesh; Paul, A; Bharati, J; Rajesh, G; Gm, Vidyalakshmi; Chouhan, V S; Bhure, S K; Maurya, V P; Singh, G; Sarkar, M

    2017-09-15

    Buffalo, the most important livestock species in tropical India, remains to be a poor breeder mainly due to embryonic mortality (65%) occurring mostly between 16 and 18 days of pregnancy. Early and accurate diagnosis of pregnancy can thus become a boon for successful herd management in buffalo. However, most of the currently available methods allow diagnosis only after 30 days post AI. Interferon tau (IFNT), the first pregnancy recognition signal in ruminants is one such molecule, which stimulates expression of various Interferon stimulated genes (ISGs) in the peripheral blood mononuclear cells (PBMC's) concomitant with IFNT signaling which occurs around maternal recognition of pregnancy (MRP). Hence, the study was planned to demonstrate the expression dynamics of ISGs (OAS1, MX1, MX2 and ISG15) in PBMCs during peri-implantation period in buffalo and also molecular cloning and expression of suitable ISG coded protein (s) in suitable host. Blood was collected from two groups of multiparous buffaloes: Group1: (n = 10) inseminated/pregnant (Experimental) and Group2: (n = 10) anestrous/non pregnant (Control). The expression profile of ISGs was then analyzed using real time qPCR. Expression profile of most ISGs was observed to increase through day 14 to day 20 post AI and declined thereafter. On the basis of differential gene expression at day 18 post AI, OAS1 and MX2 were identified as suitable ISG candidate biomarkers for accurate pregnancy diagnosis within 18 days post AI. Molecular cloning and expression of selected ISGs in a suitable prokaryotic expression vector was done thereafter. Bulk expression of the recombinant proteins was done and purified by affinity chromatography and confirmed by Western blot using Mouse Monoclonal His-probe antibodies. To conclude, as OAS1 and MX2, showed distinct differential expression at day 18 post AI, they may serve as ideal biomarkers for detection of early pregnancy in buffalo. Copyright © 2017 Elsevier Inc. All rights

  10. Experimental transmission of foot-and-mouth disease among Indian buffalo (Bubalus bubalis) and from buffalo to cattle.

    PubMed

    Maroudam, V; Nagendrakumar, S B; Madhanmohan, M; Santhakumar, P; Thiagarajan, D; Srinivasan, V A

    2008-01-01

    Indian buffalo and cattle were infected experimentally with a serotype O strain of foot-and-mouth disease virus of buffalo origin. Whereas intradermolingual inoculation of buffalo produced largely sub-clinical infection, inoculation in the dental pad produced vesicles in the mouth and on the feet. A buffalo infected via the dental pad transmitted infection to cattle and buffalo by direct contact with them for 24h. The contact-exposed buffalo developed (1) delayed-onset clinical signs, and (2) shedding of virus from the nose, commencing before the appearance of vesicles and continuing until the experiment was terminated 10 weeks after exposure. The covert nature of the disease in Indian buffalo, coupled with the prolonged shedding of virus, suggests that this species represents a host of epidemiological importance.

  11. Seasonal variations in developmental competence and relative abundance of gene transcripts in buffalo (Bubalus bubalis) oocytes.

    PubMed

    Abdoon, Ahmed S; Gabler, Christoph; Holder, Christoph; Kandil, Omaima M; Einspanier, Ralf

    2014-11-01

    Hot season is a major constraint to production and reproduction in buffaloes. The present work aimed to investigate the effect of season on ovarian function, developmental competence, and the relative abundance of gene expression in buffalo oocytes. Three experiments were conducted. In experiment 1, pairs of buffalo ovaries were collected during cold season (CS, autumn and winter) and hot season (HS, spring and summer), and the number of antral follicles was recorded. Cumulus oocyte complexes (COCs) were aspirated and evaluated according to their morphology into four Grades. In experiment 2, Grade A and B COCs collected during CS and HS were in vitro matured (IVM) for 24 hours under standard conditions at 38.5 °C in a humidified air of 5% CO2. After IVM, cumulus cells were removed and oocytes were fixed, stained with 1% aceto-orcein, and evaluated for nuclear configuration. In vitro matured buffalo oocytes harvested during CS or HS were in vitro fertilized (IVF) using frozen-thawed buffalo semen and cultured in vitro to the blastocyst stage. In experiment 3, buffalo COCs and in vitro matured oocytes were collected during CS and HS, and then snap frozen in liquid nitrogen for gene expression analysis. Total RNA was extracted from COCs and in vitro matured oocytes, and complementary DNA was synthesized; quantitative Reverse Transcription-Polymerase Chain Reaction was performed for eight candidate genes including GAPDH, ACTB, B2M, GDF9, BMP15, HSP70, and SOD2. The results indicated that HS significantly (P < 0.01) decreased the number of antral follicles and the number of COCs recovered per ovary. The number of Grade A, B, and C COCs was lower (P < 0.05) during HS than CS. In vitro maturation of buffalo oocytes during HS significantly (P < 0.01) reduced the number of oocytes reaching the metaphase II stage and increased the percentage of degenerated oocytes compared with CS. Oocytes collected during HS also showed signs of cytoplasmic degeneration. After IVF

  12. Pregnancy rates following AI with sexed semen in Mediterranean Italian buffalo heifers (Bubalus bubalis).

    PubMed

    Campanile, G; Gasparrini, B; Vecchio, D; Neglia, G; Senatore, E M; Bella, A; Presicce, G A; Zicarelli, L

    2011-08-01

    The use of sexed semen in farm animal production and genetic improvement has been shown to be feasible with variable degree of efficiency in a number of species, and proved to be economically viable in cattle. In the last two decades, various newly developed reproductive technologies applicable in buffaloes have mushroomed. Recently, following the birth of the first buffalo calves using AI with sexed semen, commercial interest to exploit sexing of semen in this species too is aroused. In order to verify the successful adoption of this technology in the buffalo, the present study on the use of sexed semen for AI was carried out and compared with conventional artificial insemination using nonsexed semen. A total of 379 buffalo heifers were used for synchronization of ovulation using the Presynch protocol in the South of Italy. Selected animals at the time of AI were randomly allocated to three different experiment groups: (1) 102 animals subjected to AI in the body of the uterus with sexed semen (SS body); (2) 104 animals subjected to AI in the horn of the uterus with sexed semen (SS horn); and (3) 106 animals subjected to AI in the body of the uterus with conventional nonsexed semen (NSS body). Semen of three buffalo bulls was sexed by a collaborating company and commercially distributed in 0.25 mL straws with a total of 2 million sexed spermatozoa. Pregnancy rates were first assessed at Day 28 following AI, and rechecked at Day 45 by ultrasound. Pregnancy rates were nonsignificantly different between animals inseminated with sexed or nonsexed semen: 80/206 (38.8%) and 40/106 (37.7%), respectively (P = 0.85). However, site of insemination of sexed semen affected pregnancy rate significantly as higher pregnancy rates were obtained when sexed semen was deposited into the body rather than the horn of the uterus: 46/101 (45.5%) and 34/105 (32.3%), respectively (P = 0.05). In conclusion, the use of sexed semen in buffalo heifers gave satisfactory and similar pregnancy

  13. Plasma hormonal and electrolyte alterations in cycling buffaloes ( Bubalus bubalis) during hot summer months

    NASA Astrophysics Data System (ADS)

    Singh, Narinder; Chaudhary, K. C.

    1992-09-01

    Plasma levels of progesterone, prolactin, luteinizing hormone, and electrolytes were monitored by radioimmunoassay in ten cycling buffaloes maintained at Punjab Agricultural University, Ludhiana during the hot summer months of June July. The plasma progesterone concentration ranged from 0.28±0.04 to 3.09±0.03 ng/ml at various stages of the oestrous cycle. Prolactin values ranged from 319±23 to 371±25 ng/ml and LH levels from 0.95±0.05 to 1.35±0.08 ng/ml. Concentrations differed significantly ( P⩽0.05) at various stages of the cycle. Levels of electrolytes, viz. Ca+ +, Na+ and K+, were well within the normal range. The high levels of prolactin, progesterone and LH during the hot summer were assessed in relation to poor reproductive efficiency in buffaloes.

  14. Trace elements in raw milk of buffaloes (Bubalus bubalis) from Campania, Italy.

    PubMed

    Esposito, Mauro; Miedico, Oto; Cavallo, Stefania; Pellicanò, Roberta; Rosato, Guido; Baldi, Loredana; Chiaravalle, A Eugenio

    2017-10-15

    The profile of 18 trace elements was traced in 68 milk samples collected from buffalo farms in the territory known as the "Land of Fires" in the Campania region (Italy). This area has been polluted by the illegal dumping in fields of industrial or domestic waste, wich is sometimes then burned spreading toxic contaminants. Milk from buffaloes raised on rural farms might be a good indicator of environmental contamination risk in the human food chain. Trace element analysis in milk was performed using mass spectrometry. One milk sample was found to be non-compliant due to high Pb concentration. In the absence of threshold values for the elements, established through legislation, the results were compared with similar studies from other countries, and in most cases the content determined in this study was in agreement with values reported elsewhere and do not represent a risk to human health. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Reproductive characteristics and thyroidal function in relation with season in Khuzestan buffalo (Bubalus bubalis) bulls

    PubMed Central

    Mayahi, Sadegh; Mamouei, Morteza; Tabatabaei, Saleh; Mirzadeh, Khalil

    2014-01-01

    High ambient temperature is the major constraint on Buffalo productivity. The aim of this study was to evaluate the reproductive performance and thyroid gland function in winter and summer seasons in Khuzestan buffalo bulls. Six male indigenous buffaloes of Khuzestan with nearly the same age (2-3 years old) and weight were used. Semen and blood samples through jugular vein were collected, every two weeks throughout the summer and winter seasons. The thyroid hormones and thyrotropin stimulating hormone (TSH) concentration in blood serum were measured by radioimmunoassay method. Semen quality was determined, using computer assisted sperm analyzer (CASA) and routine methods. The concentration of thyroxin (T4) was lower in winter than summer (p ≤ 0.05). The level of T3 uptake was higher in cold season than that of in hot season (p ≤ 0.05). The differences of tri-iodotyronine (T3) and TSH concentrations, as well as free thyroxin index were not significant between seasons. The semen volume and spermatozoa parameters including concentration, progressive motility, linear velocity, mean velocity, beat cross frequency, linear coefficient and straightness coefficient were higher in winter than summer (p ≤ 0.05). Semen pH and amplitude of lateral head displacement of spermatozoa were higher in summer than winter (p ≤ 0.05). In winter, there was positive correlation between spermatozoa concentration and T3 value of blood serum (p ≤ 0.05). There were positive correlations between values of semen volume and T4, progressive spermatozoa motility percent and TSH, as well as, total motility of spermatozoa and TSH in summer (p ≤ 0.05). In general, thyroid function and semen quality of Khuzestan buffaloes may be affected by seasons. PMID:25568719

  16. Physicochemical properties and oxidative inactivation of soluble lectin from water buffalo (Bubalus bubalis) brain.

    PubMed

    Rizvi, Sabika; Banu, Naheed

    2008-03-01

    Lectins are carbohydrate-binding proteins present in a wide variety of plants and animals, which serve various important physiological functions. A soluble beta-galactoside binding lectin has been isolated and purified to homogeneity from buffalo brain using ammonium sulphate precipitation (40-70%) and gel permeation chromatography on Sephadex G50-80 column. The molecular weight of buffalo brain lectin (BBL) as determined by SDS-PAGE under reducing and non-reducing conditions was 14.2 kDa, however, with gel filtration it was 28.5 kDa, revealing the dimeric form of protein. The neutral sugar content of the soluble lectin was estimated to be 3.3%. The BBL showed highest affinity for lactose and other sugar moieties in glycosidic form, suggesting it to be a beta-galactoside binding lectin. The association constant for lactose binding as evidenced by Scatchard analysis was 6.6 x 10(3) M(-1) showing two carbohydrate binding sites per lectin molecule. A total inhibition of lectin activity was observed by denaturants like guanidine HCl, thiourea and urea at 6 M concentration. The treatment of BBL with oxidizing agent destroyed its agglutination activity, abolished its fluorescence, and shifted its UV absorption maxima from 282 to 250 nm. The effect of H2O2 was greatly prevented by lactose indicating that BBL is more stable in the presence of its specific ligand. The purified lectin was investigated for its brain cell aggregation properties by testing its ability to agglutinate cells isolated from buffalo and goat brains. Rate of aggregation of buffalo brain cells by purified protein was more than the goat brain cells. The data from above study suggests that the isolated lectin may belong to the galectin-1 family but is glycosylated unlike those purified till date.

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

    PubMed

    Ali, A; Fahmy, S

    2008-06-01

    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.

  18. Molecular identification of methanogenic archaea from surti buffaloes (bubalus bubalis), reveals more hydrogenotrophic methanogens phylotypes.

    PubMed

    Singh, K M; Pandya, P R; Parnerkar, S; Tripathi, A K; Rank, D N; Kothari, R K; Joshi, C G

    2011-01-01

    Methane emissions from ruminant livestock are considered to be one of the more potent forms of greenhouses gases contributing to global warming. Many strategies to reduce emissions are targeting the methanogens that inhabit the rumen, but such an approach can only be successful if it targets all the major groups of ruminant methanogens. Therefore, a thorough knowledge of the diversity of these microbes in breeds of buffaloes, as well as in response to geographical location and different diets, is required. Therefore, molecular diversity of rumen methanogens in Surti buffaloes was investigated using 16S rRNA gene libraries prepared from pooled rumen contents from three Surti buffaloes. A total of 171 clones were identified revealing 23 different sequences (phylotypes). Of these 23 sequences, twelve sequences (12 OTUs, 83 clones) and 10 sequences (10 OTUs, 83 clones) were similar to methanogens belonging to the orders Methanomicrobiales and Methanobacteriales, and the remaining 1 phylotype (5 clones) were similar to Methanosarcina barkeri. These unique sequences clustered within a distinct and strongly supported phylogenetic group. Further studies and effective strategies can be made to inhibit the growth of Methanomicrobiales and Methanobacteriales phylotypes to reduce the methane emission from rumen and thus help in preventing global warming.

  19. Hemato-biochemical and hormonal profiles in post-partum water buffaloes (Bubalus bubalis)

    PubMed Central

    Kumar, Sunil; Balhara, A. K.; Kumar, Rajesh; Kumar, Naresh; Buragohain, Lukumoni; Baro, Daoharu; Sharma, R. K.; Phulia, S. K.; Singh, Inderjeet

    2015-01-01

    Aim: The objective of the present study was to compare serum as well as follicular fluid (FF) biochemical and hormonal profiles along with hematological parameters in postpartum estrus, anestrus, and cystic buffaloes. Materials and Methods: Postpartum buffaloes were selected in three different groups (within 40-60 days of parturition at estrus-Group-I, postpartum >90 days at anestrum-Group-II, and postpartum cystic buffaloes in Group III). The animals selected were examined for follicular wave dynamics by routine trans-rectal ultrasonography and FF was collected by transvaginal ultrasound-guided ovum pick up technique. All hematological and biochemical parameters were analyzed by automatic analyzers while hormonal profiles analyzed by commercially available ELISA kits. Results: In the present investigation, estrum and anestrum animal differ significantly in hemoglobin levels. Serum estradiol differs significantly in estrus and anestrus while no significant difference in progesterone concentration was noted among all three stages. The results of our study suggest that significant higher increase in total protein (TP), calcium and glucose values in estrum while urea, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase significantly higher in anestrum animals. Conclusion: The conclusion of the present study is that TP and albumin, calcium, urea, glucose affects oocyte development and quality. PMID:27047125

  20. A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction.

    PubMed

    Girish, P S; Haunshi, S; Vaithiyanathan, S; Rajitha, R; Ramakrishna, C

    2013-02-01

    Buffalo (Bubalus bubalis) meat is a major item of export from India but export of beef i.e. meat from cattle (Bos indicus) is prohibited. Also, adulteration of buffalo meat with that of beef (meat from cattle) is a common fraudulent practice because of prohibition on cow slaughter in most states of India. Food analysts require precise identification techniques to implement such regulations. In the present study, a method of DNA extraction by Alkaline lysis from meat samples and speciation of buffalo meat using species specific Polymerase Chain Reaction (PCR) has been reported. Alkaline lysis technique is a rapid method which involves triturating meat with four volumes of 0.2N NaOH, dilution of resultant liquid extract with eight volumes of 0.2N NaOH, heating the mix 75 °C for 20 min followed by neutralization with eight volumes of 0.04N Tris HCl. Entire procedure of DNA extraction takes less than 30 min and it is economical as it involves less expensive chemicals. Method was successfully applied in animal byproducts also viz., liver, heart and kidney. For authentication of buffalo meat, pair of primers was designed based on mitochondrial D loop gene nucleotide sequence. PCR amplification using the designed primers gave amplicon of size 482 bp in buffalo and no amplification was detected in closely related species viz., cattle, sheep and goat meat samples. Results of the assay were highly repetitive and reliable. An export sample referred by export regulation authorities was also analyzed by using the Alkaline lysis method of DNA extraction and species specific PCR which enabled authentication of meat within 5 h.

  1. Rumen adaptation of swamp buffaloes (Bubalus bubalis) by high level of urea supplementation when fed on rice straw-based diet.

    PubMed

    Wanapat, Metha; Phesatcha, Kampanat; Kang, Sungchhang

    2016-08-01

    Four rumen-fistulated swamp buffaloes (Bubalus bubalis) were randomly allocated to investigate rumen adaptation of urea on feed intake, nutrient digestibility, fermentation efficiency, and microbial protein synthesis. Buffaloes were fed with rice straw ad libitum for a period of 2 weeks and then were shifted to a step-up diet regimen by supplementation of concentrate containing 20 and 40 g/kg urea at 5 g/kg BW for a period of 2 and 4 weeks, respectively. The results revealed that feed intake and nutrient digestibility were increased by urea supplementation (P < 0.05) both at two and four period of consumption. However, ruminal pH, temperature, and protozoal population were neither affected by urea nor adaptation period (P > 0.05) while bacterial and fungal zoospores were increased especially at 40 g/kg urea. Data from real-time PCR further showed that total bacteria and the three predominant cellulolytic bacteria (Ruminococcus albus, Fibrobacter succinogenes, Ruminococcus flavefaciens) were increased by urea supplementation both at 2 and 4 weeks of urea feeding. Furthermore, methane production was similar among treatments while microbial protein synthesis was enhanced when buffaloes were fed with urea after a period of 2 weeks especially at 40 g/kg urea (P < 0.05). It can be concluded that urea supplementation could increase feed intake, nutrient digestibility, microbial protein synthesis, and fermentation efficiency of swamp buffaloes fed on rice straw. It is suggested that buffaloes could adapt well and utilize urea as a N source effectively within a period of 2 weeks uptake without adverse effect.

  2. Domestic cats (Felis catus) are definitive hosts for Sarcocystis sinensis from water buffaloes (Bubalus bubalis).

    PubMed

    Gjerde, Bjørn; Hilali, Mosaad

    2016-08-01

    The definitive hosts of Sarcocystis sinensis in water buffaloes have hitherto been unknown, but the close similarity of this species to the cat-transmitted Sarcocystis bovifelis in cattle suggested they were felids. In a previous study, two domestic cats were fed macroscopic sarcocysts of Sarcocystis fusiformis contained within or dissected from the esophageal muscles of water buffaloes, while no microscopic sarcocysts of S. sinensis were noticed. Both cats started shedding small numbers of sporocysts 8-10 days post infection (dpi) and were euthanized 15 dpi. Using a PCR-based molecular assay targeting the mitochondrial cox1 gene of S. fusiformis, both cats were shown to act as definitive hosts for this species. In the present study, DNA samples derived from oocysts/sporocysts in the intestinal mucosa of both cats were further examined by PCR for the presence of S. sinensis using 2 newly designed primers selectively targeting the cox1 gene of this species. All 6 DNA samples examined from each cat tested positive for S. sinensis. A 1,038-bp-long portion of cox1 was amplified and sequenced as 2 overlapping fragments from 5 of these DNA samples. The 5 sequences shared 99.3-100% identity with 7 previous cox1 sequences of S. sinensis obtained from sarcocysts in water buffaloes. Additionally, amplification of the ITS1 region with primers targeting various Sarcocystis spp., yielded amplicons of 2 different lengths, corresponding to those obtained from sarcocyst isolates of S. sinensis and S. fusiformis, respectively. This is the first study to show that cats act as definitive hosts for S. sinensis.

  3. Oocyte recovery by ovum pick up and embryo production in river buffaloes (Bubalus bubalis).

    PubMed

    Manjunatha, B M; Ravindra, J P; Gupta, P S P; Devaraj, M; Nandi, S

    2008-08-01

    Ovum pick up (OPU) was conducted twice a week for 12 weeks in six cycling, non-descriptive (local breed), Indian buffaloes to study the efficiency of OPU on recovery of oocytes for embryo production. OPU was performed using an ultrasound equipment with a 5-MHz transvaginal transducer, a single-lumen, 18-gauge, 55-cm-long needle and a constant vacuum pressure of 110 mmHg. The number and size of follicles were determined before puncture. The recovered oocytes were graded, washed, matured for 24 h and then fertilized with frozen-thawed semen, followed by embryo culture on the oviductal monolayer. The mean number of follicles observed per animal per session did not differ between animals or between puncture sessions. A mean number of 3.62 +/- 0.32 mm follicles were observed, 2.90 +/- 0.15 mm follicles were punctured and 1.21 +/- 0.07 oocytes were recovered per animal per session, with an average recovery rate of 42%. Of the total oocytes recovered, 64% were suitable for in vitro embryo production (grade A + B) whereas 36% were classified to be of grades C + D. A mean number of 0.25 +/- 0.2 transferable embryos was produced in vitro per buffalo per session with a transferable embryo production rate of 32%. In conclusion, this study demonstrated that twice-a-week OPU could be applied repeatedly, without any adverse effects on the follicular growth and oocyte recovery and that recovered oocytes could be used for in vitro embryo production in buffaloes.

  4. Domestic cats (Felis catus) are definitive hosts for Sarcocystis sinensis from water buffaloes (Bubalus bubalis)

    PubMed Central

    GJERDE, Bjørn; HILALI, Mosaad

    2016-01-01

    The definitive hosts of Sarcocystis sinensis in water buffaloes have hitherto been unknown, but the close similarity of this species to the cat-transmitted Sarcocystis bovifelis in cattle suggested they were felids. In a previous study, two domestic cats were fed macroscopic sarcocysts of Sarcocystis fusiformis contained within or dissected from the esophageal muscles of water buffaloes, while no microscopic sarcocysts of S. sinensis were noticed. Both cats started shedding small numbers of sporocysts 8–10 days post infection (dpi) and were euthanized 15 dpi. Using a PCR-based molecular assay targeting the mitochondrial cox1 gene of S. fusiformis, both cats were shown to act as definitive hosts for this species. In the present study, DNA samples derived from oocysts/sporocysts in the intestinal mucosa of both cats were further examined by PCR for the presence of S. sinensis using 2 newly designed primers selectively targeting the cox1 gene of this species. All 6 DNA samples examined from each cat tested positive for S. sinensis. A 1,038-bp-long portion of cox1 was amplified and sequenced as 2 overlapping fragments from 5 of these DNA samples. The 5 sequences shared 99.3–100% identity with 7 previous cox1 sequences of S. sinensis obtained from sarcocysts in water buffaloes. Additionally, amplification of the ITS1 region with primers targeting various Sarcocystis spp., yielded amplicons of 2 different lengths, corresponding to those obtained from sarcocyst isolates of S. sinensis and S. fusiformis, respectively. This is the first study to show that cats act as definitive hosts for S. sinensis. PMID:27075117

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

    NASA Astrophysics Data System (ADS)

    Mehta, S. N.; Gangwar, P. C.

    1983-03-01

    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.

  6. Osteopontin improves sperm capacitation and in vitro fertilization efficiency in buffalo (Bubalus bubalis).

    PubMed

    Boccia, Lucia; Di Francesco, Serena; Neglia, Gianluca; De Blasi, Marina; Longobardi, Valentina; Campanile, Giuseppe; Gasparrini, Bianca

    2013-08-01

    The aim of this study was to evaluate the effect of osteopontin (OPN), an ubiquitous acid glycoprotein, on in vitro sperm capacitation and on in vitro embryo production (IVEP) efficiency in buffalo. In experiment 1, after swim-up separation the sperm were incubated in Tyrode albumin lactate pyruvate medium in the absence of capacitating agents (control), with the standard concentration of heparin (0.01 mM) and three different concentrations of OPN (0.1, 1, and 10 mcg/mL), both in the presence and absence of heparin, for 2 and 4 hours. Capacitation was assessed indirectly by estimating the percentage of acrosome-reacted sperm after incubation with lysophosphatidylcholine. In order to determine the effect of OPN, in the presence of heparin, on fertilization (Experiment 2) and in vitro embryo development (experiment 3), in vitro-matured buffalo oocytes were fertilized in the presence of 0, 0.1, 1, and 10 mcg/mL of OPN. After IVF, the presumptive zygotes were dezonated, fixed, stained, and then evaluated microscopically. At Days 5 and 7 of culture, the cleavage and blastocyst rates were evaluated, respectively. Two hours of treatment with OPN at the two higher concentrations (1 and 10 mcg/mL) promoted in vitro capacitation of buffalo sperm (experiment 1). A synergic action of OPN with heparin was also done for all OPN concentrations tested. At 4 hours incubation, all treatments, including heparin (20.4%), improved (P < 0.01) capacitation compared with the control (16.2%). Interestingly, the best results were reported in all groups treated with OPN + heparin (40.8%, 38.6%, and 33.8%, respectively; P < 0.01). The addition of OPN to the IVF medium had a positive influence on total penetration, synchronous pronuclei formation (experiment 2), and IVEP efficiency (experiment 3). In particular, the two lower concentrations of OPN (0.1 and 1 mcg/mL), compared with the control, gave higher synchronous pronuclei formation (73.5%, 75.0%, and 46.5%, respectively; P < 0.01) and

  7. Molecular Characterization and Comparison of Phospholipase C zeta (PLCZ1) Gene Between Swamp (Bubalus carabanensis) and Riverine (Bubalus bubalis) Buffaloes: Its Implications and Future Perspectives.

    PubMed

    Atabay, Eufrocina P; Tadeo, Roseline D; Atabay, Edwin C; Venturina, Emma V; Fissore, Rafael A; Mingala, Claro N

    2017-08-11

    Phospholipase C zeta, a novel sperm-specific protein which is widely known to induce oocyte activation following fertilization, had already been characterized in various mammalian species, but not in water buffaloes thus far. The present study was conducted to initially characterize and compare the sequences of PLCZ1 gene of swamp and riverine buffaloes. Semen samples were collected; total RNA was extracted and reverse-transcribed. PLCZ1 cDNA was then amplified, and submitted for sequencing. Buffalo PLCZ1 gene yielded a sequence of 1905 base pair nucleotides translated into 634 bp amino acids. In general, the buffalo PLCZ1 gene was found to have high sequence identity with cattle and other domestic species. Similarly, significant residues and motifs in PLCZ1 gene sequence are found conserved in water buffaloes. However, there are variations in sequences identified between types of water buffaloes that may play a role in species-specific differences in terms of gene and protein expression, physiological mechanisms, and biological functions. The molecular information on buffalo PLCZ1 gene is highly valuable in subsequent works such as correlation studies on the identified gene variations with semen quality and fertility, and the development of biomarkers for bull fertility.

  8. Clinicopathological and ultrasonographic findings in 40 water buffaloes (Bubalus bubalis) with traumatic pericarditis.

    PubMed

    Mohamed, T

    2010-11-20

    Forty buffaloes with traumatic pericarditis were examined to characterise the ultrasonographic findings in buffaloes with traumatic pericarditis, determine the extent of the lesions and assess the prognosis. The most noticeable clinical presentations were presternal oedema (73 per cent) and jugular and mammary vein distension (88 per cent). Laboratory findings included neutrophilic leucocytosis, elevated total protein concentration, hypoalbuminaemia, hypergammaglobulinaemia and increased concentration of free fatty acids. Ultrasonographically, fluid in the pericardium appeared as either mild or massive anechoic accumulations containing fibrin threads or were imaged as homogenous, echogenic pericardial effusions. Moderate to severe corrugation of the reticular wall was observed. Deposits of fibrinous tissue interspersed with fluid pockets were seen between the reticulum, dorsal ruminal sac and diaphragm. Perireticular and mediastinal abscesses were imaged and appeared as echogenic lines with anechoic, echogenic, homogenous or heterogeneous contents. Additional ultrasonographic findings included hepatomegaly, dilation of the caudal vena cava, hepatic and portal veins, ascites, echogenic pleural effusions and vegetations of the tricuspid, mitral and pulmonary valves. The ultrasonographic findings were confirmed at postmortem examination.

  9. Transplacental transfer of iron in the water buffalo (Bubalus bubalis): uteroferrin and erythrophagocytosis.

    PubMed

    Pereira, F T V; Braga, F C; Burioli, K C; Kfoury, J R; Oliveira, L J; Papa, P C; Carvalho, A F; Ambrósio, C E; Bazer, F W; Miglino, M A

    2010-10-01

    The objectives of this investigation were to understand transplacental transport of iron by secreted uteroferrin (UF) and haemophagous areas of water buffalo placenta and clarify the role(s) of blood extravasation at the placental-maternal interface. Placentomes and interplacentomal region of 51 placentae at various stages of gestation were fixed, processed for light and transmission electron microscopy, histochemistry and immunohistochemistry. Haemophagous areas were present in placentomes collected between 4 and 10 months of pregnancy. Perl's reaction for ferric iron was negative in placentomes, but positive in endometrial glands. Positive staining for UF indicated areas in which it was being taken up by phagocytosis and/or fluid phase pinocytosis in areolae of the interplacentomal mesenchyme, with little staining in endometrial stroma. Imunohistochemistry detected UF in trophectoderm of haemophagous regions of placentomes and in other parts of the foetal villous tree, but the strongest immunostaining was in the epithelial cells and lumen of uterine glands. Ultrastructural analyses indicated that erythrophagocytosis was occurring and that erythrocytes were present inside cells of the chorion that also contained endocytic vesicles and caveolae. Results of this study indicate that both the haemophagous areas of placentomes and the areolae at the interface between chorion and endometrial glands are important sites for iron transfer from mother to foetal-placental tissues in buffalo throughout pregnancy. © 2009 Blackwell Verlag GmbH.

  10. Inhibitory potential of Buffalo (Bubalus bubalis) colostrum immunoglobulin G on Klebsiella pneumoniae.

    PubMed

    L S, Mamatha Bhanu; Nishimura, S-I; H S, Aparna

    2016-07-01

    The unique components of colostrum like free oligosaccharides and glycoconjugates are known to offer resistance to enzymatic digestion in the gastrointestinal tract and have the ability to inhibit the localized adherence of enteropathogens to the digestive tract of the neonates. In this context, we have evaluated the in vitro effect of buffalo colostrum immunoglobulin G on human pathogen Klebsiella pneumoniae, a predominant multidrug resistant pathogen associated with nasocomial infections. The investigation revealed growth inhibitory potential of immunoglobulin G in a dose dependent manner supported by scanning electron microscopic studies. The N-glycan enriched fraction of immunoglobulin G after PNGase treatment was found more effective, comparable to ampicillin than native immunoglobulin G supporting the fact that colostrum derived oligosaccharides is crucial and act as ideal substrates for undesirable and pathogenic bacteria. The MALDI TOF/TOF analysis confirmed the glycostructures of abundant N-glycans of immunoglobulin G exerting antibacterial activity. The proteomic analysis revealed variations between control and treated cells and expression of chemotaxis-CheY protein (14kDa) was evidenced in response to immunoglobulin G treatment. Hence, it would be interesting to investigate the mode of inhibition of multidrug-resistant K. pneumoniae by buffalo colostrum immunoglobulin G with the identification of a newly expressed signalling protein.

  11. Subcutaneous pharmacokinetics and dosage regimen of cefotaxime in buffalo calves (Bubalus bubalis)

    PubMed Central

    Srivastava, Anil Kumar

    2006-01-01

    The pharmacokinetics and dosage regimen of cefotaxime following its single subcutaneous administration (10 mg/kg) were investigated in buffalo calves. Plasma and urine samples were collected over 10 and 24 h post administration, respectively. Cefotaxime in plasma and urine was estimated by microbiological assay technique using E. coli as test organism. The pharmacokinetic profiles fitted one-compartment open model. The peak plasma levels of cefotaxime were 6.48 ± 0.52 µg/ml at 30 min and the drug was detected upto 10 h. The absorption half-life and elimination half-life were 0.173 ± 0.033 h and 1.77 ± 0.02 h, respectively. The apparent volume of distribution and total body clearance were 1.17 ± 0.10 l/kg and 0.45 ± 0.03 l/kg/h, respectively. The urinary excretion of cefotaxime in 24 h, was 5.36 ± 1.19 percent of total administrated dose. A satisfactory subcutaneous dosage regimen for cefotaxime in buffalo calves would be 13 mg/kg repeated at 12 h intervals. PMID:16645334

  12. Utility of ultrasonography for diagnosis of superficial swellings in buffalo (Bubalus bubalis)

    PubMed Central

    ABOUELNASR, Khaled; EL-SHAFAEY, El-Sayed; MOSBAH, Esam; EL-KHODERY, Sabry

    2016-01-01

    We studied 72 buffalo with superficial swellings in the head (n=4), neck (n=5), chest wall (n=4), abdominal wall (n=28), limbs (n=16), gluteal region (n=8), perineal region (n=6) and udder (n=1). Ultrasonographically, the swellings varied according to type, duration, content and location. The clinical use of ultrasound to assess these superficial swellings allowed diagnosis of abscesses (n=21), hematomas (n=11), hernias (n=17), bursitis (n=13), urethral diverticula (n=6) and tumors (n=4). Ultrasonography could precisely discriminate each lesion type (sensitivity, 71–100%; specificity, 75–100%; odds ratio, 1.0–8.4; Confidence Interval, 74.2–20; and P value 0.001). The specificity for ultrasonographic evaluation of superficial swellings was 100% for hernias, urethral diverticula and tumors, whilst the lowest specificity was recorded for hematomas (75%) and abscesses (92%). In conclusion, ultrasonography provides a precise, non-invasive and fast technique for the evaluation, classification and subsequent treatment of a variety of superficial swellings in buffalo. PMID:27181085

  13. Effect of melatonin on maturation capacity and fertilization of Nili-Ravi buffalo (Bubalus bubalis) oocytes

    PubMed Central

    Nagina, G.; Asima, A.; Nemat, U.; Shamim, A.

    2016-01-01

    This study evaluated the effect of melatonin supplementation of in vitro maturation media on in vitro maturation (IVM) and in vitro fertilization (IVF) rate of buffalo oocytes. Cumulus oocytes complexes (COCs) were aspirated from follicles of 2-8 mm diameter. In experiment I, COCs were matured in IVM medium supplemented with 0 (control), 250, 500, and 1000 μM melatonin for 22-24 hours in CO2 incubator at 38.5°C with 5% CO2 and at 95% relative humidity. The maturation rate did not differ in media supplemented with melatonin at 250 μM, 500 μM, 1000 μM and control (0 μM). In experiment II, the matured oocytes were fertilized in 50 μl droplets of Tyrode’s Albumin Lactate Pyruvate (TALP) medium having 10 ug/ml heparin for sperm (2 million/ml) capacitation. The fertilization droplets were then kept for incubation at 5% CO2, 39°C and at 95% relative humidity for 18 hours. The fertilization rate was assessed by sperm penetration and pronuclear formation. Fertilization rate was improved when maturation medium was supplemented with 250 μM melatonin compared to control. In conclusion, melatonin supplementation to serum free maturation media at 250 μM improved the fertilization rate of buffalo oocytes. PMID:27540514

  14. Genetic parameters for milk, fat and protein yields in Murrah buffaloes (Bubalus bubalis Artiodactyla, Bovidae)

    PubMed Central

    2010-01-01

    The objective of the present study was to estimate genetic parameters for test-day milk, fat and protein yields and 305-day-yields in Murrah buffaloes. 4,757 complete lactations of Murrah buffaloes were analyzed. Co-variance components were estimated by the restricted maximum likelihood method. The models included additive direct genetic and permanent environmental effects as random effects, and the fixed effects of contemporary group, milking number and age of the cow at calving as linear and quadratic covariables. Contemporary groups were defined by herd-year-month of test for test-day yields and by herd-year-season of calving for 305-day yields. The heritability estimates obtained by two-trait analysis ranged from 0.15 to 0.24 for milk, 0.16 to 0.23 for protein and 0.13 to 0.22 for fat, yields. Genetic and phenotypic correlations were all positive. The observed population additive genetic variation indicated that selection might be an effective tool in changing population means in milk, fat and protein yields. PMID:21637608

  15. Production of hand-made cloned buffalo (Bubalus bubalis) embryos from non-viable somatic cells.

    PubMed

    Duah, E K A; Mohapatra, S K; Sood, T J; Sandhu, A; Singla, S K; Chauhan, M S; Manik, R S; Palta, P

    2016-12-01

    Use of non-viable somatic cells for hand-made cloning (HMC) can enable production of cloned animals from tissues obtained from elite or endangered dead animals. Buffalo skin fibroblast cells were rendered non-viable by heat treatment and used for HMC. Although fusion (93.6 ± 1.72 vs 67.1 ± 2.83%) and cleavage (90.3 ± 1.79 vs 65.8 ± 1.56%) rate was lower (P < 0.001) than that for controls, blastocysts could be successfully produced. However, blastocyst rate (34.1 ± 2.43 vs 6.9 ± 2.18%, P < 0.001) and total cell number of blastocysts (TCN, 221.3 ± 25.14 vs 151.1 ± 21.69, P < 0.05) were lower and apoptotic index (4.8 ± 1.06 vs 10.9 ± 1.21) was higher (P < 0.001) than that of controls. In another experiment, ear tissue of slaughterhouse buffaloes was preserved in mustard oil at room temperature for 48 h following which somatic cells were harvested by enzymatic digestion and used for HMC. Although fusion (96.8 ± 1.48 vs 84.2 ± 3.19%), cleavage (89.6 ± 3.59 vs 77.2 ± 3.99%), and blastocyst rate (36.9 ± 7.45 vs 13.1 ± 6.87%) were lower (P < 0.01), TCN (223.0 ± 27.89 vs 213.3 ± 28.21) and apoptotic index (3.97 ± 0.67 vs 5.22 ± 0.51) of blastocysts were similar to those of controls. In conclusion, HMC can be successfully used for production of blastocysts from non-viable cells and from cells obtained from freshly slaughtered buffaloes. This can pave the way for the restoration of farm or wild animals by HMC if somatic cells could be obtained within a few hours after their death.

  16. Comparative therapeutic effect of toltrazuril, sulphadimidine and amprolium on Eimeria bovis and Eimeria zuernii given at different times following infection in buffalo calves (Bubalus bubalis).

    PubMed

    Ghanem, Mohamed M; Radwaan, Mervat E; Moustafa, Abdel Moneim M; Ebeid, Mohamed H

    2008-04-17

    We compared the therapeutic effect of three anticoccidial drugs (toltrazuril, sulphadimidine and amprolium) in buffalo (Bubalus bubalis) calves experimentally infected with Eimeria bovis (E. bovis) and E. zuernii oocysts (3 x 104oocyst/calf). Buffalo calves (1.5-4 month old, 70-kg body weight) were randomly allocated into 3 groups (9 calves each). Group T was experimentally infected with oocysts and treated with toltrazuril (20 mg/kg BW twice orally at a 1-week interval). Group S was experimentally infected with oocysts and treated with sulphadimidine (125 mg/kg injected IM followed by half dose for 4 successive days). Group A was experimentally infected with oocysts and treated with amprolium (50 mg/kg orally for 7 successive days). Each group had three subgroups (three calves/subgroup) to represent timing of the drug administration: 1st day of coccidia infection (FD), onset of clinical signs of coccidiosis (CC), and onset of oocyst shedding into the faeces (OS). Clinical signs, body-weight gain (BWG) and number of oocysts per gram feces (OPG) were monitored daily for 35 days post-infection (DPI). The OPG were reduced (but the BWG was not different) in the T calves compared to S and A calves. Within the same group, treatment from the 1st day of infection reduced the OPG and increased the BWG compared to the later treatment timings.

  17. Cholesterol-loaded cyclodextrins prevent cryocapacitation damages in buffalo (Bubalus bubalis) cryopreserved sperm.

    PubMed

    Longobardi, Valentina; Albero, Giuseppe; De Canditiis, Carolina; Salzano, Angela; Natale, Antonio; Balestrieri, Anna; Neglia, Gianluca; Campanile, Giuseppe; Gasparrini, Bianca

    2017-02-01

    The aim of this study was to investigate the effect of cholesterol-loaded cyclodextrins (CLC) on motility, viability, capacitation status, and in vivo fertility of buffalo frozen-thawed sperm. After the initial semen assessment, buffalo sperm were diluted in BULLXcell extender containing 0- (control), 1.5-, and 3-mg/mL CLC and cryopreserved. At thawing, sperm motility was evaluated by phase contrast microscopy, and viability-capacitation status was assessed by Hoechst 33258-chlortetracycline (CTC) assay. Capacitation status was also evaluated by an indirect immunofluorescence assay to localize phosphotyrosine-containing proteins. Moreover, buffaloes were artificial inseminated to assess the in vivo-fertilizing potential of CLC-treated semen. No differences among control, 1.5-, and 3-mg/mL CLC-treated groups were recorded in both sperm motility (66.5 ± 5.6, 68.8 ± 4.8, and 68.8 ± 4.8, respectively) and viability (86.5 ± 1.9, 87.6 ± 1.5, 88.4 ± 2.3, respectively). However, the extender supplementation with CLC significantly reduced sperm cryocapacitation. Indeed, CLC treatment decreased (P < 0.01) the proportion of sperm showing the CTC pattern B (capacitated sperm) compared with the control (69.6 ± 3.4, 37.8 ± 1.5, and 51.3 ± 4.7, respectively, with 0, 1.5-, and 3-mg/mL CLC; P < 0.01). Furthermore, the percentage of sperm displaying tyrosine-phosphorylated pattern EA (i.e. high capacitation level) was reduced (P < 0.01) in both CLC-treated groups (10.8 ± 3.3 and 5.6 ± 1.6, respectively, with 1.5- and 3-mg/mL CLC) compared with the control (37.3 ± 6.9), reaching values similar to those recorded in fresh semen (11.0 ± 3.5). In addition, treating sperm with 3-mg/mL CLC increased (P < 0.01) the percentage of nonfluorescent (pattern NF), i.e., non-capacitated sperm (41.8 ± 3.6) compared with fresh semen (11.0 ± 6.9). No differences were recorded in pregnancy rates at 60 days post-artificial insemination among

  18. Use of peripheral blood for production of buffalo (Bubalus bubalis) embryos by handmade cloning.

    PubMed

    Jyotsana, Basanti; Sahare, Amol A; Raja, Anuj K; Singh, Karn P; Nala, Narendra; Singla, S K; Chauhan, M S; Manik, R S; Palta, P

    2016-09-15

    Buffalo embryos were produced by handmade cloning using peripheral blood-derived lymphocytes as donor cells. Although the blastocyst rate was lower (P < 0.01) for lymphocyte- than control skin fibroblast-derived embryos (6.6 ± 0.84% vs. 31.15 ± 2.97%), the total cell number (152.6 ± 23.06 vs. 160.1 ± 13.25) and apoptotic index (6.54 ± 0.95 vs. 8.45 ± 1.32) were similar. The global level of H3K9ac was higher (P < 0.05) in lymphocyte- than that in skin-derived blastocysts; whereas in IVF blastocysts, the level was not significantly different from the two cloned groups. The level of H3K27me3 was similar among the three groups. The expression level of DNMT1, DNMT3a, HDAC1, and IGF-1R was higher (P < 0.01) in lymphocytes than that in skin fibroblasts. The expression level of CDX2 was higher (P < 0.05) than that of DNMT3a, IGF-1R, OCT4, and NANOG was lower (P < 0.05) in lymphocyte-derived than in IVF blastocysts; that of DNMT1 and HDAC1 was similar in the two groups. The expression level of all these genes, except that of NANOG, was lower (P < 0.05) in lymphocyte- than in skin fibroblast-derived blastocysts. It is concluded that, peripheral blood-derived lymphocytes can be used for producing handmade cloning embryos in bubaline buffaloes. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Effect of supplemental light on growth, prolactin, progesterone and luteinizing hormone in water buffalo ( Bubalus bubalis)

    NASA Astrophysics Data System (ADS)

    Perera, K. S.; Gwazdauskas, F. C.; Akers, R. M.; McGilliard, M. L.

    1989-06-01

    Fifty non-pregnant Surti buffalo heifers aged between 17 and 42 months ( n=24, <24 months; n=26, >24 months) were randomly assigned to groups subject to either natural daylight +4h supplemental light ( n=25) or natural day light ( n=25), to study changes in growth, serum prolactin (Prl), progesterone (P4) and luteinizing hormone (LH) to supplemental lighting. Ambient temperatures (T) and relative humidity (RH) generally were >27° C and <70% during the day-time, respectively. Light-supplemented heifers had 16.2 kg net body weight (BW) gain at 9 weeks compared to 20.8 kg for controls, but higher mean Prl after 6.5 weeks ( P<0.01), and higher P4 (0.41 vs 0.19 ng/ml; P<0.06) than control heifers. Older heifers had 39.7% greater BW ( P<0.01), but a net 4.3% BW gain compared to a 10.1% gain for younger heifers at 10 weeks. Older, light-supplemented heifers had higher mean P4 (0.63 vs 0.19 ng/ml; P<0.07) than the other groups. These weight and hormonal changes suggest that 4 h supplemental light can alter growth and endocrine function in buffaloes under similar planes of nutrition. While light supplementation did not have a positive effect on body wieght during the 10 week study, body weight and endocrine changes due to supplemental light may be important factors for initiation of reproductive cyclicity.

  20. The in vitro effect of leptin on semen quality of water buffalo (Bubalus bubalis) bulls

    PubMed Central

    Khaki, Amir; Batavani, Rooz Ali; Najafi, Gholamreza

    2013-01-01

    The purpose of this study was to evaluate the probable effects of leptin addition in different levels to the semen extender on sperm quality (motility and motility parameters, viability, sperm membrane integrity, and DNA damage). Semen specimens were evaluated immediately after leptin addition, equilibration time and after thawing the frozen semen. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 10, 20, 50, 100, and 200 ng mL-1 leptin. The diluted semen was kept 4 hr in refrigerator to reach to the equilibration time and then packed in 0.5 mL French straws and frozen in liquid nitrogen. Our results showed that, in the fresh semen, no significant difference was observed in all sperm quality parameters evaluated among all of the examined leptin concentrations. Addition of 10 ng mL-1 leptin into semen extender significantly preserved sperm motility, all of the motility parameters, and viability in equilibrated semen compared to that of control group. However, in vitro addition of 200 ng mL-1 leptin, significantly decreased theses parameters. In the frozen thawed semen, all leptin concentrations decreased sperm motility and viability, but significant decrease was observed in concentrations of 100 and 200 ng mL-1. Adding leptin to semen extender did not have any significant influence on sperm DNA damage and sperm membrane integrity in all examined groups. These findings suggest that in vitro addition of 10 ng mL-1 leptin could preserve sperm motility and viability in cooled semen of buffaloes. PMID:25593679

  1. Genetic Evaluation of Dual-Purpose Buffaloes (Bubalus bubalis) in Colombia Using Principal Component Analysis

    PubMed Central

    Agudelo-Gómez, Divier; Pineda-Sierra, Sebastian; Cerón-Muñoz, Mario Fernando

    2015-01-01

    Genealogy and productive information of 48621 dual-purpose buffaloes born in Colombia between years 1996 and 2014 was used. The following traits were assessed using one-trait models: milk yield at 270 days (MY270), age at first calving (AFC), weaning weight (WW), and weights at the following ages: first year (W12), 18 months (W18), and 2 years (W24). Direct additive genetic and residual random effects were included in all the traits. Maternal permanent environmental and maternal additive genetic effects were included for WW and W12. The fixed effects were: contemporary group (for all traits), sex (for WW, W12, W18, and W24), parity (for WW, W12, and MY270). Age was included as covariate for WW, W12, W18 and W24. Principal component analysis (PCA) was conducted using the genetic values of 133 breeding males whose breeding-value reliability was higher than 50% for all the traits in order to define the number of principal components (PC) which would explain most of the variation. The highest heritabilities were for W18 and MY270, and the lowest for AFC; with 0.53, 0.23, and 0.17, respectively. The first three PCs represented 66% of the total variance. Correlation of the first PC with meat production traits was higher than 0.73, and it was -0.38 with AFC. Correlations of the second PC with maternal genetic component traits for WW and W12 were above 0.75. The third PC had 0.84 correlation with MY270. PCA is an alternative approach for analyzing traits in dual-purpose buffaloes and reduces the dimension of the traits. PMID:26230093

  2. Production of nuclear transfer embryos by using somatic cells isolated from milk in buffalo (Bubalus bubalis).

    PubMed

    Golla, K; Selokar, N L; Saini, M; Chauhan, M S; Manik, R S; Palta, P; Singla, S K

    2012-10-01

    Somatic cells in milk are a potential source of nuclei for nuclear transfer to produce genetically identical animals; this is especially important in animals that are susceptible to risks of bacterial infection on biopsy collection. In this study, a minimum of 10 milk samples were collected from each of the three buffaloes representing Murrah breed. All the samples were processed immediately and cell colonies were obtained. Cell colonies from one buffalo (MU-442) survived beyond 10 passages and were evaluated by fluorescence microscopy and used in nuclear transfer experiments. In culture, these cells expressed vimentin, indicating they were of fibroblast origin similar to ear cells. We compared the effectiveness of cloning using those milk-derived fibroblast (MDF) cells and fibroblast cells derived from the ear derived fibroblast (EDF). Fusion and cleavage rates of MDF-NT and EDF-NT embryos were found to be similar (92.43 ± 1.28% vs 94.98 ± 1.24%, and 80.27 ± 1.75% vs 84.56 ± 3.73%, respectively; p > 0.01); however, development to blastocyst stage and total cell number was higher for EDF-NT embryos (50.24 ± 2.54%, 227.14 ± 13.04, respectively, p < 0.01), than for MDF-NT embryos (16.44 ± 0.75%, 170.57 ± 4.50 respectively). We conclude that somatic cells from milk can be cultured effectively and used as nucleus donor to produce cloned blastocyst-stage embryos.

  3. The in vitro effect of leptin on semen quality of water buffalo (Bubalus bubalis) bulls.

    PubMed

    Khaki, Amir; Batavani, Rooz Ali; Najafi, Gholamreza

    2013-01-01

    The purpose of this study was to evaluate the probable effects of leptin addition in different levels to the semen extender on sperm quality (motility and motility parameters, viability, sperm membrane integrity, and DNA damage). Semen specimens were evaluated immediately after leptin addition, equilibration time and after thawing the frozen semen. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 10, 20, 50, 100, and 200 ng mL(-1) leptin. The diluted semen was kept 4 hr in refrigerator to reach to the equilibration time and then packed in 0.5 mL French straws and frozen in liquid nitrogen. Our results showed that, in the fresh semen, no significant difference was observed in all sperm quality parameters evaluated among all of the examined leptin concentrations. Addition of 10 ng mL(-1) leptin into semen extender significantly preserved sperm motility, all of the motility parameters, and viability in equilibrated semen compared to that of control group. However, in vitro addition of 200 ng mL(-1) leptin, significantly decreased theses parameters. In the frozen thawed semen, all leptin concentrations decreased sperm motility and viability, but significant decrease was observed in concentrations of 100 and 200 ng mL(-1). Adding leptin to semen extender did not have any significant influence on sperm DNA damage and sperm membrane integrity in all examined groups. These findings suggest that in vitro addition of 10 ng mL(-1) leptin could preserve sperm motility and viability in cooled semen of buffaloes.

  4. Genetic Evaluation of Dual-Purpose Buffaloes (Bubalus bubalis) in Colombia Using Principal Component Analysis.

    PubMed

    Agudelo-Gómez, Divier; Pineda-Sierra, Sebastian; Cerón-Muñoz, Mario Fernando

    2015-01-01

    Genealogy and productive information of 48621 dual-purpose buffaloes born in Colombia between years 1996 and 2014 was used. The following traits were assessed using one-trait models: milk yield at 270 days (MY270), age at first calving (AFC), weaning weight (WW), and weights at the following ages: first year (W12), 18 months (W18), and 2 years (W24). Direct additive genetic and residual random effects were included in all the traits. Maternal permanent environmental and maternal additive genetic effects were included for WW and W12. The fixed effects were: contemporary group (for all traits), sex (for WW, W12, W18, and W24), parity (for WW, W12, and MY270). Age was included as covariate for WW, W12, W18 and W24. Principal component analysis (PCA) was conducted using the genetic values of 133 breeding males whose breeding-value reliability was higher than 50% for all the traits in order to define the number of principal components (PC) which would explain most of the variation. The highest heritabilities were for W18 and MY270, and the lowest for AFC; with 0.53, 0.23, and 0.17, respectively. The first three PCs represented 66% of the total variance. Correlation of the first PC with meat production traits was higher than 0.73, and it was -0.38 with AFC. Correlations of the second PC with maternal genetic component traits for WW and W12 were above 0.75. The third PC had 0.84 correlation with MY270. PCA is an alternative approach for analyzing traits in dual-purpose buffaloes and reduces the dimension of the traits.

  5. Effect of egg yolk powder on freezability of Murrah buffalo (Bubalus bubalis) semen.

    PubMed

    Kumar, N; Lone, S A; Prasad, J K; Jan, M H; Ghosh, S K

    2016-06-01

    The aim of this study was to investigate the effect of commercial egg yolk powder as an alternative to fresh egg yolk on freezability of Murrah buffalo semen. Semen samples (12) from 3 Murrah buffaloes (4 from each bull) with mass motility (≥3+) and total motility (70% and above) were utilized in this study. Immediately after collection, each sample was divided into four groups. Groups I was diluted up to 60×10(6) sperm/ml with tris extender containing 10% fresh egg yolk and Groups II, III, and IV were diluted up to 60×10(6) sperm/ml with tris extender containing 2%, 4%, and 6% egg yolk powder, respectively. Semen samples were processed and cryopreserved followed by examination of frozen semen samples after 24 h. Semen samples from each group were evaluated for total motility, viability, acrosomal integrity, abnormality, and hypo-osmotic swelling test (HOST) response after dilution, pre-freeze, and post-thaw stage. Pre-freeze total motility was significantly (p<0.05) higher in Groups III and IV as compared to Groups I and II, and post-thaw total motility was significantly (p<0.01) higher in Group III as compared to other three groups. Viability was significantly (p<0.05) higher in Groups II, III, and IV than Group I at the pre-freeze stage. Significantly (p<0.01) higher viability and acrosomal integrity were recorded in Group III as compared to other three groups at the post-thaw stage. Abnormality was significantly (p<0.05) higher in Group IV than other three groups. HOST response was significantly (p<0.05) higher in Groups II and III than Groups I and IV at the pre-freeze and post-thaw stages. Addition of egg yolk powder at 4% level yielded significantly better results in terms of post-thaw semen quality as compared to the fresh egg yolk and other concentrations of egg yolk powder (2% and 6%).

  6. Effect of egg yolk powder on freezability of Murrah buffalo (Bubalus bubalis) semen

    PubMed Central

    Kumar, N.; Lone, S. A.; Prasad, J. K.; Jan, M. H.; Ghosh, S. K.

    2016-01-01

    Aim: The aim of this study was to investigate the effect of commercial egg yolk powder as an alternative to fresh egg yolk on freezability of Murrah buffalo semen. Materials and Methods: Semen samples (12) from 3 Murrah buffaloes (4 from each bull) with mass motility (≥3+) and total motility (70% and above) were utilized in this study. Immediately after collection, each sample was divided into four groups. Groups I was diluted up to 60×106 sperm/ml with tris extender containing 10% fresh egg yolk and Groups II, III, and IV were diluted up to 60×106 sperm/ml with tris extender containing 2%, 4%, and 6% egg yolk powder, respectively. Semen samples were processed and cryopreserved followed by examination of frozen semen samples after 24 h. Semen samples from each group were evaluated for total motility, viability, acrosomal integrity, abnormality, and hypo-osmotic swelling test (HOST) response after dilution, pre-freeze, and post-thaw stage. Results: Pre-freeze total motility was significantly (p<0.05) higher in Groups III and IV as compared to Groups I and II, and post-thaw total motility was significantly (p<0.01) higher in Group III as compared to other three groups. Viability was significantly (p<0.05) higher in Groups II, III, and IV than Group I at the pre-freeze stage. Significantly (p<0.01) higher viability and acrosomal integrity were recorded in Group III as compared to other three groups at the post-thaw stage. Abnormality was significantly (p<0.05) higher in Group IV than other three groups. HOST response was significantly (p<0.05) higher in Groups II and III than Groups I and IV at the pre-freeze and post-thaw stages. Conclusion: Addition of egg yolk powder at 4% level yielded significantly better results in terms of post-thaw semen quality as compared to the fresh egg yolk and other concentrations of egg yolk powder (2% and 6%). PMID:27397983

  7. Effect of presence of a dominant follicle on the superovulatory response in buffalo (Bubalus bubalis).

    PubMed

    Manik, R S; Singla, S K; Palta, P; Madan, M L

    1998-10-15

    Ten buffalo were superovulated by administration of 8 doses of FSH in a descending schedule spread over 4 d (5.5/5.5, 4.5/4.5, 3.5/3.5 and 2.5/2.5 mL, i.m.; total dose of 64 AU in 32 mL) beginning on Day 10 of an unstimulated estrous cycle, and 30 and 20 mg Lutalyse was given alongwith the 5th and 6th injections of FSH, respectively, to induce luteolysis. The number of corpora lutea (CL) was determined on 6 d post estrus. The ovaries were examined daily by ultrasonography from Day -5 to Day 5 (Day 0 = day of start of superovulation). The animals were retrospectively classified into 2 groups depending upon the presence (n = 4) or absence of a dominant follicle (n = 6). The mean diameter of the largest follicle (F1) increased from 8.25 +/- 0.48 mm on Day -5 to 10.75 +/- 0.25 mm on Day 0 in the dominant group, whereas in the nondominant group the F1 follicle exhibited a progressive decrease from 9.00 +/- 0.45 mm to 7.00 +/- 0.65 mm during the same period, the difference in profiles between the 2 groups was significant (P = 0.042). The profile of the diameter of the second largest follicle (F2) and the difference in diameters between largest and second largest follicles (F1-F2) were not significantly different between the 2 groups. The profile of mean number of large (> or = 10 mm diameter), but not small (2 to 5 mm diameter) or medium (6 to 9 mm diameter) follicles differed significantly (P = 0.001) between the 2 groups from Day -5 to Day 5 (P = 0.030). The number of CL was not significantly different between nondominant (4.00 +/- 0.97) and dominant groups (3.25 +/- 1.31). The number of CL was positively correlated (P < 0.01) with the number of medium follicles and the total number of follicles on the day of initiation of superovulation, but not with follicles of any size category or total number of follicles on any previous day. The results of this study indicate that following the use of morphological criteria based on the size of the largest follicle alone, the

  8. Expression profiling of major heat shock protein genes during different seasons in cattle (Bos indicus) and buffalo (Bubalus bubalis) under tropical climatic condition.

    PubMed

    Kumar, Anil; Ashraf, Syma; Goud, T Sridhar; Grewal, Anita; Singh, S V; Yadav, B R; Upadhyay, R C

    2015-07-01

    Heat shock proteins consist of highly conserved stress proteins, expressed in response to stress and play crucial roles in environmental stress tolerance and adaptation. The present study was conducted to identify major types of genes under the HSP70 family and other HSPs and to evaluate their expression pattern in Sahiwal and Tharparkar breeds of zebu cattle (Bos indicus) and Murrah buffalo (Bubalus bubalis) with respect to different seasons. Quantitative real time polymerase chain reaction was performed to analyze the transcript variants of three HSP70 family genes (HSPA1A, HSPA1B, and HSPA8) and HSP10, HSP60, HSP90 and HSF1 in each breed. The major finding of this study was the higher abundance of all the studied HSP genes during summer and winter compared to spring season, but the magnitude of increase was higher during summer as compared to winter. HSPA1A and HSPA1B genes showed maximal induction (P<0.001) during summer and winter while HSP60 and HSP10 were found to be the second most abundantly expressed HSPs. The relative mRNA abundance of HSF1 significantly increased (P<0.001) in Murrah buffalo compared to Tharparkar and Sahiwal cattle during summer and winter. Expression pattern of heat shock protein genes indicated that amongst the breeds, the expression was higher in Murrah buffalo compared to Sahiwal and Tharparkar cattle, thereby indicating the more adaptive capacity of later during periods of stress. Hence, this study suggests that heat shock protein genes may be conveniently used as biomarkers for assessing stress response in cattle and buffalo and the expression is species and breed-specific. Furthermore, the variation in expression is associated with heat tolerance and adaptation to different climatic conditions.

  9. Evidence for bovine besnoitiosis in Egypt-first serosurvey of Besnoitia besnoiti in cattle and water buffalo (Bubalus bubalis) in Egypt.

    PubMed

    Ashmawy, Karam Imam; Abu-Akkada, Somaia Saif

    2014-03-01

    The present study evaluated the presence of specific antibodies against Besnoitia besnoiti in cattle and water buffalo (Bubalus bubalis) in Egypt. Sera from cattle (n = 216) and water buffaloes (n = 133) collected from five different provinces of Egypt (Behera, Alexandria, Assuit, Gharbia, and Matrouh) were analyzed. Testing for B. besnoiti antibodies by PrioCHECK® Besnoitia Ab 2.0 ELISA initially identified 13.75 % (48 out of 349) of individual sera as positive at the manufacturer's suggested cutoff threshold, 15 percent positivity (PP). Statistically significant associations between B. besnoiti prevalence, species, sex, age, and geographical distribution were observed. Seropositive animals were distributed in all of the provinces from which animals were sampled except Gharbia province. Assuit province showed the highest percentage of infection (30.76 %) followed by Matrouh, Alexandria, and Behera provinces (25, 16.29, and 9.6 %, respectively). The highest infection rate of B. besnoiti was significantly higher in cattle (17.13 %) than in water buffaloes (9.02 %). Positive cases were observed in all age categories. While the highest infection rate (17.13 %) was recorded in the age group 5-10 years followed by the age group 1-5 years (15.38 %), and only one positive case (1.58 %) was recorded in the age group less than 1 year. The highest infection rate of B. besnoiti infection was recorded in the female animals (14.95 %) followed by the male animals (8.33). This is the first report on the detection of B. besnoiti in cattle and water buffaloes in Egypt.

  10. Sustained delivery of exogenous melatonin influences biomarkers of oxidative stress and total antioxidant capacity in summer-stressed anestrous water buffalo (Bubalus bubalis).

    PubMed

    Kumar, Ashok; Mehrotra, S; Singh, G; Narayanan, K; Das, G K; Soni, Y K; Singh, Mahak; Mahla, A S; Srivastava, N; Verma, M R

    2015-06-01

    High ambient temperature during summer in tropical and subtropical countries predisposes water buffaloes (Bubalus bubalis) to develop oxidative stress having antigonadotropic and antisteroidogenic actions. Melatonin is a regulator of seasonal reproduction in photoperiodic species and highly effective antioxidant and free radical scavenger. Therefore, a study was designed to evaluate the effect of sustained-release melatonin on biomarkers of oxidative stress i.e., the serum malondialdehyde (MDA) and nitric oxide (NO), and the total antioxidant capacity (TAC). For the study, postpartum buffaloes diagnosed as summer anestrus (absence of overt signs of estrus, concurrent rectal examination, and RIA for serum progesterone) were grouped as treated (single subcutaneous injection of melatonin at 18 mg/50 kg body weight dissolved in sterilized corn oil as vehicle, n = 20) and untreated (subcutaneous sterilized corn oil, n = 8). Blood sampling for estimation of serum TAC and MDA (mmol/L) and NO (μmol/L) was carried out at 4 days of interval from 8 days before treatment till 28 days after treatment or for the ensuing entire cycle length. Results showed serum TAC concentration was higher in the treatment group with a significant (P < 0.05) increasing trend, whereas MDA and NO revealed a significant (P < 0.05) decline. Serum MDA and NO were higher in control compared with those of treatment group. Moreover, buffaloes in the treatment group showed 90% estrus induction with 18.06 ± 1.57 days mean interval from treatment to the onset of estrus. These results report that melatonin has a protective effect by elevating antioxidant status and reducing oxidative stress resulting in the induction of cyclicity in summer-stressed anestrous buffaloes. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Assessment of DNA damage during in vitro development of buffalo (Bubalus bubalis) embryos: effect of cysteamine.

    PubMed

    Mukherjee, A; Kumar, D; Singh, K P; Chauhan, M S; Singla, S K; Palta, P; Manik, R S

    2010-12-01

    Comet assay was used in the present study to examine DNA damage to buffalo oocytes and embryos during in vitro culture. Embryos were produced in vitro from oocytes obtained from slaughterhouse ovaries in presence of cysteamine (IVM and IVC media supplemented with 50 and 100 μM, respectively) or in its absence (controls). Compared to controls, cysteamine supplementation increased (p < 0.01) cleavage rate and proportion of oocytes that developed to 8- to 16-cell stage. The incidence of DNA damage was lower (p < 0.01) in cysteamine group than that in controls at 8- to 16- (19.3 ± 4.24 vs 72.0 ± 5.22%) but not in 2-cell stage embryos (11.7 ± 5.63 vs 20.8 ± 5.49%) or in mature oocytes (5.3 ± 3.43 vs 10.3 ± 4.73%). The tail length, which indicates magnitude of DNA damage, was shorter (p < 0.01) in cysteamine group than in controls in mature oocytes (25.5 ± 0.5 vs 36.0 ± 0.71 pixels) and 8- to 16-cell stage (49.2 ± 1.64 vs 152.7 ± 1.28 pixels) but not in 2-cell stage embryos (36.3 ± 1.54 vs 36.4 ± 0.75 pixels). Also, exposure of oocytes/embryos to UV radiation or H2O2 caused extensive DNA damage. In conclusion, these results suggest that oocytes/embryos suffer from DNA damage during progress of in vitro culture, which can be partly ameliorated by cysteamine supplementation of culture media.

  12. Modulatory role of leptin on ovarian functions in water buffalo (Bubalus bubalis).

    PubMed

    Reshma, R; Mishra, S R; Thakur, N; Parmar, M S; Somal, A; Bharti, M K; Pandey, S; Chandra, V; Chouhan, V S; Verma, M R; Singh, G; Sharma, G T; Maurya, V P; Sarkar, M

    2016-10-15

    The aim of the present study was to demonstrate the modulatory role of leptin on bubaline granulosa cells (GCs) and luteal cells (LCs) functions using an in vitro cell culture system and to establish a cross talk between leptin and insulin-like growth factor-1 (IGF-1). GCs were collected from group IV follicles (>13 mm size) and LCs from mid-luteal phase corpus luteum and were grown in serum-containing media supplemented with leptin at three different dose rates (0.1, 1, and 10 ng/mL) and time durations (24, 48, and 72 hours). We evaluated the production and secretion of estradiol (E2) and progesterone (P4) using RIA and the mRNA expression of steroidogenic acute regulatory protein (STARD1), cytochrome P450 cholesterol side-chain cleavage (CYP11A1), 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450 aromatase (CYP19A1), sterol regulatory element-binding protein 1 (SREBP1), steroidogenic factor-1 (SF1), anti-apoptotic gene PCNA, pro-apoptotic gene caspase 3 and endothelial cell marker, Von Willebrand factor (vWF), using quantitative real-time polymerase chain reaction. The results depicted a direct inhibitory action of leptin on GCs steroidogenesis in a time-dependent manner (P < 0.05), whereas in the presence of IGF-1 the inhibitory effect was reverted. Furthermore, leptin augmented both cellular proliferation (PCNA) and apoptosis (caspase 3). On the other hand, in LCs, leptin alone showed an apparent stimulatory effect on steroidogenesis (P < 0.05); however, in the presence of IGF-1, an antagonistic effect was witnessed. Moreover, leptin had an inhibitory effect on apoptosis while promoted cellular proliferation and angiogenesis. These findings were further strengthened by immunocytochemistry. To conclude, these observations for the first time reported that in buffaloes leptin has a direct dose-, time-, and tissue-dependent effect on ovarian steroidogenesis, angiogenesis, and cytoprotection, and furthermore, it can regulate the effect of systemic

  13. Phylogenetic relationship among all living species of the genus Bubalus based on DNA sequences of the cytochrome b gene.

    PubMed

    Tanaka, K; Solis, C D; Masangkay, J S; Maeda, K; Kawamoto, Y; Namikawa, T

    1996-12-01

    The cytochrome b genes of all living species of Bubalus, including the river type and the swamp type of domestic buffaloes (Bubalus bubalis), were sequenced to clarify their phylogenetic relationships. These sequences were compared together with the African buffalo (Syncerus caffer) and banteng (Bos javanicus) sequences as an outgroup. Phylogenetic trees of Bubalus species based on the DNA sequences of the cytochrome b gene demonstrated that the tamaraw (Bubalus mindorensis), endemic to the Philippines, could be classified into the subgenus Bubalus, not the subgenus Anoa. The divergence time between the lowland anoa (B. depressicornis) and the mountain anoa (B. quarlesi) was estimated at approximately 2.0 million years (Myr), which is almost the same as the coalescence time for the Bubalus sequences. This large genetic distance supports the idea that the lowland anoa and the mountain anoa are different species. An unexpectedly large genetic distance between the river and the swamp type of domestic buffaloes suggests a divergence time of about 1.7 Myr, while the swamp type was noticed to have the closest relationship with the tamaraw (1.5 Myr). This result implies that the two types of domestic buffaloes have differentiated at the full species level.

  14. Evaluation of sperm functional attributes in relation to in vitro sperm-zona pellucida binding ability and cleavage rate in assessing frozen thawed buffalo (Bubalus bubalis) semen quality.

    PubMed

    Selvaraju, S; Ravindra, J P; Ghosh, J; Gupta, P S P; Suresh, K P

    2008-07-01

    The objective of this study was to evaluate sperm functional attributes in relation to in vitro sperm-zona binding ability and cleavage rate in assessing frozen thawed buffalo (Bubalus bubalis) semen quality. Frozen-thawed forty-eight ejaculates from eight Surti buffalo bulls (six ejaculates/bull) obtained by artificial vagina were used. Frozen semen from each bull was thawed, pooled, and subjected for sperm functional (six replicates) and in vitro fertilization (four replicates) tests. The progressive forward motility, plasmalemma functional integrity assessed by fluorogenic [6-carboxyfluorescein diacetate (CFDA), and propidium iodide (PI)], hypoosmotic swelling (HOS), and hypoosmotic swelling-Giemsa (HOS-G) test, mitochondrial membrane potential, sperm nuclear morphology, the number of sperm bound to zona and cleavage rate differed significantly (P<0.05) between bulls. When the animals were grouped based on cleavage rate (group I, >40% cleavage rate, n=5, and group II, <40% cleavage rate, n=3), in vitro fertility parameters and all the sperm functional attributes except sperm nuclear morphology differed significantly (P<0.05). The proportions of sperm with functional plasmalemma in the tail and intact acrosome assessed by HOS-G test (25.33, range: 17.48-40.27) were significantly (P<0.001) lower than the functional plasmalemma in the tail assessed by HOS test (39.80, range: 27.85-54.67). The number of sperm bound to zona had significant correlations with the mitochondrial membrane potential (r=0.90, P<0.01) and plasmalemma integrity (fluorogenic, r=0.74 and HOS, r=0.79, P<0.05) and HOS-G, r=0.87, P<0.01). The cleavage rate had significant (P<0.05) correlations with the mitochondrial membrane potential (r=0.70) and plasmalemma integrity measured by HOS-G test (r=0.68). The present study indicates that these attributes could represent important determinants of buffalo sperm quality influencing cleavage rate.

  15. Effect of breeding season on in vivo oocyte recovery and embryo production in non-descriptive Indian river buffaloes (Bubalus bubalis).

    PubMed

    Manjunatha, B M; Ravindra, J P; Gupta, P S P; Devaraj, M; Nandi, S

    2009-04-01

    The present study was carried out to examine the effect of season on in vivo oocyte recovery and embryo production in non-descriptive, Indian river buffaloes (Bubalus bubalis). Ovum pick up (OPU) was conducted twice a week for 8 weeks during peak (October-March) and low (April-September) breeding season in live buffaloes (n=6). OPU was performed using ultrasound equipment with a 5MHz transvaginal transducer, a single lumen 18-gauge, 55-cm long needle and a constant vacuum pressure of 110mmHg. The number and size of follicles was determined before puncture. The recovered oocytes were graded and only grade A and grade B oocytes were used for in vitro production (IVP) of embryos. The mean number of follicles observed per animal per session did not differed (P<0.05) between animals or between puncture sessions in both low and peak breeding seasons. Higher (P<0.05) number of follicles were observed (4.8+/-0.2 versus 3.1+/-0.3) and punctured (4.0+/-0.2 versus 2.4+/-0.2) during peak breeding season when compared to low breeding season. Oocytes recovered (1.6+/-0.1 versus 1.0+/-0.3) per animal per session were higher (P<0.05) in peak breeding season than low breeding season. During the peak breeding season, the blastocyst yield per animal per session (0.3+/-0.4 versus 0.18+/-0.4) was higher (P<0.05) than the low breeding season. However, season did not significantly affect the percentage of oocytes suitable for IVP (grade A+B) and blastocyst production rate. In conclusion, the efficiency of OPU combined with IVP was higher during the peak breeding season than the low breeding season in buffaloes.

  16. Cellular conservation of endangered midget buffalo (Lowland Anoa, Bubalus quarlesi) by establishment of primary cultured cell, and its immortalization with expression of cell cycle regulators.

    PubMed

    Fukuda, Tomokazu; Iino, Yuuka; Eitsuka, Takahiro; Onuma, Manabu; Katayama, Masafumi; Murata, Koichi; Inoue-Murayama, Miho; Hara, Kumiko; Isogai, Emiko; Kiyono, Tohru

    2016-10-01

    Lowland Anoa has become endangered due to hunting and human activity. Protection and breeding of endangered species in a controlled environment is the best way of conservation. However, it is not possible to adopt this approach for all endangered species because of the cost involved and the ever-increasing number of critically endangered species. In consideration of these limitations to the conventional conservation methods, we established a primary cell culture of endangered buffalo (Lowland Anoa, Bubalus quarlesi), for the preservation of this biological resource. In addition, we introduced human derived, mutant cyclin dependent kinase 4 (CDK4), Cyclin D, and telomerase reverse transcriptase (TERT) into the primary cells. The successful introduction of these three genes was confirmed by western blot with specific antibodies, and enzymatic activity. We also showed that the expression of mutant CDK4, Cyclin D, and TERT allows us to efficiently establish an immortalized cell line, with an intact chromosome pattern, from Lowland Anoa. To the best of our knowledge, this study is the first investigation that established an immortalized cell line of an endangered wild animal species.

  17. Comparison of α1-Antitrypsin, α1-Acid Glycoprotein, Fibrinogen and NOx as Indicator of Subclinical Mastitis in Riverine Buffalo (Bubalus bubalis)

    PubMed Central

    Guha, Anirban; Guha, Ruby; Gera, Sandeep

    2013-01-01

    Mastitis set apart as clinical and sub clinical is a disease complex of dairy cattle, with sub clinical being the most important economically. Of late, laboratories showed interest in developing biochemical markers to diagnose sub clinical mastitis (SCM) in herds. Many workers reported noteworthy alternation of acute phase proteins (APPs) and nitric oxide, (measured as nitrate+nitrite = NOx) in milk due to intra-mammary inflammation. But, the literature on validation of these parameters as indicators of SCM, particularly in riverine milch buffalo (Bubalus bubalis) milk is inadequate. Hence, the present study focused on comparing several APPs viz. α1- anti trypsin, α1- acid glycoprotein, fibrinogen and NOx as indicators of SCM in buffalo milk. These components in milk were estimated using standardized analytical protocols. Somatic cell count (SCC) was done microscopically. Microbial culture was done on 5% ovine blood agar. Of the 776 buffaloes (3,096 quarters) sampled, only 347 buffaloes comprising 496 quarters were found positive for SCM i.e. milk culture showed growth in blood agar with SCC≥2×105 cells/ml of milk. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. It was observed that α1- anti trypsin and NOx had a highly significant (p<0.01) increase in SCM milk, whereas, the increase of α1- acid glycoprotein in infected milk was significant (p<0.05). Fibrinogen was below detection level in both healthy and SCM milk. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and SCC≥2×105 cells/ml of milk as the benchmark. Udder profile correlation coefficient was also used. Allowing for statistical and epidemiological analysis, it was concluded that α1- anti trypsin indicates SCM irrespective of etiology, whereas α1- acid glycoprotein better diagnosed SCM caused by gram positive bacteria. NOx did not prove to be a good

  18. Cryopreservation of Nili-Ravi buffalo (Bubalus bubalis) semen in AndroMed(®) extender; in vitro and in vivo evaluation.

    PubMed

    Ansari, M S; Rakha, B A; Akhter, S

    2017-06-28

    The study was designed to evaluate AndroMed(®) for the freezability and fertility of Nili-Ravi buffalo semen. Semen was collected from four adult Nili-Ravi buffalo (Bubalus bubalis) bulls for 3 weeks (replicate). Semen ejaculates from each buffalo bull were divided into three aliquots. One aliquot was used for evaluation of motility, plasma membrane integrity, livability, viability, DNA integrity and normal apical ridge. Remaining two aliquots were diluted (37°C; 50 × 10(6) spermatozoa/ml) in tris-citric egg yolk or AndroMed(®) extender and cryopreserved in 0.5 ml French straws. After thawing, per cent post-thaw motility (47.9 ± 0.8, 49.2 ± 1.7), plasma membrane integrity (44.4 ± 1.2, 46.8 ± 1.8) and normal apical ridge (81.4 ± 0.3, 83.2 ± 0.3) were recorded similar (p > .05) in tris-citric egg yolk and AndroMed(®) extender. Higher (p < .05) percentage of sperm livability (70.5 ± 1.4 and 64.4 ± 1.0), viability (67.5 ± 1.5 and 61.5 ± 0.6) and DNA integrity (97.0 ± 0.3 and 93.4 ± 0.21) were recorded in AndroMed(®) compared to tris-citric egg yolk post-thaw. Values for all the aforementioned spermatozoal quality parameters were observed lower (p < .05) in frozen-thawed compared to fresh semen irrespective of the experimental extenders. Fertility rates of buffalo semen did not differ (p > .05) either cryopreserved in tris-citric egg yolk or AndroMed(®) extender (45.5% vs. 49%). It is concluded that AndroMed(®) is capable in protecting the buffalo bull sperm during freeze-thawing process and can be adopted safely for routine use replacing the tris-citric egg yolk extender in artificial insemination programme. © 2017 Blackwell Verlag GmbH.

  19. Comparison of α1-Antitrypsin, α1-Acid Glycoprotein, Fibrinogen and NOx as Indicator of Subclinical Mastitis in Riverine Buffalo (Bubalus bubalis).

    PubMed

    Guha, Anirban; Guha, Ruby; Gera, Sandeep

    2013-06-01

    Mastitis set apart as clinical and sub clinical is a disease complex of dairy cattle, with sub clinical being the most important economically. Of late, laboratories showed interest in developing biochemical markers to diagnose sub clinical mastitis (SCM) in herds. Many workers reported noteworthy alternation of acute phase proteins (APPs) and nitric oxide, (measured as nitrate+nitrite = NOx) in milk due to intra-mammary inflammation. But, the literature on validation of these parameters as indicators of SCM, particularly in riverine milch buffalo (Bubalus bubalis) milk is inadequate. Hence, the present study focused on comparing several APPs viz. α1- anti trypsin, α1- acid glycoprotein, fibrinogen and NOx as indicators of SCM in buffalo milk. These components in milk were estimated using standardized analytical protocols. Somatic cell count (SCC) was done microscopically. Microbial culture was done on 5% ovine blood agar. Of the 776 buffaloes (3,096 quarters) sampled, only 347 buffaloes comprising 496 quarters were found positive for SCM i.e. milk culture showed growth in blood agar with SCC≥2×10(5) cells/ml of milk. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. It was observed that α1- anti trypsin and NOx had a highly significant (p<0.01) increase in SCM milk, whereas, the increase of α1- acid glycoprotein in infected milk was significant (p<0.05). Fibrinogen was below detection level in both healthy and SCM milk. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and SCC≥2×10(5) cells/ml of milk as the benchmark. Udder profile correlation coefficient was also used. Allowing for statistical and epidemiological analysis, it was concluded that α1- anti trypsin indicates SCM irrespective of etiology, whereas α1- acid glycoprotein better diagnosed SCM caused by gram positive bacteria. NOx did not prove to be a

  20. The effect of a high-roughage diet on the metabolism of aromatic compounds by rumen microbes: a metagenomic study using Mehsani buffalo (Bubalus bubalis).

    PubMed

    Prajapati, Vimalkumar S; Purohit, Hemant J; Raje, Dhananjay V; Parmar, Nidhi; Patel, Anand B; Jones, Oliver A H; Joshi, Chaitanya G

    2016-02-01

    In developing countries, livestock are often fed a high-lignin, low-nutrient diet that is rich in aromatic compounds. It is therefore important to understand the structure of the microbial community responsible for the metabolism of these substances. A metagenomic analysis was therefore carried out to assess the microbial communities associated with the liquid and solid fractions of rumen biomaterial from domestic Mehsani buffalo (Bubalus bubalis) fed with varying proportions of roughage. The experimental design consisted of three feeding regimes (50, 75 and 100 % roughage) and two roughage types (green and dry). Genes associated with aromatic compound degradation were assessed via high-throughput DNA sequencing. A total of 3914.94 Mb data were generated from all treatment groups. Genes coding for functional responses associated with aromatic compound metabolism were more prevalent in the liquid fraction of rumen samples than solid fractions. Statistically significant differences (p < 0.05) were also observed between treatment groups. These differences were dependent on the proportion of roughage fed to the animal, with the type of roughage having little effect. The genes present in the highest abundance in all treatment groups were those related to aromatic compound catabolism. At the phylum level, Bacteroidetes were dominant in all treatments closely followed by the Firmicutes. This study demonstrates the use of feed type to selectively enrich microbial communities capable of metabolizing aromatic compounds in the rumen of domestic buffalo. The results may help to improve nutrient utilization efficiency in livestock and are thus of interest to farming industries, particularly in developing countries, worldwide.

  1. Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis).

    PubMed

    Dorostkar, Kamran; Alavi-Shoushtari, Sayed Mortaza; Mokarizadeh, Aram

    2012-01-01

    The aim of the present study was to investigate the effect of in vitro supplementation of selenium on fresh and frozen spermatozoa quality of buffalo (Bubalus bubalis) bulls. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 0.5, 1, 2, 4 and 8 µg mL(-1) sodium selenite and the sperm motility and viability were evaluated at 0 (T0) (immediately after dilution), 60 (T1) and 120 (T2) min after diluting semen. In the second step, semen samples were diluted with tris-egg yolk-glycerol extender containing the same amounts of sodium selenite, cooled to 4 ˚C, equilibrated and semen parameters (motility, viability, membrane integrity and DNA damage) were estimated. Then, the semen was packed in 0.5 mL French straws and frozen in liquid nitrogen. Later, the semen was thawed and analyzed for the same parameters, as well as total antioxidant capacity. Results showed that addition of 1 and 2 µgmL(-1) selenium to the semen extender significantly increased the sperm motility of fresh and equilibrated semen compared to the control without affecting other parameters. However, in frozen-thawed semen, extenders containing 1 and 2 µg mL(-1) selenium significantly improved sperm motility, viability, membrane integrity and semen total antioxidant capacity and also resulted in lower DNA damaged sperms. In this study selenium supplementation of semen extender of 4 and 8 µg mL(-1) had deleterious effects on sperm parameters as early as the samples were prepared for freezing.

  2. Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis)

    PubMed Central

    Dorostkar, Kamran; Alavi-Shoushtari, Sayed Mortaza; Mokarizadeh, Aram

    2012-01-01

    The aim of the present study was to investigate the effect of in vitro supplementation of selenium on fresh and frozen spermatozoa quality of buffalo (Bubalus bubalis) bulls. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 0.5, 1, 2, 4 and 8 µg mL-1 sodium selenite and the sperm motility and viability were evaluated at 0 (T0) (immediately after dilution), 60 (T1) and 120 (T2) min after diluting semen. In the second step, semen samples were diluted with tris-egg yolk-glycerol extender containing the same amounts of sodium selenite, cooled to 4 ˚C, equilibrated and semen parameters (motility, viability, membrane integrity and DNA damage) were estimated. Then, the semen was packed in 0.5 mL French straws and frozen in liquid nitrogen. Later, the semen was thawed and analyzed for the same parameters, as well as total antioxidant capacity. Results showed that addition of 1 and 2 µgmL-1 selenium to the semen extender significantly increased the sperm motility of fresh and equilibrated semen compared to the control without affecting other parameters. However, in frozen-thawed semen, extenders containing 1 and 2 µg mL-1 selenium significantly improved sperm motility, viability, membrane integrity and semen total antioxidant capacity and also resulted in lower DNA damaged sperms. In this study selenium supplementation of semen extender of 4 and 8 µg mL-1 had deleterious effects on sperm parameters as early as the samples were prepared for freezing. PMID:25653769

  3. Myxobolus ictiobus n. sp. and Myxobolus minutus n. sp. (Cnidaria: Myxobolidae) from the gills of the smallmouth buffalo Ictiobus bubalus Rafinesque (Cypriniformes: Catostomidae).

    PubMed

    Rosser, Thomas G; Griffin, Matt J; Quiniou, Sylvie M A; Alberson, Neely R; Woodyard, Ethan T; Mischke, Charles C; Greenway, Terrence E; Wise, David J; Pote, Linda M

    2016-07-01

    The smallmouth buffalo Ictiobus bubalus Rafinesque (Catostomidae) is native to North American waterways and occasionally grown in pond aquaculture. Species of Myxobolus Bütschli, 1882 have been reported from the gills, integument, and intestinal tract of buffalo fish, although there is ambiguity in some host records. In the summer of 2013, thirteen adult smallmouth buffalo were seined from a 0.1-acre (0.04-hectare) experimental research pond at the Thad Cochran National Warmwater Aquaculture Center in Stoneville, Mississippi, USA, and examined for the presence of parasitic infection. Two previously unknown species of Myxobolus were observed parasitising the gills. Plasmodia of the two species differed from each other in both size and shape. Morphologically the two species were distinct from one another and from other Myxobolus spp. previously reported from buffalo fish. Myxospores of Myxobolus ictiobus n. sp. were spherical and measured 12.7-14.5 (13.9 ± 0.4) µm in length and 10.7-13.6 (12.5 ± 0.7) µm in width with a thickness of 10.3-14.8 (12.6 ± 2.3) µm. Polar capsules measured 5.6-7.4 (6.6 ± 0.4) µm in length and 3.7-4.9 (4.5 ± 0.8) µm in width and each contained a coiled polar filament with 5-6 turns. Myxospores of Myxobolus minutus n. sp. were circular in shape and measured 7.4-9.6 (8.6 ± 0.7) µm in length and 7.5-9.9 (8.8 ± 0.7) µm in width with a thickness of 6.5-7.3 (6.7 ± 0.3) µm. Polar capsules measured 3.6-4.9 (4.3 ± 0.3) µm in length and 2.8-3.8 (3.3 ± 0.3) µm and each contained a coiled polar filament with 5-6 turns. Supplemental 18S rRNA gene sequencing identified unique sequences for each isolate. Phylogenetic analysis of 18S rRNA sequences demonstrated a strong clustering of both isolates with other species of Myxobolus from cypriniform fish.

  4. Morphological Studies of the Tongue of the Egyptian Water Buffalo (Bubalus bubalis) and Their Lingual Papillae Adaptation for Its Feeding Habits.

    PubMed

    El-Bakary, N E R; Abumandour, M M A

    2017-10-01

    This work was conducted to describe the morphological characters of the tongue of Egyptian water buffalo (Bubalus bubalis). The lingual root and the dorsal middle region of apex and body in addition to the dorsal and ventral surface of lingual tip were devoided from any fungiform papillae. The lingual tip contains conical papillae only. The ventral surface of lingual apex was divided into two portions by the U-shaped fungiform line into papillary and non-papillary region. Histological investigation on the lingual surface epithelium and lamina propria submucosa reflects differences in these layers in different parts of the tongue. By SEM, there are two subtypes of filiform papillae: caudally directed papillae on dorsal surface and rostrally directed papillae on the lateral region of ventral surface of lingual apex. There are two subtypes of conical papillae: small slightly rostrally directed papillae on dorsal and ventral surface of lingual tip and large posteromedially directed papillae on dorsal surface of lingual root. The rounded circumvallate papillae consisted of round bulb surrounded by deep circular groove, which surrounded by circular pad. Higher magnification of filiform papillae indicates the presence of microcrests separated by microgrooves, and these microgrooves consisted of microrodes. The fungiform surface having micropores on the tip of elevated tubercle for taste buds pores. All these observed structures (microcrests, microgrooves, microrodes, tubercles, microridges) in a higher magnification allow animals to transport food particles through the oral cavity and help in the defensive behaviour. There are strong correlations between the tongue anatomical characteristics and its functions. © 2017 Blackwell Verlag GmbH.

  5. Molecular Mining of Alleles in Water Buffalo Bubalus bubalis and Characterization of the TSPY1 and COL6A1 Genes

    PubMed Central

    Kumar, Sudhir; Ali, Sher

    2011-01-01

    Background Minisatellites are an integral part of eukaryotic genomes and show variation in the complexity of their organization. Besides their presence in non-coding regions, a small fraction of them are part of the transcriptome, possibly participating in gene regulation, expression and silencing. We studied the minisatellite (TGG)n tagged transcriptome in the water buffalo Bubalus bubalis across various tissues and the spermatozoa, and characterized the genes TSPY1 and COL6A1 discovered in the process. Results Minisatellite associated sequence amplification (MASA) conducted using cDNA and oligonucleotide primer (TGG)5 uncovered 38 different mRNA transcripts from somatic tissues and gonads and 15 from spermatozoa. These mRNA transcripts corresponded to several known and novel genes. The majority of the transcripts showed the highest level of expression either in the testes or spermatozoa with exception of a few showing higher expression levels in the lungs and liver. Transcript SR1, which is expressed in all the somatic tissues and gonads, was found to be similar to the Bos taurus collagen type VI alpha 1 gene (COL6A1). Similarly, SR29, a testis-specific transcript, was found to be similar to the Bos taurus testis-specific Y-encoded protein-1 representing cancer/testis antigen 78 (CT78). Subsequently, full length coding sequences (cds) of these two transcripts were obtained. Quantitative PCR (q-PCR) revealed 182-202 copies of theTSPY1 gene in water buffalo, which localized to the Y chromosome. Conclusions The MASA approach enabled us to identify several genes, including two of clinical significance, without screening an entire cDNA library. Genes identified with TGG repeats are not part of a specific family of proteins and instead are distributed randomly throughout the genome. Genes showing elevated expression in the testes and spermatozoa may prove to be potential candidates for in-depth characterization. Furthermore, their possible involvement in fertility or

  6. Cytogenetic analysis of the tamaraw (Bubalus mindorensis): a comparison of R-banded karyotype and chromosomal distribution of centromeric satellite DNAs, telomeric sequence, and 18S-28S rRNA genes with domestic water buffaloes.

    PubMed

    Tanaka, K; Matsuda, Y; Masangkay, J S; Solis, C D; Anunciado, R V; Kuro-o, M; Namikawa, T

    2000-01-01

    The karyotype of the tamaraw (Bubalus mindorensis, 2n = 46) was investigated by RBG-banding technique and compared with those of the river and the swamp cytotypes of domestic water buffalo (B. bubalis). The tamaraw karyotype consisted of 6 submetacentric and 16 acrocentric autosome pairs (NAA = 56), and X and Y chromosomes. The RBG-banded karyotype of the three taxa had a high degree of homology, and the tamaraw karyotype could be explained by a Robertsonian translocation between chromosomes 7 and 15 and by a telomere-centromere tandem fusion between chromosomes 4p and 12 of the standardized river buffalo cytotype (2n = 50, NAA = 58). The buffalo satellite I and II DNAs were localized to the centromeric regions of all the tamaraw chromosomes. The biarmed chromosome 2 of the tamaraw resulting from the fusion between chromosomes 7 and 15 of the standard contained much larger amounts of the satellite I DNA than the other biarmed chromosomes, suggesting that this chromosome was formed by a relatively recent Robertsonian fusion. The (TTAGGG)n telomeric sequence was specifically localized to the telomeric region of all the buffalo chromosomes. The 18S + 28S rDNA was localized to the telomeric regions of the chromosomes 5p, 7, 19, 21, and 22 of the tamaraw and of their homologous chromosomes in the river and swamp buffalo cytotypes.

  7. Investigation of anti-Toxoplasma gondii antibodies in water buffaloes (Bubalus bubalis) in Samsun and Afyon provinces.

    PubMed

    Beyhan, Yunus Emre; Babür, Cahit; Yılmaz, Oktay

    2014-12-01

    The present study was conducted to investigate the seroprevalence of Toxoplasma gondii infection in water buffaloes. The research was conducted on 131 water buffaloes from Samsun and Afyon provinces. Serum samples of buffaloes were tested for toxoplasmosis with the Sabin Feldman Dye Test (SFDT), using live tachyzoites and methylene-blue dye. 87.79% of buffaloes had anti-Toxoplasma gondii antibodies titered 1:16 and more. No statistically significant difference was observed between the age and gender of buffaloes with infection (p>0.05). These results show that toxoplasmosis is quite common in water buffaloes in the regions of study.

  8. Assessment of genetic variability and structuring of riverine buffalo population (Bubalus bubalis) of Indo-Gangetic basin.

    PubMed

    Joshi, Jyoti; Salar, R K; Banerjee, Priyanka; Sharma, Upasna; Tantia, M S; Vijh, R K

    2015-01-01

    The buffalo population of Uttar Pradesh (UP) constitutes 26.1% of the total buffalo population of India, yet this population has not been classified into distinct breeds or subpopulations due to lack of systematic study. Genetic variation at 30 microsatellite loci was examined and statistical analysis was carried out to reveal genetic diversity, demographic parameters of these buffaloes and to investigate the existence of population substructures underlying geographical distribution. The mean number of alleles per locus was 13.26 and mean effective number of alleles was 3.74, whereas mean observed and expected heterozygosities were found to be 0.57 and 0.67 in UP buffaloes. Principal component analysis (PCA) based on allele frequency data revealed subclustering of UP buffalo population. Bayesian analysis result also revealed clear membership of individuals into five clusters indicating a genetic subdivision within the UP buffalo population. The buffaloes of Western and Central regions of UP were subtly separated while buffaloes of Tarai area and Bhadawari buffaloes revealed distinctive population structure. The buffaloes of Mau, Ballia and Ghazipur districts of Eastern region also had a distinctive genetic structure. The analysis of data on buffaloes of Indo-Gangetic plains revealed that population was in mutation drift equilibrium. The observed mean M ratio in the population was above the critical significance value (Mc) suggesting that it has not suffered any severe reduction in effective population size. The statistical tests revealed a historical constancy of size of buffalo in this geographical area. The high level of genetic variability indicates UP buffalo population is a vast reservoir of genetic diversity and this shall help in taking informed conservation decisions and sustainable utilization.

  9. Investigation into omocysteine, vitamin E and malondialdehyde as indicators of successful artificial insemination in synchronized buffalo cows (Bubalus bubalis).

    PubMed

    Barbato, Olimpia; Chiaradia, Elisabetta; Barile, Vittoria Lucia; Pierri, Francesca; de Sousa, Noelita Melo; Terracina, Luigi; Canali, Claudio; Avellini, Luca

    2016-02-01

    The aim of this study was to describe modifications in plasma homocysteine (Hcy), vitamin E (VitE) and malondialdehyde (MDA) concentrations in the first 56 days after artificial insemination (AI) in buffalo. Thirty-five buffalo cows were divided, ex post, into three groups on the basis of pregnancy diagnosis: pregnant, not pregnant, with embryonic mortality. Pregnancy was diagnosed by ultrasonography and plasma concentrations of pregnancy-associated glycoproteins (PAGs). Our results showed that, in pregnant buffaloes, included those with embryonic mortality, MDA increased progressively while VitE decreased. In non-pregnant buffaloes, MDA and Vit E were unchanged. Hcy concentrations also remained unchanged within each group throughout the study period, but were lower in non-pregnant buffaloes than in the pregnant ones and in those with embryonic mortality. In conclusion, present data suggest that successful pregnancy in buffalo cows might be linked to Hcy metabolism and oxidative stress involvement.

  10. Activation and Inhibition of The Wnt3A Signaling Pathway in Buffalo (Bubalus bubalis) Embryonic Stem Cells: Effects of WNT3A, Bio and Dkk1.

    PubMed

    Zandi, Mohammad; Shah, Syed Mohamad; Muzaffar, Musharifa; Kumar Singh, Manoj; Palta, Prabhat; Kumar Singla, Suresh; Sham Manik, Radhey; Chauhan, Manmohan Singh

    2015-01-01

    This research studies the effects of activation and inhibition of Wnt3A signaling pathway in buffalo (Bubalus bubalis) embryonic stem (ES) cell-like cells. To carry on this experimental study, the effects of activation and inhibition of Wnt3A signaling in buffalo ES cell-like cells were examined using Bio (0.5 mM) combined with WNT3A (200 ng/ml), as an activator, and Dickkopf-1 (Dkk1, 250 ng/ml), as an inhibitor, of the pathway. ES cells were cultured up to three weeks in ES cell medium without fibroblast growth factor-2 (FGF-2) and leukemia inhibitory factor (LIF), but in the presence of Bio, WNT3A, Bio+WNT3A and Dkk1. The effects of these supplements were measured on the mean area of ES cell colonies and on the expression levels of a number of important genes related to pluripotency (Oct4, Nanog, Sox2 and c-Myc) and the Wnt pathway (β-catenin). ES cell colonies cultured in ES cell medium that contained optimized quantities of LIF and FGF-2 were used as the control. Data were collected for week-1 and week-3 treated cultures. In addition, WNT3A-transfected ES cells were compared with the respective mock-transfected colonies, either alone or in combination with Dkk1 for expression of β-catenin and the pluripotency-related genes. Data were analyzed by ANOVA, and statistical significance was accepted at P<0.05. Among various examined concentrations of Bio (0.5-5 mM), the optimum effect was observed at the 0.5 mM dose as indicated by colony area and expressions of pluripotency-related genes at both weeks-1 and -3 culture periods. At this concentration,the expressions of Nanog, Oct3/4, Sox2, c-Myc and β-catenin genes were nonsignificantly higher compared to the controls. Expressions of these genes were highest in the Bio+WNT3A treated group, followed by the WNT3A and Bio-supplemented groups, and lowest in the Dkk1-treated group. The WNT-transfected colonies showed higher expressions compared to both mock and Dkk1-treated mock transfected colonies. WNT3A functions to

  11. Controlled breeding and reproductive management in water buffaloes (Bubalus bubalis) using Eazi Breed controlled internal drug release.

    PubMed

    Hiremath, Shivayogi; Ramesha, Kerekoppa P

    2015-06-04

    Buffalo reproduction is considerably affected by late maturity, poor oestrus symptoms and long postpartum periods. This study was undertaken to evaluate the efficiency of Eazi Breed controlled internal drug release (CIDR), an intravaginal progesterone-releasing device, in relation to oestrus and fertility. Five hundred true anoestrus buffalo cows, in the age group 4-6 years in 10 villages of Dharwad district in Karnataka state in India, were randomly selected and treated with CIDR for 9 days. Two mL of Cidirol (1 mg oestradiol benzoate) was administered intramuscularly to all animals on day 10. Forty-two buffaloes (8.4%) that failed to show oestrus signs (1.6%) or showed weak signs of oestrus (6.8%) after the first treatment were treated again 72 h after the Cidriol injection with a new device, and inseminated after the expression of oestrus. After the second treatment all the animals showed oestrus signs. The percentage of buffaloes showing intense oestrus was 67.40%, intermediate oestrus was shown by 25.80%, whilst 6.80% buffaloes showed weak oestrus even after the second treatment. The buffaloes showing oestrus signs were inseminated twice with an interval of 12 h, starting 12 h after the start of the oestrus signs. In 86 buffaloes showing prolonged oestrus signs a third insemination was done. The conception rates were 85.16%, 60.47% and 44.11% respectively in buffaloes showing intense, intermediate and weak oestrus. Transrectal palpation of the genital tract was performed 45-60 days post-insemination to diagnose pregnancy status, and in doubtful cases pregnancy was reconfirmed at 90 days after insemination. Out of 500 buffaloes treated in this way 380 animals became pregnant and the pregnancy rate was 76%. This study revealed the usefulness of Eazi Breed CIDR along with Cidirol treatment in buffaloes to improve their reproductive performance.

  12. Production of interspecies handmade cloned embryos by nuclear transfer of cattle, goat and rat fibroblasts to buffalo (Bubalus bubalis) oocytes.

    PubMed

    Selokar, N L; George, A; Saha, A P; Sharma, R; Muzaffer, M; Shah, R A; Palta, P; Chauhan, M S; Manik, R S; Singla, S K

    2011-02-01

    The possibility of producing interspecies handmade cloned (iHMC) embryos by nuclear transfer from donor cells of cattle, goat and rat using buffalo oocytes as recipient cytoplasts was explored. Zona-free buffalo oocytes were enucleated by protrusion cone-guided bisection with a microblade. After electrofusion with somatic cells, reconstructed oocytes were activated by calcimycin A23187, treated with 6-dimethylaminopurine and were cultured in K-RVCL-50® medium for 8 days. Although the cleavage rate was not significantly different when buffalo, cattle, goat or rat cells were used as donor nuclei (74.6 ± 3.8, 82.8 ± 5.3, 86.0 ± 4.9 and 82.3 ± 3.6%, respectively), the blastocyst rate was significantly higher (P<0.01) for buffalo (51.4 ± 2.6) than for cattle (3.5 ± 1.0) or the goat (2.2 ± 0.9), whereas none of the embryos crossed the 32-cell stage when rat cells were used. However, the total cell number was similar for buffalo-buffalo (175.0 ± 5.07) and cattle-buffalo embryos (178.0 ± 11.84). Following transfer of 3 buffalo-buffalo embryos each to 6 recipients, 3 were found to be pregnant, though the pregnancies were not carried to full term. These results suggest that interspecies blastocyst stage embryos can be produced by iHMC using buffalo cytoplasts and differentiated somatic cells from cattle and goat and that the source of donor nucleus affects the developmental competence of interspecies embryos.

  13. Seroprevalence and risk factors associated with exposure of water buffalo (Bubalus bubalis) to Neospora caninum in northeast Thailand.

    PubMed

    Kengradomkij, Chanya; Inpankaew, Tawin; Kamyingkird, Ketsarin; Wongpanit, Kannika; Wongnakphet, Sirichai; Mitchell, Thomas J; Xuan, Xuenan; Igarashi, Ikuo; Jittapalapong, Sathaporn; Stich, Roger W

    2015-01-15

    Water buffalo are important draft animals for agriculture in resource-restricted areas worldwide. Water buffalo were shown to be experimentally susceptible to infection with Neospora caninum, potentially affected by neosporosis, and naturally exposed to the parasite in Asia. Although enzootic to Thailand, the distribution of N. caninum among Thai water buffalo is unclear. The objectives of this study were to determine the seroprevalence of N. caninum among water buffalo of northeast Thailand and to identify risk factors associated with their exposure to N. caninum. Sera from 628 water buffalo from 288 farms were tested with an indirect fluorescent antibody test (IFAT). A total of 57 samples from 48 herds contained antibodies to N. caninum, indicating overall seroprevalence of 9.1% and 16.7% among individual animals and herds, respectively. The overall seroprevalence was highest in provinces located in the Khorat Basin in the southern part of the region tested. Host age was also associated with seroprevalence, with the greatest seroprevalence (16.1%) among buffalo over 10 years of age, followed by 5-10 years of age (13.4%), 3-5 years (9.2%), and less than 3 years (1.2%). These results collectively suggested that horizontal transmission from canine definitive hosts was an important route of water buffalo exposure to N. caninum. These results also verified the importance of risk factor analysis for effective bovine neosporosis control strategies at the local level. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Association analysis of polymorphism in thyroglobulin gene promoter with milk production traits in riverine buffalo (Bubalus bubalis)

    PubMed Central

    Dubey, P.K.; Goyal, S.; Mishra, S.K.; Yadav, A.K.; Kathiravan, P.; Arora, R.; Malik, R.; Kataria, R.S.

    2015-01-01

    Polymorphism within the promoter region of bovine thyroglobulin has been reported to be associated with milk and meat quality. In this study, we investigated the genetic variation within thyroglobulin promoter region of swamp and riverine buffaloes using PCR–SSCP technique and sequencing, and also analyzing association of polymorphism with the milk production traits. The study revealed four conformational patterns, A, B, C, and D among 323 buffaloes of two riverine breeds and different swamp populations. The frequency of SSCP variant ‘A’ was found to be invariably high among all buffalo populations. Variant ‘C’ was found to be absent in pure swamp population and present with higher frequency among riverine dairy breeds Mehsana and Nili Ravi. Frequency of D variant was observed to be highest in buffalo population, representing riverine and hybrid types. Sequencing of three representative PCR products of each of the SSCP variants, revealed three polymorphic sites responsible, 33C > T, 176G > T and 221C > T, in the buffalo TG promoter region. Further, association studies of SSCP variants with various milk production and milk quality traits indicated significant effect on fat percentage in buffaloes belonging to Mehsana and Nili Ravi dairy breeds. The preliminary results also showed the substantial variations in the distribution of SSCP variants' frequencies across swamp and riverine buffaloes, two distinct populations being reared for meat and milk production, respectively. PMID:26273563

  15. Effect of Terminalia arjuna bark powder on some diagnostic enzymes in buffalo (Bubalus bubalis) ingesting arsenic contaminated water and fodder

    PubMed Central

    Dash, Subrat Kumar; Nayyar, Shashi; Jindal, Rajesh

    2016-01-01

    Aim: The study investigated the effect of Terminalia arjuna bark powder on some diagnostic enzymes related to hepatic and muscle function in buffaloes ingesting arsenic contaminated water and fodder in an arsenic affected area. Materials and Methods: A total of 25 samples of tube well water, fodder and buffalo blood were collected through a survey from arsenic contaminated areas and 20 samples from the uncontaminated, i.e., control areas of Ludhiana district, Punjab for determination of arsenic concentration. A total of 30 buffaloes (selected from above 45 animals) were divided into three groups of 10 each on the basis of blood arsenic level, viz., control group: Clinically healthy buffaloes from the uncontaminated area with the blood arsenic level within the normal limit (0-0.05 ppm); Arsenic exposed group: Buffaloes exposed to arsenic through intake of contaminated water and fodder in the arsenic affected area with the blood arsenic level above the normal limit of 0-0.05 ppm; treatment group: Arsenic exposed buffaloes treated with T. arjuna bark powder orally at 42 mg/kg b.w. OD for 30 days. Single blood samples were collected from control and arsenic exposed groups. Blood samples from the treatment group were collected on 0, 15th, and 30th day of treatment along with one sample on the 45th day, i.e., after withdrawal of treatment. Activities of alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), and creatine kinase (CK) were assayed in plasma. Results: Significantly (p<0.05) higher arsenic concentration was observed in tube well water, fodder and buffalo blood samples collected from the arsenic contaminated area. A significant positive correlation was noticed between arsenic concentrations of tube well water, fodder and untreated buffalo blood samples, collected from the arsenic affected area. ALP, GGT, LDH, and CK activities were significantly (p<0.05) increased in the arsenic exposed buffaloes compared to control

  16. Effect of Terminalia arjuna bark powder on some diagnostic enzymes in buffalo (Bubalus bubalis) ingesting arsenic contaminated water and fodder.

    PubMed

    Dash, Subrat Kumar; Nayyar, Shashi; Jindal, Rajesh

    2016-10-01

    The study investigated the effect of Terminalia arjuna bark powder on some diagnostic enzymes related to hepatic and muscle function in buffaloes ingesting arsenic contaminated water and fodder in an arsenic affected area. A total of 25 samples of tube well water, fodder and buffalo blood were collected through a survey from arsenic contaminated areas and 20 samples from the uncontaminated, i.e., control areas of Ludhiana district, Punjab for determination of arsenic concentration. A total of 30 buffaloes (selected from above 45 animals) were divided into three groups of 10 each on the basis of blood arsenic level, viz., control group: Clinically healthy buffaloes from the uncontaminated area with the blood arsenic level within the normal limit (0-0.05 ppm); Arsenic exposed group: Buffaloes exposed to arsenic through intake of contaminated water and fodder in the arsenic affected area with the blood arsenic level above the normal limit of 0-0.05 ppm; treatment group: Arsenic exposed buffaloes treated with T. arjuna bark powder orally at 42 mg/kg b.w. OD for 30 days. Single blood samples were collected from control and arsenic exposed groups. Blood samples from the treatment group were collected on 0, 15(th), and 30(th) day of treatment along with one sample on the 45(th) day, i.e., after withdrawal of treatment. Activities of alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), and creatine kinase (CK) were assayed in plasma. Significantly (p<0.05) higher arsenic concentration was observed in tube well water, fodder and buffalo blood samples collected from the arsenic contaminated area. A significant positive correlation was noticed between arsenic concentrations of tube well water, fodder and untreated buffalo blood samples, collected from the arsenic affected area. ALP, GGT, LDH, and CK activities were significantly (p<0.05) increased in the arsenic exposed buffaloes compared to control. Treatment with T. arjuna bark powder

  17. Pregnancy-associated glycoproteins (PAGs) concentrations in water buffaloes (Bubalus bubalis) during gestation and the postpartum period.

    PubMed

    Barbato, O; Menchetti, L; Sousa, N M; Malfatti, A; Brecchia, G; Canali, C; Beckers, J F; Barile, V L

    2017-07-15

    For the first time in literature this study describes the pregnancy-associated glycoprotein (PAG) profile of buffalo cows during gestation and the post-partum period using antiserum raised against PAG-molecules purified from buffalo placenta (AS#860). Ninety-eight buffalo cows, belonging to a buffalo herd subjected to a synchronization and artificial insemination (AI) program, were enrolled in this study. Blood samples were taken on days 0 (AI), 23, 25, 28, 30 and then biweekly until the end of pregnancy. Pregnancy was confirmed by ultrasonography on days 28 and 45, and by rectal palpation from day 60 onwards. Blood samples were suspended for the non-pregnant cows on day 45, while the blood of 20 buffaloes that had calved was tested every five days from the day of calving until day 50 post-calving. A cut-off value of 1.0 ng/mL was used in order to discriminate between pregnant and non-pregnant buffaloes. We used Linear Mixed models after Log(x+1) transformation to analyse the PAG concentrations. Fifty-two buffalo cows had become pregnant out of 98 synchronized (53%) and 46 remained non-pregnant (47%) as shown by ultrasonography and the PAG analysis. Significant differences (P < 0.001) in PAG concentrations were observed between the pregnant and non-pregnant buffaloes from day 23 as the PAG of the non-pregnant cows was always close to zero. Conversely, the PAG of the pregnant cows increased progressively from day AI until day 105 post-insemination and then stabilized until the end of pregnancy. Regarding pregnancy diagnosis, the sensitivity of PAG-RIA 860 system (ability of the test to correctly identify pregnant buffalo) ranged from 23% on day 23-98% on day 28 post AI; the specificity (ability to correctly identify non-pregnant buffaloes) was 100% throughout the sampling period. PAG progressively decreased from parturition to day 25 post-partum; from day 30 post-partum, the concentrations fell below 1 ng/mL and were close to 0 on the last day of observation (50

  18. Alterations in follicular fluid estradiol, progesterone and insulin concentrations during ovarian acyclicity in water buffalo (Bubalus bubalis).

    PubMed

    Khan, F A; Das, G K; Pande, Megha; Sarkar, M; Mahapatra, R K; Shankar, Uma

    2012-01-01

    Ovarian acyclicity is one of the most important causes of infertility in water buffalo. Recent studies have indicated alterations in the composition of follicular fluid during the condition. The aim of this study was to determine the changes in follicular fluid concentrations of estradiol, progesterone and insulin during ovarian acyclicity in water buffalo. Ovaries were collected from 50 acyclic and 95 cyclic (control) buffaloes and follicular fluid was aspirated from small (5.0-6.9 mm), medium (7.0-9.9 mm) and large (≥10.0 mm) sized follicles. Estradiol concentration was lower (P<0.0001) in acyclic (1.4 ± 0.09 ng/ml) than in cyclic (3.3 ± 0.18 ng/ml) buffaloes. Regardless of the ovarian cyclic status, there was an increase (P<0.01) in estradiol concentration with the increase in follicle size; the mean concentrations were 2.4 ± 0.16 ng/ml, 2.8 ± 0.29 ng/ml and 3.5 ± 0.41 ng/ml in small, medium and large follicles, respectively. A higher (P<0.001) progesterone concentration was recorded in acyclic (24.3 ± 2.61 ng/ml) compared to the cyclic (7.6 ± 0.79 ng/ml) group. Furthermore, acyclic buffaloes had a lower (P<0.05) concentration of insulin in the follicular fluid than that of cyclic buffaloes (15.2 ± 1.55 μIU/ml versus 25.9 ± 2.78 μIU/ml, respectively). In conclusion, acyclic buffaloes have lower concentrations of estradiol and insulin concurrent with higher concentrations of progesterone in the follicular fluid. These hormonal changes in the follicular microenvironment are possibly a manifestation of the disturbances in the normal follicular development leading to anovulation and anestrus in acyclic buffaloes.

  19. Comparison of non-linear models to describe the lactation curves for milk yield and composition in buffaloes (Bubalus bubalis).

    PubMed

    Ghavi Hossein-Zadeh, N

    2016-02-01

    In order to describe the lactation curves of milk yield (MY) and composition in buffaloes, seven non-linear mathematical equations (Wood, Dhanoa, Sikka, Nelder, Brody, Dijkstra and Rook) were used. Data were 116,117 test-day records for MY, fat (FP) and protein (PP) percentages of milk from the first three lactations of buffaloes which were collected from 893 herds in the period from 1992 to 2012 by the Animal Breeding Center of Iran. Each model was fitted to monthly production records of dairy buffaloes using the NLIN and MODEL procedures in SAS and the parameters were estimated. The models were tested for goodness of fit using adjusted coefficient of determination (Radj(2)), root means square error (RMSE), Durbin-Watson statistic and Akaike's information criterion (AIC). The Dijkstra model provided the best fit of MY and PP of milk for the first three parities of buffaloes due to the lower values of RMSE and AIC than other models. For the first-parity buffaloes, Sikka and Brody models provided the best fit of FP, but for the second- and third-parity buffaloes, Sikka model and Brody equation provided the best fit of lactation curve for FP, respectively. The results of this study showed that the Wood and Dhanoa equations were able to estimate the time to the peak MY more accurately than the other equations. In addition, Nelder and Dijkstra equations were able to estimate the peak time at second and third parities more accurately than other equations, respectively. Brody function provided more accurate predictions of peak MY over the first three parities of buffaloes. There was generally a positive relationship between 305-day MY and persistency measures and also between peak yield and 305-day MY, calculated by different models, within each lactation in the current study. Overall, evaluation of the different equations used in the current study indicated the potential of the non-linear models for fitting monthly productive records of buffaloes.

  20. Modelling lactation curve for milk fat to protein ratio in Iranian buffaloes (Bubalus bubalis) using non-linear mixed models.

    PubMed

    Hossein-Zadeh, Navid Ghavi

    2016-08-01

    The aim of this study was to compare seven non-linear mathematical models (Brody, Wood, Dhanoa, Sikka, Nelder, Rook and Dijkstra) to examine their efficiency in describing the lactation curves for milk fat to protein ratio (FPR) in Iranian buffaloes. Data were 43 818 test-day records for FPR from the first three lactations of Iranian buffaloes which were collected on 523 dairy herds in the period from 1996 to 2012 by the Animal Breeding Center of Iran. Each model was fitted to monthly FPR records of buffaloes using the non-linear mixed model procedure (PROC NLMIXED) in SAS and the parameters were estimated. The models were tested for goodness of fit using Akaike's information criterion (AIC), Bayesian information criterion (BIC) and log maximum likelihood (-2 Log L). The Nelder and Sikka mixed models provided the best fit of lactation curve for FPR in the first and second lactations of Iranian buffaloes, respectively. However, Wood, Dhanoa and Sikka mixed models provided the best fit of lactation curve for FPR in the third parity buffaloes. Evaluation of first, second and third lactation features showed that all models, except for Dijkstra model in the third lactation, under-predicted test time at which daily FPR was minimum. On the other hand, minimum FPR was over-predicted by all equations. Evaluation of the different models used in this study indicated that non-linear mixed models were sufficient for fitting test-day FPR records of Iranian buffaloes.

  1. Novel polymorphisms of the IGF1R gene and their association with average daily gain in Egyptian buffalo (Bubalus bubalis).

    PubMed

    El-Magd, M A; Abbas, H E; El-kattawy, A M; Mokhbatly, A

    2013-08-01

    The objective of this study was to detect insulin-like growth factor 1 receptor (IGF1R) polymorphisms, their allele, and genotype frequencies and to determine associations between these polymorphisms and growth traits in Egyptian water buffalo. Three loci of the IGF1R coding region were amplified by RT-PCR and, subsequently, subjected to sequence analysis, followed by single-strand conformation polymorphism to identify different allelic patterns. A total of 11 novel polymorphisms were detected; 6 SNPs among Egyptian water buffaloes and 5 polymorphisms compared with Indian buffalo (Y12700). Three of those polymorphisms; GAG Indel polymorphism, C261G, and G263C SNPs, were nonsynonymous mutations. The GAG Indel polymorphism led to deletion of E (glutamic) amino acid (aa) in the IGF1R of Egyptian water buffaloes compared with Indian buffalo. However, C261G SNP, which replaced A (alanine) by G (glycine) aa, and G263C SNP, which changed A (alanine) to P (proline) aa, were detected among Egyptian water buffaloes. Three different single-strand conformation polymorphism patterns were observed in exon 21: CC/CC, GG/GG, and CG/GC with frequencies of 0.291, 0.253, and 0.556, respectively. The heterozygous animals (CG/GC) had a higher ADG than homozygous animals (CC/CC and GG/GG) from birth to 6 mo of age. We conclude that the heterozygous haplotype, C261G/G263C, in exon 21 of the IGF1R gene is associated with the ADG during the early stages of life (from birth to 6 mo of age) and could be used as a genetic marker for selection of growth traits in Egyptian buffalo.

  2. Loss of heat shock protein 70 from apical region of buffalo (Bubalus bubalis) sperm head after freezing and thawing.

    PubMed

    Varghese, Tincy; Divyashree, Bannur C; Roy, Sudhir C; Roy, Kajal S

    2016-03-15

    The post-thaw fertility of frozen-thawed mammalian spermatozoa is substantially low as compared with that of fresh sperm. Furthermore, the post-thaw fertility of the cryopreserved buffalo sperm has been reported to be poor as compared with that of cattle sperm. Recently, heat shock protein 70 (HSP70) has been found to play a critical role in mammalian fertilization and early embryonic development in boar and cattle. However, the presence of such fertility-related HSP70 in buffalo sperm and its status after cryopreservation has not been reported so far. Thus, a study was conducted to determine the effect of cryopreservation on the level and distribution pattern of HSP70 molecule in buffalo sperm after cryopreservation. Buffalo semen samples, after dilution in semen extender, were aliquoted in straws and divided into two groups. One group was not cryopreserved, and the other group was cryopreserved for 60 days. Sperm proteins were extracted from both non-cryopreserved (NC) and cryopreserved (C) sperm and subjected to Western blot analysis for detection of HSP70 using a monoclonal anti-HSP70 antibody. The distribution pattern of these proteins in buffalo sperm was also monitored before and after cryopreservation using indirect immunofluorescence technique. A prominent 70-kDa protein band of HSP70 protein was detected in protein extracts of both NC and C buffalo sperm. Densitometry analysis revealed that the intensity of 70-kDa HSP70 protein band of cryopreserved sperm decreased significantly (P < 0.05) compared with that of NC sperm. However, the level of HSP70 in cryopreserved extended seminal plasma (ESP) did not change as compared with that of NC samples indicating a possible degradation of HSP70 in the spermatozoa itself rather than leakage of the protein into the ESP. Furthermore, Western blot also confirmed that several HSP70 immunoreactive protein bands detected in the ESP were contributed by the egg yolk that was added to the extender. Immunocytochemistry

  3. Development of a sequential multicolor-FISH approach with 13 chromosome-specific painting probes for the rapid identification of river buffalo (Bubalus bubalis, 2n = 50) chromosomes.

    PubMed

    Pauciullo, Alfredo; Perucatti, Angela; Iannuzzi, Alessandra; Incarnato, Domenico; Genualdo, Viviana; Di Berardino, Dino; Iannuzzi, Leopoldo

    2014-08-01

    The development of new molecular techniques (array CGH, M-FISH, SKY-FISH, etc.) has led to great advancements in the entire field of molecular cytogenetics. However, the application of these methods is still very limited in farm animals. In the present study, we report, for the first time, the production of 13 river buffalo (Bubalus bubalis, 2n = 50) chromosome-specific painting probes, generated via chromosome microdissection and the DOP-PCR procedure. A sequential multicolor-FISH approach is also proposed on the same slide for the rapid identification of river buffalo chromosome/arms, namely, 1p-1q, 2p-2q, 3p-3q, 4p-4q, 5p-5q, 18, X, and Y, using both conventional and late-replicating banded chromosome preparations counterstained by DAPI. The provided 'bank' of chromosome-specific painting probes is useful for any further cytogenetic investigation not only for the buffalo breeds, but also for other species of the family Bovidae, such as cattle, sheep, and goats, for chromosome abnormality diagnosis, and, more generally, for evolutionary studies.

  4. Anatomic reference for computed tomography of paranasal sinuses and their communication in the Egyptian buffalo (Bubalus bubalis).

    PubMed

    Alsafy, M A M; El-Gendy, S A A; El Sharaby, A A

    2013-06-01

    The purpose of this work was to present an anatomic reference for computed tomography (CT) for the paranasal sinuses of adult buffalo fit the use of anatomists, radiologists, clinicians and veterinary students. CT images with the most closely corresponding cross sections of the head were selected and studied serially in a rostral to caudal progression from the level of the interdental space to the level of the nuchal line. The anatomical features were compared with the dissected heads and skulls. The paranasal sinuses of buffalo comprise dorsal conchal, middle conchal, maxillary, frontal, palatine, sphenoidal (inconstant, small and shallow when present), lacrimal and ethmoidal that were identified and labelled according to the premolar and molar teeth as landmarks. The topographic description of all the compartments, diverticula, septa and communication of the paranasal sinuses in buffalo has been presented. The relationship between the various air cavities and paranasal sinuses was easily visualized.

  5. 208 PRODUCTION OF TRANSGENIC CLONED BUFFALO (BUBALUS BUBALIS) EMBRYOS CONTAINING HUMAN INSULIN GENE THROUGH HAND-GUIDED CLONING.

    PubMed

    Mehta, P; Kaushik, R; Chauhan, M S; Palta, P; Singla, S; Singh, M K; Manik, R S

    2016-01-01

    Diabetes is a growing disease worldwide and has emerged as a major healthcare problem in India. Insulin is an essential medicine for the treatment of diabetes. Large dairy animals, such as buffaloes and cows, may be used as bioreactors for cost-effective production of human insulin. The present study was aimed to produce transgenic buffalo embryos containing the human insulin gene through hand-guided cloning for production of transgenic animals. Buffalo female fetal fibroblast cells at passage number 3 were transfected using mammary gland- specific expression vector containing the human insulin gene under buffalo β-lactoglobulin promoter by nucleofection method and cultured with G418 drug for 3 weeks to obtain positive transgenic cell clones. Transgene integration into buffalo female fetal fibroblast genome was confirmed by PCR and Southern blotting. Nontransfected and transgene integrated cells were used as nuclear donors to produce embryos by the hand-guided cloning technique. The developmental competence and quality of embryos as judged by total cell number and TUNEL assay were compared among transgenic and nontransgenic (control) embryos. The blastocyst rate was lower (P<0.05) for transgenic embryos than that of nontransgenic cloned embryos (35.97±2.16v. 45.80±4.11, respectively). The apoptotic index was found to be lower (P<0.05) for control blastocysts than that for transgenic blastocysts. However, the total cell number was similar (P<0.05) among transgenic and control cloned blastocysts. Thus, transgenic cells, and subsequently transgenic embryos containing the human insulin gene, were successfully produced and transferred in recipients. In the future, these may be used for production of transgenic buffalo expressing human insulin in its milk and thus can be further utilised in large-scale production of human insulin.

  6. In vitro embryo production in buffalo (Bubalus bubalis) using sexed sperm and oocytes from ovum pick up.

    PubMed

    Liang, X W; Lu, Y Q; Chen, M T; Zhang, X F; Lu, S S; Zhang, M; Pang, C Y; Huang, F X; Lu, K H

    2008-04-15

    The objective was to explore the use of sexed sperm and OPU-derived oocytes in an IVP system to produce sex-preselected bubaline embryos. Oocytes were recovered from 20 fertile Murrah and Nili-Ravi buffalo cows by repeated (twice weekly) ultrasound-guided transvaginal ovum pick up (OPU), or by aspiration of abbatoir-derived bubaline ovaries, and subjected to IVF, using frozen-thawed sexed or unsexed bubaline semen. On average, 4.6 oocytes were retrieved per buffalo per session (70.9% were Grades A or B). Following IVF with sexed sperm, oocytes derived from OPU had similar developmental competence as those from abattoir-derived ovaries, in terms of cleavage rate (57.6 vs. 50.4%, P=0.357) and blastocyst development rate (16.0 vs. 23.9%, P=0.237). Furthermore, using frozen-thawed sexed versus unsexed semen did not affect rates of cleavage (50.5 vs. 50.9%, P=0.978) or blastocyst development (15.3 vs. 19.1%, P=0.291) after IVF using OPU-derived oocytes. Of the embryos produced in an OPU-IVP system, 9 of 34 sexed fresh embryos (26.5%) and 5 of 43 sexed frozen embryos (11.6%) transferred to recipients established pregnancies, whereas 7 of 26 unsexed fresh embryos (26.9%) and 6 out of 39 unsexed frozen embryos (15.4%) transferred to recipients established pregnancies. Eleven sex-preselected buffalo calves (10 females and one male) and 10 sexed buffalo calves (six females and four males) were born following embryo transfer. In the present study, OPU, sperm sexing technology, IVP, and embryo transfer, were used to produce sex-preselected buffalo calves. This study provided proof of concept for further research and wider field application of these technologies in buffalo.

  7. Follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in water buffalo (Bubalus bubalis).

    PubMed

    Khan, Firdous Ahmad; Das, Goutam Kumar

    2012-01-01

    The objective of this study was to examine the follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in buffaloes. Ovaries were collected from 56 acyclic and 95 cyclic buffaloes at slaughter, surface follicle number was counted and follicles were classified into small (5.0-6.9 mm), medium (7.0-9.9 mm), and large (≥ 10.0 mm) size categories based on their diameter. Follicular fluid was aspirated and assayed for nitric oxide, ascorbic acid, estradiol, and progesterone. Acyclic buffaloes had a higher (P<0.05) number of medium-sized follicles and a lower (P<0.001) number of large follicles than the cyclic ones. In acyclic animals, the number of large follicles was lower (P<0.01) than in medium size category which in turn was lower (P<0.001) than the number of small follicles. In contrast, the number of medium and large follicles was not different (P>0.05) in the cyclic control. However, the number of small-sized follicles was higher (P<0.001) compared to the other two categories. The incidence of large-sized follicles was lower (P<0.05) in acyclic buffalo population compared to the cyclic control. Evaluation of estrogenic status demonstrated that all the follicles of acyclic buffaloes are estrogen-inactive (E (2)/P (4) ratio<1). Small- and medium-sized follicles of acyclic buffaloes had higher concentrations of nitric oxide (P<0.05 and P<0.001, respectively) and lower concentrations of ascorbic acid (P<0.05 and P<0.01, respectively) than the corresponding size estrogen-active follicles of their cyclic counterparts. In conclusion, this study indicates that follicular development continues during acyclicity in buffaloes. Although follicles in some acyclic buffaloes attain a size corresponding to morphological dominance, they are unable to achieve functional dominance, perhaps due to an altered balance of intrafollicular nitric oxide and ascorbic acid and, as a result, these follicles instead of

  8. Influence of season on corpus luteum structure and function and AI outcome in the Italian Mediterranean buffalo (Bubalus bubalis).

    PubMed

    Di Francesco, S; Neglia, G; Vecchio, D; Rossi, P; Russo, M; Zicarelli, L; D'Occhio, M J; Campanile, G

    2012-11-01

    The aim was to ascertain whether relationships between corpus luteum (CL) vascularization, CL function, and pregnancy outcome in AI in buffaloes were consistent across the breeding season and transition period to the nonbreeding season in a Mediterranean environment. Stage of the estrous cycle in Italian Mediterranean buffaloes was synchronized using the Ovsynch with timed AI program and buffaloes were mated by AI in both the breeding season (N = 131) and transition period (N = 125). Detailed investigation of CL structure and function was undertaken in 39 buffaloes at each of the respective times using realtime B-mode/color-Doppler ultrasonography on Days 10 and 20 after AI. Progesterone (P4) concentrations were determined by RIA in all buffaloes. Pregnancy rate on Day 45 after AI was greater (P < 0.05) during the breeding season (58.0%) than the transitional period (45.6%) and this was primarily the result of a lower (P < 0.05) late embryonic mortality during the breeding season (7.3%) compared with the transition period (23%). Circulating concentrations of P4 on Days 10 and 20 after AI were greater (P < 0.01) during the breeding season (4.6 ± 0.3 and 3.4 ± 0.2, respectively) than during the transition period (1.6 ± 0.12 and 1.8 ± 0.2, respectively), and this was independent of reproductive status as there was no interaction between pregnancy and season. Corpus luteum time average medium velocity at Day 10 after AI was greater (P < 0.01) during the breeding season (19.3 ± 1.5) than in the transitional period (8.3 ± 0.7). There were positive correlations in pregnant buffaloes between CL time average medium velocity and P4 concentrations on Day 10 (r = 0.722; P < 0.01) and Day 20 (r = 0.446; P < 0.01) after AI. The findings were interpreted to indicate that relationships between CL vascularization, CL function, and pregnancy outcome in AI in buffaloes are consistent across the breeding season and transition period to the nonbreeding season. The distinction

  9. Investigation of the prion protein gene (PRNP) polymorphisms in Anatolian, Murrah, and crossbred water buffaloes (Bubalus bubalis).

    PubMed

    Yaman, Yalçın; Karadağ, Orhan; Ün, Cemal

    2017-02-01

    Bovine spongiform encephalopathy (BSE) of the cattle is the outstanding disease among other transmissible spongiform encephalopathy (TSEs). It can be transmitted from the cattle to a human and causes a new variant of the Creutzfeldt-Jakob disease (CJD). It is known that prion protein coding gene (PRNP) plays a major role in the TSE susceptibility or resistance in some species. Recent researches demonstrated that the insertion (in) and deletion (del) polymorphisms within promoter and intron 1 region of the PRNP related to BSE susceptibility in cattle. In contrast to cattle, BSE has never been reported in water buffalo; hence, PRNP polymorphisms may be an explanation for buffalo resistance to BSE. The aim of this study was to evaluate allele, genotype, and haplotype frequencies of the PRNP promoter and intron 1 insertion/deletion (indel) polymorphism in healthy Anatolian, Murrah, and Murrah × Anatolian crossbred buffaloes. According to our findings, there were no deletion alleles at two mentioned loci. All studied buffaloes were monomorphic and have carried in/in haplotypes which are considered as the most resistant genotype to BSE.

  10. Isolation of bacteria in semen and evaluation of antibiotics in extender for cryopreservation of buffalo (Bubalus bubalis) bull spermatozoa.

    PubMed

    Andrabi, S M H; Khan, L A; Shahab, M

    2016-12-01

    This study was designed to investigate the occurrence of bacterial species in water buffalo semen at the time of collection/processing and to assess the efficacy of some selected antibiotics (GTLS; gentamycin, tylosin and linco-spectin or SP; streptomycin and penicillin) in cryodiluent on bacterial control and quality including in vivo fertility of buffalo spermatozoa. For this purpose, four experiments were conducted. In experiment 1, a total of 11 bacterial species were isolated from buffalo ejaculates. In experiment 2, total aerobic bacterial counts at post dilution and thawing were lower (P < 0.05) in GTLS than in SP or control. The majority of the bacterial isolates from ejaculates were more susceptible to GTLS than SP. In experiment 3, sperm acrosome integrity was higher (P < 0.05) in GTLS and SP compared to control. In experiment 4, the in vivo fertility results for GTLS were higher (P < 0.05) than that for SP. In conclusion, a number of bacterial species were isolated from the bubaline semen, which requires an efficient control before its use in artificial insemination program. The GTLS combination of antibiotics may be incorporated into a freezing extender/protocol without compromising the post-thaw quality and in vivo fertility of buffalo bull spermatozoa.

  11. Comparative study of the gut microbiome potentially related to milk protein in Murrah buffaloes (Bubalus bubalis) and Chinese Holstein cattle

    PubMed Central

    Zhang, Jiachao; Xu, Chuanbiao; Huo, Dongxue; Hu, Qisong; Peng, Qiannan

    2017-01-01

    Previous studies suggested a close relationship between ruminant gut microbes and the mammary gland. In this study, shotgun metagenomic sequencing was used to reveal the differences in the intestinal microbiome potentially related to milk components in Murrah buffaloes and Chinese Holstein cattle. A PCoA based on the weighted Unifrac distances showed an apparent clustering pattern in the structure of intestinal microbiota between buffalo and cattle. We could attribute the structural difference to the genera of Sutterella, Coprococcus and Dorea. A further analysis of microbial functional features revealed that the biosynthesis of amino acids (including lysine, valine, leucine and isoleucine), lipopolysaccharide biosynthesis and cofactor/vitamin biosynthesis were enriched in the buffalo. In contrast, dairy cattle had higher levels of pyruvate metabolism and carbon fixation in photosynthetic organisms. A further correlation analysis based on different milk components and the typical microbiome uncovered a significant positive correlation between milk protein and the microbial biosynthesis of amino acids, which was also positively correlated in the genera of Parabacteroides, Dorea and Sutterella. This study will expand our understanding of the intestinal microbiome of buffalo and cattle as representative ruminants, as well as provide new views about how to improve the production and nutritional qualities of animal milk. PMID:28176851

  12. A possible case of caprine-associated malignant catarrhal fever in a domestic water buffalo (Bubalus bubalis) in Switzerland

    PubMed Central

    2011-01-01

    Background Malignant catarrhal fever (MCF) is a fatal herpesvirus infection, affecting various wild and domestic ruminants all over the world. Water buffaloes were reported to be particularly susceptible for the ovine herpesvirus-2 (OvHV-2) causing the sheep-associated form of MCF (SA-MCF). This report describes the first case of possibly caprine-associated malignant catarrhal fever symptoms in a domestic water buffalo in Switzerland. Case presentation The buffalo cow presented with persistent fever, dyspnoea, nasal bleeding and haematuria. Despite symptomatic therapy, the buffalo died and was submitted to post mortem examination. Major findings were an abomasal ulceration, a mild haemorrhagic cystitis and multifocal haemorrhages on the epicardium and on serosal and mucosal surfaces. Eyes and oral cavity were not affected. Histopathology revealed a mild to moderate lymphohistiocytic vasculitis limited to the brain and the urinary bladder. Although these findings are typical for MCF, OvHV-2 DNA was not detected in peripheral blood lymphocytes or in paraffin-embedded brain, using an OvHV-2 specific real time PCR. With the aid of a panherpesvirus PCR, a caprine herpesvirus-2 (CpHV-2) sequence could be amplified from both samples. Conclusions To our knowledge, this is the first report of malignant catarrhal fever in the subfamily Bovinae, where the presence of CpHV-2 could be demonstrated. The etiological context has yet to be evaluated. PMID:22132808

  13. Chromium Supplements in the Feed for Lactating Murrah Buffaloes (Bubalus bubalis): Influence on Nutrient Utilization, Lactation Performance, and Metabolic Responses.

    PubMed

    Deka, Rijusmita Sarma; Mani, Veena; Kumar, Muneendra; Shiwajirao, Zade Satish; Kaur, Harjit

    2015-12-01

    This study was conducted to evaluate the effect of different levels of inorganic chromium (Cr) on nutrient intake, nutrient utilization, milk mineral contents, and blood biochemical in lactating Murrah buffaloes. Twenty-four multifarious Murrah buffaloes were blocked into four groups having six animals in each group and fed for 150 days post-partum. Feeding regimen was same in all the groups except that treatment groups were supplemented with 0.0, 0.5, 1.0, and 1.5 mg inorganic Cr/kg dry matter (DM) in the four respective groups. Group fed on basal diet supplemented with 0.0 mg Cr/kg DM served as a control. Adding inorganic Cr to the diet of lactating buffaloes increased feed intake, milk yield, and nutrient digestibility. Nitrogen (N) and Cr balance were significantly increased (P<0.05) by adding Cr. Milk and plasma Cr concentration could be increased by adding Cr (P<0.001), while no significant differences were found between the four treatments in milk and plasma concentration of calcium (Ca), copper (Cu), and zinc (Zn). Feeding of diet supplemented with inorganic Cr decreased plasma insulin concentration (P=0.016). Concentration of blood glucose and the ratio of blood glucose to insulin were increased by adding Cr (P=0.037 and P=0.011, respectively). Adding Cr to the diet of lactating buffaloes did not show any effect on blood hemoglobin (Hb), while plasma concentration of progesterone and antioxidant status was increased (P<0.05). It could be concluded that adding Cr to the diet of lactating buffaloes improved nutrient intake, nutrient utilization, and lactation performance.

  14. The effects of high temperature and roof modification on physiological responses of swamp buffalo ( Bubalus bubalis) in the tropics

    NASA Astrophysics Data System (ADS)

    Khongdee, Titaporn; Sripoon, S.; Vajrabukka, C.

    2013-05-01

    The objective of the experiments reported here was to measure the effects of cooling techniques (Modified roof vs Normal roof) on the performance and physiology of 12 young male buffaloes with a similar live weight of 160 kg. The study was conducted at Chainat Agriculture and Technology College, Chainat Province, Thailand. The animals were divided randomly into two groups, each group comprising six buffaloes, and the two groups were studied to evaluate the effects of modified roofing (normal roof fitted with woven polypropylene shade cloth) on the subjects' physiological responses to heat stress under hot humid conditions. The modified roof resulted in lowered heat stress in buffaloes compared to those under a standard roof. The difference was shown by the buffaloes having a significantly lower mean rectal temperature (39.14 ± 0.07 vs 40.00 ± 0.10°C) and plasma cortisol (2.14 ± 0.24 vs 3.38 ± 0.37 ng/ml). The average daily water consumption was significantly lower in the MR group (MR, 29.71 ± 0.86 vs NR, 34.14 ± 1.06 L head -1 day-1), while there was a tendency for the roughage intake to be higher in the MR group compared to that of the NR group (MR, 5.88 ± 0.18 vs NR, 6.44 ± 0.19 kg head-1 -1 day-1; P = 0.0508). It was concluded that roof modification facilitated a reduction in heat load from roof re-radiation, and was an effective means of alleviating thermal stress in young buffaloes.

  15. Effect of antibiotics in extender on bacterial and spermatozoal quality of cooled buffalo (Bubalus bubalis) bull semen.

    PubMed

    Akhter, S; Ansari, M S; Andrabi, S M H; Ullah, N; Qayyum, M

    2008-06-01

    The present study was designed to study the effect of traditional antibiotic combination (streptomycin and penicillin; SP) and relatively modern combination of antibiotics (gentamycin, tylosin, lincomycin and spectinomycin; GTLS) in extender on bacterial control and spermatozoal quality of liquid buffalo bull semen stored at 5 degrees C. Semen collected from Nili-Ravi buffalo bulls (n = 10) was diluted with skim milk extender containing either SP (streptomycin 1000 microg/ml and penicillin 1000 IU/ml), GTLS (gentamycin 500 microg/ml, tylosin 100 microg/ml, lincomycin 300 microg/ml and spectinomycin 600 microg/ml) or negative control with no antibiotics (NA). Liquid semen was stored at 5 degrees C for 5 days. Aerobic bacteria isolated from buffalo semen were Pseudomonas aeruginosa and Staphylococcus aureus. The only facultative anaerobic bacterium isolated was Klebsiella pneumoniae. In vitro antibiotic sensitivity test revealed that Ps. aeruginosa and Staph. aureus were susceptible to gentamycin. Staphylococcus aureus and K. pneumoniae were susceptible to tylosin and linco-spectinomycin. Total aerobic bacterial count was significantly lower in semen samples treated with GTLS than those of SP on third and fifth day of storage at 5 degrees C. There was no difference (p > 0.05) in sperm motility, longevity and plasma membrane integrity (PMI) in extender containing SP or GTLS combination until the third day of storage at 5 degrees C. On fifth day of storage sperm motility, longevity and PMI was significantly better in extender containing SP compared with GTLS and NA. Intact acrosomes, and sperm head, mid piece and tail abnormalities remained similar (p > 0.05) because of antibiotics up to 5 days of storage. In conclusion, GTLS is more capable than SP for bacterial control of buffalo bull semen. Moreover, GTLS and SP are equally efficient in preserving spermatozoal quality of extended buffalo bull semen for 3 days at 5 degrees C.

  16. Molecular cloning, sequence characterization, and gene expression profiling of a novel water buffalo (Bubalus bubalis) gene, AGPAT6.

    PubMed

    Song, S; Huo, J L; Li, D L; Yuan, Y Y; Yuan, F; Miao, Y W

    2013-10-01

    Several 1-acylglycerol-3-phosphate-O-acyltransferases (AGPATs) can acylate lysophosphatidic acid to produce phosphatidic acid. Of the eight AGPAT isoforms, AGPAT6 is a crucial enzyme for glycerolipids and triacylglycerol biosynthesis in some mammalian tissues. We amplified and identified the complete coding sequence (CDS) of the water buffalo AGPAT6 gene by using the reverse transcription-polymerase chain reaction, based on the conversed sequence information of the cattle or expressed sequence tags of other Bovidae species. This novel gene was deposited in the NCBI database (accession No. JX518941). Sequence analysis revealed that the CDS of this AGPAT6 encodes a 456-amino acid enzyme (molecular mass = 52 kDa; pI = 9.34). Water buffalo AGPAT6 contains three hydrophobic transmembrane regions and a signal 37-amino acid peptide, localized in the cytoplasm. The deduced amino acid sequences share 99, 98, 98, 97, 98, 98, 97 and 95% identity with their homologous sequences from cattle, horse, human, mouse, orangutan, pig, rat, and chicken, respectively. The phylogenetic tree analysis based on the AGPAT6 CDS showed that water buffalo has a closer genetic relationship with cattle than with other species. Tissue expression profile analysis shows that this gene is highly expressed in the mammary gland, moderately expressed in the heart, muscle, liver, and brain; weakly expressed in the pituitary gland, spleen, and lung; and almost silently expressed in the small intestine, skin, kidney, and adipose tissues. Four predicted microRNA target sites are found in the water buffalo AGPAT6 CDS. These results will establish a foundation for further insights into this novel water buffalo gene.

  17. In-vitro indicators of natural resistance and milk-producing ability in dairy buffaloes (Bubalus bubalis).

    PubMed

    Miarelli, Maria; Signorelli, Federica

    2015-01-01

    The aim of this study was to explore the possibility of detecting novel phenotypes of natural resistance at the molecular level through the in-vitro stimulation of monocyte-derived macrophages (MDMs). This study was conducted with 16 healthy buffaloes who were reared for milk production and for whom data on milk-producing ability were available for several lactations. MDMs from circulating monocytes were activated with interferon-gamma and lipopolysaccharide. The response was evaluated using Western blotting to detect the presence of 2 types of proteins separated by electrophoresis: tyrosine-phosphorylated proteins, which are indicators of the dynamic control of biochemical pathways, and IkB-alpha (Kappa light polipeptide gene enhancer in B-cells Inhibitor, alpha) protein, which controls the activity of nuclear factor kappa-light-chain-enhancer of activated B cells-a transcription factor that is responsible for the expression of proinflammatory cytokines. The results showed that the buffaloes who were positive for IkB-alpha proteins had a significantly higher milk-producing ability than the buffaloes who did not express IkB-alpha. On the contrary, no significant difference was detected between the high and low milk-producing buffaloes with regard to the presence of tyrosine-phosphorylated proteins. This preliminary study indicated that it may be possible to identify the more disease-resistant nonhuman animals on a molecular level. The results, therefore, indicate that an intense selection toward the increase of milk yield could impair natural disease resistance in future dairy buffalo generations.

  18. Supplementation of Slow-Release Melatonin Improves Recovery of Ovarian Cyclicity and Conception in Summer Anoestrous Buffaloes (Bubalus bubalis).

    PubMed

    Kumar, A; Mehrotra, S; Singh, G; Maurya, V P; Narayanan, K; Mahla, A S; Chaudhari, R K; Singh, M; Soni, Y K; Kumawat, B L; Dabas, S K; Srivastava, N

    2016-02-01

    The role of melatonin as a protective neurohormone against restoring cyclicity in summer anoestrous animals in photoperiod species has gained wider acceptance. This study was designed to uncover the evidence the slow-release melatonin (MLT) has on initiation of ovarian cyclicity and conception rate (CR) in summer anoestrous buffaloes. Thus, buffaloes diagnosed as summer anoestrous (absence of overt signs of oestrus, concurrent rectal examination and radioimmunoassay for serum progesterone at 10 days interval) were grouped as untreated (Group I, sterilized corn oil, n = 8) and treated (Group II, single subcutaneous injection of MLT @18 mg/50 kg bwt in sterilized corn oil, n = 20). Animals treated and detected in oestrus were artificially inseminated (AI) followed by division into Group III (second dose of MLT on 5th day post-AI, n = 8) and Group IV (no melatonin administration, n = 10). Blood samples were collected at 4 days interval for estimation of serum MLT, progesterone and oestrogen using radioimmunoassay kit. Mean oestrous induction rate (OIR), oestrous induction interval (OII), interoestrous interval (IOI) and CR were estimated. Compared to control, concentration of melatonin was significantly (p < 0.05) higher in treated group ranging from 14.34 ± 1.72 to 412.31 ± 14.47 pg/ml whereas other two hormones did not show any concentration difference. Melatonin-administered buffaloes showed significantly (p < 0.05) higher (90%) OIR with OII of 18.06 ± 1.57 days. Results showed improvement in conception rate in buffaloes administered with post-insemination melatonin. It can be concluded from the study that slow-release melatonin supplementation restored cyclicity in summer anoestrous animals resulting in improvement in conception rate in buffaloes. © 2015 Blackwell Verlag GmbH.

  19. The effects of high temperature and roof modification on physiological responses of swamp buffalo (Bubalus bubalis) in the tropics.

    PubMed

    Khongdee, Titaporn; Sripoon, S; Vajrabukka, C

    2013-05-01

    The objective of the experiments reported here was to measure the effects of cooling techniques (Modified roof vs Normal roof) on the performance and physiology of 12 young male buffaloes with a similar live weight of 160 kg. The study was conducted at Chainat Agriculture and Technology College, Chainat Province, Thailand. The animals were divided randomly into two groups, each group comprising six buffaloes, and the two groups were studied to evaluate the effects of modified roofing (normal roof fitted with woven polypropylene shade cloth) on the subjects' physiological responses to heat stress under hot humid conditions. The modified roof resulted in lowered heat stress in buffaloes compared to those under a standard roof. The difference was shown by the buffaloes having a significantly lower mean rectal temperature (39.14 ± 0.07 vs 40.00 ± 0.10°C) and plasma cortisol (2.14 ± 0.24 vs 3.38 ± 0.37 ng/ml). The average daily water consumption was significantly lower in the MR group (MR, 29.71 ± 0.86 vs NR, 34.14 ± 1.06 L head (-1) day(-1)), while there was a tendency for the roughage intake to be higher in the MR group compared to that of the NR group (MR, 5.88 ± 0.18 vs NR, 6.44 ± 0.19 kg head-1 (-1) day(-1); P = 0.0508). It was concluded that roof modification facilitated a reduction in heat load from roof re-radiation, and was an effective means of alleviating thermal stress in young buffaloes.

  20. Clinical outcomes and molecular genotyping of Staphylococcus aureus isolated from milk samples of dairy primiparous Mediterranean buffaloes (Bubalus bubalis).

    PubMed

    Guccione, J; Cosandey, A; Pesce, A; Di Loria, A; Pascale, M; Piantedosi, D; Steiner, A; Graber, H U; Ciaramella, P

    2014-12-01

    Staphylococcus aureus is one of the most important pathogens causing mastitis in dairy cows and in Mediterranean buffaloes. Genotype B (GTB) is contagious in dairy cows and may occur in up to 87% of cows of a dairy herd. It was the aim of this study to evaluate genotypes present, clinical outcomes, and prevalence of Staph. aureus in milk samples of primiparous Mediterranean dairy buffaloes. Two hundred composite milk samples originating from 40 primiparous buffaloes were collected from May to June 2012, at d 10, 30, 60, 90, and 150 d in milk (DIM) to perform somatic cell counts and bacteriological cultures. Daily milk yields were recorded. Before parturition until 40 to 50 DIM, all primiparous animals were housed separated from the pluriparous animals. Milking was performed in the same milking parlor, but the primiparous animals were milked first. After 50 DIM, the primiparous were mixed with the pluriparous animals, including the milking procedure. Individual quarter samples were collected from each animal, and aliquots of 1 mL were mixed and used for molecular identification and genotyping of Staph. aureus. The identification of Staph. aureus was performed verifying the presence of nuc gene by nuc gene PCR. All the nuc-positive isolates were subjected to genotype analysis by means of PCR amplification of the 16S-23S rRNA intergenic spacer region and analyzed by a miniaturized electrophoresis system. Of all 200 composite samples, 41 (20.5%) were positive for Staph. aureus, and no genotype other than GTB was identified. The prevalence of samples positive for Staph. aureus was 0% at 10 DIM and increased to a maximum of 22/40 (55%) at 90 DIM. During the period of interest, 14 buffaloes tested positive for Staph. aureus once, 6 were positive twice, and 5 were positive 3 times, whereas 15 animals were negative at every sampling. At 90 and 150 DIM, 7 (17.5%) and 3 buffaloes (7.5%), respectively, showed clinical mastitis (CM), and only 1 (2.5%) showed CM at both

  1. Genetic analysis of river, swamp and hybrid buffaloes of north-east India throw new light on phylogeography of water buffalo (Bubalus bubalis).

    PubMed

    Mishra, B P; Dubey, P K; Prakash, B; Kathiravan, P; Goyal, S; Sadana, D K; Das, G C; Goswami, R N; Bhasin, V; Joshi, B K; Kataria, R S

    2015-12-01

    This study analysed buffaloes from north-east India and compared their nuclear and mitochondrial DNA variations with buffaloes of mainland India, China, Mediterranean and South-East Asia. Microsatellite genotypes of 338 buffaloes including 210 from six north-east Indian buffalo populations and three mainland Indian breeds were analysed to evaluate their genetic structure and evolutionary relationships. Phylogenetic analysis and multidimensional scaling plot of pairwise FST revealed the clustering of all swamp-type buffaloes of north-east India with Lower Assamese (significantly hybrid type) buffaloes in one plane and all the mainland river buffaloes in another plane while the upper Assamese buffaloes being distinct from both these clusters. Analysis of mtDNA D-loop region of 530-bp length was performed on 345 sequences belonging to 23 buffalo populations from various geographical regions to establish the phylogeography of Indian water buffalo. The swamp buffaloes of north-east India clustered with both the lineages of Chinese swamp buffalo. Multidimensional scaling display of pairwise FST derived from mitochondrial DNA data showed clustering of upper Assamese, Chilika and Mediterranean buffaloes distinctly from all the other Indian buffalo populations. Median-joining network analysis further confirmed the distinctness and ancestral nature of these buffaloes. The study revealed north-east region of India forming part of the wider hybrid zone of water buffalo that may probably extend from north-east India to South-East Asia. © 2015 Blackwell Verlag GmbH.

  2. Minimum number of spermatozoa per dose in Mediterranean Italian buffalo (Bubalus bubalis) using sexed frozen semen and conventional artificial insemination.

    PubMed

    Gaviraghi, A; Puglisi, R; Balduzzi, D; Severgnini, A; Bornaghi, V; Bongioni, G; Frana, A; Gandini, L M; Lukaj, A; Bonacina, C; Galli, A

    2013-05-01

    In buffaloes, AI with sexed semen is not fully optimized, and the procedure has only been performed using the approach currently in use for cattle. The objective of the present work was to compare the pregnancy rates in Mediterranean Italian buffalo cows inseminated with sexed frozen-thawed semen at 2, 4, 6, and 8 million sperm per dose, using the Ovsynch protocol and conventional AI at a fixed time. Fresh ejaculates from three buffalo bulls were processed according to Beltsville sperm sorting technology, and packaged in 0.25-mL straws with two total concentrations of 2 and 4 million live sorted sperm per straw. After thawing, semen was evaluated for total motility, forward motility, average path velocity, membrane and DNA integrity, and membrane fluidity. Sorting efficiency was estimated using a real time polymerase chain reaction method developed and validated in our laboratory. The artificial inseminations were conducted during the breeding season on 849 Italian Mediterranean buffalo heifers and cows distributed in 13 farms in northern and central Italy. No significant difference in quality parameters was reported between nonsexed and sexed straws produced with 2 and 4 million sperm. Lower pregnancy rate (P < 0.001) was reported when inseminating doses of sexed semen at 2 million were used (53/170; 31.2%), with respect to conventional nonsexed (78/142; 54.9%), and sexed doses at 4, 6, and 8 million spermatozoa (102/205, 49.8%; 84/175, 48.0%; and 74/157, 47.1%, respectively). No differences were evident using conventional doses and sexed semen with sperm numbers equal or higher than 4 million per dose. Pregnancies were not affected by the sire; 39/82 (47.6%), 120/270 (44.4%), and 151/355 (42.5%), respectively, for the three bulls. Variability in pregnancy rates observed in different herds was not significant. Furthermore, no significant difference was reported between pregnancies obtained with sexed semen in heifers and multiparous, respectively, 179/407 (44

  3. Changes in some blood parameters, milk composition and yield of buffaloes (Bubalus bubalis) during the transition period.

    PubMed

    Fiore, Enrico; Giambelluca, Sonia; Morgante, Massimo; Contiero, Barbara; Mazzotta, Elisa; Vecchio, Domenico; Vazzana, Irene; Rossi, Pasquale; Arfuso, Francesca; Piccione, Giuseppe; Gianesella, Matteo

    2017-08-04

    In this study changes in hematochemical parameters, milk composition and yield were investigated in buffaloes during the transition period. A total of 93 buffaloes 113.9 ± 8.03 months old and 535 ± 50 kg average body weight were used. Parity was recorded, blood samples were collected from 80 days pre-partum until 70 days post-partum; milk samples were collected from 5 days to 70 days post-partum. On serum samples, the values of non-esterified fatty acids, β-hydroxybutyrate, glucose, cholesterol, triglycerides, aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, urea, total proteins were evaluated. On milk, percentage of fat, protein and lactose, along with the somatic cell count (SCC), milk yield and daily milk production (DMP) were assessed. The peripartum period significantly (P ≤ 0.01) influenced all studied parameters with the exception of glucose. Milk fat percentage showed decreasing trend from 10 until 40 days post-partum; DMP significantly (P ≤ 0.01) increased from 1 day post-partum until 40 days post-partum. Milk yield significantly (P ≤ 0.01) decreased in animals over the sixth lactation. Our results confirmed the importance of transit period in buffaloes. Blood parameters and milk composition alterations are crucial to predict the energy balance status of buffaloes in order to improve their management and feed intake during the transition period. © 2017 Japanese Society of Animal Science.

  4. Hormone variations in serum and milk of buffaloes (Bubalus bubalis) as potential indicators of treatment with recombinant bovine somatotropin.

    PubMed

    Castigliego, Lorenzo; Li, Xiao Ning; Armani, Andrea; Grifoni, Goffredo; Boselli, Carlo; Rosati, Remo; Gianfaldoni, Daniela; Guidi, Alessandra

    2011-11-01

    Recombinant bovine somatotropin (rbST) is used to increase milk yield in cows, but it has been forbidden in some countries and in the EU. However, rbST misuse represents a concern in both bovine and buffalo dairy production. A number of studies on rbST treatment have been performed on bovines, but there are few data on buffaloes. In this study, we treated eight lactating buffaloes with biweekly injections of a slow-release formulation of rbST, for five cycles of administration, and analysed total ST and insulin-like growth factor 1 (IGF-1) variations in serum and IGF-1 in milk. The aim was to assess their power as potential indicators of rbST-treatment. Blood was collected on days 2, 5, 9 and 14 of each cycle, and milk on days 2, 9 and 14 of cycles 2 and 5. Results showed an extraordinary increase in ST levels on day 2 in treated animals, followed by a rapid decrease over the following days, while a significant increase in IGF-1 was observed both in serum and in milk throughout most of each cycle. These results suggest that serum ST levels are a good indicator of treatment. However, the rapid decrease after the peak limits the useful period of sample collection.

  5. Transvaginal follicle aspiration in Thai swamp buffalo heifers using different vacuum pressures after FSH pretreatment (Bubalus bubalis).

    PubMed

    Techakumphu, Mongkol; Promdireg, Akachart; Phutikanit, Nawapen; Nachiengmai, Anchalee; Thongjan, Sak

    2004-08-01

    The objective of the experiment was to study oocyte recovery by transvaginal, ultrasound-guided, follicle aspiration, from Thai swamp buffalo using different vacuum pressures. Six adult buffalo heifers, aged 2.5-3.0 yrs were treated with a total dose of 280 mg FSH, given twice a day in a divided doses over a three day period (60/60 mg, 50/50 mg, 30/30 mg) at d7 after progesterone implant. Three vacuum pressures were used; 100 (n=12), 80 (n=12) and 60 mmHg (n=12) and all of the pressures were performed in each animal. The animals were treated repeatedly and collection took place using 2 sets of each pressure every 2 months, giving a total of 36 collections from each animal. The oocyte recovery rates from each pressure were 81.2% (69/85) 79.1% (53/67) and 90.3% (93/103) for 100, 80 and 60 mmHg respectively. The number of oocytes collected per donor were 5.33 +/- 3.27, 4.42 +/- 2.71 and 7.75 +/- 4.31 respectively. The quality of the oocytes did not improved with the lower vacuum pressure. In conclusion, the application of FSH pretreatment improves the yield of oocytes from Thai, swamp buffalo heifers after gonadotropin treatment when using the vacuum pressures between 60-100 mmHg.

  6. Repeated ultrasound-guided transvaginal oocyte retrieval from cyclic Murrah buffaloes (Bubalus bubalis): oocyte recovery and quality.

    PubMed

    Gupta, V; Manik, R S; Chauhan, M S; Singla, S K; Akshey, Y S; Palta, P

    2006-01-01

    The present study was undertaken to explore the potential of the Murrah breed of buffaloes as donors of oocytes and to find out the recovery rate and oocyte quality in cyclic Murrah buffaloes subjected to oocyte recovery once a week. Murrah buffaloes (n = 5) were synchronized for estrus by a single prostaglandin injection schedule. The animals were subjected to transvaginal oocyte retrieval (TVOR) once weekly for 6 weeks, starting from Day 7 of the oestrous cycle (Day 0 = day of oestrus). TVOR was performed using an ultrasound machine with a 5 MHz transvaginal transducer, single lumen 19-gauge, 60 cm long needle and a constant vacuum pressure of 50 mmHg. The number and size of follicles in each ovary was determined before puncture. The follicles were characterized on the basis of their diameter as small (3-5 mm), medium (6-9 mm) and large (> or = 10 mm). The oocytes recovered were classified as grade A, cumulus-oocytes complexes (COCs) with > or = 5 layers of cumulus cells; grade B, those with two to four layers; grade C, partially denuded oocytes; and grade D, completely denuded oocytes. The mean (+/- S.E.M) number of small, medium and large follicles, and the number of total follicles observed per animal per session, which was 2.2 +/- 0.3, 0.6 +/- 0.2, 0.9 +/- 0.1 and 3.7 +/- 0.3, respectively, did not differ between animals or between puncture sessions. Small follicles constituted a major proportion (59%) of the total observed follicles. A mean (+/- S.E.M) number of 3.0 +/- 0.3 follicles were punctured and 2.0 +/- 0.3 oocytes recovered per animal per session, with a recovery rate of 68%. Out of the total 61 oocytes recovered, 36 (59%) were of grades A + B whereas 25 (41%) were of grades C + D. In conclusion, this study describes the potential of cyclic Murrah buffaloes as donors of oocytes collected by repeated TVOR once a week, without any adverse effects on follicular growth and oocyte recovery. It also describes an efficient system for carrying out TVOR in

  7. Impact of Livestock Hygiene Education Programs on Mastitis in Smallholder Water Buffalo (Bubalus bubalis) in Chitwan, Nepal

    PubMed Central

    Ng, Linda; Jost, Christine; Robyn, Misha; Dhakal, I.P.; Bett, Bernard; Dhakal, Pramod; Khadkha, Rupak

    2010-01-01

    A project implemented from 2003–2005 trained women in Chitwan District, Nepal, in hygienic dairy production using a process of social mobilization. The aim of this research was to assess if the prevalence of mastitis in water buffalo in the households of women who were trained was lower one year after training than in untrained households, if the training influenced knowledge and practices for the prevention or control of mastitis, and if these practices and knowledge were associated with a lower prevalence of mastitis. A total of 202 households from Eastern and Western Chitwan District were included in the study. Of these, 60 households had participated in the project and 142 had not. Milk samples were collected from 129 households (33 project households and 96 non-project households). Clinical mastitis was determined using visual inspection of udders and detection of macroscopic clots and flakes in milk. The California Mastitis Test was used to diagnose sub-clinical mastitis from milk samples, and the IDEXX SNAP test to identify the presence of tetracycline residues. The prevalence of mastitis in trained households (39.4%) was 43.78% of that in untrained households (60.4%), lower but not significantly so (p = 0.08, 95% CI 0.17–1.12). Thirteen indicators of knowledge or practice for the control or prevention of mastitis were more likely to occur in trained households, four significantly so (not consuming milk from sick buffalo (p=0.001), using soap to wash hands before milking (p=0.001), discarding milk after antibiotic usage (p=0.01), and choosing appropriate flooring for their livestock (p=0.03)). Trained households that discarded milk from sick buffalo were 2.96 times more likely to have at least one animal with mastitis in the household (p=0.03, 95% CI 1.15–7.65). Trained households that knew to wash buffalos teats after milking were less likely (OR 0.25) to have mastitis in their herd ((p=0.02, 95% CI 0.08–0.80). Of the 138 buffalos tested, only one

  8. Impact of Heat Stress on Cellular and Transcriptional Adaptation of Mammary Epithelial Cells in Riverine Buffalo (Bubalus Bubalis)

    PubMed Central

    Kapila, Neha; Sharma, Ankita; Kishore, Amit; Sodhi, Monika; Tripathi, Pawan K.; Mohanty, Ashok K.

    2016-01-01

    The present study aims to identify the heat responsive genes and biological pathways in heat stressed buffalo mammary epithelial cells (MECs). The primary mammary epithelial cells of riverine buffalo were exposed to thermal stress at 42°C for one hour. The cells were subsequently allowed to recover at 37°C and harvested at different time intervals (30 min to 48 h) along with control samples (un-stressed). In order to assess the impact of heat stress in buffalo MECs, several in-vitro cellular parameters (lactate dehydrogenase activity, cell proliferation assay, cellular viability, cell death and apoptosis) and transcriptional studies were conducted. The heat stress resulted in overall decrease in cell viability and cell proliferation of MECs while induction of cellular apoptosis and necrosis. The transcriptomic profile of heat stressed MECs was generated using Agilent 44 K bovine oligonucleotide array and at cutoff criteria of ≥3-or ≤3 fold change, a total of 153 genes were observed to be upregulated while 8 genes were down regulated across all time points post heat stress. The genes that were specifically up-regulated or down-regulated were identified as heat responsive genes. The upregulated genes in heat stressed MECs belonged to heat shock family viz., HSPA6, HSPB8, DNAJB2, HSPA1A. Along with HSPs, genes like BOLA, MRPL55, PFKFB3, PSMC2, ENDODD1, ARID5A, and SENP3 were also upregulated. Microarray data revealed that the heat responsive genes belonged to different functional classes viz., chaperons; immune responsive; cell proliferation and metabolism related. Gene ontology analysis revealed enrichment of several biological processes like; cellular process, metabolic process, response to stimulus, biological regulation, immune system processes and signaling. The transcriptome analysis data was further validated by RT-qPCR studies. Several HSP (HSP40, HSP60, HSP70, HSP90, and HSPB1), apoptotic (Bax and Bcl2), immune (IL6, TNFα and NF-kβ) and oxidative

  9. Comparative therapeutic effect of steroidal and non-steroidal anti-inflammatory drugs on pro-inflammatory cytokine production in water buffalo calves (Bubalus bubalis) naturally infected with bronchopneumonia: a randomized clinical trial.

    PubMed

    Rizk, Mohamed Abdo; Mahmoud, Motamed Elsayed; El-Sayed, Shimaa Abd El-Salam; Salman, Doaa

    2017-08-22

    In the current study, we compared the therapeutic effects of a non-steroidal and a steroidal anti-inflammatory drug on the production of pro-inflammatory cytokines, interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-12p40 (IL-12p40), interferon gamma (IFNγ), and tumor necrosis factor alpha (TNF-α) in the blood of water buffalo (Bubalus bubalis) calves naturally infected by bronchopneumonia. Twenty-seven buffalo calves (7 ± 2-month-old, 163 ± 12 kg) reared in smallholder farms in El-Dakahlia province in Egypt were identified to have bronchopneumonia and randomly allocated into three equal groups. Ten clinically healthy buffalo calves with negative bronchoalveolar lavage results were served as negative control. Diseased calves were treated with tulathromycin alone, a combination of tulathromycin with dexamethasone (steroidal anti-inflammatory drug) or tulathromycin with flunixin meglumine (non-steroidal anti-inflammatory drug). The results revealed significant elevations (P < 0.05) in the production of selected cytokines in all diseased calves in comparison with healthy animals. Six days post-treatment, a significant inhibition (P < 0.05) in the production of all assessed cytokines was observed in the blood of all treated calves. Interestingly, the serum concentrations of IL-1β and IL-12p40 were returned to the normal levels in pneumonic calves treated with the combination therapy of tulathromycin and flunixin meglumine. A strong significant positive correlation (P < 0.05) was detected between clinical sum scoring and IL-12p40 and TNF-α concentrations. The obtained results indicate the selectively potent anti-inflammatory effect of flunixin meglumine on the production of pro-inflammatory cytokines in pneumonic buffalo calves and highlight the efficacy of flunixin meglumine in the treatment of bronchopneumonia in buffalo calves when used in combination with tulathromycin.

  10. Effect of low density lipoproteins in extender on freezability and fertility of buffalo (Bubalus bubalis) bull semen.

    PubMed

    Akhter, S; Ansari, M S; Rakha, B A; Andrabi, S M H; Khalid, M; Ullah, N

    2011-09-01

    This study was designed to determine whether low-density lipoporoteins (LDLs) extracted from egg yolk in extender improve the freezability and fertility of buffalo bull semen. Semen from three Nili-Ravi buffalo bulls was diluted at 37 °C with tris-citric acid extender (50 × 10(6) motile spermatozoa mL(-1)) containing LDLs 2.5%, 5%, 10%, and 15% extracted from egg yolk and extender containing 20% egg yolk was kept as control. Diluted semen was cooled to 4 °C in 2 h, equilibrated at 4 °C for 4 h, filled in 0.5 mL French straws, and kept on liquid nitrogen vapors for 10 min. Straws were then plunged and stored in liquid nitrogen (-196 °C). Sperm motility (visually; %), plasma membrane integrity (%; with supravital hypo-osmotic swelling test), and viability (%; with dual staining test using Trypan-blue Giemsa) were assessed at post-dilution, pre-freezing and post-thawing. At post-dilution and pre-freezing, sperm progressive motility, plasma membrane integrity and viability was similar (P > 0.05) in extender containing 10% LDLs or the control. However, at post-thaw the aforementioned parameters were higher (P < 0.05) in extender containing 10% LDLs compared with the control and other experimental extenders. The fertility rate of inseminations performed were higher (P < 0.05) with extender containing 10% LDLs than the control. It was concluded that LDLs (10%) in extender improved the freezability and fertility of buffalo bull spermatozoa. Copyright © 2011 Elsevier Inc. All rights reserved.

  11. Is one-wave follicular growth during the estrous cycle a usual phenomenon in water buffaloes (Bubalus bubalis)?

    PubMed

    Awasthi, M K; Khare, Abhishek; Kavani, F S; Siddiquee, G M; Panchal, M T; Shah, R R

    2006-05-01

    The pattern of growth and regression of ovarian follicles was monitored once daily for one complete estrous cycle in eight individual water buffaloes by ultrasonographic scanning of the ovaries for an entire interovulatory interval of normal cycle length. One-wave follicular growth was observed in five animals and two-wave follicular growth in three buffaloes during the estrous cycle. The first follicular wave of a two-wave cycle emerged significantly earlier (P < 0.05) than the emergence of the solitary wave of a one-wave cycle. One- and two-wave cycles differed significantly (P < 0.05) with respect to the mean interovulatory interval (21.0 +/- 0.54 days versus 22.7 +/- 0.33 days) and the mean interestrus interval (20.8 +/- 0.58 days versus 22.3 +/- 0.66 days). The overall linear growth rate of the ovulatory follicle was significantly greater (P < 0.01) in a two-wave cycle compared to that of a one-wave cycle (1.17 +/- 0.33 mm/day versus 0.32 +/- 0.01 mm/day). In a one-wave pattern, the growth profile of the solitary dominant follicle was atypical, showing three distinct phases, i.e. growth phase, regression phase and regrowth phase culminating in ovulation. The level of plasma progesterone steadily increased from day 0 of estrous cycle, attained peak level on day 14 and declined thereafter. A slower growth rate of the dominant follicle was observed in the presence of higher plasma progesterone concentration. The present study shows that one-wave follicular growth is a normal phenomenon in suckled water buffaloes.

  12. Metagenomic analysis of virulence-associated and antibiotic resistance genes of microbes in rumen of Indian buffalo (Bubalus bubalis).

    PubMed

    Singh, K M; Jakhesara, S J; Koringa, P G; Rank, D N; Joshi, C G

    2012-10-10

    A major research goal in rumen microbial ecology is to understand the relationship between community composition and its function, particularly involved in fermentation process is of a potential interest. The buffalo rumen microbiota impacts human food safety as well as animal health. Although the bacteria of bovine rumen have been well characterized, techniques have been lacking to correlate total community structure with gene function. We applied 454 next generations sequencing technology to characterize general microbial diversity present in buffalo rumen metagenome and also identified the repertoire of microbial genes present, including genes associated with antibiotic resistance and bacterial virulence. Results suggest that over six percent (6.44%) of the sequences from our buffalo rumen pool sample could be categorized as virulence genes and genes associated with resistance to antibiotic and toxic compounds (RATC), which is a higher proportion of virulence genes reported from metagenome samples of chicken cecum (5.39%), cow rumen (4.43%) and Sargasso sea (2.95%). However, it was lower than the proportion found in cow milk (11.33%) cattle faeces (8.4%), Antarctic marine derived lake (8.45%), human fecal (7.7%) and farm soil (7.79%). The dynamic nature of metagenomic data, together with the large number of RATC classes observed in samples from widely different ecologies indicates that metagenomic data can be used to track potential targets and relative amounts of antibiotic resistance genes in individual animals. In addition, these data can be also used to generate antibiotic resistance gene profiles to facilitate an understanding of the ecology of the microbial communities in each habitat as well as the epidemiology of antibiotic resistant gene transport between and among habitats. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Fluoride-induced oxidative stress in rat's brain and its amelioration by buffalo (Bubalus bubalis) pineal proteins and melatonin.

    PubMed

    Bharti, Vijay K; Srivastava, R S

    2009-08-01

    Fluoride (F) becomes toxic at higher doses and induces some adverse effects on various organs, including brain. The mechanisms underlying the neurotoxicity caused by excess fluoride still remain unknown. The aims of this study were to examine F-induced oxidative stress (OS) and role of melatonin (MEL) and buffalo pineal proteins (PP) against possible F-induced OS in brain of rats. The 24 rats were taken in present study and were divided into four groups: control, F, F + PP, and F + MEL. The F group was given 150 mg/L orally for 28 days. Combined 150 ppm F and 100 microg/kg BW (i.p.) PP and F (150 ppm) + MEL (10 mg/kg BW, i.p.) were also administered. The activities of enzymatic, viz., superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione reductase (GR), and non-enzymatic, viz., reduced glutathione (GSH) concentration, and the levels of malondialdehyde (MDA) in the brain tissue were measured to assess the OS. Fluoride administration significantly increased brain MDA compared with control group, while GSH levels were decreased in fluoride-treated groups, accompanied by the markedly reduced SOD, GPx, GR, and SOD activity. Buffalo PP and MEL administration caused brain MDA to decrease but caused SOD, GPx, GR, GSH, and CAT activities to increase to significant levels in F-treated animals. Together, our data provide direct evidence that buffalo PP and MEL may protect fluoride-induced OS in brain of rats through mechanisms involving enhancement of enzymatic and non-enzymatic antioxidant defense system. Therefore, this study suggested that PP and MEL can be useful in control of neurotoxicity induced by fluoride.

  14. Assessment of foot health and animal welfare: clinical findings in 229 dairy Mediterranean Buffaloes (Bubalus bubalis) affected by foot disorders.

    PubMed

    Guccione, Jacopo; Carcasole, Christian; Alsaaod, Maher; D'Andrea, Luigi; Di Loria, Antonio; De Rosa, Angela; Ciaramella, Paolo; Steiner, Adrian

    2016-06-14

    Lameness represents the third most important health-related cause of economic loss in the dairy industry after fertility and mastitis. Although, dairy Mediterranean Buffaloes (MB) and dairy cows share similar breeding systems predisposing to similar herd problems, published studies exploring its relevance and role in these ruminants are still rare and incomplete. The aims of this study were to describe the clinical findings of foot disorders (FDs) in dairy MB and their influence on animal welfare, determined by assessment of locomotion score (LS), body condition score (BCS) and cleanliness score (CS). Of 1297 multiparous MB submitted to routine trimming procedures, 229 buffaloes showed at least one FD. The prevalence of buffaloes affected by FDs was 17.7 %, while motility and lameness indexes were 84.1 % (1091/1297) and 15.9 % (206/1297), respectively. Overgrowth was present in 17.0 % (220/1297), corkscrew claw in 15.8 % (205/1297), interdigital phlegmon in 0.9 % (12/1297), white line abscess in 0.8 % (11/1297), digital dermatitis in 0.1 % (1/1297) and interdigital hyperplasia in 0.1 % (1/1297). Simultaneous presence of FDs was recorded in 17.0 % of MB (221/1297): overgrowth and corkscrew claw occurred together in 15.8 % of cases (205/1297), overgrowth and interdigital phlegmon in 0.3 % (4/1297), overgrowth and white line abscess in 0.8 % (11/1297), digital dermatitis and interdigital hyperplasia in 0.1 % (1/1297). The presence of FDs was always associated with lameness (LS > 2), except from 23 MB with simultaneous overgrowth and interdigital phlegmon occurrence. The majority of MB within the under-conditioned group (95.5 %, 43/45) and all those with CS > 2 (122/122) had a locomotion score above the threshold of normality (LS > 2). Furthermore, foot diseases such as interdigital hyperplasia, white line abscess and digital dermatitis or interdigital hyperplasia seemed to occur more frequently associated with decreased BCS and increased CS

  15. Effect of additional chromium supplementation on health status, metabolic responses, and performance traits in periparturient Murrah Buffaloes (Bubalus bubalis).

    PubMed

    Deka, Rijusmita Sarma; Mani, Veena; Kumar, Muneendra; Zade, Shiwajirao Satish; Upadhaya, Ramesh Chand; Kaur, Harjit

    2015-02-01

    This study was conducted to determine the effects of inorganic chromium (Cr) on body condition, metabolic responses, lactation performance, and reproductive parameters in periparturient Murrah buffaloes. Twenty-four multiparous advanced pregnant Murrah buffaloes were randomly assigned to four treatment diets. Feeding regimen was the same in all the groups, except that the animals in the four respective groups were additionally supplemented with 0.0, 0.5, 1.0, and 1.5 mg of Cr/kg dry matter (DM) from day 60 prepartum to 150 days postpartum. Dry matter intake (DMI) and milk production were recorded every day, while body condition score (BCS) and whole blood samples were collected at days -60, -45, -30, -15, -7, -3, 0, 3, 7, 15, 30, 45, 60, 90, 120, and 150 relative to actual calving. As the days to calving advanced, DMI, plasma glucose, insulin, leptin, and Cr levels decreased (P < 0.05), and the levels were minimum on the day of calving. In contrast, the concentration of nonesterified fatty acid (NEFA) increased (P < 0.05) and was found to be highest at parturition. No change in DMI as well as BCS was observed due to dietary treatments. Supplementation of Cr improved plasma concentration of glucose, leptin, and Cr levels. However, the concentration of insulin decreased (P < 0.05) with the increased level of supplemental Cr. Milk yield (kg/day) was improved significantly (P < 0.05) in groups supplemented with 1.0 or 1.5 mg Cr/DM; however, only a small change was recorded in the group fed 0.5 mg Cr/kg DM. Fat-corrected milk and energy-corrected milk were 28.78 and 42.32 % and 28.76 and 41.68 % higher in the 1.0 and 1.5 mg Cr/DM groups, respectively. Dietary Cr supplementation during the peripartum period had beneficial effects on the reproductive performance of buffaloes. These results could be interpreted as an improvement in the body condition, metabolic response, milk yield, efficiency of milk production and nutrient utilization, and

  16. Clinicophysiological and haemodynamic effects of fentanyl with xylazine, medetomidine and dexmedetomidine in isoflurane-anaesthetised water buffaloes (Bubalus bubalis).

    PubMed

    Singh, Gyan D; Kinjavdekar, Prakash; Amarpal; Aithal, Hari P; Pawde, Abhijeet M; Zama, Malik M S; Singh, Jasmeet; Tiwary, Ramesh

    2013-03-18

    The present study was undertaken to investigate the sedative, analgesic and clinical effects of xylazine, medetomidine and dexmedetomidine with fentanyl as pre-anaesthetics in water buffaloes and to compare the dose-sparing effect of xylazine, medetomidine and dexmedetomidine on thiopental for induction and isoflurane for maintenance of anaesthesia in water buffaloes. Six male water buffaloes randomly received intravenous fentanyl (5.0 µg/kg body weight) and xylazine (0.05 mg/kg body weight), fentanyl (5.0 µg/kg body weight) and medetomidine (2.5 µg/kg body weight), fentanyl (5.0 µg/kg body weight) and dexmedetomidine (5.0 µg/kg body weight) at weekly intervals in groups I1, I2 and I3, respectively. After 15 min, the animals were restrained in right lateral recumbency and anaesthesia was induced by 5% thiopental sodium administered intravenously. The intubated animal was connected to the large animal anaesthesia machine and isoflurane in 100% oxygen (5 L/min) was insufflated for 60 min. The treatments were compared by clinicophysiological, haematobiochemical and haemodynamic parameters. Fentanyl-medetomidine and fentanyl-dexmedetomidine produced more cardiovascular depression during the pre-anaesthetic period but less depression of cardio-respiratory dynamics in the post induction and maintenance period. Quicker recovery was recorded in I2 and I3 groups. A lower dose of thiopental was required in group I3 (4.33 mg/kg ± 0.66 mg/kg) than in groups I2 (4.41 mg/kg ± 0.98 mg/kg) and I1 (4.83 mg/kg ± 0.79 mg/kg). The dose of isoflurane was less in group I3 (45.50 mL ± 5.45 mL) than in group I1 and I2 (48.66 mL ± 5.10 mL and 48.00 mL ± 6.38 mL). Better anaesthesia was recorded with fentanyl-dexmedetomidine-thiopental-isoflurane (group I3) than with fentanyl-medetomidine-thiopental-isoflurane (group I2) and fentanyl-xylazine-thiopental-isoflurane (group I1). Fentanyl-medetomidine and fentanyl-dexmedetomidine were better pre-anaesthetic agents in comparison to

  17. Involvement of the nervous system following experimental infection with Pasteurella multocida B:2 in buffalo (Bubalus bubalis): A clinicopathological study.

    PubMed

    Marza, Ali Dhiaa; Jesse, Faez Firdaus Abdullah; Ahmed, Ihsan Muneer; Chung, Eric Lim Teik; Ibrahim, Hayder Hamzah; Zamri-Saad, Mohd; Omar, Abdul Rahman; Abu Bakar, Md Zuki; Saharee, Abdul Aziz; Haron, Abdul Wahid; Alwan, Mohammed Jwaid; Lila, Mohd Azmi Mohd

    2016-04-01

    Haemorrhagic septicaemia (HS) is an acute, fatal, septicaemic disease of cattle and buffaloes caused by one of two specific serotypes of Pasteurella multocida B:2 and E:2 in Asian and African, respectively. It is well known that HS affect mainly the respiratory and digestive tracts. However, involvement of the nervous system in pathogenesis of HS has been reported in previous studies without details. In this study, nine buffalo calves of 8 months old were distributed into three groups. Animals of Group 1 and 2 were inoculated orally and subcutaneously with 10 ml of 1 × 10(12) cfu/ml of P. multocida B:2, respectively, while animals of Group 3 were inoculated orally with 10 ml of phosphate buffer saline as a control. All calves in Group 1 and Group 3 were euthanised after 504 h (21 day) post-infection, while calves in Group 2 had to euthanise after 12 h post-infection as they develop sever clinical signs of HS. Significant differences were found in Group 2 in the mean scores of clinical signs, gross and histopathological changes which mainly affect different anatomic regions of the nervous system. In addition, successful bacterial isolation of P. multocida B:2 were obtained from different sites of the nervous system. On the other hand, less sever, clinical, gross and histopathological changes were found in Group 1. These results provide for the first time strong evidence of involving of the nervous system in pathogenesis of HS, especially in the peracute stage of the disease.

  18. Microbial population in the rumen of swamp buffalo (Bubalus bubalis) as influenced by coconut oil and mangosteen peel supplementation.

    PubMed

    Pilajun, R; Wanapat, M

    2013-06-01

    Four, rumen fistulated swamp buffalo bulls were used to study microbial populations in the rumen when supplemented with coconut oil and mangosteen peel. Animals were randomly assigned to a 4 × 4 Latin square design. Four treatments were un-supplemented (Control), supplementation with coconut oil at 50 g/kg (CO5), supplementation with mangosteen peel at 30 g/kg (MP3) and supplementation with CO5 and MP3 (COM), of total DM intake. Animals received concentrate at 10 g/kg of BW, and rice straw was given ad libitum. Abundance of total bacteria was increased by CO5 supplementation, whereas populations of protozoa and Fibrobacter succinogenes were reduced by CO5 and COM supplementation. Dietary supplementation did not affect methanogen, Ruminococcus flavefaciens or Ruminococcus albus abundances. Dietary treatments changed denaturing gradient gel electrophoresis (DGGE) band patterns of methanogens and protozoa when compared with the control group, especially when supplemented with MP3. Supplementation of COM resulted in the greatest difference in pattern of DGGE bands for total bacteria compared with the control. Coconut oil and mangosteen peel supplementation resulted in changing of rumen microbial abundances and communities; however, combination of them could be more benefit to improve rumen fermentation of swamp buffalo fed on rice straw. © 2012 Blackwell Verlag GmbH.

  19. Butylated hydroxytoluene inclusion in semen extender improves the post-thawed semen quality of Nili-Ravi buffalo (Bubalus bubalis).

    PubMed

    Ijaz, A; Hussain, A; Aleem, M; Yousaf, M S; Rehman, H

    2009-05-01

    The study was carried out to evaluate the potential impact of butylated hydroxytoluene (BHT) on the frozen-thawed semen quality of Nili-Ravi buffalo bulls. Ejaculated bull semen was extended in a Tris-citrate egg yolk extender containing various concentrations of BHT (0.5, 1.0, 2.0 and 3.0mM). Semen was frozen at -196 degrees C using 50 x 10(6) spermatozoa per 0.5 mL straws. Five straws from each treatment were thawed to assess the semen quality in terms of sperm motility, viability, plasma membrane integrity and acrosomal integrity. Post-thawed sperm motility was determined using a phase-contrast microscope. Viability, plasma membrane integrity and acrosomal integrity were evaluated by the supravital staining, hypo-osmotic swelling test and normal acrosomal reaction, respectively. The highest (P<0.05) motility, acrosomal integrity and hypo-osmotic swelling response of spermatozoa was achieved by addition of 1.0 and 2.0mM BHT to semen extender. However, highest (P<0.05) viability of spermatozoa was achieved by inclusion of 2.0mM BHT. The higher concentration of BHT (3.0mM) reduced the motility, acrosomal integrity, viability and hypo-osmotic swelling response of the spermatozoa compared to other concentration used. In conclusion, BHT when added in the semen extender can improve the semen quality of buffalo bulls.

  20. Equivalency of Buffalo (Bubalus Bubalis) Embryonic Stem Cells Derived From Fertilized, Parthenogenetic, and Hand-Made Cloned Embryos

    PubMed Central

    Muzaffar, Musharifa; Selokar, Naresh L.; Singh, Karn P.; Zandi, Mohammad; Singh, Manoj K.; Shah, Riaz A.; Chauhan, Manmohan S.; Singla, Suresh K.; Palta, Prabhat

    2012-01-01

    Abstract This study was aimed at establishing buffalo embryonic stem cells (ESCs) from in vitro fertilized (IVF), parthenogenetic, and hand-made cloned (HMC) embryos and to check their equivalency in terms of stem cell marker expression, longevity, proliferation, and differentiation pattern. ESCs derived from all three sources were found by immunofluorescence to express the pluripotency markers SSEA-4, TRA-1-60, TRA-1-81, OCT4, and SOX2 and were able to form embryoid bodies containing cells expressing genes specific to endoderm (AFP, HNF4, and GATA4), mesoderm (MSX1, BMP4, and ASA), and ectoderm (cytokeratin 8 and NF68). Reverse transcriptase PCR (RT-PCR) showed cells from all sources to be positive for pluripotency markers OCT4, SOX2, NANOG, STAT3, REX1, FOXD3, NUCLEOSTEMIN, and TELOMERASE. Pluripotency markers OCT4, SOX2, NANOG, and c-MYC were also analyzed by real-time PCR. No significant differences were observed among ESCs from all three sources for all these genes except NANOG, whose expression was higher (p<0.05) in HMC-derived ESCs (6.897±2.3) compared to that in parthenogenesis- and IVF-derived cells (1.603±0.315 and 1±0, respectively). Pluripotent, stable buffalo ESC lines derived from IVF, parthenogenesis, and HMC embryos may be genetically manipulated to provide a powerful tool for studies involving embryonic development, genomic imprinting, gene targeting, cloning, chimera formation, and transgenic animal production. PMID:22582863

  1. Effects of vitrification procedures on subsequent development and ultrastructure of in vitro-matured swamp buffalo (Bubalus bubalis) oocytes.

    PubMed

    Boonkusol, Duangjai; Faisaikarm, Tassanee; Dinnyes, Andras; Kitiyanant, Yindee

    2007-01-01

    The purpose of the present study was to investigate the effects of two vitrification procedures on developmental capacity and ultrastructural changes of matured swamp buffalo oocytes. In vitro-matured oocytes were vitrified by using 35 and 40% ethylene glycol as vitrification solution for solid surface vitrification (SSV) and in-straw vitrification (ISV), respectively. Survival rate of vitrified-warmed oocytes, evaluated on the basis of ooplasm homogeneity, oolemma integrity and zona pellucida intactness, as well as parthenogenetic blastocyst rates of vitrified-warmed oocytes were significantly higher with SSV (89.3 and 13.6%, respectively) than ISV (81.8 and 5.5%, respectively). However, they were still significantly lower than that of control oocytes (100 and 34.2%, respectively). For examining the ultrastructural changes, fresh, VS-exposed (ISV and SSV), and vitrified-warmed (ISV and SSV) oocytes were processed for transmission electron microscopy. In VS-exposed oocytes, reduction of microvilli abundance and damage of mitochondrial membrane were found only in the ISV group. In vitrified-warmed oocytes, however, it was clear that both methods of vitrification induced profound ultrastructural modifications to microvilli, mitochondria, oolemma and cortical granules as well as to the size and position of vesicles. Damaged mitochondria were, however, more abundant in ISV vitrified oocytes than in SSV vitrified oocytes, which correlated with the developmental data, showing the superiority of the SSV method. The present study demonstrated the feasibility of vitrification of in vitro-matured swamp buffalo oocytes.

  2. Seasonal variation in nuclear DNA integrity of frozen-thawed spermatozoa from Thai AI swamp buffaloes (Bubalus bubalis).

    PubMed

    Koonjaenak, S; Johannisson, A; Pongpeng, P; Wirojwuthikul, S; Kunavongkrit, A; Rodriguez-Martinez, H

    2007-09-01

    In this study, we investigated the susceptibility of frozen-thawed swamp buffalo sperm nuclear DNA to undergo controlled acid-induced denaturation in situ, as analysed by flow cytometry, and aimed to correlate the results with sperm head morphology over three seasons in tropical Thailand. Artificial insemination (AI) doses (n = 218) from 18 AI buffalo sires, prepared between 1980 and 1989 and 2003 and 2005, were tested and compared among three seasons, the rainy season, July-October; winter, November-February; and summer, March-June. The overall mean of DNA fragmentation index (DFI) (+/- SD) was 1.84 +/- 1.68%, range from 0.19 to 7.92%, with 0.221 +/- 0.021 of the x-DFI ranging from 0.190 to 0.350 and 0.023 +/- 0.009 of the SD-DFI ranging from 0.010 to 0.070. The DFI was consistently low (range 1.40 +/- 0.21% to 2.16 +/- 0.21%; LSM +/- SEM), with x-DFI ranging from 0.216 +/- 0.003 to 0.225 +/- 0.003 and SD-DFI ranging from 0.022 +/- 0.001 to 0.024 +/- 0.001 across the seasons. The DFI was low enough to be related to high fertility potential. However, DFI values varied statistically among seasons, being lower in the rainy season (1.40 +/- 0.21%, P < 0.05) than in winter (2.16 +/- 0.21%) or summer (2.00 +/- 0.20%), and were also affected by the year of semen collection and processing (P < 0.001). The proportion of morphologically abnormal sperm head shapes was low, with no significant differences between seasons. However, DFI was significantly related to the proportion of loose abnormal sperm heads (r = 0.27, P < 0.01). In conclusion, frozen-thawed swamp buffalo sperm chromatin integrity is not seriously damaged by cryopreservation or affected by the seasonal variations in temperature and humidity seen in tropical Thailand.

  3. Effect of Extender and Equilibration Time on Post Thaw Motility and Chromatin Structure of Buffalo Bull (Bubalus Bubalis) Spermatozoa

    PubMed Central

    Shahverdi, Abdolhossain; Rastegarnia, Abdolreza; Rezaei Topraggaleh, Tohid

    2014-01-01

    Objective The aim of the present study was to investigate the effects of four equilibration times (2, 4, 8 and 16 hours) and two extenders (tris or Bioxcell®) on cryopreservation of buffalo semen. Materials and Methods In this experimental study, split pooled ejaculates (n=4), possessing more than 70% visual sperm motility were divided in two aliquots and diluted in Bioxcell® and tris-citric egg yolk (TCE) extenders. Semen was cooled to 4°C within 2 hours, equilibrated at 4°C for 2, 4, 8 and 16 hours, then transferred into 0.5 ml French straws, and frozen in a programmable cell freezer before being plunged into liquid nitrogen. Postthaw motility characteristics, plasma membrane integrity, acrosome morphology and DNA integrity of the buffalo sperm were studied after thawing. Results There were significant interactions between equilibration times and extenders for sperm motility and membrane integrity. Post thaw sperm motility (PMOT), progressive motile spermatozoa (PROG), plasma membrane integrity (PMI) and normal apical ridge (NAR) measures were lower for sperm equilibrated for 2 hours in both TCE and Bioxcell® extender compared to others equilibration times. PMOT, PMI and NAR for sperm equilibrated for 4, 8 and 16 hours showed no significant differences in either extender, although PROG measures were superior in Bioxcell®compared to TCE at all equilibration times (p<0.05). Kinematic parameters such as average path velocity, curvilinear velocity and linearity in the Bioxcell®extender were superior to those in the TCE extender studied. In contrast to motility and viability, the DNA integrity of post thaw spermatozoa remained unaffected by different equilibration times. Conclusion Equilibration time is necessary for preservation of the motility and integrity of buffalo sperm membranes. Equilibration times of over than 2 hours resulted in the greatest preservation of total semen parameters during cryopreservation. There were no significant interactions between

  4. Nutrient utilisation, growth performance and blood metabolites in Murrah buffalo calves (Bubalus bubalis) divergently selected for residual feed intake.

    PubMed

    Sharma, Vijay K; Kundu, Shivlal S; Prusty, Sonali; Datt, Chander; Kumar, Muneendra

    2016-12-01

    The aim of this study was to evaluate differences in efficiency of feed utilisation between buffalo calves with low and high residual feed intake (RFI) by comparing feed intake, nutrient digestibility, growth traits and blood metabolites. Eighteen male Murrah buffalo calves (aged 4-6 months; 70 ± 1.0 kg body weight) were fed ad libitum with a total mixed ration for 120 d. Based on linear regression models involving dry matter intake (DMI), average daily gain (ADG) and mid-test metabolic body size, calves were assigned into low and high RFI groups. The RFI varied from -0.33 to +0.28 kg DM/d with an average RFI of -0.14 and 0.14 kg DM/d in low and high RFI calves, respectively. Calves had a mean DMI of 1.9 and 2.4 kg/d and an ADG of 0.5 and 0.6 kg/d in low and high RFI groups, respectively. Low RFI calves ate 19.0% less DM each day and required significantly less metabolisable energy for maintenance compared with high RFI calves (12.5 vs. 16.7 MJ/d). Nutrient digestibility and nitrogen balance did not differ among low and high RFI calves. In more efficient animals (low RFI calves) higher (p < 0.05) plasma level of growth hormone, insulin-like growth factor-1 (IGF-1), triiodothyronine (T3) and lower concentration of thyroxin hormone were detected. No significant differences in levels of insulin, hydroxyproline, plasma and urine creatinine, total protein and albumin between high and low RFI groups were found. Blood metabolites showed significant (p < 0.05) differences at initial and final stages of study in both groups. At final stage of study, RFI showed negative correlations with growth hormone, IGF-1, T3, urine creatinine and albumin. Low RFI buffalo calves are more efficient in feed utilisation and the differences in blood metabolites are probably due to differences in feed intake and body metabolism.

  5. Effect of Different Thawing Rates on Post-Thaw Viability, Kinematic Parameters and Chromatin Structure of Buffalo (Bubalus bubalis) Spermatozoa

    PubMed Central

    Rastegarnia, Abdolreza; Shahverdi, Abdolhossein; Rezaei Topraggaleh, Tohid; Ebrahimi, Bita; Shafipour, Vahid

    2013-01-01

    Objective: The aim of the present study was to evaluate three thawing rates on the post thaw motility, viability and chromatin structure of buffalo semen frozen in 0.5-ml straws. Materials and Methods: In this experimental study semen was collected with artificial vagina (42℃) from four buffalo bulls.Split pooled ejaculates (n=4) were extended at 37℃ with a Bioxcell® extender. Semen was cooled to 4℃ within 2 hours, equilibrated at 4℃ for 4 hours, then filled in 0.5 ml French straws, and frozen in programmable cell freezer before plunging into liquid nitrogen. Straws were thawed at water bath temperatures of 37, 50 or 70℃ for 30, 15 and 6 seconds, respectively. Semen was incubated at 37℃ for 2 hours and evaluated for post thaw motility, viability, acrosomal and DNA integrity of spermatozoa. Analysis of variance (ANOVA) was used for comparisons of means. When the ANOVA test showed statistical differences, the mean of the treatments were compared using Duncan’s multiple range tests. Results: The initial postthaw motility (0 hour) averaged 62.7 ± 7.2%, 73.1 ± 9.77%, and 74.9 ± 8.58% for the three thaw rates, respectively. Kinematic parameters such as average path velocity, linearity and beat/cross frequency in the thaw rate of 70℃ for 6 seconds were superior to other rates studied (p<0.05). After 2 hours of incubation, proportions of progressive motility and Kinematic parameters decreased in all groups (p>0.05). A positive correlation was detected between sperm motility and thawing rate after two hours incubation times. The percentage of viable spermatozoa and spermatozoa with an intact acrosome and plasma membrane integrity were not different between the groups of samples thawed at different temperatures (p>0.05). The percentage of spermatozoa with chromatin dispersion forthe thaw rate of 70℃ for 6 seconds was significantly higher than for the to other rates studied (p< 0.05). In contrast with motility and viability, the DNA integrity of post

  6. Cryopreservation of zona-free cloned buffalo (Bubalus Bubalis) embryos: slow freezing vs open-pulled straw vitrification.

    PubMed

    Sirisha, K; Selokar, N L; Saini, M; Palta, P; Manik, R S; Chauhan, M S; Singla, S K

    2013-08-01

    This study was carried out to compare the post-thaw cryosurvival rate and the level of apoptosis in vitro produced zona-free cloned buffalo blastocysts subjected to slow freezing or vitrification in open-pulled straws (OPS). Zona-free cloned embryos produced by handmade cloning were divided into two groups and were cryopreserved either by slow freezing or by vitrification in OPS. Cryosurvival of blastocysts was determined by their re-expansion rate following post-thaw culture for 22-24 h. The post-thaw re-expansion rate was significantly (p < 0.05) higher following vitrification in OPS (71.2 ± 2.3%) compared with that after slow freezing (41.6 ± 4.8%). For examining embryo quality, the level of apoptosis in day 8 frozen-thawed blastocysts was determined by TUNEL staining. The total cell number was not significantly different among the control non-cryopreserved cloned embryos (422.6 ± 67.8) and those cryopreserved by slow freezing (376.4 ± 29.3) or vitrification in OPS (422.8 ± 36.2). However, the apoptotic index, which was similar for embryos subjected to slow freezing (14.8 ± 2.0) or OPS vitrification (13.3 ± 1.8), was significantly (p < 0.05) higher than that for the control non-cryopreserved cloned embryos (3.4 ± 0.6). In conclusion, the results of this study demonstrate that vitrification in OPS is better than slow freezing for the cryopreservation of zona-free cloned buffalo blastocysts because it offers a much higher cryosurvival rate.

  7. Localization and expression of follicle-stimulating hormone receptor gene in buffalo (Bubalus bubalis) pre-antral follicles.

    PubMed

    Sharma, G Taru; Dubey, P K; Kumar, G Sai

    2011-02-01

    Follicle-stimulating hormone (FSH) stimulates antral follicles to grow, but its role in earlier stages (pre-antral) of follicle development, if any, is obscure. Aim of this study was to study the expression of follicle-stimulating hormone receptor (FSHR) gene in different sizes of pre-antral follicles (PFs) (<150, 200, 250, 300, 350, 400 μm) and to find out an optimum dose of FSH for better growth, development and steroidogenesis of PFs in vitro. Buffalo ovaries were collected from a local abattoir, and PFs were isolated by mechanical method. A semi-quantitative RT-PCR amplification strategy was used for mRNA expression, while FSHR protein was localized by immunohistochemistry. Isolated pre-antral follicles (80-85 μm) were cultured in TCM-199 supplemented with 10% foetal bovine serum, 1% ITS and 30 ng/ml EGF served as control medium. Addition of three different doses of FSH (0.5, 1.0, 2.0 μg/ml) in control medium was considered as treatment groups. A single 2.184-kb receptor mRNA transcript was present in all sizes (<150-400 μm) of follicles. Follicle-stimulating hormone receptor was also localized immunohistochemically in granulosa cells of all sizes of follicles. Survival and growth rate of follicles significantly (p<0.05) increased following supplementation of FSH at a concentration of 1.0 μg/ml and the culture medium also showed a significantly (p<0.05) greater accumulation of oestradiol and progesterone. In conclusion, FSHR is expressed in all sizes of PFs and in vitro survival, growth and steroidogenesis of follicles are optimally stimulated by 1.0 μg/ml FSH. These findings demonstrate that FSH has an important role during the recruitment, growth and development of buffalo ovarian PFs.

  8. Prognostic value of various spermatological attributes as predictors of zona binding and zona penetration of buffalo (Bubalus bubalis) semen.

    PubMed

    Selvaraju, S; Ghosh, J; Ravindra, J P

    2009-02-01

    Twenty-four ejaculates from six (four ejaculates each) Surti buffalo bulls aged 4-8 years were used to assess various attributes of spermatozoa influencing the zona-binding and zona-penetration tests. Ejaculates from each bulls were subjected to in vitro sperm--zona binding and sperm--zona penetration tests (four replicates per bull) using immature buffalo oocytes. The average number of spermatozoa bound per oocyte was 27.79 +/- 5.90. The average number of spermatozoa penetrated per oocyte was 3.35 +/- 0.64. The average number of zona-bound and -penetrated spermatozoa differed significantly between animals. Significant difference (p < 0.05) was observed between the plasmalemma integrity as assessed by eosin--nigrosin stain and hypo-osmotic swelling (HOS) test. Furthermore, the percentage of cells positive for the HOS test, i.e. functional membrane integrity (51.25 +/- 2.32) was significantly (p < 0.05) higher than hypo-osmotic swelling-Giemsa (HOS-G) test, i.e. the subpopulation of spermatozoa positive for functional membrane and acrosomal integrities (42.87 +/- 4.56). The HOS test had significant correlations with plasmalemma integrity as measured by the vital stain, eosin--nigrosin (r = 0.85, p < 0.05). The HOS-G test also had significant correlation with plasmalemma integrity measured by vital stains such as eosin--nigrosin (r = 0.90, p < 0.05) and fluorogenic stains [carboxyfluorescein diacetate (CFDA) and propidium iodide (PI); r = 0.92, p < 0.01] and HOS test (r = 0.93), acrosomal integrity (r = 0.86, p < 0.05) and mitochondrial membrane potential (r = 0.99, p < 0.01). The plasmalemma integrity (fluorogenic stain), functional membrane integrity (HOS test), subpopulation of spermatozoa positive for functional membrane and acrosomal integrities (HOS-G test) and mitochondrial membrane potential had significant (p < 0.05) correlation with sperm zona binding and penetration. The present study indicates that these parameters could represent important determinants

  9. Effects of the seminal plasma zinc content and catalase activity on the semen quality of water buffalo (Bubalus bubalis) bulls.

    PubMed

    Alavi-Shoushtari, S M; Rezai, S Asri; Ansari, M H Kh; Khaki, A

    2009-01-15

    In order to determine zinc and catalase content of seminal plasma in the buffalo and to study their associations with the semen characteristics, 54 semen samples were collected from 10 buffalo bulls; semen volume and sperm concentration, gross and progressive motility and viability were evaluated, seminal plasma was then harvested by centrifugation and its zinc content was estimated by atomic absorption spectrophotometer and its catalase activity determined by using a commercial kit. The zinc content of the seminal plasma (Mean +/- SEM) was recorded as 154.40 +/- 1.74 mg L(-1), while, the mean catalase value was 32.00 +/- 0.42 U mL(-1). The mean zinc values was highly correlated with sperm progressive motility and viability and with catalase values (p = 0.000 for all) and also was associated with gross motility (p = 0.020) and negatively with abnormal morphology (p = 0.049). The catalase values were highly associated with sperm progressive motility, viability and zinc content (p = 0.000 for all) and was associated with sperm gross motility (p = 0.024). For further clarification of these correlations, the samples were categorized in three groups of excellent (Ex, >90% motile, n = 33), good (Go, 80-89% motile, n = 15) and moderate (Mo, <79% motile, n = 6) according to their percentage of sperm motility. The mean progressive motility in Ex group was 92.54 +/- 0.51%, in Go group was 81.66 +/- 0.62% and in Mo group was 71.66 +/- 1.05%. The mean zinc and catalase values were recorded as 161.07 +/- 1.63 mg L(-1) and 33.41 +/- 0.34 U mL(-1) in Ex, 146.70 +/- 1.91 mg L(-1) and 31.01 +/- 0.67 in Go and 136.42 +/- 4.97 mg L(-1) and 26.51 +/- 0.87 U mL(-1) in Mo groups. The mean zinc value in Ex group was highly associated with sperm motility, viability and catalase values, in Go group was associated with catalase values and highly associated with sperm abnormal morphology and in Mo group it was highly associations with catalase values only. The mean catalase value in Ex group

  10. Potential risk factors associated with ill-thrift in buffalo calves (Bubalus bubalis) raised at smallholder farms in Egypt

    PubMed Central

    Ali, Mohamed A.; El-khodery, Sabry A.; El-said, Waleed E.

    2014-01-01

    Failure to grow (ill-thrift) in calves has a negative effect on animal production and health. The present study was carried out from November, 2009 to May, 2013 to investigate the risk factors of ill-thrift in buffalo calves. A total of 344 calves at 78 smallholder farms were selected randomly. A questionnaire was designed to include managemental, nutritional and disease risk factors. Serum selenium, copper, zinc, iron, calcium, phosphorus and magnesium were measured. Data were subjected to logistic regression analysis and results were expressed as p value, odds ratio (OR) and confidence interval (CI). Fifty-five calves (15.9%) showed ill-thrift. On animal level, the final multivariate logistic regression model showed a significant association between ill-thrift and early weaning (p < 0.01; OR: 45.755; CI: 4.35–480.25), diarrhea (p < 0.05; OR: 41.315; CI: 1.710–998.0), indoor management (p < 0.05; OR: 63.56; CI: 2.701–14.96) and low serum phosphorus (p < 0.01; 292.0; CI: 5.256–16.23). On farm level, inadequate mineral supplementation (p < 0.001; OR: 18.62; CI: 3.89–88.9) and irregular use of anthelmintics (p < 0.05; OR: 7.95; CI: 1.53–41.23) were the potential factors. Clinically, ill-thrift calves were more likely to have alopecia (p < 0.01), recumbency (p < 0.01), emaciation (p < 0.001), hypothermia (p < 0.01), inappetance (p < 0.001), lacrimation (p < 0.001), hypomotile rumen (p < 0.001), and pale mucous membrane (p < 0.001). The results of the present study indicate that ill-thrift in buffalo calves could occur as a result of interaction between management errors and disease factors. Identification of the risk factors associated with ill-thrift may provide useful information, which assist to construct the suitable preventive measures. PMID:26199751

  11. Efficiency to reach age of puberty and behaviour of buffalo heifers (Bubalus bubalis) kept on pasture or in confinement.

    PubMed

    Sabia, E; Napolitano, F; De Rosa, G; Terzano, G M; Barile, V L; Braghieri, A; Pacelli, C

    2014-11-01

    In order to evaluate the influence of rearing system (free-ranging (FR) v. confinement (C)) on buffalo heifer efficiency to reach age of puberty and on behavioural and immune functions, two experiments were conducted from September 2010 to October 2011. In Experiment I, 32 subjects aged 8 to 9 months at the start of experiment were used. A total of 16 animals (group C) were group housed in an indoor slatted floor pen (4 m2/animal) with an outdoor paddock (4 m2/animal); 16 others grazed on a Mediterranean natural pasture of 40 ha (group FR). Behavioural data were collected and organic matter digestibility, blood metabolites and progesterone were determined. At the end of the experiment, a novel object test and a skin test were conducted, and the avoidance distance (AD) at the manger was measured. Free-ranging animals were able to express natural behaviours such as wallowing and grazing. C animals devoted more time to the novel object than FR animals, whereas AD at manger was lower in group FR than in group C (P<0.01). Cellular immune response was higher in FR heifers than in C animals (P<0.01). FR animals also showed a higher digestibility of organic matter (P<0.01). Heifers from group FR had higher plasma concentrations of non-esterified fatty acids (P<0.001) and lower concentrations of glucose than heifers from group C (P<0.001). C animals showed higher daily weight gains (P<0.01) and weight at the puberty (P<0.05), but there were no differences in terms of age of puberty between the two groups. The intakes of dry matter (DM), CP and energy to reach the age of puberty were similar in both groups. In order to verify whether the results obtained in Experiment I could be replicated in different rearing conditions (reduced pasture availability, different location and altitude), a second experiment was conducted on 26 animals, where only onset of age of puberty and metabolic profile were monitored. In Experiment II, 13 heifers grazed on a natural pasture of 5 ha, other

  12. Ovarian dynamics and milk progesterone concentrations in cycling and non-cycling buffalo-cows (Bubalus bubalis) during Ovsynch program.

    PubMed

    Ali, A; Fahmy, S

    2007-07-01

    The objective was to evaluate ovarian dynamics and progesterone concentrations in cyclic (CYC, n=10) and non-cyclic (NCY, n=8) buffalo-cows during Ovsynch program. All cows received GnRH on day 0, PGF2alpha on day 7, and GnRH on day 9, and AI 14 h later. Ovarian structures were monitored by ultrasound and milk samples were collected for progesterone (P4) analysis. The first GnRH resulted in ovulation in CYC (90%) and NCY (62.5%) cows. By day 7, almost all cows had large follicle and lutein tissue. Luteolytic responses to PGF2alpha were 80 and 87.5% for CYC and NCY cows, respectively. Following second GnRH, ovulation occurred in 80% of CYC and 100% of NCY cows. Ovulation began earlier (12 h following second GnRH) and extended for longer (36 h) in NCY cows, when compared to CYC cows (36 and 12 h, respectively). The mean P4 levels increased from days 0 through 7 in CYC and NCY cows and levels were higher in CYC group. Conception rates were 60 and 37.5% in CYC and NYC cows, respectively. Early and asynchronous ovulation and luteal sub-function seemed to be a problem in NCY cows. Inseminating NCY cows twice, at 0 and 24 h of the second GnRH is recommended.

  13. Empirical and bioinformatic characterization of buffalo (Bubalus bubalis) colostrum whey peptides & their angiotensin I-converting enzyme inhibition.

    PubMed

    Ashok, N R; Aparna, H S

    2017-08-01

    Whey based peptides are well known for their nutritional and multifunctional properties. In this context, whey proteins from buffalo colostrum & milk were digested by in vitro simulation digestion and analyzed by nano-LC-MS/MS. Functional protein association networks, gene annotations and localization of identified proteins were carried out. An ACE inhibitory peptide sorted from the library was custom synthesized and an in vitro ACE assay was performed. The study led to the identification of 74 small peptides which were clustered into 5 gene functional groups and majority of them were secretory proteins. Among the identified peptides, majority of them were found identical to angiotensin I-converting enzyme (ACE) inhibitors, antioxidant, antimicrobial, immunomodulatory and opioidal peptides. An octapeptide (m/z - 902.51, IQKVAGTW) synthesized was found to inhibit ACE with an IC50 of 300±2µM. The present investigation thus establishes newer vista for food derived peptides having ACE inhibitory potential for nutraceutical or therapeutic applications.

  14. Effects of in vitro copper sulphate supplementation on the ejaculated sperm characteristics in water buffaloes (Bubalus bubalis)

    PubMed Central

    Tabassomi, Mehdi; Alavi-Shoushtari, Sayed Mortaza

    2013-01-01

    This study was carried out to investigate effects of copper sulphate (CuSO4) additive to semen extenders on sperm parameters: progressive motility, viability, membrane integrity and DNA damage, after semen dilution and cryopreservation. Semen samples of 5 buffalo bulls of 3-5 years old were collected at 5 different occasions during the autumn 2011. A total number of 25 samples were used in each examination. Sperm progressive motility and viability were measured at 0 (T0), 60 (T1) and 120 (T2) min after diluting semen in tris-citric acid extender containing 0 (control), 0.004, 0.008, 0.016, 0.032 and 0.064 mg L-1 CuSO4. Later, semen was diluted in a tris-citric acid-egg yolk-glycerol extender containing the same amounts of CuSO4, cooled to 4 ˚C and kept refrigerated for 4 hr to equilibrate, sperm progressive motility, viability, membrane integrity and DNA damage were estimated. Then, semen was packed in 0.5 mL French straws and frozen in liquid nitrogen. Later, the frozen semen was thawed in 37 ˚C water bath for 30 sec, and the same parameters as well as total antioxidant capacity (TAC) of the frozen-thawed semen were estimated. The results showed that copper additive at the rate of 0.032 mg L-1 gives a better protection of sperms through the process of dilution, equilibration and freeze-thawing than that in control and other Cu concentrations, while 0.064 mg L-1 CuSO4 had deleterious effect on the sperm. PMID:25593683

  15. Sperm surface hyaluronan binding protein (HABP1) interacts with zona pellucida of water buffalo (Bubalus bubalis) through its clustered mannose residues.

    PubMed

    Ghosh, Ilora; Datta, Kasturi

    2003-02-01

    Sperm-oocyte interaction during fertilization is multiphasic, with multicomponent events, taking place between zona pellucida (ZP) glycoproteins and sperm surface receptor. d-mannosylated glycoproteins, the major constituents of ZP are considered to serve as ligands for sperm binding. The presence of hyaluronan binding protein 1 (HABP1) on sperm surface of different mammals including cattle and its possible involvement in sperm function is already reported. Recently, we have demonstrated the specificity of clustered mannose as another ligand for HABP1 (Kumar et al., 2001: J Biosci 26:325-332). Here, we report that only N-linked mannosylated zona-glycoproteins bind to sperm surface HABP1. Labeled HABP1 interacts with ZP of intact oocyte of Bubalus bubalis, which can be competed with unlabeled HABP1 or excess d-mannosylated albumin (DMA). This data suggests the specific interaction of HABP1 with ZP, through clustered mannose residues. In order to examine the physiological significance of such an interaction, the capacity of sperm binding to oocytes under in vitro fertilization plates was examined either in presence of DMA alone or in combination with HABP1. The number of sperms, bound to oocytes was observed to reduce significantly in presence of DMA, which could be reversed by the addition of purified recombinant HABP1 (rHABP1) in the same plate. This suggests that sperm surface HABP1 may act as mannose binding sites for zona recognition. Copyright 2003 Wiley-Liss, Inc.

  16. Transcriptional status of known and novel genes tagged with consensus of 33.15 repeat loci employing minisatellite-associated sequence amplification (MASA) and real-time PCR in water buffalo, Bubalus bubalis.

    PubMed

    Srivastava, Jyoti; Premi, Sanjay; Pathak, Deepali; Ahsan, Zaid; Tiwari, Madhulika; Garg, Lalit C; Ali, Sher

    2006-01-01

    We conducted minisatellite-associated sequence amplification (MASA) with an oligo (5' CACCTCTCCACCTGCC 3') based on consensus of 33.15 repeat loci using cDNA from the testis, ovary, spleen, kidney, heart, liver, and lung of water buffalo Bubalus bubalis and uncovered 25 amplicons of six different sizes (1,263, 846/847, 602, 576, 487, and 324 base pairs). These fragments, cloned and sequenced, were found to represent several functional, regulatory, and structural genes. Blast search of all the 25 amplicons showed homologies with 43 transcribing genes across the species. Of these, the 846/847-bp fragment, having homology with the adenylate kinase gene, showed nucleotide changes at six identical places in the ovary and testis. The 1,263; 324; and 487-bp fragments showed homology with the secreted modular calcium binding protein (SMOC-1), leucine-rich repeat neuronal 6A (LRRN6A) mRNA, and human TTTY5 mRNA, respectively. Real-time PCR showed maximum expression of AKL, LRRN6A, and T-cell receptor gamma (TCR-gamma)-like genes in the testis, SMOC-1 in the liver, and the T-cell receptor-like (TCRL) gene in the spleen compared to those used as endogenous control. We construe that these genes have evolved from a common progenitor and conformed to various biological functions during the course of evolution. MASA approach coupled with real-time PCR has potentials to uncover accurate expression of a large number of genes within and across the species circumventing the screening of cDNA library.

  17. Characterization of leptin receptor gene in Bubalus bubalis and association analysis with body measurement traits.

    PubMed

    De Matteis, Giovanna; Scatà, Maria Carmela; Catillo, Gennaro; Terzano, Giuseppina Maria; Grandoni, Francesco; Napolitano, Francesco

    2015-06-01

    Leptin has a pleiotropic effect on regulating appetite, energy metabolism, growth, reproduction, body composition and immunity. This property supports leptin and its receptor as candidate genes for evaluating genetic polymorphisms to associate with growth, milk yield and other economic traits. The aim of this study is to characterize the leptin receptor gene in Bubalus bubalis, to identify single-nucleotide polymorphism (SNP) sites in different coding and non-coding regions and to analyse potential associations between SNPs identified and the body measurements traits of growing buffalo heifers. A group of 64 animals were genotyped by direct sequencing and twenty-eight SNPs were detected. A sequence analysis revealed the presence of nine interesting SNPs in gene sequence. The association analysis of polymorphisms with the body measurements traits of growing buffalo heifers shows significant statistical effects on chest depth and sacrum height. Therefore according to the results obtained from this study, the leptin receptor gene appears to have potential effects on the body measurement traits of Bubalus bubalis.

  18. Identification of tamaraw (Bubalus mindorensis) from natural habitat-derived fecal samples by PCR-RFLP analysis of cytochrome b gene.

    PubMed

    Ishihara, Shinya; Herrela, Rommel J; Ijiri, Daichi; Matsubayashi, Hisashi; Hirabayashi, Miho; Del Barrio, Arnel N; Boyles, Rodel M; Eduarte, Medardo M; Salac, Ronilo L; Cruz, Libertado C; Kanai, Yukio

    2010-12-01

    Fecal DNA analysis is a useful tool for the investigation of endangered species. Tamaraw (Bubalus mindorensis) is endemic to the Philippine island of Mindoro but knowledge of its genetic and ecological information is limited. In this study, we developed a species identification method for tamaraw by fecal DNA analysis. Eighteen feces presumed to be from tamaraw were collected in Mount Iglit-Baco National Park and species-known feces from domestic buffaloes and cattle were obtained from a farm. Additionally, one species-unknown fecal sample was obtained in Mount Aruyan Preserve, where the sighting of tamaraw has not been reported in recent years. Based on DNA sequence data previously reported, the genus Bubalus- and tamaraw-specific primers for PCR of cytochrome b gene were newly designed. The Bubalus-specific primer yielded a 976 bp fragment of cytochrome b for all fecal samples from tamaraw and domestic buffaloes, but not for cattle, whereas the tamaraw-specific primer yielded a 582 bp fragment for all tamaraw fecal samples and for one of the four domestic buffalo samples. PCR-RFLP (restriction fragment length polymorphism) analysis of the 976 bp PCR fragment with AvrII or BsaXI provided distinct differences between tamaraw and domestic buffalo. PCR-RFLP analysis also showed that the species-unknown sample obtained in Mount Aruyan Preserve, originates from tamaraw.

  19. Identification of polymorphism in fatty acid binding protein 3 (FABP3) gene and its association with milk fat traits in riverine buffalo (Bubalus bubalis).

    PubMed

    Dubey, Praveen Kumar; Goyal, Shubham; Mishra, Shailendra Kumar; Arora, Reena; Mukesh, Manishi; Niranjan, Saket Kumar; Kathiravan, Periasamy; Kataria, Ranjit Singh

    2016-04-01

    The fatty acid binding protein 3 (FABP3) gene, known to be associated with fat percentage of milk and meat in bovines, was screened among swamp and riverine buffaloes for polymorphism detection and further association with milk fat contents. An SNP g.307C > T was identified in the intron 2 (+53 exon 2) region of FABP3 gene of Indian buffaloes. The SNP identified was genotyped in 692 animals belonging to 15 riverine, swamp and hybrid (riverine × swamp) buffalo populations of diverse phenotypes and utilities, by PCR-RFLP. A marked contrast was observed between the C and T allele frequencies in three types of buffaloes. The frequency of C allele ranged from 0.67 to 0.96 in pure swamp buffalo populations, with the highest in Mizoram (0.96). Whereas the frequency of T allele was high across all the Indian riverine buffalo breeds, ranging from 0.57 to 0.96. None of the genotypes at FABP3 g.307C > T locus was found to have significant association with milk fat and other production traits in Mehsana dairy buffalo breed. Our study revealed marked differences in the allele frequencies between riverine and swamp buffaloes at FABP3 g.307C > T locus, without any significant association with different milk traits in riverine buffaloes.

  20. The efficacy and safety of alphacypermethrin as a pour-on treatment for water buffalo (Bubalus bubalis) infested with Haematopinus tuberculatus (Phthiraptera: Haematopinidae).

    PubMed

    Veneziano, Vincenzo; Neglia, Gianluca; Cimmino, Roberta; Balestrieri, Anna; Rufrano, Domenico; Bastianetto, Eduardo; Santoro, Mario; Gokbulut, Cengiz

    2013-08-01

    The sucking louse Haematopinus tuberculatus (Burmeister 1839) is an ectoparasite of buffaloes, cattle, camels, and American bison. Alphacypermethrin (ACYP) is a pyrethroid insecticide commonly used to control arthropods of veterinary and public health interest. Therapeutics, such as antiparasitic compounds, is often administered to buffaloes based on dosage and intervals recommended for cattle because very few drugs have buffalo-specific label indications. A trial was conducted on 20 louse-infested buffaloes at a farm to assess the efficacy and safety of ACYP pour-on, at the manufacturer's recommended dose for cattle, on buffaloes naturally infested by H. tuberculatus. Ten animals were assigned to ACYP-treated group (ACYP-group) and ten to untreated control group (C-group). On day 0, all ACYP-group buffaloes received alphacypermethrin pour-on. Louse counts were performed on days -1, 7, 14, 21, 28, 35, 42, 49, and 56 at eight predilection sites on the skin of each buffalo. ACYP was completely effective (100%) at day 7, highly effective (99.8%) at day 14, and completely effective (100%) from day 21 until the end of the study (day 56 post-treatment). During the trial, ACYP was well tolerated by all animals as there were no observed clinically adverse reactions. The results of this trial suggest that ACYP is an effective, safe, and user-friendly compound suitable for treatment of buffaloes with natural louse infestations.

  1. Molecular assays reveal the presence of Theileria spp. and Babesia spp. in Asian water buffaloes (Bubalus bubalis, Linnaeus, 1758) in the Amazon region of Brazil.

    PubMed

    Silveira, Júlia A G; de Oliveira, Cairo H S; Silvestre, Bruna T; Albernaz, Tatiana T; Leite, Rômulo C; Barbosa, José D; Oliveira, Carlos M C; Ribeiro, Múcio F B

    2016-07-01

    Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from

  2. Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (Bubalus bubalis).

    PubMed

    Kapila, Neha; Kishore, Amit; Sodhi, Monika; Sharma, Ankita; Kumar, Pawan; Mohanty, A K; Jerath, Tanushri; Mukesh, M

    2013-01-01

    Gene expression studies require appropriate normalization methods for proper evaluation of reference genes. To date, not many studies have been reported on the identification of suitable reference genes in buffaloes. The present study was undertaken to determine the panel of suitable reference genes in heat-stressed buffalo mammary epithelial cells (MECs). Briefly, MEC culture from buffalo mammary gland was exposed to 42 °C for one hour and subsequently allowed to recover at 37 °C for different time intervals (from 30 m to 48 h). Three different algorithms, geNorm, NormFinder, and BestKeeper softwares, were used to evaluate the stability of 16 potential reference genes from different functional classes. Our data identified RPL4, EEF1A1, and RPS23 genes to be the most appropriate reference genes that could be utilized for normalization of qPCR data in heat-stressed buffalo MECs.

  3. Influence of body condition score at calving on the metabolic status and production performance of Murrah buffaloes (Bubalus bubalis) during the transition period.

    PubMed

    Delfino, Nelson Carvalho; Bulcão, Lucas Fialho de Aragão; Alba, Henry Daniel Ruiz; Oliveira, Mauricio Xavier da Silva; Queiroz, Filipe Pinheiro Soares de; Carvalho, Gleidson Giordano Pinto de; Rennó, Francisco Palma; Freitas Júnior, José Esler

    2017-07-17

    The purpose of this study was to evaluate the influence of body condition score (BCS) at calving on the metabolic status of female Murrah buffaloes in the transition period. Thirty-seven pregnant buffaloes (multiparous) were selected and monitored during the transition period based on their body condition score and on the estimated calving date. Two groups were formed: 1) buffaloes with a BCS > 3.5 (n = 17); this group was classified and named 'high BCS at calving' (HBCS); and 2) buffaloes with a BCS ≤ 3.5 (n = 20); this group was classified and named 'low BCS at calving' (LBCS). All animals were monitored during the last 30 days of pregnancy and the first 70 days post-calving and kept in the same environment and under the same feeding and management conditions. Mean values for BCS at calving were 2.98 ± 0.9 (mean ± SD) and 4.21 ± 0.9 (mean ± SD) for the HBCS and LBCS groups, respectively. The HBCS group showed higher milk fat content (p = 0.007) and milk fat yield (p = 0.027) and a higher concentration of milk urea nitrogen (p = 0.001) than LBCS buffaloes, which in turn had a lower urine pH value (p = 0.033) than HBCS buffaloes in the pre-calving period (7.86 for HBCS vs. 7.76 for LBCS). The HBCS animals had a higher concentration of erythrocytes (p = 0.001) and hematocrit (p = 0.012) post-calving and a higher hemoglobin concentration (p = 0.004) pre-calving. Buffaloes during the transition period exhibited some variations in the oxidative stress related to their metabolic status. After calving, buffaloes with a high BCS at calving and greater lipid mobilization have a more marked alteration in oxidative status, but improved production performance.

  4. A complementary diagnosis of naturally occurring tuberculosis in water buffaloes (Bubalus bubalis) in Rio de Janeiro using a MPB70-ELISA, Brazil.

    PubMed

    Zarden, Carlos F O; Marassi, Carla D; Oelemann, Walter; Lilienbaum, Walter

    2013-06-01

    As tuberculosis is still a worldwide infection and buffalo breeding represents an important economic activity in various countries, the purpose of this study was to employ an enzyme-linked immunosorbent assay (ELISA) using MPB70 as a capture antigen for the diagnosis of naturally occurring tuberculosis in water buffaloes in Brazil. After the introduction of newly acquired cattle onto a tuberculosis (TB) free farm, an outbreak of TB was recorded in a mixed herd comprising water buffaloes (21) and cattle (46). The entire herd was tested by intradermal tuberculin injection (ITT) and positive animals were slaughtered and tested by culture, polymerase chain reaction (PCR), and ELISA. From the 21 buffaloes sampled, three were reactive by ITT. All the three had positive culture and ELISA, while PCR was positive in two of them. Besides that, one ITT-negative buffalo was slaughtered and presented positive results by both culture and ELISA, and was considered as anergic. Although there were only few animals, those findings demonstrate the diagnostic usefulness of an MPB70-ELISA to correctly detect Mycobacterium bovis tuberculosis in water buffaloes.

  5. Synchronization of follicular wave emergence following ultrasound-guided transvaginal follicle ablation or estradiol-17β administration in water buffalo (Bubalus bubalis).

    PubMed

    Honparkhe, M; Gandotra, V K; Matharoo, J S; Ghuman, S P S; Dadarwal, D; Singh, Jaswant

    2014-04-01

    The aim of this study was to assess the synchrony in follicular wave emergence and subsequent ovulation following dominant follicle ablation or estradiol-17β administration. Six cycling Murrah buffaloes were sequentially allotted to three groups, that is, control, follicular ablation, and estradiol-17β groups. For the control group, buffaloes at random stages of estrous cycle were examined daily by transrectal ultrasonography for 14 days and the day of wave emergence was recorded. Following induced luteolysis and ovulation (Day 0), these buffaloes were included in the ablation group. All follicles (>5mm) were ablated on Day 3 or 5 or 7 (n=2 each day). Seven days after the ablation, these buffaloes were administered prostaglandin F2α to induce luteolysis and ovulation. Following this, buffaloes were included in the estradiol treatment group with estradiol administered on similar days as for ablation in the ablation group. Luteolysis was induced nine days after the estradiol injection. All animals of the treatment groups were subjected to transrectal ultrasound and blood samplings daily from treatment to induced ovulation. The follicular waves emerged significantly earlier (P=0.001) in both the ablation (2.1±0.79 days) and estradiol (4.0±0.25 days) treatment groups than the control group (8.3±0.88 days). The deviation from mean day of ovulation was greater (P=0.02) for the control group buffaloes (1.66±0.3 day) than those of the treatment groups (ablation, 0.76±0.2 and estradiol, 0.58±0.2 day). In conclusion, both ablation and estradiol resulted in synchronous emergence of a new follicular wave irrespective of stage at which the treatment was given, with greater synchrony of ovulations in water buffalo. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Purification of pregnancy-associated glycoproteins from late-pregnancy Bubalus bubalis placentas and development of a radioimmunoassay for pregnancy diagnosis in water buffalo females

    PubMed Central

    2013-01-01

    Background Pregnancy-associated glycoproteins (PAGs) were first described as placental antigens present in the blood serum of the mother soon after implantation. Here, we describe the purification of several pregnancy-associated glycoproteins from water buffalo placenta (wbPAGs). A specific radioimmunoassay (RIA) was developed for early pregnancy diagnosis in buffalo species. Results Amino-terminal microsequencing of immunoreactive placental proteins allowed the identification of eleven wbPAGs sequences [Swiss-Prot accession numbers: P86369 to P86379]. Three polyclonal antisera (AS#858, AS#859 and AS#860) were raised in rabbits against distinct wbPAG fractions. A new RIA (RIA-860) was developed and used to distinguish between pregnant (n = 33) and non-pregnant (n = 26) water buffalo females. Conclusions Our results confirmed the multiplicity of PAG expression in buffalo placenta. In addition, the RIA-860 system was shown to be sensitive, linear, reproducible, accurate and specific in measuring PAG concentrations in buffalo plasma samples from Day 37 of gestation onwards. PMID:23634647

  7. Regucalcin is widely distributed in the male reproductive tract and exerts a suppressive effect on in vitro sperm capacitation in the water buffalo (Bubalus bubalis).

    PubMed

    Pillai, Harikrishna; Shende, Ashish Mukunda; Parmar, Mehtab Sing; A, Anjaneya; L, Sreela; Kumaresan, Arumugam; G, Taru Sharma; Bhure, Sanjeev Kumar

    2017-03-01

    Regucalcin is a multi-functional, calcium-binding protein with roles in calcium homeostasis, apoptosis, cell proliferation, and free radical neutralization. Regucalcin is broadly expressed in the male reproductive organs of rat and bovine; here, we report its expression in the reproductive tract of male buffalo-especially in testis, epididymis, seminal vesicle, prostate, and bulbourethral gland of buffalo-as analyzed by real-time PCR, Western blot, and immunolocalization. Regucalcin degradation in seminal plasma, despite its high abundance in vesicular fluid, was demonstrated using recombinant regucalcin co-incubated with buffalo seminal plasma. This depletion of regucalcin appears to be related to its suppressive effect on in vitro sperm capacitation, observed using the chlortetracycline assay after treating buffalo spermatozoa with recombinant protein. Indeed, addition of recombinant regucalcin to capacitating media significantly reduced (P < 0.05) the percentage of capacitated spermatozoa to 6.1 ± 0.6 from 36.4 ± 1.8 in the untreated group. Taken together, the wide distribution of regucalcin in male buffaloes, versus its degradation in the seminal plasma and suppressive effects on in vitro capacitation of spermatozoa, indicate its possible anti-capacitation role in the reproductive tract. Mol. Reprod. Dev. 84: 212-221, 2017. © 2016 Wiley Periodicals, Inc.

  8. Bovine papillomavirus type 2 infects the urinary bladder of water buffalo (Bubalus bubalis) and plays a crucial role in bubaline urothelial carcinogenesis.

    PubMed

    Roperto, Sante; Russo, Valeria; Ozkul, Ayhan; Sepici-Dincel, Aylin; Maiolino, Paola; Borzacchiello, Giuseppe; Marcus, Ioan; Esposito, Iolanda; Riccardi, Marita Georgia; Roperto, Franco

    2013-02-01

    Bovine papillomavirus type 2 (BPV-2) has been shown to infect and play a role in urinary bladder carcinogenesis of buffaloes grazed on pastures with ferns from the Marmara and Black Sea Regions of Turkey. BPV-2 DNA has been found in both neoplastic and non-neoplastic lesions of the urinary bladder. Furthermore, this virus may be a normal inhabitant of the urinary bladder since BPV-2 DNA has also been detected in clinically normal buffaloes. The viral activation by fern immunosuppressant or carcinogen may trigger the urothelial cell transformation. The E5 oncoprotein was solely detected in urothelial tumours and appeared to be co-localized with the overexpressed and phosphorylated platelet derived growth factor (PDGF) β receptor in a double-colour immunofluorescence assay. Our results indicate that the E5-PDGF β receptor interaction also occurs in spontaneous tumours of the bubaline urinary bladder, revealing an additional role of BPV-2 in bladder carcinogenesis of buffaloes.

  9. Identification, molecular characterization, and tissue expression of parathyroid hormone-related protein gene (PTHrP) from water buffalo (Bubalus bubalis).

    PubMed

    Liu, J; Qian, L D; Huo, J L; Bi, B L; Li, D L; Wang, S F; Chen, T; Li, L J; Mao, H M; Miao, Y W

    2015-03-27

    Parathyroid hormone-related protein (PTHrP) is involved in the deposition of milk calcium in mammal lactation, but its role in buffalo is unclear. In this study, the full-length coding sequence of the water buffalo PTHrP gene was first isolated using reverse transcription-polymerase chain reaction. The protein was then subjected to molecular characterization using bioinformatic methods, and the tissue expression pattern was further assayed by semi-quantitative reverse-transcription polymerase chain reaction. The water buffalo PTHrP gene contains an open reading frame of 534 base pairs encoding a polypeptide of 177 amino acid residues, a theoretical molecular weight of 20.32 kDa, and an isoelectric point of 10.00. In addition, water buffalo PTHrP was predicted to contain a signal peptide, a typical hydrophobic region with no hydrophobic transmembrane regions, and to exert its function in the cell nucleus. A conserved domain of parathyroid superfamily from amino acids 34-114 was observed in the polypeptide. Sequence comparison and the phylogenetic analysis showed that the sequence of the water buffalo PTHrP protein shared high homology with that of other mammals, particularly cattle and goat. Among the 16 tissues examined, the PTHrP gene was only expressed in adipose tissue, placenta, uterine wall, hypophysis, and mammary gland tissue, but gene expression levels were higher in the uterus wall and adipose tissue. The results of this study suggest that the PTHrP gene plays an important role in the deposition of milk calcium of water buffalo.

  10. Dietary Inorganic Chromium in Summer-Exposed Buffalo Calves (Bubalus bubalis): Effects on Biomarkers of Heat Stress, Immune Status, and Endocrine Variables.

    PubMed

    Kumar, Muneendra; Kaur, Harjit; Deka, Rijusmita Sarma; Mani, Veena; Tyagi, Amrish Kumar; Chandra, Gulab

    2015-09-01

    The aim of this study was to evaluate the effect of different levels of inorganic chromium (Cr) on heat stress, immune response, and hormonal variation in Murrah buffalo calves during the summer season. Twenty-four growing Murrah buffalo calves were randomly allocated into four treatments for a period of 120 days. Feeding regimen was same in all the groups, except the buffalo calves in treatment groups were additionally supplemented with 0.5, 1.0, and 1.5 mg of inorganic Cr/kg dry matter. Buffalo calves were monitored daily for physiological variables and dry matter intake (DMI) and fortnightly for body weight change. Blood samples were collected at day 0, 15, 30, 45, 60, 75, 90, 105, and 120 and analyzed for heat shock protein 70 (Hsp 70), lymphocyte proliferation, neutrophil phagocytic activity, immunoglobulin, ferric reducing antioxidant power (FRAP) assay, insulin, cortisol and thyroid hormones, and Cr levels. Dietary Cr supplementation did not have any effect on DMI, growth performance, and physiological variables. However, lymphocyte proliferation, neutrophil phagocytic activity, plasma immunoglobulin, FRAP value, and plasma Cr concentration increased significantly (P < 0.05) with increase in levels of Cr. Adding Cr to the diet of summer-exposed buffalo calves did not show any effect on plasma levels of thyroid hormone, while concentration of insulin, cortisol, and Hsp 70 decreased (P < 0.05). Supplementation of inorganic Cr to the diet of buffalo calves reared under high ambient temperature improved heat tolerance, immune status without affecting nutrient intake, and growth performance.

  11. Impact of Buserelin Acetate or hCG Administration on the Day of First Artificial Insemination on Subsequent Luteal Profile and Conception Rate in Murrah Buffalo (Bubalus bubalis).

    PubMed

    Pandey, A K; Ghuman, Sps; Dhaliwal, G S; Agarwal, S K; Phogat, J B

    2016-08-01

    This study was designed to investigate the impact of buserelin acetate (BA) or human chorionic gonadotropin (hCG) administration on the day of first artificial insemination (AI) on subsequent luteal profile (diameter of corpus luteum (CL) and plasma progesterone) and conception rate in Murrah buffalo. The present experiment was carried out at two locations in 117 buffalo that were oestrus-synchronized using cloprostenol (500 μg) administered (i.m.) 11 days apart followed by AI during standing oestrus. Based on treatment (i.m.) at the time of AI, buffalo were randomly categorized (n = 39 in each group) into control (isotonic saline solution, 5 ml), dAI-BA (buserelin acetate, 20 μg) and dAI-hCG (hCG, 3000 IU) group. Out of these, 14 buffalo of each group were subjected to ovarian ultrasonography on the day of oestrus to monitor the preovulatory follicle and on days 5, 12, 16 and 21 post-ovulation to monitor CL diameter. On the day of each sonography, jugular vein blood samples were collected for the estimation of progesterone concentrations. All the buffalo (n = 117) were confirmed for pregnancy on day 40 post-ovulation. The conception rate was better (p < 0.05) in dAI-BA (51.3%) and dAI-hCG (66.7%) groups as compared to their control counterparts (30.8%). Furthermore, the buffalo of dAI-hCG group had improved (p < 0.05) luteal profile, whereas the buffalo of dAI-BA group failed (p > 0.05) to exhibit stimulatory impact of treatment on luteal profile when compared to control group. In brief, buserelin acetate or hCG treatment on the day of first AI leads to an increase in conception rate; however, an appreciable impact on post-ovulation luteal profile was observed only in hCG-treated Murrah buffalo. © 2016 Blackwell Verlag GmbH.

  12. Effects of climate on the thermoregulatory responses of male buffalo ( Bubalus-bubalis) calves supplemented with different levels of testosterone and TDN

    NASA Astrophysics Data System (ADS)

    Gangwar, P. C.

    1982-03-01

    The present work was undertaken to study the changes in the thermoregulatory responses due to changing climate, androgen and TDN percent in 12 buffalo male calves from 4 months of age for a period of two years. The results obtained were: (1) The amplitudes of the diurnal cycles in rectal temperature (Tre) in each month of the experiment were very similar but longer in summer months as compared to winter months, (2) The correlation between Tre and ambient temperature (Ta) was high in all months. (3) Buffalo calves without testosterone supplementation on 70% TDN showed maximum rise in Tre by 0.076 0‡C per degree of ambient temperature, indicating more strain, (4) Highest respiratory frequencies occurred at 10:00 h during summer months. Subgroup 2 of group 1 showed the maximum rise by 311% in respiratory frequency in summer over winter due to least acclimatization, (5) The mean daily increase by 9.94‡C in ambient temperature doubled the respiratory frequency of male buffalo calves from January to April, and (6) Androgen therapy and higher TDN percent level helped the male buffalo calves to adjust better and sooner than the other calves to changing climates.

  13. Cholesterol and fatty acid composition of longissimus thoracis from water buffalo (Bubalus bubalis) and Brahman-influenced cattle raised under savannah conditions.

    PubMed

    Giuffrida-Mendoza, Maria; Arenas de Moreno, Lilia; Huerta-Leidenz, Nelson; Uzcátegui-Bracho, Sojan; Valero-Leal, Kutchynskaya; Romero, Sonia; Rodas-González, Argenis

    2015-08-01

    Male (n=66) water buffalo (Buffalo) and Brahman-influenced cattle (Brahman) were born, raised, weaned, fattened on grazing savannah and harvested at two different ages (19 and 24months) to compare lipid composition of the longissimus thoracis muscle. Half of the animals were castrated at seven months of age (MOA) to examine the castration effects. At 24 MOA Brahman steers showed the highest content of total lipids (P<0.05). No significant variation was detected in cholesterol content for either the main or interaction effects in the age groups. Some individual fatty acids varied with the species (P<0.05), however, interspecific similarities were found in fatty acid ratios. For health-related indices, only atherogenic index (AI) showed lower values in favor of Buffalo meat (P<0.05) at both harvesting ages. Although, meat derived from both bovid groups was leaner and showed lower cholesterol level, AI indicates that Buffalo meat might be beneficial from a human health standpoint. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Spermatogonia-specific proteins expressed in prepubertal buffalo (Bubalus bubalis) testis and their utilization for isolation and in vitro cultivation of spermatogonia.

    PubMed

    Goel, Sandeep; Reddy, Niranjan; Mandal, Suman; Fujihara, Mayako; Kim, Sung-Min; Imai, Hiroshi

    2010-10-15

    Buffalo is an economically important livestock species in Asia. Little is known about male germ line technology owing to lack of sufficient understanding regarding expression of germ- and somatic-cell specific-proteins in the testis. In this study, we identified UCHL-1 (PGP 9.5) and lectin- Dolichos biflorus agglutinin (DBA) as specific markers for spermatogonia in buffalo testis. Expression of germ-cell and pluripotency-specific proteins such as DDX4 (VASA) and POU5F1 (OCT3/4) were also present in spermatogonia. Interestingly, the expression of somatic cell-specific proteins such as VIMENTIN and GATA4 were also detected in germ cells. Using two-step enzymatic digestion followed by differential plating and Percoll density-gradient centrifugation, an approximately 55% spermatogonia-enriched cell population could be obtained from the prepubertal buffalo testis. Isolated spermatogonia could survive and proliferate in vitro in DMEM/F12 medium containing 10% fetal bovine serum in the absence of any specific growth factors for a week. Cultured spermatogonia showed DBA affinity and expressed DDX4 and POU5F1. These results may help to establish a long-term culture system for buffalo spermatogonia.

  15. The effect of addition of equine chorionic gonadotropin to a progesterone-based estrous synchronization protocol in buffaloes (Bubalus bubalis) under tropical conditions.

    PubMed

    Murugavel, K; Antoine, D; Raju, M S; López-Gatius, F

    2009-04-15

    Poor estrus expression and anestrus decrease the reproductive efficiency of buffaloes. The objective of this study was to determine whether the addition of equine chorionic gonadotropin (eCG) to an estrous synchronization protocol and timed insemination could improve ovulation and pregnancy rates of anestrous buffalo cows under tropical conditions. The study population comprised 65 lactating Murrah buffalo cows which were assigned to CIDR (n=33) or CIDR+eCG (n=32) treatment groups. Cows in the CIDR group were fitted for 8d with a controlled intravaginal drug release (CIDR) device containing 1.38 g progesterone, received GnRH (10 microg i.m.) on D 0, PGF(2alpha) (750 microg i.m.) on D 7, and GnRH (10 microg i.m.) on D 9; whereas cows in the CIDR+eCG group received the same treatment plus eCG (500 IU, i.m.) at the time of PGF(2alpha) treatment. All cows were inseminated 16-20 h after the second GnRH treatment. Blood samples were obtained 10d before the start of synchronization treatment (Day -10) and at the onset of treatment (Day 0). Cows with plasma progesterone concentrations <1 ng/mL recorded in both samples (Low-Low levels of P4) were classified as non-cyclic cows. Similarly, when either one or both of the sample pair contained concentrations of serum progesterone >/=1 ng/mL (High-High, Low-High, or High-Low levels of P4), the buffaloes were classified as cyclic cows. Ovulation rate, defined as the number of buffaloes with at least one corpus luteum 10 days after insemination, was significantly higher (P=0.018) in the CIDR+eCG (84.4%) cows than in the CIDR cows (57.6%). Pregnancy rate was numerically lower in CIDR (27.3%) than CIDR+eCG (40.6%) cows, though differences were not significant (P=0.25). Pregnancy rates for CIDR+eCG cows were similar to that of cows inseminated after natural estrus (40.9%; 29/71). In the non-cyclic animals, higher ovulation rates (P=0.026) were recorded for the CIDR+eCG (81%) than for the CIDR cows (47.4%). Our results indicate that

  16. Effect of feeding inorganic chromium on growth performance, endocrine variables, and energy metabolites in winter-exposed buffalo calves (Bubalus bubalis).

    PubMed

    Kumar, Muneendra; Kaur, Harjit; Tyagi, Amrish Kumar; Kewalramani, Neelam J; Mani, Veena; Deka, Rijusmita Sarma; Sharma, Vijay Kumar; Chandra, Gulab; Dang, Ajay Kumar

    2013-12-01

    We investigated the effect of chromium (Cr) supplementation on the growth performance, energy metabolites, and hormonal variation in winter-exposed buffalo calves. Twenty-four female buffalo calves were randomly allotted to four dietary treatments (n = 6) for a period of 120 days. Feeding regimen was the same in all the groups, except the animals in the four respective groups were additionally supplemented with 0.0, 0.5, 1.0, and 1.5 mg of Cr/kg DM in the form of CrCl3.6H2O. Calves were monitored daily for physiological variables and dry matter intake (DMI). Blood samples were collected at fortnightly intervals from each buffalo calves to measure concentrations of hormones (insulin, cortisol, and growth hormone), energy metabolites (glucose and non-esterified fatty acids), and plasma mineral levels. After 120 days of feeding trial, buffalo calves fed with Cr had lower (P < 0.05) circulating plasma concentrations of glucose, insulin, and cortisol hormones, whereas plasma thyroid hormone and non-esterified fatty acids concentrations were found similar (P > 0.05) among all the treatments. The results suggested that dietary Cr supplementation influenced plasma Cr levels without affecting the plasma concentrations of other trace minerals. However, physiological variables, nutrient intake, and growth performance of buffalo calves did not differ among all treatments (P > 005). In summary, the current study showed that supplementation of Cr at the level of 1.0 and 1.5 mg of Cr/kg DMI was more effective in improving glucose utilization by increasing potency of insulin hormone and reducing concentration of cortisol hormone. Results also suggested that supplemental Cr also improves blood plasma Cr levels.

  17. Serological profile of buffalo (Bubalus bubalis) female calves vaccinated with standard Brucella abortus strain 19 vaccine using rose bengal, 2-mercaptoethanol and complement fixation tests.

    PubMed

    Nardi, G Júnior; Ribeiro, M G; Jorge, A M; Megid, J; Silva, L M P

    2012-03-01

    The serological profiles of 21 female buffaloes vaccinated between 3 and 8 months of age using Brucella abortus strain 19 (S19) were evaluated by rose bengal (RBT), 2-mercaptoethanol (2ME) and complement fixation (CFT) tests. The serum strains were collected in day zero, 15, 30, 45, 60th days and subsequently to each 30 months, until 720th day after vaccination. No animal showed reaction in day zero. In 15th day above 95% of animals revealed reaction in all tests. All the animals presented absence of reactions in CFT, RBT and 2ME tests at 270, 300 and 360 days after vaccination, respectively. Our finding highlighted early response in CFT compared than other conventional agglutination tests. None of animals presented oscillation of titers or reactions in any test after 360 day of study, which enables the use of these tests after this period without interference of antibodies from S19 vaccine origin between 3 and 8 months in buffalo heifers.

  18. l-Cysteine improves antioxidant enzyme activity, post-thaw quality and fertility of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa.

    PubMed

    Iqbal, S; Riaz, A; Andrabi, S M H; Shahzad, Q; Durrani, A Z; Ahmad, N

    2016-11-01

    The effects of l-cysteine in extender on antioxidant enzymes profile during cryopreservation, post-thaw quality parameters and in vivo fertility of Nili-Ravi buffalo bull spermatozoa were studied. Semen samples from 4 buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of l-cysteine (0.0, 0.5, 1.0, 2.0 and 3.0 mm) and frozen in 0.5-ml French straws. The antioxidative enzymes [catalase, super oxide dismutase and total glutathione (peroxidase and reductase)] were significantly higher (P < 0.05) at pre-freezing and post-thawing in extender containing 2.0 mm l-cysteine as compared to other groups. Post-thaw total motility (%), progressive motility (%), rapid velocity (%), average path velocity (μm s(-1) ), straight line velocity (μm s(-1) ), curvilinear velocity (μm s(-1) ), beat cross frequency (Hz), viable spermatozoa with intact plasmalemma (%), acrosome and DNA integrity (%) were higher with the addition of 2.0 mm l-cysteine as compared to other groups (P < 0.05). The fertility rates (59 versus 43%) were higher (P < 0.05) in buffaloes inseminated with doses containing 2.0 mm of l-cysteine than in the control. In conclusion, the addition of 2.0 mm l-cysteine in extender improved the antioxidant enzymes profile, post-thaw quality and in vivo fertility of Nili-Ravi buffalo bull spermatozoa.

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

    PubMed

    Swelum, Ayman Abdel-Aziz; Amin, Said Elsayed; Eidaroos, Abdel-Salam; Hazzaa, Abo-Bakr Maher

    2012-09-15

    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. Copyright © 2012 Elsevier Inc. All rights reserved.

  20. Impact of buserelin acetate or hCG administration on day 5 post-ovulation on subsequent luteal profile and conception rate in Murrah buffalo (Bubalus bubalis).

    PubMed

    Pandey, A K; Dhaliwal, G S; Ghuman, S P S; Agarwal, S K

    2015-11-01

    The present study aimed to establish the impact of buserelin acetate or hCG administration on day 5 post-ovulation on subsequent luteal profile and conception rate in buffalo. The buffalo (n=45) were subjected to an estrous synchronization protocol (synthetic analog of PGF2α administered, through intramuscular route, 11 days apart), followed by artificial insemination (AI) during mid to late estrus. On day 5 post-ovulation, buffalo were administered (i.m.) normal saline (Control, n=14), buserelin acetate (20μg, d5-BA, n=14) or human chorionic gonadotropin (3000IU, d5-hCG, n=17). Ovarian ultrasonography was conducted on the day of induced estrus and on days 0, 5, 12, 16 and 21 post-ovulation to assess preovulatory follicle or corpus luteum (CL) diameter. Also, on these days, jugular vein blood sampling was conducted for the estimation of plasma progesterone. First service conception rate was greater (χ(2)=5.18, P>0.05) in d5-BA and d5-hCG groups (71.4% and 47.1%, respectively) as compared to control (28.6%). Both treatment groups had a greater (P<0.05) CL diameter and plasma progesterone during the post-treatment period in comparison to that control treatment group. Treatment-induced accessory CL formation was observed in 92.9% and 76.5% buffalo of d5-BA and d5-hCG groups, respectively. In conclusion, buserelin acetate and hCG administration on day 5 post-ovulation leads to accessory CL formation that may have a role in enhancing conception rate. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Effects of adding taurine and trehalose to a tris-based egg yolk extender on buffalo (Bubalus bubalis) sperm quality following cryopreservation.

    PubMed

    Shiva Shankar Reddy, N; Jagan Mohanarao, G; Atreja, S K

    2010-06-01

    Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. Under the present study, the cryoprotective effect of taurine and trehalose on buffalo sperm quality parameters after freeze-thaw process was studied. Buffalo semen was cryopreserved in tris-based egg yolk extender along with cryoprotectants like taurine (50 mM) or trehalose (100 mM) and used for the assessment of sperm quality parameters like motility, viability, plasma membrane integrity, total antioxidant status and the extent of cryocapacitation. The results were compared to semen cryopreserved in tris-based egg yolk extender only as control. Post-thaw semen evaluation clearly indicated that the addition of taurine or trehalose significantly improved (P<0.05) the motility, viability and membrane integrity compared to control spermatozoa. The extent of sperm cells underwent cryocapacitation was significantly lowered (P<0.05) in presence of taurine or trehalose. Moreover, the percentage of in vitro capacitated cells in the treated samples was comparable to the control spermatozoa along with maintaining other sperm quality parameters. Finally, compared to the control and trehalose treated sample, addition of taurine to the freezing extender showed more positive effect on the total antioxidant power of seminal plasma and spermatozoa. It is concluded that the addition of taurine or trehalose to the freezing extender led to the reduction of cryodamage to the buffalo spermatozoa. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  2. Liposome-based semen extender is suitable alternative to egg yolk-based extender for cryopreservation of buffalo (Bubalus bubalis) semen.

    PubMed

    Kumar, Pradeep; Saini, Monika; Kumar, Dharmendra; Balhara, A K; Yadav, S P; Singh, P; Yadav, P S

    2015-08-01

    Demand for alternative of egg yolk in freezing extenders have increased in recent years due to variability in egg yolk composition, risk of microbial contamination and presence of steroid hormones. The alternative to egg yolk-based extender (EY) can be soya lecithin-based extender (SL) and liposome-based extender (LP). However, the efficacy of SL is still a matter of debate. Few studies have been performed on the effect of LP but to date evaluation of buffalo semen cryopreserved in LP has not been studied. Therefore, this study was designed to compare SL and LP with conventional EY for evaluation of post-thaw quality of buffalo semen. Results showed that total, progressive and rapid sperm motility were found significantly higher (P<0.05) in LP among these extenders. In vitro assessment of post-thaw sperm longevity has also resulted in better maintenance of sperm kinetics and motility in LP in comparison to other extenders. Furthermore, sperm cryopreserved in LP travelled significantly more (P<0.05) distance in cervical mucus as compared to SL and EY. Therefore, it can be concluded that the LP is more efficient than SL and EY for the cryopreservation of buffalo semen. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Optimization of embryo culture conditions for increasing efficiency of cloning in buffalo (Bubalus bubalis) and generation of transgenic embryos via cloning.

    PubMed

    Wadhwa, Neerja; Kunj, Neetu; Tiwari, Shuchita; Saraiya, Megha; Majumdar, Subeer S

    2009-09-01

    Cloning in bovine species is marred by low efficiency of blastocyst formation. Any increase in the efficiency of blastocyst formation upon nuclear transfer will greatly enhance the efficiency of cloning. In the present study, the effect of various media, protein sources, and growth factors on the development of cloned buffalo embryos was evaluated. Among various combinations tested, culture of cloned embryos in TCM-199 media on the feeder layer of Buffalo Oviductal Epithelial Cells (BOEC) in the presence of bovine serum albumin-free fatty acid (BSA-FFA) and leukemia inhibitory factor (LIF) provided most suitable environment for efficient development of cloned blastocysts. Under these conditions, we achieved a blastocyst formation rate of 43%, which is better than those reported previously. Because preimplantation embryonic development, in vivo, occurs in an environment of oviductal cells, the blastocysts generated by this method may presumably be more suitable for implantation and further development. Additionally, we generated green blastocysts from enucleated oocytes by transfer of nuclei from cells transfected with EGFP transgene, showing possibility of transgenesis via cloning in this species. To our knowledge, this is the first report regarding the production of transgenic cloned buffalo embryos and their developmental competence with respect to various media, cocultures, and supplements.

  4. Stage specific expression of ATP-binding cassette and solute carrier superfamily of transporter genes in mammary gland of riverine buffalo (Bubalus bubalis).

    PubMed

    Sharma, Ankita; Aggarwal, Jigyasa; Sodhi, Monika; Kishore, Amit; Mishra, B P; Mohanty, A K; Kataria, R S; Kaushik, Jai K; Mukesh, Manishi

    2014-01-01

    In the present study, expression level of various ATP-binding cassette (ABC) viz., ABCA1, ABCA7, ABCG1, ABCG2, and ABCG5; associated transcription factors viz., SREBF1, LXRα (NR1H3), PPARA, and Solute Carriers (SLC); or Glucose transporters (GLUT) viz., SLC2A1(GLUT1), SLC2A4 (GLUT4), SLC2A8 (GLUT8), and SLC2A12 (GLUT12) superfamily of transporters were compared across physiological stages of buffalo mammary gland. The relative expression of ABCA1, and ABCG1 was significantly (p < 0.05) higher in mammary gland of heifer followed by involution and lactation stages. Similarly, ABCA7 gene expression was highest in heifer mammary gland followed by lactation and involution stages. ABCG2 gene expression was significantly (p < 0.05) high in lactating mammary gland in comparison to involution and heifer stages. On the other hand, ABCG5 gene expression was highest in involuting mammary gland followed by lactation and involution stages. Additionally, the expression of LXRα SREBF1, and PPARA which are known to regulate some of the ABC tranporters were also analyzed. The expression of LXRα gene was high in involuting as compared to lactating mammary gland. In contrast, SREBF1 and PPARA expression was significantly (p < 0.05) high in lactating mammary gland. Among the several SLC transporters studied, SLC2A1, SLC2A4, and SLC2A8 showed significant (p < 0.05) higher expression during lactation stage, whereas SLC2A12 expression was greater during heifer stage suggesting SLC2A1, SLC2A4, and SLC2A8 to be the major transporters associated with glucose uptake in buffalo mammary gland. The expression profile of (lactoferrin) LTF, known to be expressed at high level in mammary gland during involution was also studied. As expected, its expression was significantly (p < 0.05) higher during involution in comparison to lactating mammary gland.in buffaloes as well. The inclusion of LTF as a control gene further provided the confidence in the buffalo mammary gland expression data generated

  5. Evaluation of α-linolenic acid for freezability and in vivo fertility of Nili Ravi (Bubalus bubalis) buffalo semen.

    PubMed

    Ejaz, R; Ansari, M S; Rakha, B A; Qadeer, S; Husna, A U; Akhter, S

    2017-07-27

    Alpha linolenic acid (ALA) is integral component of cell membrane that protects the cell in stressful events and involves in many metabolic pathways. It was hypothesized that ALA have the ability to protect the structural and functional integrity of buffalo spermatozoa during freeze-thawing. Therefore, study was designed to evaluate ALA supplementation (0, 5, 10 and 20 ng/mL) in extender on freezability and in vivo fertility of buffalo bull spermatozoa. Semen from three adult Nili-Ravi buffalo bulls of similar age was collected with artificial vagina (42 °C) for five weeks (replicates; N = 30). Qualified semen ejaculates (>1 mL volume, >60% motility; >0.5 billion/mL concentration) were diluted with tris-citric acid extender containing 0.0 (control), 5.0, 10.0 and 20.0 ng/mL ALA at 37 °C and cryopreserved following established protocol. Sperm motility and plasma membrane integrity were recorded higher (P < 0.05) in extender containing 5.0 ng/mL of ALA compared to control. Nevertheless, sperm viability, live dead ratio and chromatin integrity were observed higher (P < 0.05) in all experimental extenders with ALA compared to control. The number of abnormal sperm reduced significantly in all experimental extenders having ALA. A total of 539 artificial inseminations were performed with the best evolved extender having ALA (5.0 ng/mL; 272 inseminations) and control (267 inseminations). In vivo fertility rates of buffalo semen were recorded higher (P < 0.05) with extender containing ALA (5.0 ng/mL) (58%) compared to control (46%). In conclusion, supplementing 5.0 ng/mL ALA in extender improved the post-thaw quality and in vivo fertility of cryopreserved Nili-Ravi buffalo bull semen. Copyright © 2017. Published by Elsevier Inc.

  6. Effect of sexual excitation on testosterone and nitric oxide levels of water buffalo bulls (Bubalus bubalis) with different categories of sexual behavior and their correlation with each other.

    PubMed

    Swelum, Ayman Abdel-Aziz; Saadeldin, Islam M; Zaher, Hany A; Alsharifi, Sawsan A M; Alowaimer, Abdullah N

    2017-06-01

    We studied the effect of sexual excitation on serum testosterone and nitric oxide (NO) levels in water buffalo bulls with different categories of sexual behavior and their correlation with each other. Buffalo bulls were classified according to their sexual behavior (including reaction time, sexual aggressiveness and mating ability): acceptable (good to excellent) (n=5), fair (n=5), and unacceptable (poor) (n=5) sexual behavior. Blood samples were collected from all animals immediately before and after sexual teasing and/or mounting to estimate the testosterone and NO levels using a commercial radioimmunoassay kit and Griess reaction test, respectively. Comparisons among groups were evaluated using a mixed-design analysis of variance. Pearson's correlation coefficients were calculated to determine the relationship between testosterone and NO levels before and after sexual excitation besides sexual behavior. The level of testosterone before sexual excitation was higher (p≤0.05) in bulls with acceptable and fair sexual behavior than in bulls with unacceptable sexual behavior (0.86±0.01, 0.69±0.02, and 0.29±0.02ng/mL, respectively). The level of NO was higher (p≤0.05) in bulls with acceptable and fair sexual behavior than in bulls with unacceptable sexual behavior (8.00±0.03, 7.66±0.19, and 6.29±0.33μM, respectively). Sexual excitation significantly (p<0.05) increase testosterone and NO levels in bulls with acceptable (1.45±0.01ng/mL and 19.04±0.32μM, respectively) or fair (0.92±0.02ng/mL and 14.95±0.34μM, respectively) sexual behavior, but not in bulls with unacceptable sexual behavior. The unacceptable sexual behavior bulls had significantly lower testosterone and NO levels than the other bulls. There was a strong correlation and association between serum testosterone and NO levels besides sexual behavior of buffalo bulls. In conclusion, the alteration in the testosterone and NO levels after sexual excitation depends on the sexual behavior category

  7. Basic fibroblast growth factor is critical to reprogramming buffalo (Bubalus bubalis) primordial germ cells into embryonic germ stem cell-like cells.

    PubMed

    Wang, Caizhu; Deng, Yanfei; Chen, Feng; Zhu, Peng; Wei, Jingwei; Luo, Chan; Lu, Fenghua; Yang, Sufang; Shi, Deshun

    2017-03-15

    Primordial germ cells (PGCs) are destined to form gametes in vivo, and they can be reprogrammed into pluripotent embryonic germ (EG) cells in vitro. Buffalo PGC have been reported to be reprogrammed into EG-like cells, but the identities of the major signaling pathways and culture media involved in this derivation remain unclear. Here, the effects of basic fibroblast growth factor (bFGF) and downstream signaling pathways on the reprogramming of buffalo PGCs into EG-like cells were investigated. Results showed bFGF to be critical to buffalo PGCs to dedifferentiate into EG-like cells (20 ng/mL is optimal) with many characteristics of pluripotent stem cells, including alkaline phosphatase (AP) activity, expression of pluripotency marker genes such as OCT4, NANOG, SOX2, SSEA-1, CDH1, and TRA-1-81, and the capacity to differentiate into all three embryonic germ layers. After chemically inhibiting pathways or components downstream of bFGF, data showed that inhibition of the PI3K/AKT pathway led to significantly lower EG cell derivation, while inhibition of P53 activity resulted in an efficiency of EG cell derivation comparable to that in the presence of bFGF. These results suggest that the role of bFGF in PGC-derived EG-like cell generation is mainly due to the activation of the PI3K/AKT/P53 pathway, in particular, the inhibition of P53 function. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Exploring genetic polymorphism in innate immune genes in Indian cattle (Bos indicus) and buffalo (Bubalus bubalis) using next generation sequencing technology.

    PubMed

    Patel, Shreya M; Koringa, Prakash G; Nathani, Neelam M; Patel, Namrata V; Shah, Tejash M; Joshi, Chaitanya G

    2015-02-01

    Activation of innate immunity initiates various cascades of reactions that largely contribute to defense against physical, microbial or chemical damage, prompt for damage repair and removal of causative organisms as well as restoration of tissue homeostasis. Genetic polymorphism in innate immune genes plays prominent role in disease resistance capabilities in various breeds of cattle and buffalo. Here we studied single nucleotide variations (SNP/SNV) and haplotype structure in innate immune genes viz CHGA, CHGB, CHGC, NRAMP1, NRAMP2, DEFB1, BNBD4, BNBD5, TAP and LAP in Gir cattle and Murrah buffalo. Targeted sequencing of exonic regions of these genes was performed by Ion Torrent PGM sequencing platform. The sequence reads obtained corresponding to coding regions of these genes were mapped to reference genome of cattle BosTau7 by BWA program using genome analysis tool kit (GATK). Further variant analysis by Unified Genotyper revealed 54 and 224 SNPs in Gir and Murrah respectively and also 32 SNVs was identified. Among these SNPs 43, 36, 11,32,81,21 and 22 variations were in CHGA, CHGB, CHGC, NRAMP1, NRAMP2, DEFB1 and TAP genes respectively. Among these identified 278 SNPs, 24 were found to be reported in the dbSNP database. Variant analysis was followed by structure formation of haplotypes based on multiple SNPs using SAS software revealed a large number of haplotypes. The SNP discovery in innate immune genes in cattle and buffalo breeds of India would advance our understanding of role of these genes in determining the disease resistance/susceptibility in Indian breeds. The identified SNPs and haplotype data would also provide a wealth of sequence information for conservation studies, selective breeding and designing future strategies for identifying disease associations involving samples from distinct populations.

  9. Effect of boron as an antidote on dry matter intake, nutrient utilization and fluorine balance in buffalo (Bubalus bubalis) exposed to high fluoride ration.

    PubMed

    Bharti, Vijay K; Gupta, Meenakshi; Lall, D

    2008-12-01

    It is well known that excessive accumulation of fluorides can exert toxic effects on various tissues and organs so as to severely damage the health and production of animals. The aim of this study was to determine beneficial effect of boron on nutrient utilization in buffalo calves exposed to high fluoride (F) ration. For this purpose, we used three groups of four male Murrah buffalo calves (body weight 98-100 kg, aged 6-8 month) each. Control animal was given only basal diet and concentrate mixture. However, treatment I animals were fed basal diet, concentrate mixture, and F [as NaF, 60 ppm of dry matter (DM)]. The treatment II animals were fed basal diet, concentrate mixture, F (as NaF, 60 ppm of DM), and B (as sodium tetraborate, 140 ppm of DM). After 90 days of experimental feeding, a metabolism trial of 7 days duration was conducted to study the treatment effect on nutrient utilization of proximate nutrients, absorption, excretion, and retention of N, Ca, P, Fe, Zn, Cu, and F. Dietary F significantly (p < 0.05) depressed the dry matter intake and increased the apparent digestibility, absorption, and retention of F. However, boron supplementation significantly (p < 0.05) decreased the apparent digestibility, absorption, and retention of F and improved the dry matter intake, fecal excretion, and percent of absorbed F excreted via urine. Apparent digestibility of proximate nutrients (viz. DM, crude protein, crude fiber, ether extract, and nitrogen free extract) was unaffected on either F or F+B treatment. However, absorption and excretion of N, Ca, P, Fe, Zn, and Cu were affected significantly (p < 0.05) on F or F+B treatment. These findings suggest that fluoride-containing diet for short duration has effect on nutrient utilization, and boron at 140-ppm dose level, in general, antagonized the absorption and retention of F and also improved the feed intake in buffalo calves.

  10. Seasonal and Ageing-Depending Changes of Aquaporins 1 and 9 Expression in the Genital Tract of Buffalo Bulls (Bubalus bubalis).

    PubMed

    Arrighi, S; Bosi, G; Accogli, G; Desantis, S

    2016-08-01

    The presence of Aquaporins 1 (AQP1) and 9 (AQP9), integral membrane water channels that facilitate rapid passive movement of water and solutes, was immunohistochemically detected in the excurrent ducts collected from sexually mature buffalo bulls of proven fertility during the mating (late autumn-winter) and non-mating (late spring to the beginning of autumn) seasons. Furthermore, the research was performed also on the epididymal cauda of a senile buffalo bull with inactive testis. Aquaporins 1 and 9 were immunolocalized at distinct levels. In the efferent ducts, AQP1 immunoreactivity was strongly evidenced at the apical surface of the non-ciliated cells and weakly along the basal membrane of the epithelial cells. The latter reactivity disappeared during the non-mating season. No AQP1 immunoreactivity was detected in the epithelium of epididymis and vas deferens, whereas AQP1 was expressed in the smooth muscle layer of the vas deferens. Aquaporin 1 was present in the blood vessels and in small nerve bundles all along the genital tract. The supranuclear zone of the epididymal principal cells was AQP9 immunoreactive, limited to the corpus and cauda regions, and vas deferens. The samples collected in the two reproductive seasons showed a weaker AQP9 immunoreactivity during the non-mating season. A typical AQP9 immunoreactivity was noticed in the old buffalo examined. The tested AQP molecules showed a different expression pattern in comparison with laboratory mammals, primates, equine, dog and cat. In addition, seasonal differences were noticed which are possibly useful in regard to the comprehension of the morphophysiology of reproduction in the bubaline species, which are still a matter of debate.

  11. Effect of incorporation of additives in tris-based egg yolk extender on buffalo (Bubalus bubalis) sperm tyrosine phosphorylation during cryopreservation.

    PubMed

    Kumar, R; Atreja, S K

    2012-06-01

    Phosphorylation of tyrosine residues on sperm protein is a known indicator of capacitation and a major intracellular signalling event. There is evidence that sperm cryopreservation promotes tyrosine phosphorylation and is associated with reduced fertility of spermatozoa. Under this study, cryoprotective role of different additives namely taurine, trehalose, catalase and 4-bromophenacyl bromide on buffalo sperm quality was evaluated. Buffalo semen was cryopreserved in tris-based egg yolk extender supplemented with additives like taurine (50 mm) or trehalose (100 mm) or 4-bromophenacyl bromide (200 μm) or catalase (100 U/ml) and used for assessment of levels of tyrosine phosphorylation in frozen-thawed spermatozoa. The results obtained were compared with the level of protein tyrosine phosphorylation of semen cryopreserved in tris-based egg yolk extender without additives. Proteins were extracted from a total number of nine ejaculates from three individual buffalo bulls chosen at random and analysed for tyrosine phospho-proteins using SDS-PAGE followed by immunoblotting. Monoclonal anti-phosphotyrosine antibody (Clone pT-154) was used as primary antibody followed by treatment with HRP-conjugated secondary antibody. Signals were detected on X-ray film using chemiluminescence. Nine proteins (p20, p30, p32, p38, p49, p56, p59, p72 and p86) were found to be tyrosine phosphorylated in cryopreserved spermatozoa. Supplementation of additives significantly (p<0.05) reduced the level of protein tyrosine phosphorylation in spermatozoa. Moreover, this study showed improved (p<0.05) post-thaw motility, viability and membrane integrity of spermatozoa on addition of these additives. The results obtained clearly indicate reduced level of capacitation like changes on supplementation of additives in terms of protein tyrosine phosphorylation.

  12. Ovum pick-up interval in buffalo (Bubalus bubalis) managed under wetland conditions in Argentina: Effect on follicular population, oocyte recovery, and in vitro embryo development.

    PubMed

    Konrad, J; Clérico, G; Garrido, M J; Taminelli, G; Yuponi, M; Yuponi, R; Crudeli, G; Sansinena, M

    2017-08-01

    The excellent adaptation of water buffalo (Bubalis bubalis) to swampy environments means that animals are frequently managed in areas with restricted access for reproductive procedures. The objective of this study was to evaluate the effect of the ovum pick-up (OPU) interval on follicular population, oocyte recovery, oocyte quality and in vitro embryo production. Twelve Murrah buffaloes were subjected to two consecutive dominant follicle reductions, and randomly assigned to either 7-day (n=6) or 14-day (n=6) OPU interval groups. Although there was no significant difference in the average number of small (<3mm) and large (>8mm) diameter follicles available per OPU, a higher proportion of medium-sized follicles (3-8mm) were observed in the 14-day interval group (5.129 vs 3.267; p<0.05). The number of recovered oocytes per donor was also significantly higher (4.51 vs. 2.8; p<0.05) in the 14-day interval group, although this was attributed to an increase in the proportion of lower quality oocytes (grades III and IV). After in vitro fertilization, embryo developmental competence from grade I and II oocytes was superior to that from grade III and IV oocytes, irrespective of OPU interval group. There was no significant difference in the proportion of grade I and II oocytes cleaved after sperm co-incubation; however, there was a higher proportion of blastocysts produced in 14-day interval group (28 vs. 6%, p<0.05). No blastocysts were produced from grade III and IV oocytes. This study indicates it is possible to use a 14-day interval for oocyte collection in water buffalo; this approach could be considered as an alternative when access to animals is restricted. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Characterization of β-lactoglobulin from buffalo (Bubalus bubalis) colostrum and its possible interaction with erythrocyte lipocalin-interacting membrane receptor.

    PubMed

    Chougule, Rohit A; Aparna, Huligerepura S

    2011-09-01

    Lipocalins form a widespread class of proteins involved in the transport of weakly soluble vitamins, hormones or hydrophobic molecules. β-lactoglobulin (BLG-col), a major lipocalin present in whey was purified and characterized from buffalo colostrum. The molecular weight of BLG-col as determined by Liquid chromatography -electrospray ionization mass spectrometry (LC-ESI-MS) was 18.257 kDa and the peptide mass fingerprint of the purified protein revealed 67% sequence homology to buffalo milk β-lg. The N-terminal-IIVTQ and LC-ESI-collision-induced dissociation-Electron transfer dissociation mass spectrometry/mass spectrometry analyses of doubly (m/z 1156(+2)) and triply (m/z 546(+3)) charged ion pairs corresponding to VYVEELKPTPEGDLEILLQK (41-60) and TPEVDDEALEKFDK (125-138) sequences confirmed the identity of BLG-col. Using these peptide sequences, the location of a gene encoding for BLG-col was identified on chromosome 11 at 11q28 loci of bovine genome. The unique property of the BLG-col isolated from buffalo colostrum was its strong and specific haemagglutinating activity with 'O' blood of human erythrocytes with 10,309 HAU/mg protein. The cell surface localization of BLG-col on human erythrocytes was confirmed by immunocytochemistry and the specificity of interaction was established by immunoblot analysis of human erythrocyte membrane proteins. Based on these observations, we suggest the presence of lipocalin receptor (70 kDa) on human erythrocyte membrane and the multiple sequence alignment supported structural diversity among lipocalin receptors.

  14. Comparative quality assessment of buffalo (Bubalus bubalis) semen chilled (5°C) in egg yolk- and soya milk-based extenders.

    PubMed

    Singh, A K; Singh, V K; Narwade, B M; Mohanty, T K; Atreja, S K

    2012-08-01

    Egg yolk-Tris is most commonly used semen extender; however, its use involves hygienic risk, interference with fertility and poor microscopic examination. Therefore, replacement of egg yolk with a plant-based component with protective effects on spermatozoa would be advantageous. In present study, we observed effect of soya milk-based extenders on dilution and liquid preservation of Murrah buffalo bull semen at 5°C up to 72 h in comparison with conventional egg yolk-Tris extender (Ext.1). In experiment one, a total of 32 buffalo semen ejaculates from four animals were extended and preserved at 5°C for 72 h in soya milk-based extender (Ext.2) with different percentages (10%, 15%, 20%, 25% and 30%) of soya milk for optimization of soya milk concentration. Semen quality was assessed for individual motility, viability, membrane integrity and acrosome integrity at 0, 24, 48 and 72 h of liquid preservation. The results of experiment one indicated that 25% soya milk is an optimum concentration for buffalo bull semen extender preparation. A modified method was used to prepare another soya milk-based extender (Ext.3). In the second experiment, two soya extenders (Ext.2 and 3) with optimized concentration (25%) of soya milk were comparatively assessed with egg yolk-Tris extender (Ext.1) for semen quality parameters at 0, 24, 48 and 72 h of liquid preservation. The individual sperm motility at 0 and 24 h following dilution were found non-significant among extenders. However, after 48 h of dilution, individual motility in Ext.3 was observed significantly (p < 0.05) higher than Ext.1. After 24, 48 and 72 h of dilution sperm membrane integrity in Ext.3 was found significantly (p < 0.05) higher than Ext.1. Overall, comparative evaluation of sperm parameters obtained revealed that Ext.3 containing 25% soya milk can be used as a substitute of egg yolk-based extender for buffalo semen liquid preservation. © 2011 Blackwell Verlag GmbH.

  15. Analysis of genetic variations across regulatory and coding regions of kappa-casein gene of Indian native cattle (Bos indicus) and buffalo (Bubalus bubalis)

    PubMed Central

    Kishore, Amit; Mukesh, M.; Sobti, R.C.; Kataria, R.S.; Mishra, B.P.; Sodhi, Monika

    2014-01-01

    The promoter region of kappa-casein (κ-CN) gene in Indian native cattle and buffalo breeds was sequenced and analyzed for nucleotide variations. Sequence comparison across breeds of Indian cattle revealed a total of 7 variations in the promoter region, of which − 515 G/T, − 427 C/T, − 385 C/T, − 283 A/G and − 251 C/T were located within consensus binding sites for octamer-binding protein (OCT1)/pregnancy specific mammary nuclear factor (PMF), activator protein-2 (AP2), hepatocyte nuclear factor (HNF-1) and GAL4 transcription factors (TFs), respectively. These variations might be involved in gain or loss of potential transcription factor binding sites (TFBSs). Unlike the other 4 variants, the − 283 (A/G) variant located within HNF-1 TFBS was specific to Indian cattle as this change has not been observed in the Bos taurus sequence. Other TFBSs viz., MGF, TBP, NF-1, milk box and C/EBP were conserved across species. For the Indian native buffalo breeds, only 3 changes were identified in the promoter region; − 305 (A/C), − 160 (T/C) and − 141 (A/G) and most of the TFBSs were found to be conserved. However, deletion of two adjacent nucleotides located in and around binding site for C/EBP TF was identified in buffalo when compared with promoter sequence of bovine κ-CN. For κ-CN of Indian native cattle, a strong linkage disequilibrium (LD) was observed for variations 515 G/T, − 427 C/T and − 385 C/T in the promoter region; and for variations at codons 136 and 148 of exon-IV. Further, among intragenic haplotypes, variation − 427 C/T was found to be in LD with variations at codons 136 and 148. The information generated in the present work provides comprehensive characterization of κ-CN gene promoter and coding regions in Indian cattle and buffaloes and reported variations could become important candidates for carrying out further research in dairy traits. PMID:25606460

  16. Analysis of genetic variations across regulatory and coding regions of kappa-casein gene of Indian native cattle (Bos indicus) and buffalo (Bubalus bubalis).

    PubMed

    Kishore, Amit; Mukesh, M; Sobti, R C; Kataria, R S; Mishra, B P; Sodhi, Monika

    2014-12-01

    The promoter region of kappa-casein (κ-CN) gene in Indian native cattle and buffalo breeds was sequenced and analyzed for nucleotide variations. Sequence comparison across breeds of Indian cattle revealed a total of 7 variations in the promoter region, of which - 515 G/T, - 427 C/T, - 385 C/T, - 283 A/G and - 251 C/T were located within consensus binding sites for octamer-binding protein (OCT1)/pregnancy specific mammary nuclear factor (PMF), activator protein-2 (AP2), hepatocyte nuclear factor (HNF-1) and GAL4 transcription factors (TFs), respectively. These variations might be involved in gain or loss of potential transcription factor binding sites (TFBSs). Unlike the other 4 variants, the - 283 (A/G) variant located within HNF-1 TFBS was specific to Indian cattle as this change has not been observed in the Bos taurus sequence. Other TFBSs viz., MGF, TBP, NF-1, milk box and C/EBP were conserved across species. For the Indian native buffalo breeds, only 3 changes were identified in the promoter region; - 305 (A/C), - 160 (T/C) and - 141 (A/G) and most of the TFBSs were found to be conserved. However, deletion of two adjacent nucleotides located in and around binding site for C/EBP TF was identified in buffalo when compared with promoter sequence of bovine κ-CN. For κ-CN of Indian native cattle, a strong linkage disequilibrium (LD) was observed for variations 515 G/T, - 427 C/T and - 385 C/T in the promoter region; and for variations at codons 136 and 148 of exon-IV. Further, among intragenic haplotypes, variation - 427 C/T was found to be in LD with variations at codons 136 and 148. The information generated in the present work provides comprehensive characterization of κ-CN gene promoter and coding regions in Indian cattle and buffaloes and reported variations could become important candidates for carrying out further research in dairy traits.

  17. Fatal Theileria orientalis N2 genotype infection among Asian water buffaloes (Bubalus bubalis) in a commercial dairy farm in Kerala, India.

    PubMed

    Vinodkumar, Kulangara; Shyma, Varikkottil; Justin, Davis Kollannur; Ashok, Sivasailam; Anu, Joseph Parassery; Mini, Kattilveetil; Muhammedkutty, Varikkottil; Sasidharan, Suchithra; Chullipparambil, Sunanda

    2016-01-01

    Fifteen dairy buffaloes of a farm in the state of Kerala, India developed fatal oriental theileriosis within 2 months of their procurement. Typical piroplasms of Theileria orientalis were observed in the erythrocytes of all affected animals by Giemsa-Leishman staining of blood smears. Case fatality rate was 87·5% (seven out of eight) in the clinically progressed cases. Therapeutic management with anti-theilerial drugs buparvaquone and oxytetracycline led to recovery of seven other animals in less advanced stages of the disease. The aim of this study was to determine the reasons for increased virulence of this pathogen, hitherto considered to be benign. Acute haemolytic anaemia was the predominant haematological finding in the affected animals. Lymphocytic infiltration and degeneration of vital organs leading to functional derangement was the cause of the high mortality. Identification of T. orientalis was confirmed by polymerase chain reaction (PCR). DNA sequencing of the PCR products revealed close identity with already reported sequences of T. orientalis/buffeli N2 genotype. The sequences were deposited in GenBank with accession number KM609973 and KM043772. Rhipicephalus ticks, previously not reported as vectors for oriental theileriosis, were identified as the potential vectors. This is the first report of fatal oriental theileriosis in Asian water buffaloes.

  18. Estimation of endogenous levels of osteopontin, total antioxidant capacity and malondialdehyde in seminal plasma: Application for fertility assessment in buffalo (Bubalus bubalis) bulls.

    PubMed

    Kumar, P; Saini, M; Kumar, D; Bharadwaj, A; Yadav, P S

    2017-04-01

    This study was attempted to identify subfertile bulls by quantifying the endogenous levels of osteopontin (OPN), total antioxidant capacity (TAC) and malondialdehyde (MDA) in seminal plasma of buffalo bulls. On the basis of conception rate, buffalo bulls were classified into two groups: high-fertile (conception rate >50%) and subfertile bulls (conception rate <40%). A total of 100 ejaculates (10 ejaculates from each bull) were collected through artificial vagina method. The concentration of OPN, TAC and catalase (CAT) of high-fertile bulls was found to be higher (p < .05) than that of subfertile bulls. Further, MDA level in seminal plasma was found to be lower (p < .05) in high-fertile bulls compared with subfertile bulls. The fertility status had no effect on the superoxide dismutase (SOD) concentration in seminal plasma of both the groups. The levels of OPN (r = .678, p = 0.013) and TAC (r = .648, p = .042) were found to be positively correlated with bull fertility and the level of MDA (r = -.718, p = .019) was found to be negatively correlated with bull fertility. However, the fertility of bulls was not found to be significantly correlated with SOD, CAT and sperm motility. In conclusion, seminal OPN, TAC and MDA tended to be more realistic in identification of subfertile bulls from breeding herds. © 2016 Blackwell Verlag GmbH.

  19. Use of post-thaw semen quality parameters to predict fertility of water buffalo (Bubalus bubalis) bull during peak breeding season.

    PubMed

    Ahmed, H; Andrabi, S M H; Anwar, M; Jahan, S

    2017-05-01

    This study was designed to predict the fertility of water buffalo bull using post-thaw semen quality parameters during peak breeding season. Thirty ejaculates were collected from five bulls with artificial vagina and cryopreserved. At post-thaw, semen was analysed for motility parameters, velocity distribution, kinematics, DNA integrity/fragmentation, viability, mitochondrial transmembrane potential, morphology, plasma membrane and acrosome integrity. Data of 514 inseminations were collected for estimation of in vivo fertility. Pearson's correlation coefficients showed that progressive motility (PM), rapid velocity, average path velocity, straight line velocity, straightness, supravital plasma membrane integrity, viable spermatozoon with intact acrosome or with high mitochondrial activity were correlated with in vivo fertility (r = .81, p < .01; r = .85, p < .01; r = .64, p < .05; r = .73, p < .05; r = .57, p < .05; r = .88, p < .01; r = .84, p < .01 and r = .81, p < .01 respectively). Step forward multiple regression analysis showed that the best single predictor of fertility was PM. However, combinations of semen quality parameters to predict fertility were better as compared to single parameter. In conclusion, fertility of buffalo bull can be predicted through some of the post-thaw in vitro semen quality tests during peak breeding season. © 2016 Blackwell Verlag GmbH.

  20. Improvement of liquid and frozen-thawed semen quality of Nili-Ravi buffalo bulls (Bubalus bubalis) through supplementation of fat.

    PubMed

    Adeel, M; Ijaz, A; Aleem, M; Rehman, H; Yousaf, M S; Jabbar, M A

    2009-05-01

    The aim of the study was to investigate the effects of dietary fat on quality of liquid and frozen-thawed semen of Nili-Ravi buffalo bulls. Adult bulls (n=21) were fed a balanced ration (Con; n=7) or the same ration either containing sunflower oil (SF-O; n=7) or whole sunflower seeds (SF-S; n=7) for 63 days. Body weight and body condition score of each bull was recorded on days 0, 30 and 60 of the experiment. Semen was collected on days 39, 46, 53 and 60, frozen by a fast method and stored at -196 degrees C for 24h. Sperm motility was assessed using a bright field microscope. Plasma membrane integrity of fresh and frozen-thawed spermatozoa was assessed using a hypo-osmotic swelling (HOS) assay. The concentration of spermatozoa and volume of semen was not different among groups on various days of collection. Sunflower-enriched diets did not affect the motility and number of HOS-positive spermatozoa in the fresh semen. Motility and HOS of post-thawed spermatozoa were higher (p<0.05) in bulls fed the sunflower-enriched diets. Similarly, diets did not affect the body condition score and body weight of bulls. In conclusion, feeding of sunflower oil or sunflower seed as fat sources can improve the quality of buffalo bull spermatozoa.

  1. Use of real-time PCR technique in determination of major fibrolytic and non fibrolytic bacteria present in Indian Surti buffaloes (Bubalus bubalis).

    PubMed

    Singh, Krishna M; Tripathi, Ajai K; Pandya, Paresh R; Parnerkar, Subhash; Rank, Dharamshibhai N; Kothari, Ramesh K; Joshi, Chaitanya G

    2013-01-01

    In the milk industry in India, buffalo breeds are most commonly used for milk production. Efficiency of fiber digestion in ruminants is critical for animal productivity. Bacteria play an important role in fiber digestion and utilization. Absolute quantification real-time PCR was used to quantify ten bacterial species in rumen fluid of Surti buffalo fed green fodder, dry roughage and compound concentrate mixture. Abundance of each target taxon was calculated as a fraction of the total 16S rRNA gene copies in the samples, using taxon-specific primers. Bacterial populations showed a clear predominance of Ruminococcus albus, which comprised 5.66% of the bacterial rRNA gene copies in the samples. However, only 0.9% to 4.24% of the bacterial rRNA gene copies were represented by the ruminal Fibrobacter succinogenes, Ruminococcus flavefaciens and Prevotella species. The proportion of rRNA gene copies attributable to Selenomonas ruminantium, Streptococcus bovis, Ruminobacter amylophilus, Treponema bryantii and Anaerovibrio lipolytica was even less abundant, each comprising < 0.11% of the bacterial rRNA gene copies. The data suggest that the aggregate abundance of the most intensively studied ruminal bacterial species is relatively low and that a large fraction of the uncultured population represents a single bacterial genus.

  2. Expression pattern of pluripotent markers in different embryonic developmental stages of buffalo (Bubalus bubalis) embryos and putative embryonic stem cells generated by parthenogenetic activation.

    PubMed

    Singh, Karn P; Kaushik, Ramakant; Garg, Veena; Sharma, Ruchi; George, Aman; Singh, Manoj K; Manik, Radhey S; Palta, Prabhat; Singla, Suresh K; Chauhan, Manmohan S

    2012-12-01

    In this study, we describe the production of buffalo parthenogenetic blastocysts and subsequent isolation of parthenogenetic embryonic stem cell (PGESC)-like cells. PGESC colonies exhibited dome-shaped morphology and were clearly distinguishable from the feeder layer cells. Different stages of development of parthenogenetic embryos and derived embryonic stem cell (ESC)-like cells expressed key ESC-specific markers, including OCT-4, NANOG, SOX-2, FOXD3, REX-1, STAT-3, TELOMERASE, NUCLEOSTEMIN, and cMYC. Immunofluorescence-based studies revealed that the PGESCs were positive for surface-based pluripotent markers, viz., SSEA-3, SSEA-4, TRA 1-80, TRA 1-60, CD-9, and CD-90 and exhibited high alkaline phosphatase (ALP) activity. PGEC cell-like cells formed embryoid body (EB)-like structures in hanging drop cultures and when cultured for extended period of time spontaneously differentiated into derivatives of three embryonic germ layers as confirmed by RT-PCR for ectodermal (CYTOKERATIN8, NF-68), mesodermal (MSX1, BMP-4, ASA), and endodermal markers (AFP, HNF-4, GATA-4). Differentiation of PGESCs toward the neuronal lineage was successfully directed by supplementation of serum-containing media with retinoic acid. Our results indicate that the isolated ESC-like cells from parthenogenetic blastocyst hold properties of ESCs and express markers of pluripotency. The pluripotency markers were also expressed by early cleavage-stage of buffalo embryos.

  3. Molecular characterization and expression of buffalo (Bubalus bubalis) DEAD-box family VASA gene and mRNA transcript variants isolated from testis tissue.

    PubMed

    Kaushik, Ramakant; Singh, Karn Pratap; Bahuguna, Vivek; Rameshbabu, K; Singh, Manoj Kumar; Manik, Radhey Shyam; Palta, Prabhat; Singla, Suresh Kumar; Chauhan, Manmohan Singh

    2015-11-01

    VASA is a member of the DEAD-box protein family that plays an indispensable role in mammalian spermatogenesis, particularly during meiosis. In the present study, we isolated, sequenced, and characterized VASA gene in buffalo testis. Here, we demonstrated that VASA mRNA is expressed as multiple isoforms and uses four alternative transcriptional start sites (TSSs) and four different polyadenylation sites. The TSSs identified by 5'-RNA ligase-mediated rapid amplification of cDNA ends (RLM-5'-RACE) were positioned at 48, 53, 85, and 88 nucleotides upstream relative to the translation initiation codon. 3'-RACE experiment revealed the presence of tandem polyadenylation signals, which lead to the expression of at least four different 3'-untranslated regions (209, 233, 239 and 605 nucleotides). The full-length coding region of VASA was 2190 bp, which encodes a 729 amino acid (aa) protein containing nine consensus regions of the DEAD box protein family. VASA variants are highly expressed in testis of adult buffalo. We found five variants, one full length VASA (729 aa) and four splice variants VASA 2, 4, 5, 6 (683, 685, 679, 703 aa). The expression level of VASA 1 was significantly higher than rest of all (P < 0.05) except VASA 6. The relative ratio for VASA 1:2:4:5:6 was 100:1.0:1.6:0.9:48.

  4. Chicken egg yolk plasma in tris-citric acid extender improves the quality and fertility of cryopreserved water buffalo (Bubalus bubalis) spermatozoa.

    PubMed

    Hussain Shah, S Aftab; Hassan Andrabi, S Murtaza; Ahmed, Hussain; Qureshi, Irfan Zia

    2017-02-01

    This study was primarily designed to evaluate the effect of different concentrations of ultraviolet (UV)-C-irradiated chicken egg yolk plasma (EYP; v:v; 10%, P1; 15%, P2; 20%, P3) or 20% (v:v) of whole chicken egg yolk (WCEY) in tris-citric acid (TCA) extender on water buffalo sperm quality during cryopreservation (postdilution, PD; postequilibration, PE; post-thawing, PT). Also the effect of best evolved concentration of UV-C-irradiated EYP in extender on in vivo fertility of buffalo spermatozoa was evaluated. At PE and PT, computer-assisted sperm analysis progressive motility (PM, %) was significantly higher in P3 compared with P1 and WCEY. Rapid velocity (RV, %) was higher (P < 0.05) in P3 compared with P1 and WCEY during cryopreservation (PD, PE, and PT). Average path velocity (μm/s) and straight line velocity (μm/s) were higher (P < 0.05) in P2 and P3 than WCEY at PE and PT. The decline percentage (%, longevity) in PM and RV was lower (P < 0.05) in P3 compared with WCEY during 2 hours incubation under in vitro condition at PT. Supravital plasma membrane integrity (%) was higher (P < 0.05) in P2 and P3 compared with control at different stages (PE and PT). Mitochondrial transmembrane potential (%) was higher (P < 0.05) in P2 and P3 compared with P1 and WCEY at different stages (PD and PT). Percentage of viable sperm with intact acrosome, and sperm DNA integrity (%) were higher (P < 0.05) in P2 and P3 compared with WCEY at PT. The in vivo fertility rate (%) was significantly higher with P3 compared with WCEY (76.61 vs. 64.49). In conclusion, WCEY (20%) can be replaced with UV-C-irradiated chicken EYP (20%) in TCA extender for cryopreservation of water buffalo spermatozoa. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Caspase-9 inhibitor Z-LEHD-FMK enhances the yield of in vitro produced buffalo (Bubalus bubalis) pre-implantation embryos and alters cellular stress response.

    PubMed

    Mullani, N; Singh, M K; Sharma, A; Rameshbabu, K; Manik, R S; Palta, P; Singla, S K; Chauhan, M S

    2016-02-01

    The present investigation was done to study the effect of caspase-9 inhibitor Z-LEHD-FMK, on in vitro produced buffalo embryos. Z-LEHD-FMK is a cell-permeable, competitive and irreversible inhibitor of enzyme caspase-9, which helps in cell survival. Buffalo ovaries were collected from slaughterhouse and the oocytes were subjected to in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC). The culture medium was supplemented with Z-LEHD-FMK at different concentrations i.e. 0 μM (control), 10 μM, 20 μM, 30 μM and 50 μM during IVM and IVC respectively. After day-2 post-insemination, the cleavage rate was significantly higher (74.20 ± 5.87% at P<0.05) in the group treated with 20 μM of Z-LEHD-FMK than at any other concentration. Same trend was observed in the blastocyst production rate which was higher at 20 μM (27.42 ± 2.94% at P<0.05). The blastocysts obtained at day-8 of the culture at different concentrations were subjected to TUNEL assay, to determine the level of apoptosis during the culture medium supplied with 20 μM Z-LEHD-FMK which showed apoptotic index significantly lower (1.88 ± 0.87 at P<0.05). There was a non-significant increase in total cell number in all Z-LEHD-FMK treated blastocysts. The quantitative gene expression of CHOP and HSP10 genes showed significant increase (P<0.05) in the group treated with 50 μM Z-LEHD-FMK, while, HSP40 showed significant increase (P<0.05) at 30 μM and 50 μM Z-LEHD-FMK concentrations. From the afore mentioned results we conclude that, Z-LEHD-FMK at 20 μM increased the cleavage and blastocyst rate of buffalo pre-implantation embryos also affecting the rate of apoptosis and cellular stress at various concentrations. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Effects of recipient oocyte age and interval from fusion to activation on development of buffalo (Bubalus bubalis) nuclear transfer embryos derived from fetal fibroblasts.

    PubMed

    Lu, F; Jiang, J; Li, N; Zhang, S; Sun, H; Luo, C; Wei, Y; Shi, D

    2011-09-15

    The objective was to investigate the effect of recipient oocyte age and the interval from activation to fusion on developmental competence of buffalo nuclear transfer (NT) embryos. Buffalo oocytes matured in vitro for 22 h were enucleated by micromanipulation under the spindle view system, and a fetal fibroblast (pretreated with 0.1 μg/mL aphidicolin for 24 h, followed by culture for 48 h in 0.5% fetal bovine serum) was introduced into the enucleated oocyte, followed by electrofusion. Both oocytes and NT embryos were activated by exposure to 5 μM ionomycin for 5 min, followed by culture in 2 mM 6-dimethyl-aminopurine for 3 h. When oocytes matured in vitro for 28, 29, 30, 31, or 32 h were activated, more oocytes matured in vitro for 30 h developed into blastocysts in comparison with oocytes matured in vitro for 32 h (31.3 vs 19.9%, P < 0.05). When electrofusion was induced 27 h after the onset of oocyte maturation, the cleavage rate (78.0%) was higher than that of electrofusion induced at 28 h (67.2%, P < 0.05), and the blastocyst yield (18.1%) was higher (P < 0.05) than that of electrofusion induced at 25 or 26 h (7.4 and 8.5%, respectively). A higher proportion of NT embryos activated at 3 h after electrofusion developed to the blastocyst stage (18.6%) in comparison with NT embryos activated at 1 h (6.0%), 2 h (8.3%), or 4 h (10.6%) after fusion (P < 0.05). No recipient was pregnant 60 d after transfer of blastocysts developed from NT embryos activated at 1 h (0/8), 2 h (0/10), or 4 h (0/9) after fusion. However, 3 of 16 recipients were pregnant following transfer of blastocysts developed from the NT embryos activated at 3 h after fusion, and two of these recipients maintained pregnancy to term. We concluded that the developmental potential of buffalo NT embryos was related to recipient oocyte age and the interval from fusion to activation. Copyright © 2011 Elsevier Inc. All rights reserved.

  7. Molecular differentiation of Sarcocystis buffalonis and Sarcocystis levinei in water buffaloes (Bubalus bubalis) from Sarcocystis hirsuta and Sarcocystis cruzi in cattle (Bos taurus).

    PubMed

    Gjerde, Bjørn; Hilali, Mosaad; Abbas, Ibrahim E

    2016-06-01

    The purpose of the present study was to obtain sarcocysts of Sarcocystis buffalonis and Sarcocystis levinei from water buffaloes and characterize the isolates by molecular methods in order to determine whether the two species were genetically different from Sarcocystis hirsuta and Sarcocystis cruzi, respectively, from cattle, which had been characterized before. About 35 macroscopically visible (3-4 × 1-2 mm) and 20 barely visible (1-3 × 0.2 mm) sarcocysts were excised from the esophagus of 18 naturally infected and freshly slaughtered adult water buffaloes at three slaughterhouses in Egypt. Genomic DNA was extracted from the sarcocysts, and all isolates were first characterized at the mitochondrial cytochrome c oxidase subunit I gene (cox1) gene through PCR amplification and direct sequencing. Selected isolates were subsequently further characterized at the 18S and 28S ribosomal (r) RNA genes and the internal transcribed spacer 1 (ITS1) region of the nuclear rDNA unit by direct sequencing or cloning. Only six of the isolated macroscopic sarcocysts belonged to S. buffalonis, whereas the others belonged to Sarcocystis fusiformis. Twelve of the smaller cysts belonged to S. levinei and seven to Sarcocystis sinensis. The characterization of the sarcocysts of S. sinensis and some of the sarcocysts of S. fusiformis have been reported before. Fifteen additional sarcocyst isolates of S. fusiformis were characterized at cox1 in the present study and found to be identical or closely similar to previous isolates. At cox1, the sequence identity between the six isolates of S. buffalonis was 99.8-100 % (two haplotypes), whereas the identity between the 12 isolates of S. levinei was 99.0-100 % (10 haplotypes). The identity between cox1 sequences of S. buffalonis and S. hirsuta (n = 56) was 92.9-93.6 % (on average 93.4 %), and the identity between cox1 sequences of S. levinei and S. cruzi (n = 22) was 92.9-94.0 % (on average 93.5 %). The phylogenetic

  8. Effect of different seasons on concentration of plasma luteinizing hormone and seminal quality vis-à-vis freezability of buffalo bulls ( Bubalus bubalis)

    NASA Astrophysics Data System (ADS)

    Bahga, C. S.; Khokar, B. S.

    1991-12-01

    Seasonal variations in semen quality, freezability and plasma luteinizing hormone (LH) levels were studied between summer and spring. Semen volume, density and initial sperm motility did not differ significantly between different seasons. Plasma LH decreased between summer and spring but the differences were, however, not significant. Pre-freezing motility did not differ significantly but post-freezing motility varied significantly ( P<0.01) between seasons. Post-freezing motility was lowest during summer and highest during winter. It can be concluded that summer spermatozoa may be fragile and cannot withstand freezing stress. To increase reproductive efficiency in buffalo during summer, semen should be frozen during winter and spring and used during hot weather conditions. Seasonal variations in plasma LH levels were insignificant.

  9. Computational studies on receptor-ligand interactions between novel buffalo (Bubalus bubalis) nucleotide-binding oligomerization domain-containing protein 2 (NOD2) variants and muramyl dipeptide (MDP).

    PubMed

    Brahma, Biswajit; Patra, Mahesh Chandra; Mishra, Purusottam; De, Bidhan Chandra; Kumar, Sushil; Maharana, Jitendra; Vats, Ashutosh; Ahlawat, Sonika; Datta, Tirtha Kumar; De, Sachinandan

    2016-04-01

    Nucleotide binding and oligomerization domain 2 (NOD2), a member of intracellular NOD-like receptors (NLRs) family, recognizes the bacterial peptidoglycan, muramyl dipeptide (MDP) and initiates host immune response. The precise ligand recognition mechanism of NOD2 has remained elusive, although studies have suggested leucine rich repeat (LRR) region of NOD2 as the possible binding site of MDP. In this study, we identified multiple transcripts of NOD2 gene in buffalo (buNOD2) and at least five LRR variants (buNOD2-LRRW (wild type), buNOD2-LRRV1-V4) were found to be expressed in buffalo peripheral blood mononuclear cells. The newly identified buNOD2 transcripts were shorter in lengths as a result of exon-skipping and frame-shift mutations. Among the variants, buNOD2-LRRW, V1, and V3 were expressed more frequently in the animals studied. A comparative receptor-ligand interaction study through modeling of variants, docking, and molecular dynamics simulation revealed that the binding affinity of buNOD2-LRRW towards MDP was greater than that of the shorter variants. The absence of a LRR segment in the buNOD2 variants had probably affected their affinity toward MDP. Notwithstanding a high homology among the variants, the amino acid residues that interact with MDP were located on different LRR motifs. The binding free energy calculation revealed that the amino acids Arg850(LRR4) and Glu932(LRR7) of buNOD2-LRRW, Lys810(LRR3) of buNOD2-LRRV1, and Lys830(LRR3) of buNOD2-LRRV3 largely contributed towards MDP recognition. The knowledge of MDP recognition and binding modes on buNOD2 variants could be useful to understand the regulation of NOD-mediated immune response as well as to develop next generation anti-inflammatory compounds.

  10. Birth of cloned calves from vitrified-warmed zona-free buffalo (Bubalus bubalis) embryos produced by hand-made cloning.

    PubMed

    Saha, Ambikaprasanna; Panda, Sudeepta K; Chauhan, Manmohan S; Manik, Radhey S; Palta, Prabhat; Singla, Suresh K

    2013-01-01

    The availability of techniques for the vitrification of cloned blastocysts can improve their effective use. The present study compared the developmental competence of buffalo cloned embryos derived from adult (BAF), newborn (BNF) and fetal fibroblast (BFF) before and after vitrification. Despite similar cleavage rates among the three groups, the blastocyst rate was lower for BAF- than BNF- and BFF-derived embryos (30.2±2.2% vs 41.7±1.7% and 39.1±2.1%, respectively; P<0.01). The total cell number of BNF-derived blastocysts was significantly higher (P<0.01) than that of BFF-derived blastocysts, which, in turn, was higher (P<0.01) than that of BAF-derived blastocysts. Following transfer of vitrified-warmed blastocysts to recipients, no pregnancy was obtained with fresh (n=8) or vitrified-warmed (n=18) BAF-derived blastocysts, whereas transfer of fresh BNF- (n=53) and BFF-derived (n=32) blastocysts resulted in four and three pregnancies, respectively, which aborted within 90 days of gestation. The transfer of vitrified-warmed BNF-derived blastocysts (n=39) resulted in the live birth of a calf weighing 41kg, which is now 23 months old and has no apparent abnormality, whereas the transfer of vitrified-warmed BFF-derived blastocysts (n=18) resulted in one live birth of a calf that died within 6h. These results demonstrate that cloned buffalo embryos cryopreserved by vitrification can be used to obtain live offspring.

  11. Using real-time PCR to identify pregnancy-associated glycoprotein 2 (PAG-2) in water buffalo (Bubalus bubalis) blood in early pregnancy.

    PubMed

    Barbato, O; Guelfi, G; Barile, V L; Menchetti, L; Tortiello, C; Canali, C; Brecchia, G; Traina, G; Beckers, J F; Melo de Sousa, N

    2017-02-01

    This study investigates for the first time mRNA pregnancy-associated glycoprotein 2 (PAG-2) expression in blood cells during early pregnancy in water buffalo. The PAGs constitute a large family of glycoproteins expressed in the outer epithelial layer of the placenta in eutherian species. All PAGs are not concomitantly expressed throughout pregnancy; some of them are expressed in the earlier phases, whereas others appear later and are expressed over a shorter period. Twenty-one lactating buffaloes were analyzed-17 females were synchronized with PRID and artificially inseminated (AI), whereas four females were synchronized but not inseminated (control group). Blood was collected at Days 0, 18, 28, 40, and 75 from AI (AI = Day 0). Expression of PAG-2 mRNA in blood samples was measured with real-time polymerase chain reaction. Pregnancy diagnosis was performed on Day 28 (D28) and Day 40 (D40) after AI by ultrasonography (US) and by PAG-1 RIA method. The females diagnosed pregnant at D28 and confirmed pregnant at D40 were defined as D28(+)D40(+) group; the females diagnosed pregnant at D28 but not confirmed pregnant at D40 were defined as D28(+)D40(-) group; and the females that were diagnosed as nonpregnant on either days were defined as D28(-)D40(-) group. PAG-2 mRNA at Day 0 was not observed in any groups. The D28(+)D40(+) group showed the highest expression, starting on Day 18 and increasing progressively up to Day 75. PAG-2 mRNA was also expressed on Day 18 in both D28(+)D40(-) and D28(-)D40(-) groups, but their levels were lower than those of D28(+)D40(+) group and almost constant over time. PAG-2 mRNA was never detected in the control group. The significant difference in the expression of PAG-2 mRNA between the D28(+)D40(+) group and the D28(-)D40(-) group, starting from Day 18, suggests that these animals might have conceived, but have experienced early embryonic loss; therefore, the PAG-2 mRNA was still present in blood circulation although at lower levels

  12. Roscovitine treatment improves synchronization of donor cell cycle in G0/G1 stage and in vitro development of handmade cloned buffalo (Bubalus bubalis) embryos.

    PubMed

    Selokar, Naresh L; Saini, Monika; Muzaffer, Mushariffa; Krishnakanth, G; Saha, Ambika P; Chauhan, Manmohan S; Manik, Radheysham; Palta, Prabhat; Madan, Pavneesh; Singla, Suresh K

    2012-04-01

    This study investigated the effects of serum-starvation, total confluence, and roscovitine treatment on cell-cycle synchronization of buffalo ear skin fibroblasts to the G0/G1 stage and on the developmental competence of cloned embryos. Serum starvation of total confluence cultures for 24 h had a higher (p<0.05) proportion of cells at G0/G1 stage (94.4%) compared with serum starved cyclic and nonstarved confluent cultures (76.8 and 86.0%, respectively), whereas differences between cyclic cells with or without serum starvation were not significant. The proportion of cells at G0/G1 was higher (p<0.05) with 20 and 30 μM roscovitine treatment than that with 10 μM (94.4, 96.4, and 86.6%, respectively), which was similar to that for total confluence (86.0%). MTT assay showed that cell viability decreased as dose of roscovitine increased. The blastocyst rate was significantly higher (p<0.05) when nuclear transfer embryos were reconstructed using donors cells from total confluence, confluence serum starved, and roscovitine-treated (20 and 30 μM) groups (48.8, 48.9, 57.9, and 62.9%, respectively) compared to nontreated cyclic cells (20.2%). However, the cleavage rate and total cell number of cloned embryos were similar for all the groups. The number of ICM cells was improved by 30 μM roscovitine treatment (45.25 ± 2.34). The cryosurvival rate of blastocysts derived from cells synchronized with 20 or 30 μM roscovitine was higher compared to that for total confluence group (33.6, 37.8 vs. 23.8%). In conclusion, treatment with 30 μM roscovitine is optimal for harvesting G0/G1 stage cells for producing high quality cloned buffalo embryos, and that it is better than serum-starvation or total confluence for cell synchronization.

  13. Molecular characterisation of three regions of the nuclear ribosomal DNA unit and the mitochondrial cox1 gene of Sarcocystis fusiformis from water buffaloes (Bubalus bubalis) in Egypt.

    PubMed

    Gjerde, Bjørn; Hilali, Mosaad; Mawgood, Sahar Abdel

    2015-09-01

    A total of 33 macroscopically visible (3-11 × 1-5 mm) sarcocysts of Sarcocystis fusiformis were excised from the oesophagus of 12 freshly slaughtered water buffalos in Giza, Egypt. Genomic DNA was extracted from the sarcocysts, and all isolates were characterised at the mitochondrial cytochrome c oxidase subunit I (cox1) gene through PCR amplification and direct sequencing, whereas a few selected isolates were characterised at the 18S and 28S ribosomal (r) RNA genes and the internal transcribed spacer 1 (ITS1) region of the nuclear rDNA unit following cloning. Among the 33 cox1 sequences (1,038-bp long), there was a total of 13 haplotypes, differing from each other by one to seven substitutions and sharing an identity of 99.3-99.9 %. In comparison, the sequence identity was 98.8-99.0 % among eight complete 18S rRNA gene sequences (1,873-1,879-bp long), 98.1-100 % among 28 complete ITS1 sequences (853-864-bp long) and 97.4-99.6 % among five partial 28S rRNA gene sequences (1,607-1,622 bp). At the three nuclear loci, the intraspecific (and intra-isolate) sequence variation was due to both substitutions and indels, which necessitated cloning of the PCR products before sequencing. Some additional clones of the 18S and 28S rRNA genes were highly divergent from the more typical clones, but the true nature of these aberrant clones could not be determined. Sequence comparisons and phylogenetic analyses based on either 18S rRNA gene or cox1 nucleotide sequences, placed S. fusiformis closest to Sarcocystis cafferi from the African buffalo, but only the analyses based on cox1 data separated the two taxa clearly from each other and showed that they were separate species (monophyletic clusters and 93 % sequence identity at cox1 versus interleaved sequences and 98.7-99.1 % sequence identity at the 18S rRNA gene). Two cats experimentally infected with sarcocysts of S. fusiformis started shedding small numbers of sporocysts 8-10 days post-infection (dpi) and were euthanized 15

  14. Effect of season, late embryonic mortality and progesterone production on pregnancy rates in pluriparous buffaloes (Bubalus bubalis) after artificial insemination with sexed semen.

    PubMed

    Campanile, Giuseppe; Vecchio, Domenico; Neglia, Gianluca; Bella, Antonino; Prandi, Alberto; Senatore, Elena M; Gasparrini, Bianca; Presicce, Giorgio A

    2013-03-01

    The use of sexed semen technology in buffaloes is nowadays becoming more and more accepted by farmers, to overcome the burden of unwanted male calves with related costs and to more efficiently improve production and genetic gain. The aim of this study was to verify the coupling of some variables on the efficiency of pregnancy outcome after deposition of sexed semen through AI. Pluriparous buffaloes from two different farms (N = 152) were screened, selected, and subjected to Ovsynch protocol for AI using nonsexed and sexed semen from four tested bulls. AI was performed in two distinct periods of the year: September to October and January to February. Neither farms nor bulls had a significant effect on pregnancy rates pooled from the two periods. The process for sexing sperm cells did not affect pregnancy rates at 28 days after AI, for nonsexed and sexed semen, respectively 44/73 (60.2%) and 50/79 (63.2%), P = 0.70, and at 45 days after AI, for nonsexed and sexed semen, respectively 33/73 (45.2%) and 33/79 (49.3%), P = 0.60. Pregnancy rate at 28 days after AI during the transitional period of January to February was higher when compared with September to October, respectively 47/67 (70.1%) versus 47/85 (55.2%), P = 0.06. When the same pregnant animals were checked at Day 45 after AI, the difference disappeared between the two periods, because of a higher embryonic mortality, respectively 32/67 (47.7%) versus 40/85 (47.0%), P = 0.93. Hematic progesterone concentration at Day 10 after AI did not distinguish animals pregnant at Day 28 that would or would not maintain pregnancy until Day 45 (P = 0.21). On the contrary, when blood samples were taken at Day 20 after AI, the difference in progesterone concentration between pregnant animals that would maintain their pregnancy until Day 45 was significant for both pooled (P = 0.00) and nonsexed (P = 0.00) and sexed semen (P = 0.09). A similar trend was reported when blood samples were taken at Day 25, being highly significant

  15. Extra and intracellular calcium signaling pathway(s) differentially regulate histamine-induced myometrial contractions during early and mid-pregnancy stages in buffaloes (Bubalus bubalis).

    PubMed

    Sharma, Abhishek; Nakade, Udayraj P; Choudhury, Soumen; Yadav, Rajkumar Singh; Garg, Satish Kumar

    2017-04-01

    This study examines the differential role of calcium signaling pathway(s) in histamine-induced uterotonic action during early and mid-pregnancy stages in buffaloes. Compared to mid pregnancy, tonic contraction, amplitude and mean-integral tension were significantly increased by histamine to produce myometrial contraction during early pregnancy with small effects on phasic contraction and frequency. Although uterotonic action of histamine during both stages of pregnancy is sensitive to nifedipine (a L-type Ca(2+) channels blocker) and NNC55-0396 (T-type Ca(2+) channels blocker), the role of extracellular calcium seems to be more significant during mid-pregnancy as in this stage histamine produced only 9.38±0.96% contraction in Ca(2+) free-RLS compared to 21.60±1.45% in uteri of early pregnancy stage. Intracellular calcium plays major role in histamine-induced myometrial contraction during early pregnancy as compared to mid pregnancy, as in the presence of cyclopiazonic acid (CPA) Ca(2+)-free RLS, histamine produced significantly higher contraction in myometrial strips of early-pregancy in comparison to mid-pregnancy (10.59±1.58% and 3.13±0.46%, respectively). In the presence of U-73122, the DRC of histamine was significantly shifted towards right with decrease in maximal effect (Emax) only in early pregnancy suggesting the predominant role of phospholipase-C (PL-C) in this stage of pregnancy. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Identification and evaluation of reference genes for accurate gene expression normalization of fresh and frozen-thawed spermatozoa of water buffalo (Bubalus bubalis).

    PubMed

    Ashish, Shende; Bhure, S K; Harikrishna, Pillai; Ramteke, S S; Muhammed Kutty, V H; Shruthi, N; Ravi Kumar, G V P P S; Manish, Mahawar; Ghosh, S K; Mihir, Sarkar

    2017-04-01

    The quantitative real time PCR (qRT-PCR) has become an important tool for gene-expression analysis for a selected number of genes in life science. Although large dynamic range, sensitivity and reproducibility of qRT-PCR is good, the reliability majorly depend on the selection of proper reference genes (RGs) employed for normalization. Although, RGs expression has been reported to vary considerably within same cell type with different experimental treatments. No systematic study has been conducted to identify and evaluate the appropriate RGs in spermatozoa of domestic animals. Therefore, this study was conducted to analyze suitable stable RGs in fresh and frozen-thawed spermatozoa. We have assessed 13 candidate RGs (BACT, RPS18s, RPS15A, ATP5F1, HMBS, ATP2B4, RPL13, EEF2, TBP, EIF2B2, MDH1, B2M and GLUT5) of different functions and pathways using five algorithms. Regardless of the approach, the ranking of the most and the least candidate RGs remained almost same. The comprehensive ranking by RefFinder showed GLUT5, ATP2B4 and B2M, MDH1 as the top two stable and least stable RGs, respectively. The expression levels of four heat shock proteins (HSP) were employed as a target gene to evaluate RGs efficiency for normalization. The results demonstrated an exponential difference in expression levels of the four HSP genes upon normalization of the data with the most stable and the least stable RGs. Our study, provides a convenient RGs for normalization of gene-expression of key metabolic pathways effected during freezing and thawing of spermatozoa of buffalo and other closely related bovines.

  17. Developmental Competence of Buffalo (Bubalus bubalis) Pluripotent Embryonic Stem Cells Over Different Homologous Feeder Layers and the Comparative Evaluation with Various Extracellular Matrices

    PubMed Central

    Sharma, Manjinder; Dubey, Pawan K.; Kumar, Rajesh; Nath, Amar; Kumar, G. Sai; Sharma, G. Taru

    2013-01-01

    Background and Objectives Use of somatic cells as a feeder layer to maintain the embryonic stem cells (ESCs) in undifferentiated state limits the stem cell research design, since experimental data may result from a combined ESCs and feeder cell response to various stimuli. Therefore, present study was designed to evaluate the developmental competence of the buffalo ESCs over different homogenous feeders and compare with various extracellular matrices using different concentrations of LIF. Methods and Results Inner cell masses (ICMs) of in vitro hatched blastocysts were cultured onto homologous feeders viz. fetal fibroblast, granulosa and oviductal cell feeder layers and synthetic matrices viz. fibronectin, collagen type I and matrigel in culture medium. Developmental efficiency was found higher for ESCs cultured on fetal fibroblast and granulosa layers (83.33%) followed by fibronectin (77.78%) at 30 ng LIF. Oviductal feeder was found to be the least efficient feeder showing only 11.11% undifferentiated primary ESC colonies at 30 ng LIF. However, neither feeder layer nor synthetic matrix could support the development of primary colonies at 10 ng LIF. Expression of SSEA- 4, TRA-1-60 and Oct-4 were found positive in ESC colonies from all the feeders and synthetic matrices with 20 ng and 30 ng LIF. Conclusions Fetal fibroblast and granulosa cell while, amongst synthetic matrices, fibronectin were found to be equally efficient to support the growth and maintenance of ESCs pluripotency with 30 ng LIF. This well-defined culture conditions may provide an animal model for culturing human embryonic stem cells in the xeno-free or feeder-free conditions for future clinical applications. PMID:24298371

  18. Molecular characterisation of Sarcocystis bovifelis, Sarcocystis bovini n. sp., Sarcocystis hirsuta and Sarcocystis cruzi from cattle (Bos taurus) and Sarcocystis sinensis from water buffaloes (Bubalus bubalis).

    PubMed

    Gjerde, Bjørn

    2016-04-01

    About 200 individual sarcocysts were excised from 12 samples of cattle beef from five countries (Argentina, Brazil, Germany, New Zealand, Uruguay) and tentatively identified to species or cyst type on the basis of their size and shape and cyst wall morphology. Genomic DNA was extracted from 147 of these sarcocysts and used initially for PCR amplification and sequencing of the partial mitochondrial cytochrome c oxidase subunit I gene (cox1) in order to identify the sarcocysts to species and/or sequence type. In addition, seven Sarcocystis sinensis-like sarcocysts collected from the oesophagus of water buffaloes in Egypt were examined at cox1 for comparative purposes. Based on the results from the cox1 marker, selected sarcocyst isolates from both hosts were further characterised at one to three regions of the nuclear ribosomal (r) DNA unit, i.e. the complete 18S rRNA gene, the complete internal transcribed spacer 1 (ITS1) region and the partial 28S rRNA gene. This was done in order to compare the results with previous molecular identifications based on 18S rRNA gene sequences and to evaluate the utility of these regions for species delimitations and phylogenetic inferences. On the basis of sarcocyst morphology and molecular data, primarily the cox1 sequences, four Sarcocystis spp. were identified in the samples of cattle beef. Twenty-two microscopic sarcocysts (1 × 0.1 mm) with hair-like protrusions were assigned to Sarcocystis cruzi, 56 macroscopic sarcocysts (3-8 × 0.5 mm) with finger-like protrusions were assigned to Sarcocystis hirsuta and 45 and 24 microscopic sarcocysts (1-3 × 0.1-0.2 mm) with finger-like protrusions were assigned to Sarcocystis bovifelis and Sarcocystis bovini n. sp., respectively. Sarcocysts of S. cruzi were identified in samples of beef from Argentina and Uruguay; sarcocysts of S. hirsuta in samples from Argentina, Brazil, Germany and New Zealand; sarcocysts of S. bovifelis in samples from Argentina and Germany; and

  19. Transmission of foot-and-mouth disease virus from experimentally infected Indian buffalo (Bubalus bubalis) to in-contact naïve and vaccinated Indian buffalo and cattle.

    PubMed

    Madhanmohan, M; Yuvaraj, S; Nagendrakumar, S B; Srinivasan, V A; Gubbins, Simon; Paton, David James; Parida, Satya

    2014-09-03

    This study investigated the transmission of foot-and-mouth disease virus (FMDV) from experimentally infected Indian buffalo to in-contact naïve and vaccinated cattle and buffalo. In each of six rooms, two donor buffalo that had been inoculated with FMDV were housed for five days with four recipient animals, comprising one vaccinated buffalo, one vaccinated calf, one unvaccinated buffalo and one unvaccinated calf. Vaccination was carried out with current Indian vaccine strain (O/IND/R2/75) and challenged on 28 days post-vaccination with an antigenically similar strain (O/HAS/34/05). All 12 donor buffalo and the six unvaccinated cattle and six unvaccinated calves developed clinical signs of foot-and-mouth disease (FMD). In contrast, all six vaccinated cattle (100%) and four out of six vaccinated buffalo (66.6%) were protected from disease but all became infected with FMDV. This confirms that buffalo have the potential to spread FMD by direct contact and that vaccination can block this spread. The numbers of animals in the study were too small to determine if the differences in clinical protection afforded by vaccination of cattle and buffalo are significant and warrant a different dose regime. Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

  20. Satellite-tagged transcribing sequences in Bubalus bubalis genome undergo programmed modulation in meiocytes: possible implications for transcriptional inactivation.

    PubMed

    Chattopadhyay, M; Gangadharan, S; Kapur, V; Azfer, M A; Prakash, B; Ali, S

    2001-09-01

    We cloned and sequenced a 1378 bp BamHI satellite DNA fraction from the water buffalo Bubalus bubalis and have studied its expression in different tissues. The GC-rich sequences of the resultant contig pDS5 crosshybridize only with bovid DNA and are not conserved evolutionarily. Typing of buffalo genomic DNA using pDS5 with several restriction enzymes revealed multilocus monomorphic bands. Similar typing of cattle, buffalo, goat, sheep, and gaur genomic DNA revealed variations in copy number and allele length giving rise to species-specific band patterns. Expression study of pDS5 in bubaline samples by RNA slot-blot, Northern blot, and RT-PCR showed various levels of signal in all the somatic tissues and germline cells except heart. A GenBank database search revealed homology of pDS5 sequences in the 5' region from nt 1-1261 with collagen gene. An AluI typing analysis of DNA from bubaline semen samples showed consistent loss of two bands. The presence of corresponding bands in somatic tissues suggests a sequence modulation within the pDS5 array in meiocytes during spermatogenesis, which is restored in the somatic cells after fertilization. Modulation of the satellite-tagged transcribing sequence in the meiocytes may be a mechanism of its inactivation.

  1. Carboxypeptidase-B from Bubalus bubalis pancreas: purification, properties and MALDI-TOF monitored activation of proinsulin.

    PubMed

    Nadeem, Muhammad Shahid; Murtaza, Bibi Nazia; Ahmad, Habib

    2013-09-01

    Carboxypeptidase-B (E.C 3.4.17.2) catalyzes the hydrolysis of peptides and esters at C-terminus of arginine and lysine residues. Our study describes the large scale purification, N-terminal sequence analysis and physiochemical properties of pancreatic enzyme from river buffalo (Bubalus bubalis). The enzyme was purified up to 71 folds by anion-exchange chromatography with 21% final recovery. Purified enzyme displayed two bands on SDS-PAGE with molecular weights of 9 kDa and 26 kDa respectively, the N-terminal sequence of later was EFLDKLDFYV. The enzyme has shown optimum activity at pH 9.0 and 40◦C. The KM, Kcat and Kcat/KM values of purified carboxypeptidase-B with Hippuryl-L-Arg are 30μM, 72sec(-1) and 2.4x10(5) M(-1) sec(-1) respectively. A computer based model for the structure of enzyme was proposed by chromatographic studies of component fragments and N-terminal sequence. The enzyme purified in the present study was free of carboxypeptidase A and endoprotease contamination. It was efficiently used in the processing of recombinant buffalo proinsulin, in combination with trypsin. Activation of proinsulin was monitored by MALDI-TOF analysis of peptides before and after the action of enzymes.

  2. Characterization of Smoc-1 uncovers two transcript variants showing differential tissue and age specific expression in Bubalus bubalis

    PubMed Central

    Srivastava, Jyoti; Premi, Sanjay; Kumar, Sudhir; Parwez, Iqbal; Ali, Sher

    2007-01-01

    Background Secreted modular calcium binding protein-1 (Smoc-1) belongs to the BM-40 family which has been implicated with tissue remodeling, angiogenesis and bone mineralization. Besides its anticipated role in embryogenesis, Smoc-1 has been characterized only in a few mammalian species. We made use of the consensus sequence (5' CACCTCTCCACCTGCC 3') of 33.15 repeat loci to explore the buffalo transcriptome and uncovered the Smoc-1 transcript tagged with this repeat. The main objective of this study was to gain an insight into its structural and functional organization, and expressional status of Smoc-1 in water buffalo, Bubalus bubalis. Results We cloned and characterized the buffalo Smoc-1, including its copy number status, in-vitro protein expression, tissue & age specific transcription/translation, chromosomal mapping and localization to the basement membrane zone. Buffalo Smoc-1 was found to encode a secreted matricellular glycoprotein containing two EF-hand calcium binding motifs homologous to that of BM-40/SPARC family. In buffalo, this single copy gene consisted of 12 exons and was mapped onto the acrocentric chromosome 11. Though this gene was found to be evolutionarily conserved, the buffalo Smoc-1 showed conspicuous nucleotide/amino acid changes altering its secondary structure compared to that in other mammals. In silico analysis of the Smoc-1 proposed its glycoprotein nature with a calcium dependent conformation. Further, we unveiled two transcript variants of this gene, varying in their 3'UTR lengths but both coding for identical protein(s). Smoc-1 evinced highest expression of both the variants in liver and modest to negligible in other tissues. The relative expression of variant-02 was markedly higher compared to that of variant-01 in all the tissues examined. Moreover, expression of Smoc-1, though modest during the early ages, was conspicuously enhanced after 1 year and remained consistently higher during the entire life span of buffalo with gradual

  3. Protein Modeling and Molecular Dynamics Simulation of Cloned Regucalcin (RGN) Gene from Bubalus bubalis.

    PubMed

    Pillai, Harikrishna; Yadav, Brijesh Singh; Chaturvedi, Navaneet; Jan, Arif Tasleem; Gupta, Girish Kumar; Baig, Mohammad Hassan; Bhure, Sanjeev Kumar

    2017-01-01

    Regucalcin (RGN), a calcium regulating protein having anti-prolific, antiapoptotic functions, plays important part in the biosynthesis of ascorbic acid. It is a highly conserved protein that has been reported from many tissue types of various vertebrate species. Employing its effect of regulating enzyme activities through reaction with sulfhydryl group (-SH) and calcium, structural level study believed to offer a better understanding of binding properties and regulatory mechanisms of RGN, was performed. Using sample from testis of Bubalus bubalis, amplification of regucalcin (RGN) gene was subjected to characterization by performing digestion using different restriction endonucleases (RE). Alongside, cDNA was cloned into pPICZαC vector and transformed in DH5α host for custom sequencing. To get a better insight of its structural characteristics, three dimensional (3D) structure of protein sequence was generated using in silico molecular modelling approach. The full trajectory analysis of structure was achieved by the Molecular Dynamics (MD) that explains the stability, flexibility and robustness of protein during simulation in a time of 50ns. Molecular docking against 1,5-anhydrosorbitol was performed for functional characterization of RGN. Preliminary screening of amplified products on Agarose gel showed expected size of ~893 bp of PCR product corresponding to RGN. Following sequencing, BLASTp search of the target sequence revealed that it shares 91% similarity score with human senescence marker protein-30 (pdb id: 3G4E). Molecular docking of 1,5-anhydrosorbitol reveals information regarding important binding site residues of RGN. 1,5-anhydrosorbitol was found to interact with binding free energy of - 6.01 Kcal/mol. RMSD calculation of subunits A, B and D-F might be responsible for functional and conserved regions of modeled protein. Three dimensional structure of RGN was generated and its interactions with 1,5- anhydrosorbitol, demonstrates the role of key

  4. Peripheral blood mononuclear cells: a potential cellular system to understand differential heat shock response across native cattle (Bos indicus), exotic cattle (Bos taurus), and riverine buffaloes (Bubalus bubalis) of India.

    PubMed

    Kishore, Amit; Sodhi, Monika; Kumari, Parvesh; Mohanty, A K; Sadana, D K; Kapila, Neha; Khate, K; Shandilya, Umesh; Kataria, R S; Mukesh, M

    2014-09-01

    Circulating leukocytes can be used as an effective model to understand the heat stress response of different cattle types and buffaloes. This investigation aimed to determine the temporal profile of HSPs (HSP40, HSP60, HSP70, and HSP90) expression in circulating peripheral blood mononuclear cells (PBMCs) of Murrah buffaloes, Holstein-Friesian (HF), and Sahiwal cows in response to sublethal heat shock at 42 °C. The viability data indicated HF PBMCs to be the most affected to the heat shock, whereas Sahiwal PBMCs were least affected, indicating its better survivability during the heat stress condition. The qRT-PCR expression data showed significant increase in mRNA expression of the analyzed HSPs genes after heat stimuli to the PBMCs under in vitro condition. In each case, the HSPs were most upregulated at 2 h after the heat stress. Among the HSPs, HSP70 was relatively more expressed followed by HSP60 indicating the action of molecular chaperones to stabilize the native conformation of proteins. However, PBMCs from different cattle types and buffaloes showed difference in the extent of transcriptional response. The level of expression of HSPs throughout the time period of heat stress was highest in buffaloes, followed by HF and Sahiwal cows. The higher abundance of HSP70 mRNA at each time point after heat stress showed prolonged effect of heat stress in HF PBMCs. The data presented here provided initial evidence of transcriptional differences in PBMCs of different cattle types and buffaloes and warrant further research.

  5. Assessment of chromium content of feedstuffs, their estimated requirement, and effects of dietary chromium supplementation on nutrient utilization, growth performance, and mineral balance in summer-exposed buffalo calves (Bubalus bubalis).

    PubMed

    Kumar, Muneendra; Kaur, Harjit; Tyagi, Amrish; Mani, Veena; Deka, Rijusmita Sarma; Chandra, Gulab; Sharma, Vijay Kumar

    2013-10-01

    This study was conducted to determine the chromium content of different feedstuffs, their estimated requirement, and effect of dietary Cr supplementation on nutrient intake, nutrient utilization, growth performance, and mineral balance in buffalo calves during summer season. Levels of Cr was higher in cultivated fodder, moderate in cakes and cereal grains, while straw, grasses, and non-conventional feeds were poor in Cr content. To test the effect of Cr supplementation in buffalo calves, 0, 0.5, 1.0, and 1.5 ppm of inorganic Cr were fed to 24 buffalo calves. Buffalo calves were randomly assigned to four treatments (n = 6) and raised for 120 days. A metabolic trial for a period of 7 days was conducted after 3 months of dietary treatments. Blood samples were collected at fortnight interval for plasma mineral estimation. The results suggested that dietary Cr supplementation in summer did not have any affects (P > 0.05) on feed consumption, growth performance, nitrogen balance, and physiological variables. However, dietary Cr supplementation had significant effect (P < 0.05) on balance and plasma Cr (ppb) levels without affecting (P > 0.05) balance and plasma levels of other trace minerals. The estimated Cr requirement of buffalo calves during summer season was calculated to be 0.044 mg/kg body mass and 10.37 ppm per day. In conclusion, dietary Cr supplementation has regardless effect on feed consumption, mass gain, and nutrient utilization in buffalo calves reared under heat stress conditions. However, supplementation of Cr had positive effect on its balance and plasma concentration without interacting with other trace minerals.

  6. A new extinct dwarfed buffalo from Sulawesi and the evolution of the subgenus Anoa: An interdisciplinary perspective

    NASA Astrophysics Data System (ADS)

    Rozzi, Roberto

    2017-02-01

    The fossil and extant faunas of Sulawesi, the largest island within the Wallacea biogeographic region, exhibit a high degree of endemism. The lowland anoa Bubalus depressicornis and the mountain anoa Bubalus quarlesi, two closely-related dwarfed buffaloes, are among the most peculiar endemic mammals of the region. Here, I describe a new species, Bubalus grovesi, from the Late Pleistocene/Holocene of South Sulawesi and I give a revised diagnosis of Anoa. Bubalus grovesi sp. nov. differs from all previously described Bubalus in both the size and proportions of the skeleton and in possessing a unique combination of discrete character states. Body mass estimates suggest an average mass of 117 kg for Bubalus grovesi sp. nov. and a body size reduction of about 90% with respect to a typical water buffalo. A comprehensive overview of body mass estimates of dwarfed buffaloes and differences in their dental and postcranial features is included. Finally, new evidence on the taxonomy and island dwarfing of the anoas and available data from different disciplines are used to discuss the timing and mode of their evolution. The representatives of the subgenus Anoa would be dwarfed forms of the Asian water buffalo that arose following dispersal to Sulawesi during the Middle/Late Pleistocene.

  7. Temporal leakage of Cu,Zn superoxide dismutase and loss of two low-molecular-weight forms of glutathione peroxidase-1 from buffalo (Bubalus bubalis) sperm after freezing and thawing.

    PubMed

    Kar, Senjuti; Divyashree, Bannur C; Roy, Sudhir C

    2015-03-01

    The postthaw motility and fertility of frozen-thawed buffalo spermatozoa are substantially low as compared with those of cattle sperm. The sperm motility and fertility have been positively correlated with the antioxidant enzyme activities of human and canine sperm. However, the extent of antioxidant enzyme loss during cryopreservation, although reported for human and cattle sperm, is still not clear for buffalo sperm. Thus, in the present study, an attempt was made to determine the activities of various antioxidant enzymes in buffalo spermatozoa cryopreserved for various durations (0, 30, and 60 days) and the mechanism of antioxidant enzyme loss, if any, during the process. Total superoxide dismutase (SOD) activity of cryopreserved sperm decreased and that of extended seminal plasma increased progressively with the increase in duration of cryopreservation indicating the possible time-dependent leakage of these enzymes from cryopreserved sperm into the extended seminal plasmas. The catalase and glutathione peroxidase (GPx) enzyme activities could not be detected in buffalo sperm but could be detected in fresh and extended seminal plasmas. Total GPx activities of extended seminal plasma decreased progressively with the increase in duration of cryopreservation. To confirm the presence of these enzymes at protein levels, specific antioxidant enzymes such as Cu,Zn SOD of 16 kDa and three molecular weight forms (57.7, 40.9, and 26.05 kDa) of GPx-1 were detected in buffalo sperm by Western blot. Furthermore, the intensities of 16-kDa Cu,Zn SOD in 60-day cryopreserved sperm and those of two low-molecular-weight forms of GPx-1 (40.9 and 26.05 kDa) in 30-day cryopreserved sperm decreased significantly (P < 0.05) as compared with those of noncryopreserved (0-day cryopreserved) sperm indicating selective and temporal leakage of only low-molecular-weight antioxidant proteins in the initial phase. However, all the mentioned GPx-1 forms disappeared in 60-day

  8. Pregnancy-specific protein B (PSPB), progesterone and some biochemical attributes concentrations in the fetal fluids and serum and its relationship with fetal and placental characteristics of Iraqi riverine buffalo (Bubalus bubalis).

    PubMed

    Abdulkareem, T A; Eidan, S M; Ishak, M A; Al-Sharifi, S A M; Alnimer, M A; Passavant, C W; Branen, J R; Sasser, R G

    2012-01-01

    This study was carried out to demonstrate the pregnancy-specific protein B (PSPB), progesterone and some biochemical parameters concentrations in amniotic fluid, allantoic fluid and fetal serum collected from slaughtered Iraqi riverine pregnant buffaloes at three different months of gestation (6th, 7th and 8th). Ten out of 22 adult buffaloes of 4.6 ± 0.97 years old were used in this study. The buffaloes were mated naturally by monitoring the estrus cycles via appearance of vaginal fluids and mounting by bulls. Pregnancy was checked for these buffaloes by non-returning to estrus for three estrus cycles and assured by rectal palpation on day 61 post-mating (PM). Buffaloes were slaughtered at three different periods of gestation (three at 6th month, four at 7th month and three at 8th month of gestation) to verify the progesterone and PSPB as well as some blood attributes levels (glucose, cholesterol, total protein, albumin, globulins and albumin: globulins ratio) in amniotic fluid (AF), allantoic fluid (LF) and fetal serum (FS). Progesterone was higher (P<0.01) in LF at the 8th month of gestation and lower in FS during the 7th and 8th months of pregnancy. PSPB concentrations were greater in FS (6th and 8th months in particular) than in both AF and LF. The overall mean of cholesterol concentration was higher in FS (P<0.05) followed by AF and LF that had the lowest concentration. The FS exhibited higher total protein during the three gestation periods. Most of fetal and placental measurements increased as the pregnancy advanced. In conclusion, these results described, for the first time, the PSPB and progesterone concentrations and blood characteristics in fetal fluids and serum in water riverine buffaloes during different stages of pregnancy. Progesterone concentrations were greater in allantoic fluid than in other fluids. In contrast, PSPB and other blood attributes were higher in fetal serum than other fluids of Iraqi riverine buffaloes. These findings reflect the

  9. Novel polymorphisms in UTR and coding region of inducible heat shock protein 70.1 gene in tropically adapted Indian zebu cattle (Bos indicus) and riverine buffalo (Bubalus bubalis).

    PubMed

    Sodhi, M; Mukesh, M; Kishore, A; Mishra, B P; Kataria, R S; Joshi, B K

    2013-09-25

    Due to evolutionary divergence, cattle (taurine, and indicine) and buffalo are speculated to have different responses to heat stress condition. Variation in candidate genes associated with a heat-shock response may provide an insight into the dissimilarity and suggest targets for intervention. The present work was undertaken to characterize one of the inducible heat shock protein genes promoter and coding regions in diverse breeds of Indian zebu cattle and buffaloes. The genomic DNA from a panel of 117 unrelated animals representing 14 diversified native cattle breeds and 6 buffalo breeds were utilized to determine the complete sequence and gene diversity of HSP70.1 gene. The coding region of HSP70.1 gene in Indian zebu cattle, Bos taurus and buffalo was similar in length (1,926 bp) encoding a HSP70 protein of 641 amino acids with a calculated molecular weight (Mw) of 70.26 kDa. However buffalo had a longer 5' and 3' untranslated region (UTR) of 204 and 293 nucleotides respectively, in comparison to Indian zebu cattle and Bos taurus wherein length of 5' and 3'-UTR was 172 and 286 nucleotides, respectively. The increased length of buffalo HSP70.1 gene compared to indicine and taurine gene was due to two insertions each in 5' and 3'-UTR. Comparative sequence analysis of cattle (taurine and indicine) and buffalo HSP70.1 gene revealed a total of 54 gene variations (50 SNPs and 4 INDELs) among the three species in the HSP70.1 gene. The minor allele frequencies of these nucleotide variations varied from 0.03 to 0.5 with an average of 0.26. Among the 14 B. indicus cattle breeds studied, a total of 19 polymorphic sites were identified: 4 in the 5'-UTR and 15 in the coding region (of these 2 were non-synonymous). Analysis among buffalo breeds revealed 15 SNPs throughout the gene: 6 at the 5' flanking region and 9 in the coding region. In bubaline 5'-UTR, 2 additional putative transcription factor binding sites (Elk-1 and C-Re1) were identified, other than three common sites

  10. Functional involvement of protein kinase C, Rho-kinase and TRPC3 decreases while PLC increases with advancement of pregnancy in mediating oxytocin-induced myometrial contractions in water buffaloes (Bubalus bubalis).

    PubMed

    Sharma, Abhishek; Nakade, Udayraj P; Choudhury, Soumen; Garg, Satish Kumar

    2017-04-01

    Present study unravels the involvement of different calcium signaling pathways in oxytocin-induced contractions in myometrium of non-pregnant and pregnant buffaloes during early and mid-pregnancy stages. Uteri of pregnant animals were more sensitive than of non-pregnant buffaloes. Phasic contractions and frequency of contraction significantly increased with advancement of pregnancy, while tonic contractions non-significantly and amplitude significantly decreased from six months pregnancy onward. Oxytocin produced concentration-dependent-contraction on isolated myometrial strips of pregnant and non-pregnant buffaloes and the dose response curves (DRCs) of oxytocin were significantly (P < 0.05) shifted to right in the presence of nifedipine (1 μM), in Ca(2+)-free Ringer Locke solution (RLS), ruthenium red (30 μM), ruthenium red + nifedipine, cyclopiazonic acid (CPA; Ca(2+) free RLS as well as RLS), CPA (10 μM)+nifedipine, U-73122 (1 μM) + nifedipine and SKF96365 (25 μM) on uteri of non-pregnant and pregnant (early and mid) animals. The DRCs were also significantly shifted towards right in the presence of Y-27632 (10 μM), GF109203X (5 μM) and Pyr3 (10 μM) on uteri of non-pregnant and early pregnancy stage buffaloes while only in the presence of U-73122 (1 μM) on uteri of mid-pregnancy stage buffaloes. Our finding suggest that and L-type Ca(2+) channels, IP3-RyR-gated, and store-operated calcium channels including transient receptor potential channel (TRPC) pathways play significant role in mediating oxytocin-induced contractions in myometrium of pregnant and non-pregnant buffaloes. SERCA plays major role only during early-pregnancy while functional role of protein kinase C (PKC), Rho-kinase and TRPC3 pathways decreased and role of G-protein coupled receptor-phospholipase C (GPCR-PLC) pathway increased with advancement of pregnancy. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. A comparative analysis of sperm selection procedures prior to cryopreservation for Nili-Ravi buffalo bull (Bubalus bubalis) semen-: Assessment of its impact on post-thaw sperm functional quality.

    PubMed

    Husna, Asma Ul; Ejaz, Rabea; Qadeer, Saima; Azam, Asima; Rakha, Bushra Allah; Ansari, Muhammad Sajjad; Shahzad, Qaisar; Javed, Moazzam; Vazquez-Levin, Mónica H; Akhter, Shamim

    2016-11-01

    Sperm selection techniques have been developed to get sperm suspensions enriched in motile and functional cells. Studies show that selection before cryopreservation improves post-thaw quality of cryopreserved sperm but information on buffalo bull sperm is scarce. The study was aimed to 1) perform a comparative analysis of sperm selection procedures; Swim-Up (SU), Sephadex™-G15 Filtration (S-G15) or Glass Wool Filtration (GWF) for total and motile cell recovery, 2) to assess the impact of sperm selection prior to cryopreservation on sperm quality (motility, morphology, cell membrane and normal apical ridge, viability and livability, chromatin integrity) and sperm functionality (Embryo Cleavage after IVF with selected sperm) in post-thawed suspensions of buffalo bull sperm. Semen was collected from 5 Nili Ravi buffalo bulls maintained at the Semen Production Unit Qadirabad, District Sahiwal, Pakistan. Ejaculates were divided into four aliquots for SU, S-G15 and GWF and control. After sperm selection, total and motile sperm recovery was highest in GWF samples (total sperm=84.08±8.39%; motile sperm=80.42±3.57%). An improvement (P<0.05) in all post-thaw parameters was observed in S-G15-selected sperm and, in some parameters in GWF-filtered sperm suspensions compared to control. The highest (P<0.05) embryo cleavage rate (%) was achieved with frozen-thawed sperm selected with S-G15 prior to cryopreservation (44.72±4.18) compared to control (21.98±3.00). In conclusion, post thaw sperm quality was improved after sperm selection from fresh buffalo bull semen through S-G15 and GWF procedures compared to SU and control while, the fertility rate (cleavage rate) was improved with sperm processed using the S-G15 procedure. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Comparative efficacy of E-17β and GnRH administration on day 0 of a controlled internal drug release (CIDR) based protocol on synchrony of wave emergence, ovulation and conception rates in Murrah buffalos (Bubalus bubalis)

    PubMed Central

    Bhat, G. R.; Dhaliwal, G. S.; Ghuman, S.; Honparkhe, M.

    2015-01-01

    The study was undertaken to compare the efficacy of an estradiol-17β + CIDR based protocol with the conventional ovsynch + CIDR based protocol for synchrony of wave emergence and ovulation in Murrah buffalos. In group I (n=25), on day 0 (beginning of experiment), buffaloes were administered a CIDR device (1.38 g P4) and concurrently received 1.5 mg E-17β. On day 9, the CIDR was removed and a prostaglandin (PG) F2α analogue (500 µg) was administered. On day 11, buffaloes were administered a gonadotropin releasing hormone (GnRH) analogue (20 µg) and inseminated twice at 12 h and 24 h following GnRH injections. Group II (n=25) protocol was based on an ovsynch regimen plus CIDR for 7 days followed by double insemination at induced estrus. Group III (n=10) served as control and was not given any hormone on day 0 of the protocol. In groups I, II and III, the duration of new follicular wave emergences were observed on days 4.22 ± 0.12, 3.12 ± 0.33 and 5.14 ± 0.42, respectively. In group I, synchrony of wave emergence was more and the diameter of pre-ovulatory follicles was larger (P<0.05) compared to groups II and III. The first service conception rate (FSCR) was higher (P<0.05) in group I while ovulation rates were not different between groups I and II. In conclusion, more synchrony of wave emergence, larger diameter of dominant follicles and higher first service conception rate was observed following the E-17β + CIDR based protocol in buffalos. PMID:27175151

  13. Effects of glial cell line-derived neurotrophic factor, fibroblast growth factor 2 and epidermal growth factor on proliferation and the expression of some genes in buffalo (Bubalus bubalis) spermatogonial cells.

    PubMed

    Kadam, Prashant H; Kala, Sushila; Agrawal, Himanshu; Singh, Karn P; Singh, Manoj K; Chauhan, Manmohan S; Palta, Prabhat; Singla, Suresh K; Manik, Radhay S

    2013-01-01

    The present study evaluated the effects of glial cell line-derived neurotrophic factor (GDNF), fibroblast growth factor (FGF) 2 and epidermal growth factor (EGF) on proliferation and the expression of some genes in spermatogonial cells. Spermatogonial cells were isolated from prepubertal buffalo testes and enriched by double enzyme treatment, filtration through 80- and 60-μm nylon mesh filters, differential plating on lectin-coated dishes and Percoll density gradient centrifugation. Cells were then cultured on a buffalo Sertoli cell feeder layer and formed colonies within 15-18 days. The colonies were found to predominantly contain undifferentiated Type A spermatogonia because they bound Dolichos biflorus agglutinin and did not express c-kit. The colonies expressed alkaline phosphatase, NANOG, octamer-binding transcription factor (OCT)-4 and tumour rejection antigen (TRA)-1-60. Cells were subcultured for 15 days, with or without growth factor supplementation. After 15 days, colony area and the relative mRNA abundance of PLZF were higher (P<0.05) following supplementation with 40 ng mL⁻¹ GDNF + 10 ng mL⁻¹ EGF + 10 ng mL⁻¹ FGF2 than with the same concentrations of GDNF alone or GDNF plus either EGF or FGF2. Expression of TAF4B was higher (P<0.05) in the presence of FGF2, whereas the expression of THY1 was not affected by growth factor supplementation. In the Sertoli cell feeder layer, EGF and FGF2 decreased (P<0.05), whereas GDNF increased (P<0.05), the relative mRNA abundance of ETV5 compared with control. In conclusion, an in vitro culture system that incorporates various growth factors was developed for the short-term culture of buffalo spermatogonia.

  14. Buffalo (Bubalus bubalis) SCNT embryos produced from somatic cells isolated from frozen-thawed semen: effect of trichostatin A on the in vitro and in vivo developmental potential, quality and epigenetic status.

    PubMed

    Selokar, Naresh L; Saini, Monika; Agrawal, Himanshu; Palta, Prabhat; Chauhan, Manmohan S; Manik, Radheysham; Singla, Suresh K

    2016-08-01

    This study examined the effects of trichostatin A (TSA) treatment of reconstructed buffalo embryos, produced by hand-made cloning using somatic cells isolated from over a decade old frozen-thawed semen, on their in vitro and in vivo developmental competence, quality and epigenetic status. Following treatment of reconstructed embryos with TSA (0, 50 or 75 nM) for 10 h prior to culture, the cleavage (100.0 ± 0, 94.5 ± 2.3 and 96.1 ± 1.2%, respectively) and blastocyst rate (50.6 ± 2.3, 48.4 ± 2.7 and 48.1 ± 2.6%, respectively), total cell number (275 ± 17.4, 289 ± 30.1 and 317 ± 24.2, respectively) and apoptotic index (5.6 ± 0.7, 3.4 ± 0.9 and 4.5 ± 1.4, respectively) were not significantly different among the three groups. However, TSA treatment increased (P < 0.05) the global level of H4K5ac and decreased (P < 0.05) that of H3K27me3 in blastocysts whereas the global level of H3K18ac was not affected significantly. Transfer of embryos treated with 75 nM TSA (n = 10) to recipients resulted in two pregnancies (20%), one out of which was aborted in the second and the other in the third trimester whereas transfer of control embryos (n = 20) or those treated with 50 nM TSA (n = 12) did not result in any pregnancy. In conclusion, these results suggest that TSA treatment of cloned buffalo embryos produced using somatic cells isolated from frozen-thawed semen improved their epigenetic status but not the in vitro developmental potential and offspring rate.

  15. Treatment of buffalo (Bubalus bubalis) donor cells with trichostatin A and 5-aza-2'-deoxycytidine alters their growth characteristics, gene expression and epigenetic status and improves the in vitro developmental competence, quality and epigenetic status of cloned embryos.

    PubMed

    Saini, M; Selokar, N L; Agrawal, H; Singla, S K; Chauhan, M S; Manik, R S; Palta, P

    2016-04-01

    We examined the effects of treating buffalo skin fibroblast donor cells with trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, and 5-aza-2'-deoxycytidine (5azadC), a DNA methyltransferase (DNMT) inhibitor, on the cells and embryos produced by hand-made cloning. Treatment of donor cells with TSA or 5azadC resulted in altered expression levels of the HDAC1, DNMT1, DNMT3a, P53, CASPASE3 and CASPASE9 genes and global levels of acetylation of lysine at position 9 or 14 in histone 3 (H3K9/14ac), acetylation of lysine at position 5 in histone 4 (H4K5ac), acetylation of lysine at position 18 in histone 3 (H3K18ac) and tri-methylation of lysine at position 27 in histone 3 (H3K27me3). Moreover, global levels of DNA methylation and activity of DNMT1 and HDAC1 were decreased, while global acetylation of H3 and H3K9 was significantly increased in comparison to untreated cells. Simultaneous treatment of donor cells with TSA (50nM) and 5azadC (7.5nM) resulted in higher in vitro development to the blastocyst stage, reduction of the apoptotic index and the global level of H3K27 me3 and altered expression levels of HDAC1, P53, CASPASE3, CASPASE9 and DNMT3a in cloned blastocysts. Transfer of cloned embryos produced with donor cells treated with TSA led to the birth of a calf that survived for 21 days. These results show that treatment of buffalo donor cells with TSA and 5azadC improved developmental competence and quality of cloned embryos and altered their epigenetic status and gene expression, and that these beneficial effects were mediated by a reduction in DNA and histone methylation and an increase in histone acetylation in donor cells.

  16. Downregulation of DNA Methyltransferase 1 in Zona-Free Cloned Buffalo (Bubalus bubalis) Embryos by Small Interefering RNA Improves In Vitro Development But Does Not Alter DNA Methylation Level

    PubMed Central

    Selokar, Naresh L.; Saini, Monika; Agrawal, Himanshu; Palta, Prabhat; Chauhan, Manmohan S.; Manik, Radheysham

    2015-01-01

    Abstract Aberrant epigenetic reprogramming, especially genomic hypermethylation, is implicated as the primary reason behind the failure of the cloning process during somatic cell nuclear transfer (SCNT). We transfected one-cell-stage zona-free buffalo embryos produced by handmade cloning with 50 nM DNMT1 small interfering RNA (siRNA), using lipofectamine, to knockdown the DNA methyltransferase 1 (DNMT1) gene. siRNA treatment decreased (p<0.001) the expression level of DNMT1 mRNA and DNMT1 protein in the one-cell-stage embryos and increased (p<0.05) the blastocyst rate (52.3±1.3% vs. 45.3±2.5%) compared to that in the controls, but did not reduce the DNA methylation level similar to the in vitro–fertilized (IVF) embryos. It also increased (p<0.05) the relative mRNA abundance of P53 and CASPASE 3, but not that of HDAC1, DNMT1, and DNMT3a, in the blastocysts of the siRNA group compared to the controls. The global level of H3K18ac was higher (p<0.05) in the blastocysts of the siRNA group than in the controls, whereas that of H3K9ac and H3K27me3 was not significantly different between the two groups. In conclusion, lipofection can be successfully used for transfection of DNMT1 siRNA into one-cell-stage zona-free cloned buffalo embryos. It results in a concomitant decrease in the DNMT1 mRNA and protein levels in the one-cell-stage embryos. siRNA-mediated knockdown increases the blastocyst rate but does not alter the DNA methylation level. PMID:25826721

  17. Co-culture of buffalo (Bubalus bubalis) preantral follicles with antral follicles: a comparative study of developmental competence of oocytes derived from in vivo developed and in vitro cultured antral follicles.

    PubMed

    Sharma, G Taru; Dubey, Pawan K; Nath, Amar; Saikumar, G

    2013-08-01

    The present study was undertaken to examine whether the presence of antral follicles (AFs) affects the survival, growth and steroidogenesis of preantral follicles (PFs) and compare the maturation and developmental competence of buffalo oocytes derived from in vivo developed and in vitro cultured AFs. Two experiments were carried out. In experiment I, PFs (200-250 μm) were isolated and cultured with or without AFs (3-5 mm) in TCM-199 medium that contained 10% fetal bovine serum (FBS), 1% insulin transferin selenium (ITS), 20 ng/ml epidermal growth factor (EGF), 0.5 μg/ml follicle-stimulating hormone (FSH) and 100 ng/ml insulin-like growth factor (IGF)-I. In experiment II, in vitro developmental competence was compared for the cumulus-oocyte complexes (COCs) recovered from in vivo developed and in vitro cultured AFs. Survival, growth, development of antrum, accumulation of estradiol and progesterone was (P < 0.05) higher when PFs were co-cultured with AFs. Developmental competence of both types of follicular oocytes did not differ significantly in terms of maturation and cleavage rate, but morula and blastocyst production rate were (P < 0.05) higher with in vivo developed AFs as compared with the in vitro cultured antral follicular oocytes. In conclusion, co-culture of PFs with AFs supports long-term survival and growth of buffalo PFs and this co-culture system plays a dual role for in vitro production of embryos as well as understanding the relationship between developing PFs and AFs.

  18. Oxygen concentration and cysteamine supplementation during in vitro production of buffalo (Bubalus bubalis) embryos affect mRNA expression of BCL-2, BCL-XL, MCL-1, BAX and BID.

    PubMed

    Elamaran, G; Singh, K P; Singh, M K; Singla, S K; Chauhan, M S; Manik, R S; Palta, P

    2012-12-01

    This study examined the effects of O(2) concentration (5% vs 20%) during in vitro maturation (IVM), fertilization (IVF) and culture (IVC) or supplementation of IVM and IVC media with cysteamine (50 and 100 μm, respectively; IVM, IVF and IVC carried out in 20% O(2)), on blastocyst rate and relative mRNA abundance of some apoptosis-related genes measured by real-time qPCR in immature and in vitro-matured buffalo oocytes and in embryos at 2-, 4-, 8- to 16-cell, morula and blastocyst stages. The blastocyst rate was significantly higher (p < 0.05) while the percentage of TUNEL-positive cells was significantly lower (p < 0.05) under 5% O(2) than that under 20% O(2). The mRNA expression of anti-apoptotic genes BCL-2 and MCL-1 was significantly higher (p < 0.05) and that of pro-apoptotic genes BAX and BID was lower (p < 0.05) under 5% O(2) than that under 20% O(2) concentration at many embryonic stages. Following cysteamine supplementation, the blastocyst rate and the relative mRNA abundance of BCL-XL and MCL-1 was significantly higher (p < 0.05) and that of BAX but not BID was lower (p < 0.05) at many stages of embryonic development, although it did not affect the percentage of TUNEL positive cells in the blastocysts significantly. The mRNA expression pattern of these genes during embryonic development was different in 5% vs 20% O(2) groups and in cysteamine supplemented vs controls. At the 8- to 16-cell stage, where developmental block occurs in buffalo, the relative mRNA abundance of BCL-2 and MCL-1 was highest under 5% O(2) concentration and that of BAX and BID was highest (p < 0.05) under 20% O(2) concentration. These results suggest that one of the mechanisms through which beneficial effects of low O(2) concentration and cysteamine supplementation are mediated during in vitro embryo production is through an increase in the expression of anti-apoptotic and a decrease in the expression of pro-apoptotic genes.

  19. Development of a pronuclear DNA microinjection technique for production of green fluorescent protein-expressing bubaline (Bubalus bubalis) embryos.

    PubMed

    Verma, V; Gautam, S K; Palta, P; Manik, R S; Singla, S K; Chauhan, M S

    2008-04-01

    Oocytes from abattoir-derived bubaline (Bubalus bubalis) ovaries were subjected to IVM and IVF; the objective was to develop a pronuclear DNA microinjection technique to produce embryos expressing green fluorescent protein (GFP). The largest proportion (61.2%) of zygotes in which one (1 PN) or two pronuclei (2 PN) were visible was when centrifugation (14,000 x g for 15 min) was done 16 h after insemination. Centrifugation had no adverse effects on cleavage rate, development to morulae/blastocysts, and total cell number of embryos. Piercing the pronuclear but not the plasma membrane reduced (P<0.05) cleavage rate (44.0% vs. 51.0%), without affecting subsequent development. Following microinjection of a GFP-DNA construct, cleavage rate (55.9, 38.9, and 30.9%) and proportion of cleaved embryos that developed to morulae (39.9, 25.6, and 15.5%) and blastocyst stages (22.4, 13.4, and 2.8%) were higher (P<0.05) for non-injected controls than for those injected with buffer alone, which, in turn, were higher (P<0.05) than for those injected with buffer containing 5 microg/mL DNA. The cleavage rate (39.2% vs. 34.8%) and proportion of cleaved embryos that developed to morulae/blastocysts (37.5% vs. 10.9%) were higher (P<0.05) for microinjected zygotes with 2 PN than for those with 1 PN. The cleavage rate and the proportion of cleaved embryos that developed to morulae and blastocysts were higher (P<0.05) following culture of microinjected zygotes in mCR2aa medium (40.7, 32.7, and 9.1%, respectively) compared to those for mSOFaa (33.3, 26.0, and 6.5%, respectively) or after culture in TCM-199+co-culture with buffalo oviductal epithelial cells (31.2, 25.0, and 4.5%, respectively). The proportion of embryos expressing GFP was higher (P<0.01) for 2 PN than for 1 PN zygotes (15.9% vs. 13.7%). Thirty-five embryos expressed GFP; the proportion of mosaic embryos (62.8%) was higher (P<0.01) than of embryos in which all blastomeres expressed GFP (37.2%); eight and two of those embryos

  20. Structure-function studies of Bubalus bubalis lingual antimicrobial peptide analogs.

    PubMed

    Kalita, Dhruba Jyoti; Kumar, Ashok; Kumar, Satish

    2009-02-01

    Antimicrobial peptides expressed on different epithelial lining are major components of the innate immune system. Based on the deduced amino acid sequence of Bubalus bubalis lingual antimicrobial peptide (LAP) cDNA (Accession No. DQ458768), five overlapping peptides LAP(23-55), LAP(42-64), LAP(21-64), LAP(1-26) and LAP(1-64) were synthesized using solid phase fluorenylmethoxycarbonyl (Fmoc) chemistry. Circular Dichroism spectroscopy of synthesized peptides revealed predominantly beta-structure for LAP(23-55,) LAP(42-64) and LAP(21-64) with less alpha-helix in different solutions. Quantitation of secondary structure indicated the highest beta-structure for all these three peptides in membrane mimetic SDS solution. The helicogenic solvent TFE could not induce helix in LAP(23-55) however TFE induced helical propensity was observed in LAP(42-64) and LAP(21-64). The quantitation of secondary structure indicated the highest ordered structure for LAP(23-55) followed by LAP(42-64) and LAP(21-64). The antibacterial activity of LAP(23-55) was found to be more potent against Staphylococcus aureus, Listeria monocytogens, Escherichia coli and Salmonella typhimurium followed by LAP(42-64) and LAP(21-64). Minimum inhibitory concentration (MIC) also showed similar trend with lowest value for LAP(23-55) followed by LAP(42-64) and LAP(21-64). Haemolysis and cytotoxicity was observed above 3 fold for LAP(21-64,) above six fold for LAP(23-55) and LAP(42-64) of their MIC. The LAP(1-26) and LAP(1-64) could not produce any characteristic CD spectra and did not show any antimicrobial activity, indicating that N- terminal of the peptide negates the antimicrobial activity.

  1. Sarcocystis cafferi, n. sp. (Protozoa: Apicomplexa) from the African buffalo (Syncerus caffer)

    USDA-ARS?s Scientific Manuscript database

    Four species of Sarcocystis are currently recognized in the Asian water buffalo (Bubalus bubalis): Sarcocystis fusiformis with macrocysts and cats as definitive hosts, S. buffalonis also with macrocysts and cats as definitive hosts, S. levinei with microcysts and dogs as definitive hosts, and S. dub...

  2. First molecular characterization of Cryptosporidium and Giardia from bovines (Bos taurus and Bubalus bubalis) in Sri Lanka: unexpected absence of C. parvum from pre-weaned calves

    PubMed Central

    2014-01-01

    Background The genetic characterization of Cryptosporidium and Giardia has important implications for investigating their epidemiology and underpins their control. We undertook the first molecular epidemiological survey of domestic bovids in selected regions of Sri Lanka to establish whether they excreted Cryptosporidium and/or Giardia with zoonotic potential. Methods Faecal samples were collected from dairy calves (n = 340; Bos taurus; < 3 months of age; weekly sampling for six weeks) and water buffaloes (n = 297; Bubalus bubalis; <6 months and ≥6 months of age; one sampling) from seven different farms in Sri Lanka. Genomic DNAs were extracted from individual faecal samples and then tested for the presence of parasite DNA using a PCR-based mutation scanning-targeted sequencing-phylogenetic approach, employing genetic markers within the small subunit of nuclear ribosomal RNA and 60 kDa glycoprotein genes (designated pSSU and pgp60, respectively) for Cryptosporidium, and within the triose phosphate isomerise (ptpi) gene for Giardia. Results Based on pSSU sequence data, C. bovis, C. ryanae and six new genotypes that were genetically similar but not identical to C. andersoni (n = 1), C. bovis (n = 1), C. ryanae (n = 3) and C. suis (n = 1) were recorded in cattle. For pSSU, two other, new genotypes were defined in water buffalo, which were genetically most similar to Cryptosporidium genotypes recorded previously in this host species in other countries including Australia. Consistent with the findings for pSSU, no species or genotypes of Cryptosporidium with zoonotic potential were detected using pgp60. Based on ptpi sequence data, G. duodenalis assemblages A and E were detected in four and 137 samples from cattle, respectively, and assemblage E in two samples from water buffaloes. Conclusions The present study showed that C. parvum, the most commonly reported zoonotic species of Cryptosporidium recognised in bovine calves globally, was not

  3. Differential involvement of L- and T-type Ca(2+) channels, store-operated calcium channel (TRPC) and Rho-kinase signaling pathway(s) in PGF2α-induced contractions in myometrium of non-pregnant and pregnant buffaloes (Bubalus bubalis).

    PubMed

    Sharma, Abhishek; Nakade, Udayraj P; Jaitley, Pooja; Sharma, Vipin; Choudhury, Soumen; Garg, Satish Kumar

    2017-09-12

    This study unravels the differential involvement of calcium signaling pathway(s) in PGF2α-induced contractions in myometrium of non-pregnant and pregnant buffaloes. Compared to the myometrium of pregnant animals, myometrium of non-pregnant buffaloes was more sensitive to PGF2α-induced contractile effect as manifested by the changes in mean integral tension (MIT) and tonicity. However, phasic contraction was significantly more in myometrium of pregnant animals. The uterotonic effect of PGF2α was dependent on extracellular Ca(2+) and its influx through nifedipine-sensitive L-type Ca(2+) channels both in non-pregnant and pregnant animals, but T-type Ca(2+) channels play an additional role during pregnancy. Entry of extracellular Ca(2+) is triggered by enhanced functional involvement of Pyr3-sensitive TRPC3 channels and Rho-kinase pathways to regulate uterotonic action of PGF2α in myometrium of non-pregnant buffaloes while these are down-regulated during pregnancy as there was significantly reduced expression of Rho-A proteins in myometrium of pregnant buffaloes and down-regulation of these pathways facilitate uterine quiescence. Intracellular Ca(2+) plays minor role in myometrium of both the non-pregnant and pregnant buffaloes. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. First genome sequences of buffalo coronavirus from water buffaloes in Bangladesh.

    PubMed

    Lau, S K P; Tsang, A K L; Shakeel Ahmed, S; Mahbub Alam, M; Ahmed, Z; Wong, P-C; Yuen, K-Y; Woo, P C Y

    2016-05-01

    We report the complete genome sequences of a buffalo coronavirus (BufCoV HKU26) detected from the faecal samples of two domestic water buffaloes (Bubalus bubalis) in Bangladesh. They possessed 98-99% nucleotide identities to bovine coronavirus (BCoV) genomes, supporting BufCoV HKU26 as a member of Betacoronavirus 1. Nevertheless, BufCoV HKU26 possessed distinct accessory proteins between spike and envelope compared to BCoV. Sugar-binding residues in the N-terminal domain of S protein in BCoV are conserved in BufCoV HKU26.

  5. The "Linh Duong" Pseudonovibos spiralis (Mammalia, Artiodactyla) is a new buffalo

    NASA Astrophysics Data System (ADS)

    Kuznetsov, G. V.; Kulikov, E. E.; Petrov, N. B.; Ivanova, N. V.; Lomov, A. A.; Kholodova, M. V.; Poltaraus, A. B.

    2001-03-01

    The controversial phylogenetic position of the recently described South-East Asian endemic bovid, Pseudonovibos spiralis, was evaluated on the basis of phylogenetic analyses of originally obtained nearly complete 12S mitochondrial rDNA sequences for this species and Bubalus bubalis and 26 sequences of Bovidae from the Genbank using Cervus elaphus (Cervidae) as outgroup. In most of the phylogenetic analyses performed using PAUP 4.0 (maximum parsimony, maximum likelihood and neighbour-joining), Bovidae consisted of two major clades: Bovinae including the tribes Bovini, Tragelaphini and Boselaphini, and Antilopinae+Caprinae, incorporating all other bovids. In most trees P. spiralis fell within the buffalos (subtribe Bovina) between Bubalus and Syncerus. Therefore, our phylogenetic analyses of bovid mitochondrial 12S rRNA gene sequences suggest the close relationship of this enigmatic species with the buffalos and its placement within the subtribe Bovina.

  6. Ontogeny, postnatal development and ageing of endocrine pancreas in Bubalus bubalis

    PubMed Central

    LUCINI, C.; CASTALDO, L.; LAI, O.; DE VICO, G.

    1998-01-01

    The ontogenesis, postnatal development and ageing of the endocrine pancreas in mammals have not been extensively studied. In order to improve understanding of this organ, we studied the buffalo pancreas during fetal and postnatal development. Glucagon, insulin and somatostatin immunoreactive cells (i.c.) were first seen in 2-mo-old embryos. Pancreatic polypeptide (PP) i.c. were observed during the 3rd month of gestation. The early embryo pancreas was almost totally composed of endocrine tissue. The endocrine portion only slightly increased in mass with animal growth, whereas the exocrine portion noticeably increased in mass during the late fetal and postnatal periods. In adults, therefore, the exocrine portion was more evident than the endocrine portion. Three types of islet were observed in fetal and young buffalos: small, large and PP-islets. The small islets were composed of insulin, glucagon, somatostatin and PP i.c. The large islets were primarily composed of insulin i.c. and a few glucagon, somatostatin and PP i.c. The PP islets were mostly composed of PP i.c. with a few somatostatin, insulin and glucagon i.c. The number of large islets greatly diminished by adulthood. Glucagon, insulin, somatostatin and PP i.c. were also seen scattered in the exocrine parenchyma and along the duct epithelium. In the duct epithelium, these cells were either single or grouped, and they sometimes formed a protrusion projecting towards the connective tissue. These morphological features were primarily observed in fetuses and young buffalos. PMID:9688507

  7. Malignant Catarrhal Fever: An Emerging Disease in the African Buffalo (Syncerus caffer).

    PubMed

    Pfitzer, S; Last, R; Espie, I; van Vuuren, M

    2015-06-01

    Within the tribe Bovini in the subfamily Bovinae, the water buffalo (Bubalus Bubalis), American bison (Bison bison), European bison (Bubalus bonasus) and yak (Bos grunniens) are recognized as species highly susceptible to malignant catarrhal fever (MCF). In contrast, the lack of reports describing clinical MCF in the African buffalo (Syncerus caffer) whether free ranging or captive has led to a perception that African buffaloes are resistant to MCF. During the last decade, several cases of MCF in African buffaloes were confirmed in South Africa and experience with seven of these cases is described in this report. Detection of viral nucleic acid in blood or tissues was successful in six African buffaloes that suffered from clinical signs compatible with MCF. Four were positive for infection with ovine herpesvirus type 2 (the causative virus of sheep-associated MCF), and two were positive for alcelaphine herpesvirus type 1 (causative virus of wildebeest-associated MCF). Histopathological examination of tissue samples from all the animals yielded typical lesions that were consistent with those described for MCF in domestic cattle. Developments in the management of African buffaloes translocated from their traditional habitats have likely contributed to the identification of another susceptible host in the subfamily Bovinae.

  8. Retinoic acid induces differentiation of buffalo (Bubalus bubalis) embryonic stem cells into germ cells.

    PubMed

    Shah, Syed Mohmad; Singla, Suresh Kumar; Palta, Prabhat; Manik, Radhey Sham; Chauhan, Manmohan Singh

    2017-10-05

    Development of precise and reproducible culture system for in vitro differentiation of embryonic stem (ES) cells into germ cells counts as a major leap forward for understanding not only the remarkable process of gametogenesis, otherwise obscured by limited availability of precursor primordial germ cells (PGCs), but in finally treating the catastrophic infertility. Taking into account the significant role of retinoic acid (RA) during in vivo gametogenesis, we designed the present study to investigate the effects of its stimulation on directing the differentiation of ES cells into germ cells. The effects of RA were analyzed across dose-and-time upon various stages of gametogenesis like PGC induction, meiosis initiation and completion, haploid cell formation and development of the final gamete (oocyte and spermatozoa). Out of the series of RA doses (2, 4, 8, 16, 20 and 30μM), 16μM RA for 8day culture interval was found to induce highest expression of PGC- and meiosis-associated genes like DAZL, VASA, SYCP3, MLH1, TNP1/2 and PRM2, while mature germ cell genes like BOULE and TEKT1 (Spermatocyte markers), GDF9 and ZP2 (Oocyte markers) showed higher expression at 2μM RA dose, suggesting functional concentration-gradient of RA activity. Immunocytochemistry revealed expression of germ lineage-specific markers like: c-KIT, DAZL and VASA (PGC-markers); SYCP3, MLH1 and PROTAMINE1 (Meiotic-markers); ACROSIN and HAPRIN (Spermatocyte-markers); and GDF9 and ZP4 (Oocyte-markers) in optimally differentiated embryoid bodies (EBs) and adherent cultures. We observed significantly reduced (p<0.05) concentration of 5-methyl-2-deoxycytidine in RA-differentiated EBs which is suggestive of the occurrence of methylation erasure. FACS analysis of optimally differentiated cultures detected 3.07% haploid cell population, indicating completion of meiosis. Oocyte-like structures (OLS) were obtained in adherent differentiated cultures. They had a big nucleus and a zona pellucida (ZP4) coat. They showed progression through 2-cell, 4-cell, 8-cell, morula and blastocyst-like structures upon extended culture beyond 14days. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Retinoic acid induces differentiation of buffalo (Bubalus bubalis) embryonic stem cells into germ cells.

    PubMed

    Shah, Syed Mohmad; Singla, Suresh Kumar; Palta, Prabhat; Manik, Radhey Sham; Chauhan, Manmohan Singh

    2017-08-30

    Development of precise and reproducible culture system for in vitro differentiation of embryonic stem (ES) cells into germ cells counts as a major leap forward for understanding not only the remarkable process of gametogenesis, otherwise obscured by limited availability of precursor primordial germ cells (PGCs), but in finally treating the catastrophic infertility. Taking into account the significant role of retinoic acid (RA) during in vivo gametogenesis, we designed the present study to investigate the effects of its stimulation on directing the differentiation of ES cells into germ cells. The effects of RA were analyzed across dose-and-time upon various stages of gametogenesis like PGC induction, meiosis initiation and completion, haploid cell formation and development of the final gamete (oocyte and spermatozoa). Out of the series of RA doses (2, 4, 8, 16, 20 and 30μM), 16μM RA for 8day culture interval was found to induce highest expression of PGC- and meiosis-associated genes like DAZL, VASA, SYCP3, MLH1, TNP1/2 and PRM2, while mature germ cell genes like BOULE and TEKT1 (Spermatocyte markers), GDF9 and ZP2 (Oocyte markers) showed higher expression at 2μM RA dose, suggesting functional concentration-gradient of RA activity. Immunocytochemistry revealed expression of germ lineage-specific markers like: c-KIT, DAZL and VASA (PGC-markers); SYCP3, MLH1 and PROTAMINE1 (Meiotic-markers); ACROSIN and HAPRIN (Spermatocyte-markers); and GDF9 and ZP4 (Oocyte-markers) in optimally differentiated embryoid bodies (EBs) and adherent cultures. We observed significantly reduced (p<0.05) concentration of 5-methyl-2-deoxycytidine in RA-differentiated EBs which is suggestive of the occurrence of methylation erasure. FACS analysis of optimally differentiated cultures detected 3.07% haploid cell population, indicating completion of meiosis. Oocyte-like structures (OLS) were obtained in adherent differentiated cultures. They had a big nucleus and a zona pellucida (ZP4) coat. They showed progression through 2-cell, 4-cell, 8-cell, morula and blastocyst-like structures upon extended culture beyond 14days. Copyright © 2017. Published by Elsevier B.V.

  10. Strategies to overcome seasonal anestrus in water buffalo.

    PubMed

    de Carvalho, Nelcio Antonio Tonizza; Soares, Julia Gleyci; Baruselli, Pietro Sampaio

    2016-07-01

    Reproductive seasonality in buffalo (Bubalus bubalis) is characterized by behavioral, endocrine, and reproductive changes that occur over distinct periods of the year. During the nonbreeding season (spring and summer), the greater light-dark ratio (long days) suppresses estrus behavior and the occurrence of ovulation. Anestrous buffaloes have insufficient pulsatile of LH to support the final stages of follicular development, and subsequently, estrus behavior and ovulation do not occur, limiting reproductive efficiency, especially in artificial insemination (AI) programs. A number of therapeutic strategies designed to synchronize follicular wave emergence and ovulation have allowed for the use of AI throughout the year, overcoming seasonal anestrus in buffalo. These therapies also improve reproductive performance by increasing the service rate and pregnancy per AI in buffalo herds, regardless of reproductive seasonality. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Cystic echinococcosis in water buffaloes: epidemiological survey and molecular evidence of ovine (G1) and buffalo (G3) strains.

    PubMed

    Capuano, F; Rinaldi, L; Maurelli, M P; Perugini, A G; Veneziano, V; Garippa, G; Genchi, C; Musella, V; Cringoli, G

    2006-04-30

    A survey of cystic echinococcosis (CE) in the water buffalo (Bubalus bubalis) of the Italian Mediterranean breed was carried out in Campania, a region of southern Italy. In addition, a molecular study was performed on 48 hydatid cysts coming from 48 water buffaloes in order to determine the Echinococcus granulosus strain(s) present in this host. Out of a total of 722 water buffaloes examined for CE, 76 (10.5%) were found infected. The average number of cysts per buffalo was 4.3 (minimum 1, maximum 45). Seventeen buffaloes had hydatid cysts only in the liver (with an average of 5 cysts/liver), 34 only in the lungs (with an average of 1.8 cysts/lungs), and 25 buffaloes had cysts both in the liver and in the lungs. Fertile cysts were found in 10 (13.2%) out of the 76 positive buffaloes. The sequencing of the mitochondrial cytochrome C oxidase subunit 1 (CO1) gene of the 48 hydatid cysts produced sequences of 419 bp for each sample analysed. For 33 samples, alignment of the obtained sequences with those present in GenBank showed a total homology with the common domestic sheep strain G1; for 15 samples, sequences obtained showed 100% homology with buffalo strain G3. The findings of the present survey represent the first epidemiological and molecular comprehensive studies on CE in water buffalo from an endemic area for E. granulosus.

  12. Phylogenetic analysis and comparison between cow and buffalo (including Egyptian buffaloes) mitochondrial displacement-loop regions.

    PubMed

    Ramadan, Hassan A I; El-Hefnawi, Mahmoud M

    2008-08-01

    Mitochondrial DNA (mtDNA) analysis has been used extensively for phylogenetic analysis studies and systematics. The displacement loop (D-loop) region inside the mtDNA is a non-coding part whose analysis can indicate variations between closely related populations. This paper reports for the first time the characterization and analysis of the complete sequence of the D-loop region from Egyptian buffaloes and analysis in conjunction with previously published Indian and European Bubalus bubalis and Bos sub-tribe sequences. In the entire D-loop of the Egyptian buffaloes, we identified four haplotypes and nine polymorphic sites from the nine sequenced D-loop regions--while in the studied set of buffaloes we identified 28 polymorphic sites in the entire D-loop, and 49 polymorphic sites in the case of cows. Alignment between buffaloes and cows to evaluate the characteristics of the D-loop region showed that the second region of the conserved sequence block (CSB2) is apparently the most variable region in the D-loop between cows and buffaloes, with four insertions in all buffaloes and two substitutions, followed by the second region of the extended termination associated sequence (ETAS2) with a substitution rate of 1/10. The Egyptian buffaloes were shown to be closest to the Italian counterparts, exemplifying the closeness of ethnicity and the history of civilization of that region.

  13. Understanding tenderness variability and ageing changes in buffalo meat: biochemical, ultrastructural and proteome characterization.

    PubMed

    Kiran, M; Naveena, B M; Reddy, K S; Shahikumar, M; Reddy, V R; Kulkarni, V V; Rapole, S; More, T H

    2016-06-01

    Understanding of biological impact of proteome profile on meat quality is vital for developing different approaches to improve meat quality. Present study was conducted to unravel the differences in biochemical, ultrastructural and proteome profile of longissimus dorsi muscle between buffaloes (Bubalus bubalis) of different age groups (young v. old). Higher (P<0.05) myofibrillar and total protein extractability, muscle fibre diameter, and Warner-Bratzler shear force (WBSF) values was observed in old buffalo meat relative to meat from young buffaloes. Scanning electron microscopy photographs revealed reduced fibre size with increased inter-myofibrillar space in young compared with old buffalo meat. Transmission electron microscopy results revealed longer sarcomeres in young buffalo meat relative to meat from old buffaloes. Proteomic characterization using two-dimensional gel electrophoresis (2DE) found 93 differentially expressed proteins between old and young buffalo meat. Proteome analysis using 2DE revealed 191 and 95 differentially expressed protein spots after 6 days of ageing in young and old buffalo meat, respectively. The matrix assisted laser desorption ionization time-of flight/time-of flight mass spectrometry (MALDI-TOF/TOF MS) analysis of selected gel spots helped in identifying molecular markers of tenderness mainly consisting of structural proteins. Protein biomarkers identified in the present study have the potential to differentiate meat from young and old buffaloes and pave the way for optimizing strategies for improved buffalo meat quality.

  14. First report of Cryptosporidium species in farmed and wild buffalo from the Northern Territory, Australia.

    PubMed

    Zahedi, Alireza; Phasey, Jordan; Boland, Tony; Ryan, Una

    2016-03-01

    A molecular epidemiological survey of Cryptosporidium from water buffalo (Bubalus bubalis) in the Northern Territory in Australia was conducted. Fecal samples were collected from adult farmed (n = 50) and wild buffalo (n = 50) and screened using an 18S quantitative PCR (qPCR). Positives were typed by sequence analysis of 18S nested PCR products. The qPCR prevalence of Cryptosporidium species in farmed and wild buffalo was 30 and 12 %, respectively. Sequence analysis identified two species: C. parvum and C. bovis, with C. parvum accounting for ~80 % of positives typed from the farmed buffalo fecal samples compared to 50 % for wild buffalo. Subtyping at the 60 kDa glycoprotein (gp60) locus identified C. parvum subtypes IIdA19G1 (n = 4) and IIdA15G1 (n = 1) in the farmed buffalo and IIaA18G3R1 (n = 2) in the wild buffalo. The presence of C. parvum, which commonly infects humans, suggests that water buffaloes may contribute to contamination of rivers and waterways with human infectious Cryptosporidium oocysts, and further research on the epidemiology of Cryptosporidium in buffalo populations in Australia is required.

  15. Histo-morphology of the uterus and early placenta of the African buffalo (Syncerus caffer) and comparative placentome morphology of the African buffalo and cattle (Bos taurus).

    PubMed

    Schmidt, S; Gerber, D; Soley, J T; Aire, T A; Boos, A

    2006-08-01

    Differences exist in reproductive physiology between African buffalo (Syncerus caffer), cattle (Bos taurus) and water buffalo (Bubalus bubalis). The aim of this study was to histo-morphologically compare the anatomy of non-pregnant and pregnant uteri of buffalo and cattle. Two non-pregnant uteri and placentae of six pregnant African buffalo were used. Early placentome formation (fetal crown rump length (CRL): 2-17.5 cm) in S. caffer and B. taurus was compared. The endometrium of buffalo uteri comprises round to ovoid, dome-shaped and gland-free caruncles. A predominantly simple columnar epithelium of non-ciliated cells covers caruncular tissue, while, additionally, ciliated cells occur in the epithelium of the intercaruncular areas and within the simple columnar or pseudostratified epithelium of the endometrial glands. During early gestation, multiple placentomes develop. Unlike the placentomes in cattle at similar CRL, buffalo placentomes do not develop a caruncular stalk. The sessile, dome-shaped buffalo placentome has simple, slightly conical villi branching less than in cattle, thus indicating different and less complex feto-maternal interdigitation than seen in the latter. A synepitheliochorial interhaemal barrier can be expected in the buffalo placenta, as the occurrence and ultrastructure of trophoblast giant cells resemble those described in cattle.

  16. RETRACTED: Bone morphogenetic protein 4 (BMP4) induces buffalo (Bubalus bubalis) embryonic stem cell differentiation into germ cells.

    PubMed

    Shah, Syed Mohmad; Saini, Neha; Ashraf, Syma; Singh, Manoj Kumar; Manik, Radhey Sham; Singla, Suresh Kumar; Palta, Prabhat; Chauhan, Manmohan Singh

    2015-12-01

    This article has been retracted: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). This article has been retracted at the request of the Editor-in-Chief. Problems related to images published in this paper in Figure 12 were brought to the authors' attention. Unfortunately this figure contains duplicate images for ESC controls for VASA, GDF9, and ZP4, which display identical patterns superimposed on varying intensities of background. Therefore, the authors retract the paper with the agreement of the editors and deeply regret this situation and apologize for any inconvenience to the editors and readers of Biochimie. Copyright © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  17. PHYLOGEOGRAPHIC PATTERNS IN LARGE RIVER ECOSYSTEMS: GENETIC STRUCTURE OF SMALLMOUTH BUFFALO (ICTIOBUS BUBALUS) IN THE OHIO RIVER

    EPA Science Inventory

    Genetic studies on populations of large river fishes provide a potentially useful but underutilized research and assessment tool. Population genetic research on freshwater systems has provided meaningful insight into stock structure, hybridization issues, and gene flow/migration...

  18. PHYLOGEOGRAPHIC PATTERNS IN LARGE RIVER ECOSYSTEMS: GENETIC STRUCTURE OF SMALLMOUTH BUFFALO (ICTIOBUS BUBALUS) IN THE OHIO RIVER

    EPA Science Inventory

    Genetic studies on populations of large river fishes provide a potentially useful but underutilized research and assessment tool. Population genetic research on freshwater systems has provided meaningful insight into stock structure, hybridization issues, and gene flow/migration...

  19. Analysis of microsatellite markers in a Cuban water buffalo breed.

    PubMed

    Uffo, Odalys; Martínez, Nadia; Acosta, Atzel; Sanz, Arianne; Martín-Burriel, Inmaculada; Osta, Rosario; Rodellar, Clementina; Zaragoza, Pilar

    2017-08-01

    The aim of this Regional Research Communication was to validate a panel of 30 microsatellite markers recommended by FAO/ISAG for studies of biodiversity in cattle to improve the characterisation of Cuban buffalo populations. The water buffalo (Bubalus bubalis) is an economically important livestock species. Therefore, research focused on the study of the genetic relationships among water buffalo populations is useful to support conservation decisions and to design breeding schemes. Twenty-eight of the 30 tested regions were amplified, one of which (ETH10) turned out to be monomorphic. A total of 143 alleles were observed in the Cuban water buffalo population. The average number of alleles per locus was 5·04. The number of alleles per polymorphic locus ranged from two (INRA 63 and MM12) to nine (ETH185). The observed and expected heterozygosity ranged from 0·108 (HAUT24) to 0·851 (CSSM66) and 0·104 (MM12) to 0·829(INRA32), respectively. The polymorphic information content (PIC) ranged from 0·097 (MM12) to 0·806 (INRA32), and the overall value for these markers was 0·482. Within the population, inbreeding estimates (F IS) was positive in 14 of the 30 loci analysed. This study thus highlights the usefulness of heterologous bovine microsatellite markers to assess the genetic variability in Cuban water buffalo breeds. Furthermore, the results can be utilised for future breeding strategies and conservation.

  20. Profile of muscle tissue gene expression specific to water buffalo: Comparison with domestic cattle by genome array.

    PubMed

    Zhang, Yingying; Wang, Hongbao; Gui, Linsheng; Wang, Hongcheng; Mei, Chugang; Zhang, Yaran; Xu, Huaichao; Jia, Cunlin; Zan, Linsen

    2016-02-10

    In contrast with the past, the water buffalo is now not only a draft animal, but also an important food source of milk and meat. It is increasingly apparent that the water buffalo have huge potential for meat production, but its breeding needs to be investigated. Regarding the molecular mechanisms involved in the meat quality difference between the buffalo (Bubalus bulabis) and yellow cattle (Bos taurus), 12 chemical-physical characteristics related to the meat quality of longissimus thoracis muscles (LTM) have been compared at the age of 36 months. Intramuscular lipid and b* (yellowness) were greater in cattle than the buffalo, whereas a* (redness) was greater in the buffalo. Gene expression profiles were constructed by bovine genome array. A total of 8884 and 10,960 probes were detected in buffalo and cattle, respectively, with 1580 genes being differentially expressed. Over 400 probes were upregulated and nearly 1200 were downregulated in LTM of the buffalo, most being involved in ribosomal RNA (rRNA) processing, cholesterol homeostasis, regulation of transcription, response to hypoxia, and glycolysis. Quantitative real-time PCR was used to validate the microarray data. Enriched GO analyses of highly expressed genes in LTM showed that protein biosynthesis, striated muscle contraction, iron homeostasis, iron transport, glycolysis and glucose metabolism were similar between the buffalo and cattle. High protein content, low fat content and deep meat color of buffalo LTM may be closely associated with the increased expression of genes involved in cholesterol and iron homeostasis, while also reducing the expression of genes involved in ubiquitin-mediated proteolysis and protein oxidative phosphorylation. These results establish the groundwork for further studies on buffalo meat quality and will be beneficial in improving water buffalo breeding by molecular biotechnology. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. The prion protein gene polymorphisms associated with bovine spongiform encephalopathy susceptibility differ significantly between cattle and buffalo.

    PubMed

    Zhao, Hui; Du, Yanli; Chen, Shunmei; Qing, Lili; Wang, Xiaoyan; Huang, Jingfei; Wu, Dongdong; Zhang, Yaping

    2015-12-01

    Prion protein, encoded by the prion protein gene (PRNP), plays a crucial role in the pathogenesis of transmissible spongiform encephalopathies (TSEs). Several polymorphisms within the PRNP are known to be associated with influencing bovine spongiform encephalopathy (BSE) susceptibility in cattle, namely two insertion/deletion (indel) polymorphisms (a 23-bp indel in the putative promoter and a 12-bp indel in intron 1), the number of octapeptide repeats (octarepeats) present in coding sequence (CDS) and amino acid polymorphisms. The domestic buffaloes, Bubalus bubalis, are a ruminant involved in various aspects of agriculture. It is of interest to ask whether the PRNP polymorphisms differ between cattle and buffalo. In this study, we analyzed the previously reported polymorphisms associated with BSE susceptibility in Chinese buffalo breeds, and compared these polymorphisms in cattle with BSE, healthy cattle and buffalo by pooling data from the literature. Our analysis revealed three significant findings in buffalo: 1) extraordinarily low deletion allele frequencies of the 23- and 12-bp indel polymorphisms; 2) significantly low allelic frequencies of six octarepeats in CDS and 3) the presence of S4R, A16V, P54S, G108S, V123M, S154N and F257L substitutions in buffalo CDSs. Sequence alignments comparing the buffalo coding sequence to other species were analyzed using the McDonald-Kreitman test to reveal five groups (Bison bonasus, Bos indicus, Bos gaurus, Boselaphus tragocamelus, Syncerus caffer caffer) with significantly divergent non-synonymous substitutions from buffalo, suggesting potential divergence of buffalo PRNP and others. To the best of our knowledge this is the first study of PRNP polymorphisms associated with BSE susceptibility in Chinese buffalo. Our findings have provided evidence that buffaloes have a unique genetic background in the PRNP gene in comparison with cattle. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Genetic diversity analysis of buffalo fatty acid synthase (FASN) gene and its differential expression among bovines.

    PubMed

    Niranjan, S K; Goyal, S; Dubey, P K; Kumari, N; Mishra, S K; Mukesh, M; Kataria, R S

    2016-01-10

    Fatty Acid Synthase (FASN) gene seems to be structurally and functionally different in bovines in view of their distinctive fatty acid synthesis process. Structural variation and differential expression of FASN gene is reported in buffalo (Bubalus bubalis), a bovine species close to cattle, in this study. Amino acid sequence and phylogenetic analysis of functionally important thioesterase (TE) domain of FASN revealed its conserved nature across mammals. Amino acid residues at TE domain, responsible for substrate binding and processing, were found to be invariant in all the mammalian species. A total of seven polymorphic nucleotide sites, including two in coding region of TE domain were identified across the 10 buffalo populations of riverine and swamp types. G and C alleles were found almost fixed at g18996 and g19056 loci, respectively in riverine buffaloes. Principal component analysis of three SNPs (g18433, g18996 and g19056) revealed distinct classification of riverine and swamp buffalo populations. Reverse Transcription-PCR amplification of mRNA corresponding to exon 8-10 region of buffalo FASN helped in identification of two transcript variants; one transcript of 565 nucleotides and another alternate transcript of 207 nucleotides, seems to have arisen through alternative splicing. Both the transcripts were found to be expressed in most of the vital tissues of buffalo with the highest expression in mammary gland. Semi-quantitative and real-time expression analysis across 13 different buffalo tissues revealed its highest expression in lactating mammary gland. When compared, expression of FASN was also found to be higher in liver, adipose and skeletal muscle of buffalo tissues, than cattle. However, the FASN expression was highest in adipose among the three tissues in both the species. Results indicate structural and functional distinctiveness of bovine FASN. Presence of alternate splicing in buffalo FASN also seems to be a unique phenomenon to the bovines

  3. Green laser irradiation effects on buffalo semen.

    PubMed

    Abdel-Salam, Z; Dessouki, S H M; Abdel-Salam, S A M; Ibrahim, M A M; Harith, M A

    2011-04-01

    The overall objective of this paper is to develop a more sensitive and less costly technique of laser irradiation of spermatozoa at certain wavelengths and exposure times suitable for improvement of buffalo semen quality. A 532 nm continuous wave (CW) DPSS laser light has been used to irradiate buffalo semen for different time intervals. Three semen pools from three different bulls (Bubalus bubalis) were used in the experiment, each pool was divided into six groups : control (not irradiated), and the other five were exposed to laser light for 1, 2, 3, 4 and 5 minutes with fluencies of 0.076, 0.15, 0.23, 0.31, and 0.38 Joule/cm² respectively at an output power 1mW. The results show that the semen quality parameters increase under the effect of laser irradiation. Maximum improvement in the semen quality has been reached after 4 minutes of exposure. Such results indicate the possibility of adopting laser irradiation as an easy and straightforward technique for in situ improvement of the semen quality to optimize the artificial insemination conditions. Copyright © 2011 Elsevier Inc. All rights reserved.

  4. Identification of functional consequence of a novel selection signature in CYP11b1 gene for milk fat content in Bubalus bubalis

    PubMed Central

    Maryam, J.; Babar, Masroor Ellahi; Nadeem, Asif; Yaqub, Tahir; Hashmi, Abu Saeed

    2015-01-01

    Genomic selection for traits of economic importance is an emerging approach carrying tremendous potentials. Many of polygenic traits as milk fat, protein and yield have been characterize at genomic level and important selection signatures have been identified. Cytochrome P450 enzymes are potential loci for affecting many of dairy capabilities. Present study was conducted for genomic dissection of CYP11b1 gene in riverine buffaloes and seven genetic variations were identified. Out of these, one novel polymorphism (p.A313T) was found well associated with milk fat %age. AB genotyped buffaloes were found to have higher milk fat %age (8.9%) for this loci. p.A313T was further validated at larger data set by restriction digestion using CviAII enzyme. Functional consequences of this locus were also predicted by studying three dimensional structure of CYP11b1 protein. For this purpose, 3D protein model was predicted by homology modeling, secondary structural attributes were determined, signal peptide was predicted and a transmembrane helix was also identified. One of polymorphism (p.Y205L) was found in the vicinity of functionally significant F-G loop region, which is the part of protein gets attached to the inner mitochondrial membrane. But this variation could not be associated and needs further investigation. p.A30V, a popular selection marker in cattle, was found in buffaloes as well but could not be associated and might need further confirmation on larger data set. Results of this study illustrate the impending potential of this gene in determining dairy capabilities of buffaloes and might have a role in selection of superior dairy buffaloes. PMID:26629413

  5. The interval between the emergence of pharmacologically synchronized ovarian follicular waves and ovum pickup does not significantly affect in vitro embryo production in Bos indicus, Bos taurus, and Bubalus bubalis.

    PubMed

    Gimenes, Lindsay U; Ferraz, Márcio L; Fantinato-Neto, Paulo; Chiaratti, Marcos R; Mesquita, Lígia G; Sá Filho, Manoel F; Meirelles, Flávio V; Trinca, Luzia A; Rennó, Francisco P; Watanabe, Yeda F; Baruselli, Pietro S

    2015-02-01

    The aim of the present study was to determine the optimal phase of the follicular wave to perform ovum pickup (OPU) for in vitro embryo production (IVEP) in various genetic groups. For this purpose, 27 heifers-nine Bos taurus (Holstein), nine Bos indicus (Nelore), and nine Bubalus bubalis (Mediterranean)-were maintained under the same nutritional, management, and environmental conditions. Heifers within each genetic group were submitted to six consecutive OPU trials with 14-day intersession intervals, at three different phases of the pharmacologically synchronized follicular wave (Day 1, 3, or 5 after follicular wave emergence), in a 3 × 3 crossover design. When OPU was performed at different phases of the pharmacologically synchronized follicular wave (Day 1, 3, or 5), no differences were found in the percent of oocytes recovered (70.5 ± 3.1%, 75.0 ± 3.1%, 76.0 ± 3.2%, respectively; P = 0.41) or blastocyst production rates (19.4 ± 2.9%, 16.6 ± 2.9%, 15.9 ± 2.6%, respectively; P = 0.36). Comparing genetic groups, B indicus showed a higher blastocyst rate (28.3(a) ± 2.8%; P < 0.01) than B taurus and B bubalis (14.1(b) ± 2.9% and 10.2(b) ± 2.0%, respectively). However, only B indicus heifers showed a variation in the number of visualized follicles and the total and viable oocytes along consecutive OPU sessions. In conclusion, different phases of the pharmacologically synchronized ovarian follicular wave did not affect OPU-IVEP in B indicus, B taurus, and B bubalis heifers. Additionally, B indicus heifers showed greater OPU-IVEP efficiency than did the other genetic groups, under the same management conditions.

  6. Whole Mitogenomes Reveal the History of Swamp Buffalo: Initially Shaped by Glacial Periods and Eventually Modelled by Domestication.

    PubMed

    Wang, S; Chen, N; Capodiferro, M R; Zhang, T; Lancioni, H; Zhang, H; Miao, Y; Chanthakhoun, V; Wanapat, M; Yindee, M; Zhang, Y; Lu, H; Caporali, L; Dang, R; Huang, Y; Lan, X; Plath, M; Chen, H; Lenstra, J A; Achilli, A; Lei, C

    2017-07-05

    The newly sequenced mitochondrial genomes of 107 Asian swamp buffalo (Bubalus bubalis carabensis) allowed the reconstruction of the matrilineal divergence since ~900 Kya. Phylogenetic trees and Bayesian skyline plots suggest a role of the glacial periods in the demographic history of swamp buffalo. The ancestral swamp-buffalo mitogenome is dated ~232 ± 35 Kya. Two major macro-lineages diverged during the 2(nd) Pleistocene Glacial Period (~200-130 Kya), but most (~99%) of the current matrilines derive from only two ancestors (SA1'2 and SB) that lived around the Last Glacial Maximum (~26-19 Kya). During the late Holocene optimum (11-6 Kya) lineages differentiated further, and at least eight matrilines (SA1, SA2, SB1a, SB1b, SB2a, SB2b, SB3 and SB4) were domesticated around 7-3 Kya. Haplotype distributions support an initial domestication process in Southeast Asia, while subsequent captures of wild females probably introduced some additional rare lineages (SA3, SC, SD and SE). Dispersal of domestic buffaloes created local population bottlenecks and founder events that further differentiated haplogroup distributions. A lack of maternal gene flow between neighboring populations apparently maintained the strong phylogeography of the swamp buffalo matrilines, which is the more remarkable because of an almost complete absence of phenotypic differentiation.

  7. Elucidating the genetic basis of an oligogenic birth defect using whole genome sequence data in a non-model organism, Bubalus bubalis

    PubMed Central

    Whitacre, Lynsey K.; Hoff, Jesse L.; Schnabel, Robert D.; Albarella, Sara; Ciotola, Francesca; Peretti, Vincenzo; Strozzi, Francesco; Ferrandi, Chiara; Ramunno, Luigi; Sonstegard, Tad S.; Williams, John L.; Taylor, Jeremy F.; Decker, Jared E.

    2017-01-01

    Recent strong selection for dairy traits in water buffalo has been associated with higher levels of inbreeding, leading to an increase in the prevalence of genetic diseases such as transverse hemimelia (TH), a congenital developmental abnormality characterized by absence of a variable distal portion of the hindlimbs. Limited genomic resources available for water buffalo required an original approach to identify genetic variants associated with the disease. The genomes of 4 bilateral and 7 unilateral affected cases and 14 controls were sequenced. A concordance analysis of SNPs and INDELs requiring homozygosity unique to all unilateral and bilateral cases revealed two genes, WNT7A and SMARCA4, known to play a role in embryonic hindlimb development. Additionally, SNP alleles in NOTCH1 and RARB were homozygous exclusively in the bilateral cases, suggesting an oligogenic mode of inheritance. Homozygosity mapping by whole genome de novo assembly also supported oligogenic inheritance; implicating 13 genes involved in hindlimb development in bilateral cases and 11 in unilateral cases. A genome-wide association study (GWAS) predicted additional modifier genes. Although our data show a complex inheritance of TH, we predict that homozygous variants in WNT7A and SMARCA4 are necessary for expression of TH and selection against these variants should eradicate TH. PMID:28045068

  8. Effect of mastitis on luteal function and pregnancy rates in buffaloes.

    PubMed

    Mansour, Mohamed Mohsen; Hendawy, Amin O; Zeitoun, Moustafa M

    2016-09-15

    The aim of this study was to investigate the effects of mastitis on CL development and function and pregnancy rate in buffaloes. Sixty-six buffaloes (Bubalus bubalus) reared in a commercial farm at El-Beheira governorate, north of Egypt were used in this study. According to the visual observation of milk, physical examination of the udder and actual somatic cell count in milk, buffalo cows were divided into three groups: without mastitis (W), n = 23; subclinical mastitis (SC), n = 18; and clinical mastitis (C), n = 25. All buffalo cows were synchronized by double dose of PGF2α (11-day interval) and inseminated by frozen-thawed semen of fertile bull. Mean CL diameter was ultrasonically examined on Days 5, 9, 12, 16, 21, and 25 after artificial insemination (AI). Blood samples were taken on the days of ultrasonography for progesterone (P4) assay. Results indicated that pregnancy rates were lower (P < 0.05) in C (28.00%) and SC (55.56%) compared with W (69.57%) on Day 25 after first AI. Pregnancy rates reduced to 60.87%, 44.45%, and 16.00% in W, SC, and C, respectively, at Day 45 after insemination. Thus, the embryonic loss was 8.7%, 11.11%, and 12.00 % in W, SC, and C cows, respectively. Pregnancy rates decreased between 44.32% and 50.51% when mastitis occurred during Day -15 before to Day +30 after AI, compared with 59.22% in the uninfected cows. The diameter of CL was greater (P < 0.05) in W than SC and C cows starting at Day 9 postbreeding onward. Likewise, P4 concentrations on Days 9 through 25 after AI were greater (P < 0.05) in W cows as compared to SC and C cows. Positive correlations (P < 0.01) were found on Days 5, 9, 12, 16, 21, and 25 after AI between CL diameter and P4 concentrations. Similar trend was found among CL diameter, P4 concentrations, and pregnancy rate. Accordingly, incidence of mastitis revealed suppression to both CL diameter and function leading to significant reduction in pregnancy outcome of buffalo cows. Copyright © 2016 Elsevier Inc

  9. Trypanosoma vivax in water buffalo of the Venezuelan Llanos: An unusual outbreak of wasting disease in an endemic area of typically asymptomatic infections.

    PubMed

    Garcia, Herakles A; Ramírez, Oneyda J; Rodrigues, Carla M F; Sánchez, Rafael G; Bethencourt, Angelica M; Del M Pérez, Gabriela; Minervino, Antonio H H; Rodrigues, Adriana C; Teixeira, Marta M G

    2016-10-30

    Trypanosoma vivax has been associated with asymptomatic infections in African and South American buffalo. In this study, T. vivax was analyzed in water buffalo (Bubalus bubalis) from Venezuela in a molecular survey involving 293 blood samples collected from 2006 to 2015 across the Llanos region. Results demonstrated constant infections (average 23%) during the years analyzed. In general, animals were healthy carriers of T. vivax with low levels of parasitemia and were diagnosed exclusively by TviCATL-PCR. However, an outbreak of severe acute infections mostly in dairy animals was reported during a prolonged drought affecting 30.4% of a buffalo herd (115 animals examined). During the outbreak, animals exhibiting anemia and neurological disorders developed fatal infections, and 7% of the herd died within nine months before treatment against trypanosomosis. Microsatellite locus genotyping (MLG) of T. vivax samples before and during the outbreak revealed similar genotypes, but outbreak isolates exhibited the most divergent MLG. Venezuelan samples from symptomless and sick buffalo did not share the MLGs previously detected in asymptomatic Brazilian buffalo. Trypanosoma evansi was not detected in the herd examined during the outbreak. However, as expected Babesia sp. (62.6%) and Anaplasma sp. (55.6%) infections were highly prevalent in asymptomatic buffalo in the studied areas. This is the first South American outbreak of highly lethal acute T. vivax infections in water buffalo. Our results suggest that chronically infected and asymptomatic buffalo living in areas of enzootic equilibrium can develop symptomatic/lethal disease triggered by stressful scarcity of green forage and water during long droughts, inappropriate management of herds and likely concomitant anaplasmosis and babesiosis. Altogether, these factors weaken buffalo immune defenses, allowing T. vivax to proliferate and, consequently, allowing for progression to wasting disease. Copyright © 2016 Elsevier B

  10. Parentage verification in field progeny testing program of Mehsana buffalo.

    PubMed

    Jakhesara, S J; Rank, D N; Kansara, J D; Parikh, R C; Patel, V M; Vataliya, P H; Solanki, J V

    2012-12-01

    The present study was undertaken to construct a multiplex microsatellite panel for parentage testing in Mehsana buffalo (Bubalus bubalis). The study was based on a total of 212 Mehsana buffalos (100 dams, 100 daughters, and 12 sires). Genomic DNA was extracted from blood and semen samples. A panel of 10 microsatellite markers (CSSM61, ILSTS29, ILSTS17, ILSTS28, CSSM57, CSSM22, ILSTS61, CSSM8, ETH152, and ILSTS11) was amplified in a single multiplex reaction and analyzed by capillary electrophoresis on an automated DNA sequencer. The expected heterozygosity ranged from 0.642 to 0.833 (mean 0.762). The total exclusion probability using 10 microsatellite loci with 1 known parent was 0.993. Seven out of 10 microsatellite loci revealed relatively high polymorphic information content (>0.7). Eighty-one daughters out of 100 daughters qualified by compatibility according to Mendelism. The results suggest that multiplex microsatellite panel is a fast, robust, reliable, and economic tool to verify the parentage as well as to assign the putative sire to daughters under progeny testing with very high accuracy and hence can be used in routine parentage testing.

  11. Biochemical, immunological and kinetic characterization and partial sequence analysis of a thiol proteinase inhibitor from Bubalus bubalis kidney: An attempt targeting kidney disorders.

    PubMed

    Shamsi, Anas; Ahmed, Azaj; Bano, Bilqees

    2017-01-01

    In the present study a thiol proteinase inhibitor was isolated from buffalo kidney making use of ammonium sulphate precipitation and gel filtration chromatography on Sephacryl S-100HR column. Purified inhibitor is homogeneous as it displayed a single band in gel electrophoresis both under reducing and non-reducing environment and is of 65KDa as revealed by gel filtration and SDS PAGE. Kinetic studies revealed the presence of reversible accompanied with competitive mode of inhibition; showing maximum efficacy against papain (Ki=2.90×10(-4)). It was maximally active at pH 8.0 and was stable for a period of 30, 60 and 90 days at 37, 4 and -20°C respectively. Immunological studies confirmed its purity of epitopes as a single precipitin line is obtained in immunodiffusion. N-terminal analysis revealed that it shared a good homology with mouse kidney cystatin as well as with Human Cys C and Cys E thereby advocating its use as a model for various human oriented studies which targets how the kidney cystatin level varies in accordance with various drugs that are currently being used as a target for variety of diseases.

  12. Influence of cutting and deboning operations on the microbiological quality and shelf life of buffalo meat.

    PubMed

    Voloski, F L S; Tonello, L; Ramires, T; Reta, G G; Dewes, C; Iglesias, M; Mondadori, R G; Gandra, E A; da Silva, W P; Duval, E H

    2016-06-01

    Considering the specific biochemical composition of buffalo (Bubalus bubalis) meat (high iron content, high biological value proteins and essential fatty acids, low amounts of fat and cholesterol), we evaluated the influence of cutting and deboning operations on the microbiological quality and shelf-life of vacuum-packed buffalo meat stored under refrigeration. On the processing day, samples were collected from carcass, deboning room surfaces and meat cuts. Samples from meat cuts were evaluated weekly for two months. On the processing day, higher counts of Pseudomonas spp. were observed in samples from meat cuts compared with the hindquarters and the processing surfaces. For thermotolerant coliform scores, the averages were -0.5 log MPN·cm(-2), -0.4 log MPN·cm(-2) and 0.9 log MPN·g(-1), respectively. Higher counts of Pseudomonas spp. and LAB in meat cuts were observed on the processing day and after the first week of storage, respectively, remaining constant during shelf life. Listeria grayi was identified in two samples of hindquarters and meat cuts during storage. Listeria innocua was identified in one meat cut. In conclusion, cutting and deboning operations influence the microbiological quality and shelf life of vacuum-packed buffalo meat stored under refrigeration. Copyright © 2016. Published by Elsevier Ltd.

  13. Cryopreservation Of Nili-Ravi Buffalo Bull Sperm in Cryodiluent Supplemented with Lolium perenne Protein Preparations.

    PubMed

    Qadeer, S; Khan, M A; Ansari, M S; Rakha, B A; Ejaz, R; Husna, A U; Azam, A; Ullah, N; Walker, V K; Akhter, S

    Semen from the Nili-Ravi buffalo bull, Bubalus bubalis, shows poor survival after freeze storage compared to bovine (Bos taurus and Bos indicus) semen. Freeze-susceptibility distinctions in these two genera have been attributed to differences in sperm membranes. We measured the impact of protein preparations derived from a frost-resistant perennial grass, Lolium perenne, with ice recrystallization inhibition activity on the low temperature storage of B. bubalis semen. When the L. perenne preparations (0.1, 1, 10 µg/mL) were added to buffalo semen [2 ejaculates per bull (N=3) per replicate (r=3)] in Tris-citrate extender (50×10(6)sperm mL(-1)), there was no impact on semen quality, as measured by sperm motility and plasma membrane integrity, after storage at 4 degree C (P>0.05). However, when semen supplemented with the grass proteins (0.1 and 1 µg mL(-1)) was evaluated after freezing and storage in liquid nitrogen for 24 h, post-thaw sperm progressive motility and plasma membrane integrity was higher (P<0.05) than in control samples. Post-thaw sperm viability and sperm acrosome integrity was similar (P > 0.05) to controls. The improvement in cryopreserved buffalo sperm progressive motility and plasma membrane integrity suggests that the use of these easily-made preparations may improve fertility after cryopreservation and offers the prospect of improved conception rates after artificial insemination with cryopreservation.

  14. In vitro production of cattlexbuffalo hybrid embryos using cattle oocytes and African buffalo (Syncerus caffer caffer) epididymal sperm.

    PubMed

    Owiny, O D; Barry, D M; Agaba, M; Godke, R A

    2009-04-01

    Interspecies hybridization of bovids occurs between domestic cattle and at least three other species; American bison (Bison bison), yak (Bos grunniens) and banteng (Bos banteng). Birth of a cattlexbuffalo (Bubalus bubalis) hybrid has reportedly occurred in Russia and in China, but these reports were not authenticated. Such hybrids could be important in improving livestock production and management of diseases that impede production in tropical Africa. This study investigated hybridization between cattle and its closest African wild bovid relative, the African buffalo (Syncerus caffer caffer). In an attempt to produce cattlexbuffalo hybrid embryos in vitro, matured cattle oocytes were subjected to a standard in vitro fertilization (IVF) procedure with either homologous cattle (n=1166 oocytes) or heterologous African buffalo (n=1202 oocytes) frozen-thawed epididymal sperm. After IVF, 67.2% of the oocytes inseminated with the homologous cattle sperm cleaved. In contrast, fertilization with buffalo sperm resulted in only a 4.6% cleavage rate. The cleavage intervals were also slower in hybrid embryos than in the IVF-derived cattle embryos. Of the cleaved homologous cattle embryos 52.2% progressed to the morula stage compared with 12.7% for the buffalo hybrid embryos. No hybrid embryos developed beyond the early morula stage, while 40.1% of the cleaved cattlexcattle embryos developed to the blastocyst stage. Transfer of buffalo hybrid IVF embryos to domestic cattle surrogates resulted in no pregnancies at 60 days post-transfer. This study indicates that interspecies fertilization of cattle oocytes with African buffalo epididymal sperm can occur in vitro, and that a barrier to hybridization occurs in the early stages of embryonic development. Chromosomal disparity is likely the cause of the fertilization abnormalities, abnormal development and subsequent arrest impairing the formation of hybrid embryos beyond the early morula stage. Transfer of the buffalo hybrid embryos

  15. Populations of domesticated cattle and buffalo in the Western Forest Complex of Thailand and their possible impacts on the wildlife community.

    PubMed

    Chaiyarat, Rattanawat; Srikosamatara, Sompod

    2009-03-01

    The Western Forest Complex (WEFCOM) of Thailand is comprised of many protected areas and has one of the highest wildlife populations in the country. Populations of wildlife in the WEFCOM have decreased dramatically over recent years. Rapid economic development has resulted in the conversion of forest into agricultural and pastoral land, which has directly and indirectly impacted the wildlife community. This research aimed to evaluate populations of domesticated cattle (Bos indicus) and buffalo (Bubalus bubalis) in the WEFCOM and their possible impacts on the wildlife community. Domesticated cattle and buffalo keepers from 1561 (or 3.3%) of houses in and near WEFCOM were interviewed. The average number of animals per household was 15.6 cattle and 8.5 buffalo. Most villagers released domesticated cattle and buffalo to forage in the protected areas. This tended to have a high impact on the wildlife community in Huai Kha Khaeng Wildlife Sanctuary and Tungyai Naresuan Wildlife Sanctuary. The least impacted areas were Luam Khlong Ngu National Park, Thong Pha Phum National Park and Chaleam Ratanakosin National Park. With a high risk to the wildlife community, law enforcement should be used in combination with a certain level of co-management with local communities.

  16. OPTIXcell improves the postthaw quality and fertility of buffalo bull sperm.

    PubMed

    Ansari, M S; Rakha, B A; Akhter, S; Ashiq, M

    2016-02-01

    The present study was conducted to compare the liposome-containing, animal protein-free, commercially available OPTIXcell extender with the Tris-citric-egg yolk extender for postthaw quality and fertility of buffalo semen. Semen was collected from five adult Nili-Ravi buffalo (Bubalus bubalis) bulls of similar age group with an artificial vagina (at 42 °C) for 3 weeks (replicates). Semen ejaculates from each buffalo bull were divided into two aliquots and diluted (at 37 °C having 50 × 10(6) spermatozoa/mL) in the OPTIXcell or Tris-citric-egg yolk (control) extender. Diluted semen was cooled to 4 °C in 2 hours, equilibrated for 4 hours, and filled in 0.5-mL straws. The semen straws were kept over liquid nitrogen vapors (5 cm) for 10 minutes. The straws were then plunged and stored in liquid nitrogen (-196 °C). After 24 hours of storage, the semen straws were thawed at 37 °C for 30 seconds to assess postthaw quality. Percentages of sperm motility, plasma membrane integrity, viability, and acrosomal integrity were improved (P < 0.05) in the OPTIXcell extender compared to the Tris-citric-egg yolk extender. Values for DNA integrity (%) did not differ in the OPTIXcell and Tris-citric-egg yolk extenders. The overall conception rate in buffaloes was improved (P < 0.05) with semen cryopreserved in the OPTIXcell extender (59.5%) compared to semen cryopreserved in the Tris-citric-egg yolk extender (41.5%). It is concluded that the liposome-containing commercially available OPTIXcell extender is more efficient to conserve postthaw quality and resulted in higher fertility rate of buffalo in the field. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Polymorphism and gene organization of water buffalo MHC-DQB genes show homology to the BoLA DQB region.

    PubMed

    Sena, L; Schneider, M P C; Brenig, B B; Honeycutt, R L; Honeycutt, D A; Womack, J E; Skow, L C

    2011-08-01

    In cattle (Bos taurus), there is evidence of more than 50 alleles of BoLA-DQB (bovine lymphocyte antigen DQB) that are distributed across at least five DQB loci, making this region one of the most complex in the BoLA gene family. In this study, DQB alleles were analysed for the water buffalo (Bubalus bubalis), another economically important bovine species. Twelve alleles for Bubu-DQB (Bubalis bubalis DQB) were determined by nucleotide sequence analysis. A phylogenetic analysis revealed numerous trans-species polymorphisms, with alleles from water buffalo assigned to at least three different loci (BoLA-DQB1, BoLA-DQB3 and BoLA-DQB4) that are also found in cattle. These presumptive loci were analysed for patterns of synonymous (d(S)) and non-synonymous (d(N)) substitution. Like BoLA-DQB1, Bubu-DQB1 was observed to be under strong positive selection for polymorphism. We conclude that water buffalo and cattle share the current arrangement of their DQB region because of their common ancestry. © 2011 The Authors, Animal Genetics © 2011 Stichting International Foundation for Animal Genetics.

  18. Bovine herpesvirus type 1 marker vaccine induces cross-protection against bubaline herpesvirus type 1 in water buffalo.

    PubMed

    Montagnaro, Serena; De Martinis, Claudio; Iovane, Valentina; Ciarcia, Roberto; Damiano, Sara; Nizza, Sandra; De Martino, Luisa; Iovane, Giuseppe; Pagnini, Ugo

    2014-09-01

    Water buffalo (Bubalus bubalis) are susceptible to bovine herpesvirus type 1 (BoHV-1) and a species-specific herpesvirus, bubaline herpesvirus type 1 (BuHV-1). In this study, an attenuated marker BoHV-1 based vaccine against BuHV-1 challenge was evaluated to determine whether it induces protection from viral replication. One group of water buffalo calves was immunized with an attenuated BoHV-1 marker vaccine. A second group was not vaccinated and used as the control. During the post-vaccination period, we monitored the humoral immune response. The efficacy of the vaccine was tested after intranasal challenge of the calves with a BuHV-1 strain. The experiment showed that after vaccination, BuHV-1 replication was significantly reduced by approximately three titer points compared to the controls. The control animals showed high levels of viral shedding and mild signs associated with BuHV-1 infection. Therefore, our study provides evidence for the existence of cross-protection between BoHV-1 and BuHV-1 in buffalo calves.

  19. Physical mapping of 20 unmapped fragments of the btau_4.0 genome assembly in cattle, sheep and river buffalo.

    PubMed

    De Lorenzi, L; Genualdo, V; Perucatti, A; Iannuzzi, A; Iannuzzi, L; Parma, P

    2013-01-01

    The recent advances in sequencing technology and bioinformatics have revolutionized genomic research, making the decoding of the genome an easier task. Genome sequences are currently available for many species, including cattle, sheep and river buffalo. The available reference genomes are very accurate, and they represent the best possible order of loci at this time. In cattle, despite the great accuracy achieved, a part of the genome has been sequenced but not yet assembled: these genome fragments are called unmapped fragments. In the present study, 20 unmapped fragments belonging to the Btau_4.0 reference genome have been mapped by FISH in cattle (Bos taurus, 2n = 60), sheep (Ovis aries, 2n = 54) and river buffalo (Bubalus bubalis, 2n = 50). Our results confirm the accuracy of the available reference genome, though there are some discrepancies between the expected localization and the observed localization. Moreover, the available data in the literature regarding genomic homologies between cattle, sheep and river buffalo are confirmed. Finally, the results presented here suggest that FISH was, and still is, a useful technology to validate the data produced by genome sequencing programs. Copyright © 2013 S. Karger AG, Basel.

  20. Microphthalmia-associated transcription factor mutations are associated with white-spotted coat color in swamp buffalo.

    PubMed

    Yusnizar, Y; Wilbe, M; Herlino, A O; Sumantri, C; Noor, R Rachman; Boediono, A; Andersson, L; Andersson, G

    2015-12-01

    A candidate gene analysis of the microphthalmia-associated transcription factor (MITF) gene was used in an attempt to identify the genetic basis for a white-spotted coat color phenotype in the Asian swamp buffalo (Bubalus bubalis carabanensis). Ninety-three buffaloes-32 solid, 38 spotted and 23 white individuals-were Sanger-sequenced for all MITF exons as well as highly conserved intronic and flanking regions. MITF cDNA representing skin and iris tissue from six spotted, nine solid and one white buffaloes was also Sanger-sequenced to confirm detected mutations. Two independent loss-of-function mutations, a premature stop codon (c.328C>T, p.Arg110*) and a donor splice-site mutation (c.840+2T>A, p.Glu281_Leu282Ins8), both of which cause white-spotted coat color in swamp buffaloes, were identified. The nonsense mutation leads to a premature stop codon in exon 3, and likely removal of the resulting mRNA via nonsense-mediated decay pathway, whereas the donor splice-site mutation leads to aberrant splicing of exon 8 that encodes part of a highly conserved region of MITF. The resulting insertion of eight amino acid residues is expected to perturb the leucine zipper part in the basic helix-loop-helix leucine zipper (bHLH-Zip) domain and will most likely influence dimerization and DNA binding capacity. Electrophoretic mobility shift assay was performed using mutant and wild-type MITF proteins and showed that the mutant MITF protein resulting from the splice-site mutation decreased in vitro DNA binding capacity compared to wild-type MITF. White-spotted buffalo bulls are sacrificed in funeral ceremonies in Tana Toraja, Indonesia, because they are considered holy, and our results show that genetic variation causes a tie to the cultural use of these buffaloes.

  1. Low incidence of bovine leukocyte adhesion deficiency (BLAD) carriers in Indian cattle and buffalo breeds.

    PubMed

    Patel, Rajesh K; Singh, Krishna M; Soni, Kalpesh J; Chauhan, Jenabhai B; Sambasiva Rao, Krothapalli R S

    2007-01-01

    BLAD is an autosomal recessive genetic disease that affects Holstein-Friesian (HF) cattle worldwide. It is a disease characterized by a reduced expression of the adhesion molecules on neutrophils. The disease is caused by a mutation that replaces adenine at 383 with guanine, which causes an amino acid change from aspartic acid to glycine. Blood samples and a few semen samples were collected from 1250 phenotypically normal individuals, including HF (N=377), HF crossbred (N=334), Jersey (105), other breeds of cattle (N=160) and water buffalo Bubalus bubalis (N=274) belonging to various artificial insemination stations, bull mother farms (BMFs) and embryo transfer (ET) centres across the country. PCR-RFLP was performed to detect a point mutation in CD18, surface molecules of neutrophils. The results indicate that out of 1250 cattle and buffaloes tested for BLAD, 13 HF purebreds out of 377 and 10 HF crossbreds out of 334 appear to be BLAD carriers. In the HF and HF crossbred population, the percentage of BLAD carriers was estimated as 3.23%. The condition is alarming as the mutant gene has already entered the HF crossbred cattle population and therefore, the population of HF and its crossbreds needs regular screening to avoid the risk of spreading BLAD in the breeding cattle population of India.

  2. Actin Polymerization: An Event Regulated by Tyrosine Phosphorylation During Buffalo Sperm Capacitation.

    PubMed

    Naresh, S; Atreja, S K

    2015-12-01

    In the female reproductive tract, the spermatozoa undergo a series of physiological and biochemical changes, prior to gaining the ability to fertilize, that result to capacitation. However, the actin polymerization and protein tyrosine phosphorylation are the two necessary steps for capacitation. In this study, we have demonstrated the actin polymerization and established the correlation between protein tyrosine phosphorylation and actin reorganization during in vitro capacitation in buffalo (Bubalus bubalis) spermatozoa. Indirect immunofluorescence and Western blot techniques were used to detect actin polymerization and tyrosine phosphorylation. The time-dependent fluorimetric studies revealed that the actin polymerization starts from the tail region and progressed towards the head region of spermatozoa during capacitation. The lysophosphatidyl choline (LPC)-induced acrosome reaction (AR) stimulated quick actin depolymerization. The inhibitor cytochalasin D (CD) blocked the in vitro capacitation by inhibiting the actin polymerization. In addition, we also performed different inhibitor (Genistein, H-89, PD9809 and GF-109) and enhancer (dbcAMP, H(2)O(2) and vanadate) studies on actin tyrosine phosphorylation and actin polymerization. The inhibitors of tyrosine phosphorylation inhibit actin tyrosine phosphorylation and polymerization, whereas enhancers of tyrosine phosphorylation stimulate F-actin formation and tyrosine phosphorylation. These observations suggest that the tyrosine phosphorylation regulates the actin polymerization, and both are coupled processes during capacitation of buffalo spermatozoa.

  3. In vitro embryo development and blastocyst hatching rates following vitrification of river buffalo embryos produced from oocytes recovered from slaughterhouse ovaries or live animals by ovum pick-up.

    PubMed

    Manjunatha, B M; Gupta, P S P; Ravindra, J P; Devaraj, M; Nandi, S

    2008-03-03

    The present study was undertaken to determine whether the source of oocytes (ovum pick up versus slaughterhouse ovaries) affected in vitro embryo production and embryo survival (as measured by blastocyst hatching rates) following vitrification in buffaloes (Bubalus bubalis). Oocytes recovered from live buffaloes (n=6) by ovum pick up (OPU) and by manual aspiration from slaughterhouse ovaries were in vitro matured, fertilized and cultured to blastocyst stage under same culture conditions. Vitrification of blastocysts was carried out in two steps at 24 degrees C. Embryos were equilibrated in 10% EG+10% DMSO+0.3 M sucrose in base medium for 4 min. Subsequently, the embryos were transferred into 25% EG+25% DMSO+0.3 M sucrose in base medium for 45 s and then the embryos were loaded into straws and immersed in liquid nitrogen. Following warming, blastocysts were cultured in vitro for 48 h to assess hatching. Oocytes derived from live animals by OPU resulted in a significantly higher blastocyst yield then those derived from slaughterhouse ovaries (30.6+/-4.3 versus 18.5+/-1.8). Blastocyst hatching rates following vitrification of buffalo embryos produced from the oocytes collected from live animals by OPU was significantly higher than the oocytes collected from slaughterhouse ovaries (52.8+/-4.2 versus 40.2+/-4.4). In conclusion, the present study showed that source of oocytes (OPU versus slaughterhouse ovaries) affects the in vitro embryo development and blastocyst hatching rates following vitrification of embryos in buffaloes.

  4. Neospora caninum serostatus is affected by age and species variables in cohabiting water buffaloes and beef cattle.

    PubMed

    Moore, D P; Konrad, J L; San Martino, S; Reichel, M P; Cano, D B; Méndez, S; Späth, E J L; Odeón, A C; Crudeli, G; Campero, C M

    2014-07-14

    The aim of this study was to investigate how Neospora caninum serostatus may be affected by variables such as host species (water buffaloes or cattle) and age in animals cohabiting in the same ranch. A convenience cross-sectional study was performed on four ranches in the Northeast of Argentina, where water buffalo are cohabitating with beef cattle. Blood samples were collected from 1350 female water buffaloes (Bubalus bubalis) and 880 female beef cattle (Bos taurus and Bos indicus crossbreeds) from four ranches. Calving and weaning percentages at herd level for each ranch were also recorded. N. caninum antibody levels were measured by an indirect fluorescent antibody test (IFAT) (reciprocal antibody titers ≥ 100). Serological results were classified into 2 categories (0: negative; 1: positive). A logistic regression model was used to describe the relationship between N. caninum serostatus and specie (water buffalo or cattle), age or ranch and their interactions. Likelihood ratio tests were used to assess the significance of the model and their terms. Odds ratios were estimated and 95% profile likelihood (LR) and Wald confidence intervals (CI) obtained. Overall, specific antibody titers were found in 43.3% (584/1350) of water buffaloes and 28.6% (252/880) of cattle. Seropositive water buffaloes and cattle were observed on all ranches. Age was statistically significant (p=0.01) with an overall estimate of logit (log odds) of age of 0.03 for both species. This indicates that for every one year increase in age, the expected change in log odds of being seropositive increased by 0.03. On three of four ranches a water buffalo was 4.48, 1.54 and 2.25 times more likely to be seropositive than cattle for animals of the same age. The N. caninum serostatus was affected by age in the first place, but also by species on at least three of the four ranches. Calving and weaning percentages were higher in water buffaloes than in beef cattle (p<0.05). Even though the low

  5. Control of ovulation with a GnRH agonist after superstimulation of follicular growth in buffalo: fertilization and embryo recovery.

    PubMed

    Carvalho, Nelcio A T; Baruselli, Pietro S; Zicarelli, Luigi; Madureira, Edward H; Visintin, Jose A; D'Occhio, Michael J

    2002-12-01

    The potential to use a GnRH agonist bioimplant and injection of exogenous LH to control the time of ovulation in a multiple ovulation and embryo transfer (MOET) protocol was examined in buffalo. Mixed-parity buffalo (Bubalus bubalis; 4-15-year-old; 529 +/- 13 kg LW) were randomly assigned to one of five groups (n = 6): Group 1, conventional MOET protocol; Group 2, conventional MOET with 12 h delay in injection of PGF2alpha; Group 3, implanted with GnRH agonist to block the preovulatory surge release of LH; Group 4, implanted with GnRH agonist and injected with exogenous LH (Lutropin, 25 mg) 24 h after 4 days of superstimulation with FSH; Group 5, implanted with GnRH agonist and injected with LH 36 h after superstimulation with FSH. Ovarian follicular growth in all buffaloes was stimulated by treatment with FSH (Folltropin-V, 200 mg) administered over 4 days, and was monitored by ovarian ultrasonography. At the time of estrus, the number of follicles >8 mm was greater (P < 0.05) for buffaloes in Group 2 (12.8) than for buffaloes in Groups 1(8.5), 3 (7.3), 4 (6.1) and 5 (6.8), which did not differ. All buffaloes were mated by Al after spontaneous (Groups 1-3) or induced (Groups 4 and 5) ovulation. The respective number of buffalo that ovulated, number of corpora lutea, ovulation rate (%), and embryos + oocytes recovered were: Group 1 (2, 1.8 +/- 1.6, 18.0 +/- 13.6, 0.2 +/- 0.2); Group 2 (4,6.1 +/- 2.9, 40.5 +/- 17.5, 3.7 +/- 2.1); Group 3 (0, 0, 0, 0); Group4 (6, 4.3 +/- 1.2, 69.3 +/- 14.2, 2.0 +/- 0.9); and Group 5 (1, 2.5 +/- 2.5, 15.5 +/- 15.5, 2.1 +/- 2.1). All buffaloes in Group 4 ovulated after injection of LH and had a relatively high ovulation rate (69%) and embryo recovery (46%). It has been shown that the GnRH agonist-LH protocol can be used to improve the efficiency of MOET in buffalo.

  6. A distinct genetic population of Gongylonema pulchrum from water buffaloes in Nepal.

    PubMed

    Makouloutou, Patrice; Rana, Hari Bahadur; Adhikari, Bishunu; Devkota, Bhuminand; Dhakal, Ishwari Prasad; Sato, Hiroshi

    2013-08-01

    Whole-length esophagi of 111 Murrah cross water buffaloes (Bubalus bubalis) were collected in the Kathmandu and Chitwan districts of Nepal from December 2009 to February 2010. Gullet worms showing a typical epithelium-dwelling character were detected in 13 of 53 (24.5%) buffaloes in Kathmandu and in 5 of 58 (8.6%) buffaloes in Chitwan. The worms' morphology and measurements were identical to those of Gongylonema pulchrum Molin, 1857, except for the length of the left spicules relative to the body length. Scanning electron microscopy did not detect any further morphological differences regarding the collected specimen from Nepal compared with G. pulchrum . The ribosomal RNA gene (rDNA), including internal transcribed spacer (ITS) 1 and 2, and a partial region of the cytochrome c oxidase subunit I (COI) of mitochondrial DNA of the worms were characterized and compared with those of G. pulchrum collected from cattle, deer, wild boars, and monkeys in Japan and from cattle in Iran. The 18S, 5.8S, and 28S rDNA nucleotide sequences of the buffalo-collected worms had 99.8% (1,779/1,782), 100% (158/158), and 98.3-98.8% (3,494-3,507/3,551) identities, respectively, with those of G. pulchrum from the other host mammals. The ITS regions exhibited higher variations between the buffalo-collected worms and G. pulchrum from the other host mammals (85-88% identity for ITS1 and 56-80% identity for ITS2). The COI also showed lower identities (89.2-90.2%), although only a single amino acid substitution was noted compared with the majority of G. pulchrum samples collected in Japan. Based on these molecular genetic characters in the rDNA and COI mitochondrial DNA, together with a shorter left spicule length relative to body length, the gullet worms isolated from buffaloes in Nepal might belong to a distinct local or buffalo-preferring population of G. pulchrum, although its geographical distribution on the continent and host specificity remain to be clarified.

  7. Efficiency of beetle (Dendroides canadensis) recombinant antifreeze protein for buffalo semen freezability and fertility.

    PubMed

    Qadeer, S; Khan, M A; Shahzad, Q; Azam, A; Ansari, M S; Rakha, B A; Ejaz, R; Husna, A U; Duman, J G; Akhter, S

    2016-10-15

    Overwintering larvae of the beetle Dendroides canadensis produce potent antifreeze proteins to inhibit inoculative freezing and promote supercooling. We hypothesized that addition of Dendroides canadensis recombinant antifreeze proteins (DAFPs) in the extender will improve the quality and fertility of cryopreserved Nili-Ravi buffalo (Bubalus bubalis) sperm. The study was divided into two parts: (1) Evaluation of the effect of DAFPs on the quality of frozen-thawed buffalo bull sperm and (2) Examination of the fertilizing ability of frozen-thawed buffalo bull sperm. Semen was collected from three bulls using an artificial vagina (42 °C). Qualifying ejaculates from each bull were divided into four aliquots and diluted (at 37 °C, 50 × 10(6) sperm/mL) in tris-citric acid extender containing DAFP (at 0.1, 1.0, and 10 μg/mL), and the sperm were evaluated for important characteristics relative to a control without DAFP. D canadensis recombinant antifreeze proteins at any of the three concentrations did not affect sperm progressive motility or plasma membrane integrity (PMI), either before or after the semen was cooled to 4 °C in 2 hours. However, after 24 hours of cryostorage at -196 °C, followed by thawing at 37 °C for 30 seconds, sperm progressive motility and PMI were higher (P < 0.05) in extender containing DAFP at 10 μg/mL compared with control. The in vitro-fertilizing ability of cryopreserved (-196 °C) sperm supplemented with DAFP (10 μg/mL) was slightly higher (P = 0.098) compared with control, as assessed through in vitro cleavage rate of in vitro matured buffalo oocytes. Also, the in vivo fertility rate was evaluated by inseminating 100 buffaloes (50 inseminations per extender) 12 hours after standing heat. The fertility rate of cryopreserved buffalo bull sperm in terms of positive pregnancy at 90 days after insemination was clinically higher but remained statistically nonsignificant in extender containing DAFP at 10 μg/mL (52

  8. Multilocus phylogeographical analysis of Trypanosoma (Megatrypanum) genotypes from sympatric cattle and water buffalo populations supports evolutionary host constraint and close phylogenetic relationships with genotypes found in other ruminants.

    PubMed

    Garcia, Herakles A; Rodrigues, Adriana C; Martinkovic, Franjo; Minervino, Antonio H H; Campaner, Marta; Nunes, Vânia L B; Paiva, Fernando; Hamilton, Patrick B; Teixeira, Marta M G

    2011-11-01

    Species of the subgenus Trypanosoma (Megatrypanum) have been reported in cattle and other domestic and wild ruminants worldwide. A previous study in Brazil found at least four genotypes infecting cattle (Bos taurus), but only one in water buffalo (Bubalus bubalis). However, the small number of isolates examined from buffalo, all inhabiting nearby areas, has precluded evaluation of their diversity, host associations and geographical structure. To address these questions, we evaluated the genetic diversity and phylogeographical patterns of 25 isolates from water buffalo and 28 from cattle from four separate locations in Brazil and Venezuela. Multigene phylogenetic analyses of ssrRNA, internal transcribed spacer of rDNA (ITSrDNA), 5SrRNA, glycosomal glyceraldehyde 3-phosphate dehydrogenase (gGAPDH), mitochondrial cytochrome b (Cyt b), spliced leader (SL) and cathepsin L-like (CATL) sequences positioned all isolates from sympatric and allopatric buffalo populations into the highly homogeneous genotype TthIA, while the cattle isolates were assigned to three different genotypes, all distinct from TthIA. Polymorphisms in all of these sequences separated the trypanosomes infecting water buffalo, cattle, sheep, antelope and deer, and suggested that they correspond to separate species. Congruent phylogenies inferred with all genes indicated a predominant clonal structure of the genotypes. The multilocus analysis revealed one monophyletic assemblage formed exclusively by trypanosomes of ruminants, which corresponds to the subgenus T. (Megatrypanum). The high degree of host specificity, evidenced by genotypes exclusive to each ruminant species and lack of genotype shared by different host species, suggested that the evolutionary history of trypanosomes of this subgenus was strongly constrained by their ruminant hosts. However, incongruence between ruminant and trypanosome phylogenies did not support host-parasite co-evolution, indicating that host switches have occurred across

  9. Improving smallholder food security through investigations of carcass composition and beef marketing of buffalo and cattle in northern Lao PDR.

    PubMed

    Nampanya, Sonevilay; Khounsy, Syseng; Phonvisay, Aloun; Bush, Russell David; Windsor, Peter Andrew

    2015-04-01

    This study determined the carcass composition of Lao indigenous buffalo (Bubalus bubalis) and cattle (Bos indicus), then examined trends in bovine meat marketing following review of records of beef production and prices in the two major cities of Luang Prabang (LPB) and Xieng Khoung (XK) provinces in northern Laos. Samples from 41 buffalo and 81 cattle (n = 122) were collected from animals slaughtered in May-June 2014, with live weights, carcass weights and other carcass-related variables collected. The animals were classified into four age cohort groups (<2, 2-<4, 4-6 and >6 years) with quantitative and dichotomous qualitative traits determined. There were significant differences in buffalo and cattle predicted mean carcass weights between age classification categories (p = 0.003 and 0.001) but not in dressing percentages (p = 0.1 and 0.1). The carcass weight of buffalo was 104 (±23.1)-176 (±12.0) kg compared to 65 (±8.7)-84 (±6.5) kg of cattle, with dressing percentages of 37-40 and 39-42 %, respectively. Despite an average bovine meat price increase of 42-48 % between 2011 and 2013, there was a reduction in the numbers of large ruminants slaughtered in the surveyed cities of LPB (11 %) and XK (7 %), with bovine meat availability per person of 5.2-6.6 kg (LPB) and 3.0-3.8 kg (XK). Improving the sustainability of the bovine meat supply in Laos requires a systems approach involving improvements to animal health and production, livestock marketing, plus the critical development of improved slaughterhouse facilities enabling a meat-processing sector to emerge. This development pathway is of particular importance for building the capacity of Laos to reduce food insecurity and alleviate the poverty of its largely rural smallholder community.

  10. Dispositions of enrofloxacin and its major metabolite ciprofloxacin in Thai swamp buffaloes

    PubMed Central

    RUENNARONG, Nitwarat; WONGPANIT, Kannika; SAKULTHAEW, Chainarong; GIORGI, Mario; KUMAGAI, Susumu; POAPOLATHEP, Amnart; POAPOLATHEP, Saranya

    2015-01-01

    Given the limited information available in this species, the aim of this study was to investigate the pharmacokinetic characteristics of enrofloxacin (ER) and its major metabolite ciprofloxacin (CP) in buffaloes, Bubalus bubalis. ER was administered intravenously (i.v.) or subcutaneously (s.c.) to buffaloes at doses of 5.0 and 7.5 mg/kg BW, and plasma, urine and fecal samples were collected until 48 hr post-administration. The concentrations of ER and CP in the plasma, urine and feces were analyzed using high-performance liquid chromatography equipped with a fluorescence detector. The plasma concentrations of ER and CP could be determined up to 24 hr and 32 hr after i.v. and s.c. administrations at doses of 5.0 and 7.5 mg/kg BW, respectively. CP concentrations were always lower than those of parental drug. The s.c. bioavailability of ER was 52.36 ± 4.24% and 72.12 ± 5.39% at doses of 5.0 and 7.5 mg/kg BW, respectively. Both ER and CP were detectable in urine and feces up to 24 hr. ER and CP were mainly excreted via the urine. Based on the pharmacokinetic data and PK-PD indices, s.c. administration of ER at doses of 5.0 and 7.5 mg/kg BW might be appropriate for the treatment of susceptible bacterial diseases in Thai swamp buffaloes. PMID:26596287

  11. Isolation of a gene encoding a cellulolytic enzyme from swamp buffalo rumen metagenomes and its cloning and expression in Escherichia coli.

    PubMed

    Cheema, Tanzeem Akbar; Jirajaroenrat, Kanya; Sirinarumitr, Theerapol; Rakshit, Sudip K

    2012-01-01

    Ruminants are capable of hydrolyzing lignocellulosic residues to absorbable sugars by virtue of the microbial communities residing in their rumen. However, large sections of such microbial communities are not yet culturable using conventional laboratory techniques. Therefore in the present study, the metagenomic DNA of swamp buffalo (Bubalus bubalis) rumen contents was explored using culture-independent techniques. The consensus regions of glycosyl hydrolase 5 (GH5) family of cellulases were used as primers for PCR amplification. A full-length metagenomic cellulase gene, Umcel5B29, with a complete open reading frame (ORF) of 1611 bp was identified. The similarity search analysis revealed that Umcel5B29 is closely related to the cellulases (73% to 98% similarity) of ruminal unculturable microorganisms, indicating its phylogenetic origin. Further analysis indicated that Umcel5B29 does not contain a carbohydrate binding module (CBM). Subsequently, Umcel5B29 was overexpressed in Escherichia coli. The recombinant enzyme worked optimally at pH 5.5 and 45°C, a condition similar to the buffalo's rumen. However, the enzyme retained more than 70% of its maximal activity after incubation at pH 4-7 and more than 50% maximal activity after incubation at 30-60°C for 30 min. These characteristics render Umcel5B29 as a potential candidate for the bio-stoning process of denim.

  12. Advanced comparative cytogenetic analysis of X chromosomes in river buffalo, cattle, sheep, and human.

    PubMed

    Perucatti, A; Genualdo, V; Iannuzzi, A; Rebl, A; Di Berardino, D; Goldammer, T; Iannuzzi, Leopoldo

    2012-05-01

    Based on a recently generated comprehensive gene map for Ovis aries chromosome X (OARX) with an approximately even locus distribution, we assigned selected bacterial artificial chromosome (BAC) probes corresponding to these OARX loci to Bubalus bubalis (BBU) and Bos taurus (BTA) by comparative fluorescence in-situ hybridization (FISH) to improve cytogenetically the X chromosome maps in these species. Twenty-five added loci in BBUX and BTAX, respectively, contribute to a more detailed description of the cytogenetic organization of these chromosomes. Further seven loci were identified in OARX and two DNA probes were assigned to X and Y chromosomes in river buffalo, cattle, and sheep, respectively, and thus identified loci in the pseudoautosomal region. The additional assignments double the number of cytogenetic loci in BBUX and increase their number in BTAX and OARX. The larger quantity of cytogenetic anchors allows a more precise morphological comparison of bovid X chromosomes among each other and with the Homo sapiens (HSA) X chromosome. The anchor loci confirm and refine syntenic fragments in HSAX and identify several evolutionary breakpoints between the compared chromosomes. The cytogenetic assignments in BBUX, BTAX, and OARX represent useable anchors for the ongoing genome sequence assembly in Bovidae.

  13. Ovum pick-up and in vitro embryo production (OPU-IVEP) in Mediterranean Italian buffalo performed in different seasons.

    PubMed

    Di Francesco, Serena; Novoa, Maria Virginia Suarez; Vecchio, Domenico; Neglia, Gianluca; Boccia, Lucia; Campanile, Giuseppe; Zicarelli, Luigi; Gasparrini, Bianca

    2012-01-01

    This study was designed to evaluate the effect of season on in vivo oocyte recovery and embryo production in Mediterranean Italian buffalo (Bubalus bubalis). For this purpose repeated transvaginal ultrasound-guided ovum pick up (OPU) was conducted twice a week throughout autumn, mid-winter (transitional period) and spring-summer. The number and size of follicles was determined before puncture. The recovered oocytes were first classified in morphological categories and then used for in vitro embryo production (IVEP) according to standard procedures. The mean number of total follicles observed per session did not differ among the three periods we examined (on average 4.6). Although season did not considerably affect the number of oocytes recovered (on average 2.3/buffalo/session), the number of degenerated and abnormally expanded oocytes increased during autumn. Furthermore, the percentage of abnormally expanded oocytes significantly increased during autumn (6.1%) compared with both the transitional period and spring-summer (1.9 and 2.3%, respectively). Interestingly, the embryo output we recorded at day 7, in terms of tight morulae-blastocysts was higher in autumn (30.9%) compared to the other two periods (13.3% and 10.3%, respectively, in spring-summer and in the transitional period; P<0.01). The results of this trial demonstrated that the morphological features of the oocytes did not vary substantially among the considered periods, with the exception of degenerated and abnormally expanded oocytes. On the other hand, the oocyte developmental competence improved in autumn compared to spring-summer and the transitional period. This datum reflects buffalo reproductive pattern expressed in vivo at Italian latitudes. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Evaluation of thermal comfort, physiological, hematological, and seminal features of buffalo bulls in an artificial insemination station in a tropical environment.

    PubMed

    Barros, Daniel Vale; Silva, Lilian Kátia Ximenes; de Brito Lourenço, José; da Silva, Aluizio Otávio Almeida; E Silva, André Guimarães Maciel; Franco, Irving Montanar; Oliveira, Carlos Magno Chaves; Tholon, Patrícia; Martorano, Lucieta Guerreiro; Garcia, Alexandre Rossetto

    2015-06-01

    This study aimed to assess the variation over time in thermal comfort indices and the behavior of physiological parameters related to thermolysis, blood parameters, and semen in natura of buffalo bulls reared in tropical climate. The study was carried out in an artificial insemination station under a humid tropical climate (Afi according to Köppen). Ten water buffalo bulls (Bubalus bubalis) were used during the 5 months (April to August) of study. The environmental Temperature Humidity Index (THId) and the pen microclimate Temperature Humidity Index (THIp) were calculated. Every 25 days, respiratory rate (RR), heart rate (HR), rectal temperature (RT), and Benezra's thermal comfort index (BTCI) were assessed in the morning and in the afternoon. A blood assay was performed every month, while semen was collected weekly. THIp did not vary over the months (P > 0.05) and was higher in the afternoon than in the morning (77.7 ± 2.6 versus 81.8 ± 2.1, P < 0.05). RR, HR, and BTCI significantly increased over the months and were different between the periods of the day (P > 0.05) but within the physiological limits. RT varied between the periods of the day and decreased over the months, being the lowest in August (37.8 ± 0.7 °C), time-impacted hematocrit, mean corpuscular volume, hemoglobin levels, and spermatic gross motility and vigor (P < 0.05). Thus, buffalo bulls reared under a humid tropical climate may have variations in thermal comfort during the hotter periods but are able to efficiently activate thermoregulatory mechanisms and maintain homeothermy, hence preserving their physiological and seminal parameters at normal levels.

  15. Lipid peroxidation, mitochondrial membrane potential and DNA integrity of spermatozoa in relation to intracellular reactive oxygen species in liquid and frozen-thawed buffalo semen.

    PubMed

    Kadirvel, G; Kumar, Satish; Kumaresan, A

    2009-08-01

    Several factors affect sperm motility and functional integrity during preservation; the damage caused by reactive oxygen species (ROS) being an important factor. The present study investigated intracellular ROS generation and its relationship with sperm motility, lipid peroxidation (LPO), mitochondrial membrane potential (MMP) and DNA integrity during preservation (liquid preservation at 4 degrees C and cryopreservation) of buffalo semen. Fifty-six ejaculates, eight each from seven buffalo bulls (Bubalus bubalis) were utilized for the study. Intracellular ROS level was detected using 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA) and propodium iodide (PI) by flow cytometry. 3,3'-Dihexyloxacarbocyanine iodide [DiOC(6) (3)]/PI and acridine orange were used for detection of MMP and DNA integrity of spermatozoa, respectively. Results revealed that ROS and LPO level in sperm increased linearly between 0 h and 72 h of liquid preservation at 4 degrees C, with significantly (P<0.01) higher levels at 48 h and 72 h of storage compared to fresh semen. The ROS level in viable sperm in frozen-thawed semen did not differ significantly from fresh semen, but the LPO was significantly (P<0.05) higher in frozen-thawed sperm compared to fresh sperm. There was a linear reduction in the sperm with high MMP and DNA integrity in liquid semen, which was significantly (P<0.01) higher at 48 h and 72 h of storage compared to fresh semen. The intracellular ROS was strongly associated to sperm motility, LPO, MMP and DNA integrity during liquid preservation, while this association did not exist in frozen-thawed sperm. The study concluded that ROS generation and its associated effects are likely to be an important contributor to the reduced sperm motility and functional integrity during liquid preservation of buffalo semen at 4 degrees C, but ROS generation and its damage had only minor effects during freezing and thawing process.

  16. Use of real-time PCR technique in studying rumen cellulolytic bacteria population as affected by level of roughage in swamp buffalo.

    PubMed

    Wanapat, Metha; Cherdthong, Anusorn

    2009-04-01

    A real-time polymerase chain reaction approach was used in this study to determine the population of major ruminal bacterial species (Fibrobacter succinogenes, Ruminococcus albus, and Ruminococcus flavefaciens) in digesta and rumen fluid of swamp buffalo (Bubalus bubalis). Four rumen-fistulated, male swamp buffalo were randomly assigned according to a 4 x 4 Latin square design to evaluate the effect of the urea-treated rice straw (roughage source)-to-concentrate ratio on cellulolytic bacterial distribution. Animals were fed roughage-to-concentrate (R:C) ratios of 100:0, 75:25, 50:50, and 25:75, respectively. At the end of each period, rumen fluid and digesta were collected at 0 h and 4 h post-morning-feeding. It was found that feeding urea-treated rice straw solely increased these three cellulolytic bacteria numbers up to 2.65 x 10(9) and 3.54 x 10(9) copies per milliliter for F. succinogenes, 5.10 x 10(7) and 7.40 x 10(7) copies per milliliter for R. flavefaciens, and 4.00 x 10(6) and 6.00 x 10(6) copies per milliliter for R. albus in rumen fluid and digesta, respectively. The distribution of the three cellulolytic bacteria species in digesta were highest at 3.21 x 10(9), 4.55 x 10(7), and 4.56 x 10(6) copies per milliliter for F. succinogenes, R. flavefaciens, and R. albus, respectively. Moreover, at 4 h post-morning-feeding, the populations of the three cellulolytic bacteria were higher than found at 0 h post-morning-feeding. It is most notable that F. succinogenes were the highest in population in the rumen of swamp buffalo and cellulolytic bacteria mostly adhered to feed digesta in the rumen.

  17. Cloning, characterization, and expression studies in Escherichia coli of growth hormone cDNAs from Indian zebu cattle, reverine buffalo, and beetal goat.

    PubMed

    Mukhopadhyay, U K; Sahni, G

    2002-11-01

    The growth hormone cDNAs from three different economically important animal species of indian origin viz., indian zebu cattle (Bos indicus), indian reverine buffalo (Bubalus bubalis), and beetal goat (Capra hircus) were isolated by the RT-PCR technique. The amplified product was then cloned into phagemid pBluescriptIIKS- and the nucleotide sequence of the entire 573 base coding region for each product was determined. The genetic sequences as well as the translated protein sequence of these ruminant species were compared to that of closely related species like taurine cattle (Bos taurus) and sheep (Ovis aries). A very high degree of nucleotide sequence homology, ranging between 97-98%, was observed. Subsequently, the buffalo and goat cDNAs were used for expression studies in Escherichia coli. Very low levels of expression resulted when the growth hormone cDNAs were directly placed under the strong E. coli (trc) or phage (T7) promoters with the approximate level being less than 0.1% and 1% of the intracellular E. coli proteins, respectively. The nearly 10-fold enhancement of the level of expression as observed was attributable to the nature of the untranslated leader sequence donated by the individual expression element. High level (about 20% of soluble E. coli protein) expression of buffalo/goat growth hormone was achieved as a fusion protein with glutathione-s-transferase (GST) in pGEX-KT. Further, although attempts at converting the GST-GH fusion protein system to a two-cistronic gene expression system were unsuccessful, the utilization of a short synthetic first cistron in the two-cistronic mode of expression resulted in high levels (approximately 30% of soluble protein cell fraction) of GH polypeptide with a native N-terminus in E. coli for all three cDNAs.

  18. Long term effect of Ovum Pick-up in buffalo species.

    PubMed

    Neglia, Gianluca; Gasparrini, Bianca; Vecchio, Domenico; Boccia, Lucia; Varricchio, Ettore; Di Palo, Rossella; Zicarelli, Luigi; Campanile, Giuseppe

    2011-02-01

    The aim of this study was to evaluate the effect of an Ovum Pick-up (OPU) treatment carried out for 9 months in buffalo (Bubalus bubalis) species. Eight pluriparous non-lactating buffalo cows underwent OPU for 9 months. Recovered cumulus enclosed oocytes (COCs) were classified and COCs suitable for in vitro embryo production (IVEP) were in vitro matured (IVM), fertilized (IVF) and cultured (IVC) to the blastocyst (Bl) stage. Animals were monitored for a total period of 270 days, but at the summer solstice, follicular turnover decreased and at the 68-day of the trial, we decided to increase the OPU sampling interval from 3-4 to 7 days. It was therefore possible to distinguish two phases: a first phase (18 sessions), during which OPU was carried out twice weekly and a second phase (16 sessions) during which OPU sessions were performed weekly. This reduction did not modify the percentage of good quality COCs, while the incidence of grade D COCs decreased (P<0.01). Furthermore, embryo production was higher in the second phase, either if embryos were calculated on the total recovered COCs (8.3% vs. 21.4%; P<0.01) and on grade A+B COCs (13.0% vs. 32.1%; P<0.01), that supposedly should have given similar blastocyst yield. During the total period of the trial it was possible to distinguish a first period of 6 months (34 sessions) characterized by blastocyst production (0.36 blastocyst/buffalo/session), followed by an unproductive period of 3 months (12 sessions), during which embryos were not produced. During the first 6 months a higher (P<0.01) number of follicles (5.06 vs. 3.71), small follicles (3.38 vs. 2.07), total COCs (2.58 vs. 1.56) and good quality (A+B) COCs (1.51 vs. 0.94) per subject/session were recorded compared to the last 3 months. No Blastocyst were produced during the second period, even if the percentage of grade A+B COCs was similar to that recorded during the first period. In conclusion, buffalo cows submitted to repeated OPU sampling for a 9-month

  19. First report of isolation and molecular characterization of bubaline herpesvirus 1 (BuHV1) from Argentinean water buffaloes.

    PubMed

    Maidana, Silvina S; Konrad, José L; Craig, María I; Zabal, Osvaldo; Mauroy, Axel; Thiry, Etienne; Crudeli, Gustavo; Romera, Sonia A

    2014-11-01

    Herpesviruses have mainly co-evolved with their hosts for millions of years. However, bovine herpesvirus 1 (BoHV1) and related ruminant alphaherpesviruses have been reported to cross the species barrier. Bubaline herpesvirus 1 (BuHV1) is an alphaherpesvirus closely related to BoHV1 and BoHV5. According to the serological cross-relationships between ruminant alphaherpesviruses, several surveys have studied the occurrence of BoHV1-related virus infection in wild and domestic ruminant species. Recent studies in Argentina showed an increase in serological prevalence against BoHV1 related viruses in water buffaloes (Bubalus bubalis) population. The aim of this study was to investigate the presence of related ruminant alphaherpesvirus in the Argentinean water buffalo population. BuHV1 was successfully isolated from 5 out of 225 buffaloes analyzed. One isolate was obtained from nasal secretions, and the others were from vaginal swabs. The buffaloes belonged to four different farms located in northeastern Argentina. The isolates were characterized as alphaherpesvirus by direct immunofluorescence using FITC-anti-BoHV1 IgG. Restriction analysis performed with BamHI and BstEII on the complete genome showed differences between the isolates and those from BoHV1 and BoHV5 subtypes. Phylogenetic analysis on both UL27 and US6 showed similarity in tree topology. While three of the isolates grouped together with sequences of BoHV5, two other isolates clustered separately. Genetic analysis of eight concatenated sequences from all isolates and references strains showed high nucleotide sequence identity between BuHV1 and BoHV5. While three of the isolates clustered together with the BoHV5 reference strain, the last two isolates were closely related to an Australian BuHV1 strain. To our knowledge, this is the first report on the isolation and molecular characterization of BuHV1 in South America. Phylogenetic analysis suggested that two different BuHV1 lineages circulate in the

  20. Arachidic acid in extender improves post-thaw parameters of cryopreserved Nili-Ravi buffalo bull semen.

    PubMed

    Ejaz, R; Ansari, M S; Rakha, B A; Ullah, N; Husna, A U; Iqbal, R; Akhter, S

    2014-02-01

    Cryopreservation process reduces lipids and phospholipids from buffalo bull spermatozoa. It was therefore hypothesized that supplementation of fatty acid to extender may improve the post-thaw quality of buffalo semen. The objective was to evaluate the effect of arachidic acid supplementation in extender on post-thaw quality of buffalo bull (Bubalus bubalis) spermatozoa. Semen was collected from three adult Nili-Ravi buffalo bulls of similar age group with artificial vagina (42°C) for 3 weeks (replicate). Qualified semen ejaculates (n = 18) were split into four aliquots and diluted in tris-citric acid extender containing 0.0 (control), 5.0, 10.0 and 20.0 ng/ml at 37°C having approximately 50 × 10(6) spermatozoa/ml. Diluted semen was cooled to 4°C in 2 h and equilibrated for 4 h at 4°C. Cooled semen was filled in 0.5-ml straws at 4°C, kept on liquid nitrogen vapours for 10 min and plunged in liquid nitrogen for storage. Thawing of frozen semen was performed after 24 h at 37°C for 30 s. Sperm progressive motility (%) was improved in a dose-dependent manner by supplementing arachidic acid at 5.0, 10.0 and 20.0 ng/ml compared with control. Structural and functional integrity of sperm plasma membrane (%), number of acrosome-intact live sperm (%) and sperm chromatin integrity (%) were better (p < 0.05) in extender having 5.0 ng/ml of arachidic acid compared with control. At 10.0 ng/ml, these values did not vary (p > 0.05) from those at 5.0 ng/ml. Further improvement in structural and functional integrity of sperm plasma membrane, number of acrosome-intact live sperm and chromatin integrity was observed at 20.0 ng/ml of arachidic acid in extender. In conclusion, arachidic acid supplementation in extender improved the post-thaw quality parameters of cryopreserved Nili-Ravi buffalo bull spermatozoa. Among the arachidic acid concentrations studied, maximum improvement in post-thaw semen quality parameters was observed at 20.0 ng/ml. © 2013

  1. Effect of Ground Corn Cob Replacement for Cassava Chip on Feed Intake, Rumen Fermentation and Urinary Derivatives in Swamp Buffaloes

    PubMed Central

    Wanapat, M.; Pilajun, R.; Kang, S.; Setyaningsih, K.; Setyawan, A. R.

    2012-01-01

    Four Thai - rumen fistulated male swamp buffaloes (Bubalus bubalis), about four years old with 400±20 kg liveweight, were randomly assigned according to a 4×4 Latin square design to receive dietary treatments. The treatments were: ground corn cob (GCC) replacement for cassava chip (CC) in concentrate at 0% (T1); GCC replacement at 33% (T2); GCC replacement at 67% (T3); and GCC replacement at 100% (T4), respectively. During the experiment, concentrate was offered at 0.5% BW while 5% urea-treated rice straw was given at ad libitum. The result revealed that there was no effect of GCC replacement on DMI among treatments. In addition, digestibilities of DM, OM and CP were not different while aNDF linearly increased with an increasing level of GCC replacement. However, GCC replacement did not affect rumen fermentation such as ruminal pH, NH3-N and VFA concentration; except C3 proportion which was the highest at 33% replacement while the lowest was at 100% replacement. All replacements of GCC resulted in similar protozoal and bacterial populations and microbial protein synthesis (MPS). Purine derivatives (PD) concentration in urine and PD to creatinine (PDC) index were varied with time of urination and among treatments at 0 to 8 and 8 to 16 h post feeding and higher values were shown among the GCC replacement groups. However at 16 to 24 h-post feeding, it was untraceable. In addition, creatinine concentration was similar among all treatments at every sampling time. Based on the above results, GCC can be used as an energy source for swamp buffalo fed with rice straw. Spot sampling of urine can be used for purine derivatives determination. PMID:25049671

  2. Effect of feeding of blend of essential oils on methane production, growth, nutrient utilization in growing buffaloes.

    PubMed

    Yatoo, M A; Chaudhary, L C; Agarwal, Neeta; Chaturvedi, V B; Kamra, D N

    2017-02-23

    An experiment was conducted to study the effect of a blend of essential oils (BEO) on enteric methane emission and growth performance of buffaloes (Bubalus bubalis). Twenty one growing male buffaloes (average body weight of 279 ± 9.3 kg) were divided in to three groups. The animals of all the three groups were fed on a ration consisting of wheat straw and concentrate mixture targeting 500g daily live weight gain. The three dietary groups were; Group1: control without additive, Group 2 and 3: supplemented with BEO @ 0.15 and 0.30 ml/kg of dry matter intake (DMI), respectively. During six months feeding trial, the intake and digestibility of dry matter and nutrients (organic matter, crude protein, ether extract, neutral detergent fibre and acid detergent fibre) were similar in all the groups. The average body weight gain was tended to improve (P=0.084) in Group 2 and Group 3 as compared to control animals. Feeding of BEO did not affect feed conversion efficiency of the animals. The calves of all the three groups were in positive nitrogen balance with no difference in nitrogen metabolism. During respiration chamber studies the methane production (l/kg DMI and l/kg DDMI) was significantly (P<0.001) lower in Group 2 and Group 3 as compared to control animals. The results indicated that the blend of essential oils tested in the present study have shown potential to reduce enteric methane production without compromising the nutrient utilization and animal performance and could be further explored for its use as feed additive to mitigate enteric methane production livestock.

  3. Sarcocystis sinensis is an ultrastructurally distinct parasite of water buffalo that can cause foodborne illness but cannot complete its life-cycle in human beings.

    PubMed

    Chen, Xinwen; Zuo, Yangxian; Rosenthal, Benjamin M; He, Yongshu; Cui, Liwang; Yang, Zhaoqing

    2011-05-31

    In this study, we compared the morphology of Sarcocystis sinensis and Sarcocystis hominis, and assessed the infectiousness of S. sinensis for human volunteers. The cysts of S. sinensis were from water buffalo (Bubalus bubalis) and those of S. hominis were from cattle (Bos taurus). Transmission electron microscopy of S. sinensis cysts revealed that the cyst wall had leaning, finger-like protrusions measuring 1.44-5.08 μm in length and without invaginations on the tip surface of the protrusions. In contrast, the cyst wall of S. hominis had upright, finger-like protrusions measuring 9.43 μm×2.42 μm and with vesicle-like invaginations on the tip surface of the protrusions. Scanning electron microscopy revealed that surface of the protrusions was arranged as rectangles in S. sinensis, as compared to tongue-shaped in S. hominis. Other distinguishing features of S. sinensis include a thin ground substrate (GS) zone with microtubules and small, circle-like structures located at the base of the protrusions. Human volunteers, after consuming S. sinensis cysts, produced no sporocysts or oocysts in feces, suggesting that humans could not serve as definitive hosts for S. sinensis. By contrast, many sporocysts and oocysts were passed in feces of a human volunteer 11-29 days after ingestion of S. hominis cysts. These results showed that S. sinensis and S. hominis are separate species and S. sinensis cannot use human being as the definitive host.

  4. Serotyping, antibiotic susceptibility, and virulence genes screening of Escherichia coli isolates obtained from diarrheic buffalo calves in Egyptian farms.

    PubMed

    Hakim, Ashraf S; Omara, Shimaa T; Syame, Sohier M; Fouad, Ehab A

    2017-07-01

    In Egypt as in many other countries, river water buffalo (Bubalus bubalis) is considered an important source of high-quality milk and meat supply. The objective of this study was to investigate serotypes, virulence genes, and antibiotic resistance determinants profiles of Escherichia coli isolated from buffalo at some places in Egypt; noticibly, this issue was not discussed in the country yet. A number of 58 rectal samples were collected from diarrheic buffalo calves in different regions in Egypt, and bacteriological investigated for E. coli existence. The E. coli isolates were biochemically, serologicaly identified, tested for antibiotic susceptibility, and polymerase chain reaction (PCR) analyzed for the presence of antibiotic resistance determinants and virulence genes. Overall 14 isolates typed as E. coli (24.1%); 6 were belonged to serogroup O78 (10.3%), followed by O125 (4 isolates, 6.9%), then O158 (3 isolates, 5.2%) and one isolate O8 (1.7%), among them, there were 5 E. coli isolates showed a picture of hemolysis (35.7%). The isolates exhibited a high resistance to β lactams over 60%, followed by sulfa (50%) and aminoglucoside (42.8%) group, in the same time the isolates were sensitive to quinolone, trimethoprim-sulfamethoxazole, tetracycline (100%), and cephalosporine groups (71.4%). A multiplex PCR was applied to the 14 E. coli isolates revealed that all were carrying at least one gene, as 10 carried blaTEM (71.4%), 8 Sul1 (57.1%), and 6 aadB (42.8%), and 9 isolates could be considered multidrug resistant (MDR) by an incidence of 64.3%. A PCR survey was stratified for the most important E. coli virulence genes, and showed the presence of Shiga toxins in 9 isolates carried either one or the two Stx genes (64.3%), 5 isolates carried hylA gene (35.7%), and eae in 2 isolates only (14.3%), all isolates carried at least one virulence gene except two (85.7%). The obtained data displayed that in Egypt, buffalo as well as other ruminants could be a potential

  5. Determination of an Optimal Membrane-permeable Cryoprotectant Addition and Dilution Protocol for Water Buffalo Spermatozoa.

    PubMed

    Mehmood, S; Ahmed, H; Hussain Shah, S A; Wattoo, F H; Akhtar, S; Hassan Andrabi, S M

    There is a possibility to reduce the toxicity of glycerol and osmotic stress of DMSO by lowering their concentrations in freezing extenders. To investigate the effect of glycerol and DMSO in tris-citric acid based extender on post- thaw quality of buffalo (Bubalus bubalis) bull spermatozoa. Semen was collected from five adult buffalo bulls with artificial vagina. Five aliquots of semen per bull were separated for dilution with the treatment extenders. The first aliquot was diluted at 37C with 6 percent glycerol (T1). The second aliquot was diluted at 37C with extenders containing 4.5 percent glycerol and 1.5 percent DMSO (T2). The third aliquot was diluted with extenders containing 4.5 percent glycerol at 37C and 1.5 percent DMSO at 4С (T3). The fourth aliquot was diluted with extenders containing 1.5 percent DMSO at 37C and 4.5 percent glycerol at 4С (T4). The fifth aliquot was diluted with extender containing 2.5percent DMSO at 37 as well as at 4C (T5). The final concentration of spermatozoa was 50×10(6)/ml in all the treatment groups. Semen was cooled from 37 to 4C in 2 h and equilibration was done at 4 C for 4 h. Later on, packing of cooled semen was undertaken in 0.54 ml French straws and frozen in a programmable cell freezer. At post thawing, treatment groups T1 and T2 yielded significant (P < 0.05) outcome for CASA parameters, longevity, acrosomal integrity, plasma membrane integrity, mitochondrial transmembrane potential and DNA integrity. We concluded that by decreasing glycerol concentration (4.5 percent) and combining it with DMSO (1.5 percent) at 37C (T2) in tris-citric acid based extender provided similar results to those observed when glycerol (6 percent) alone is used at 37C (T1) for improving the post-thaw quality of buffalo bull spermatozoa.

  6. Maturation competence of swamp buffalo oocytes obtained by ovum pick-up and from slaughterhouse ovaries.

    PubMed

    Yindee, M; Techakumphu, M; Lohachit, C; Sirivaidyapong, S; Na-Chiangmai, A; Roelen, B A J; Colenbrander, B

    2011-10-01

    This study was designed with the final goal of improving in vitro embryo production in the Thai swamp buffalo (Bubalus bubalis carabensis). Oocytes were collected by ovum pick-up (OPU) from six non-lactating multiparous swamp buffalo twice per week for 10 consecutive sessions followed by once-weekly collection for 10 consecutive sessions without hormone stimulation. In addition, oocytes were collected from slaughterhouse ovaries that were classified as follows: ovaries from non-pregnant cows with a visible corpus luteum (NPCL); pregnant cows with a corpus luteum (P); and non-pregnant cows without a corpus luteum (NP). Follicles in each group of ovaries were categorized as small (2-4 mm), medium-sized (5-8 mm) or large follicles (≥ 9 mm). The quality of the oocytes was assessed by their capacity to undergo in vitro maturation. The total number of observed follicles per session (all sizes combined) was larger in the once-weekly OPU group compared with the twice-weekly OPU group. In particular, the numbers of small and large follicles were higher in the once-weekly OPU group (5.2 ± 0.7 and 0.9 ± 0.2, respectively) than in the twice-weekly OPU group (3.9 ± 0.5 and 0.5 ± 0.1). The number of medium-sized follicles did not differ between the groups. The percentages of oocytes with an abnormal spindle morphology were not different between oocytes from the twice-weekly (30.0%) and the once-weekly (28.6%) OPU groups. A higher percentage of oocytes obtained in vitro (49.5%) exhibited nuclear abnormalities compared with those obtained in vivo (≤34.8%) after in vitro maturation. In conclusion, oocytes can be successfully collected by OPU in the swamp buffalo, without hormonal pretreatment, and per week more good-quality oocytes can be collected by twice-weekly OPU. In addition, oocytes collected from slaughterhouse ovaries can be used with the reproductive status of the cow having no influence on the maturation competence of oocytes. © 2011 Blackwell Verlag GmbH.

  7. Effects of various plant protein sources in high-quality feed block on feed intake, rumen fermentation, and microbial population in swamp buffalo.

    PubMed

    Foiklang, Suban; Wanapat, Metha; Toburan, Wetchasit

    2011-12-01

    This study was designed to determine effect of various plant protein sources in high-quality feed block (HQFB) on feed intake, rumen fermentation, and microbial population in swamp buffalo. Four rumen-fistulated swamp buffaloes (Bubalus bubalis) were randomly assigned according to a 4 × 4 Latin square design. Four kinds of plant protein sources (coarse rice bran (CRB), cassava hay (CH), Phaseolus calcaratus hay, and mulberry hay (MH)) were mixed in the HQFB. HQFBs were allowed to be licked at free choice, and urea-lime-treated rice straw (ULRS) were fed ad libitum. It was found that bacterial population and fungal zoospores in CH-fed group tended to be higher than those in other groups. Moreover, protozoal population in CH, P. calcaratus hay, and MH were lower than those in CRB supplemented group (P < 0.05). Cellulolytic bacterial population was highest in CH-fed group while proteolytic bacteria population was highest in P. calcaratus hay-fed group (P < 0.05). CH-fed group had higher ULRS intake than those in other groups (P < 0.05). Nutrient digestibility of CP, NDF, and ADF in CH-fed group was significantly higher than those in other groups (P < 0.05). Total VFA was highest in CH-fed group (P < 0.05). N absorption was highest in CH-fed group (P < 0.05). Based on this study, it could be concluded that cassava hay, P. calcaratus hay, and mulberry hay are potential to be used as protein sources in the HQFBs especially cassava hay.

  8. Population structure and genetic variability in the Murrah dairy breed of water buffalo in Brazil accessed via pedigree analysis.

    PubMed

    Malhado, Carlos Henrique Mendes; Malhado, Ana Claudia Mendes; Carneiro, Paulo Luiz Souza; Ramos, Alcides Amorim; Ambrosini, Diego Pagung; Pala, Akin

    2012-12-01

    The objective of this study was to use pedigree analysis to evaluate the population structure and genetic variability in the Murrah dairy breed of water buffalo (Bubalus bubalis) in Brazil. Pedigree analysis was performed on 5,061 animals born between 1972 and 2002. The effective number of founders (fe) was 60, representing 6.32 % of the potential number of founders. The effective number of ancestors (fa) was 36 and the genetic contribution of the 17 most influent ancestors explained 50 % of the genetic variability in the population. The ratio fe/fa (effective number of founders/effective number of ancestors), which expresses the effect of population bottlenecks, was 1.66. Completeness level for the whole pedigree was 76.8, 49.2, 27.7, and 12.8 % for, respectively, the first, second, third, and fourth known parental generations. The average inbreeding values for the whole analyzed pedigree and for inbreed animals were, respectively, 1.28 and 7.64 %. The average relatedness coefficient between individuals of the population was estimated to be 2.05 %-the highest individual coefficient was 10.31 %. The actual inbreeding and average relatedness coefficient are probably higher than estimated due to low levels of pedigree completeness. Moreover, the inbreeding coefficient increased with the addition of each generation to the pedigree, indicating that incomplete pedigrees tend to underestimate the level of inbreeding. Introduction of new sires with the lowest possible average relatedness coefficient and the use of appropriate mating strategies are recommended to keep inbreeding at acceptable levels and increase the genetic variability in this economically important species, which has relatively low numbers compared to other commercial cattle breeds. The inclusion of additional parameters, such as effective number of founders, effective number of ancestors, and fe/fa ratio, provides better resolution as compared to the inclusion of inbreeding coefficient and may help

  9. Generation of a Novel Bacteriophage Library Displaying scFv Antibody Fragments from the Natural Buffalo Host to Identify Antigens from Adult Schistosoma japonicum for Diagnostic Development

    PubMed Central

    Hosking, Christopher G.; McWilliam, Hamish E. G.; Driguez, Patrick; Piedrafita, David; Li, Yuesheng; McManus, Donald P.; Ilag, Leodevico L.; Meeusen, Els N. T.; de Veer, Michael J.

    2015-01-01

    The development of effective diagnostic tools will be essential in the continuing fight to reduce schistosome infection; however, the diagnostic tests available to date are generally laborious and difficult to implement in current parasite control strategies. We generated a series of single-chain antibody Fv domain (scFv) phage display libraries from the portal lymph node of field exposed water buffaloes, Bubalus bubalis, 11–12 days post challenge with Schistosoma japonicum cercariae. The selected scFv-phages showed clear enrichment towards adult schistosomes and excretory-secretory (ES) proteins by immunofluorescence, ELISA and western blot analysis. The enriched libraries were used to probe a schistosome specific protein microarray resulting in the recognition of a number of proteins, five of which were specific to schistosomes, with RNA expression predominantly in the adult life-stage based on interrogation of schistosome expressed sequence tags (EST). As the libraries were enriched by panning against ES products, these antigens may be excreted or secreted into the host vasculature and hence may make good targets for a diagnostic assay. Further selection of the scFv library against infected mouse sera identified five soluble scFv clones that could selectively recognise soluble whole adult preparations (SWAP) relative to an irrelevant protein control (ovalbumin). Furthermore, two of the identified scFv clones also selectively recognised SWAP proteins when spiked into naïve mouse sera. These host B-cell derived scFvs that specifically bind to schistosome protein preparations will be valuable reagents for further development of a cost effective point-of-care diagnostic test. PMID:26684756

  10. Effect of pour-on alphacypermethrin on feed intake, body condition score, milk yield, pregnancy rates, and calving-to-conception interval in buffaloes.

    PubMed

    Bifulco, G; Veneziano, V; Cimmino, R; Esposito, L; Auletta, L; Varricchio, E; Balestrieri, A; Claps, S; Campanile, G; Neglia, G

    2015-04-01

    The aims of this study were to assess the efficacy of alphacypermethrin (ACYP) on pediculosis due to Haematopinus tuberculatus and to evaluate the influence of the treatment on productive and reproductive performance in buffaloes (Bubalus bubalis) reared in an intensive system. The trial was performed on 56 pluriparous buffaloes at 86.8 ± 8.1 d in milk. The animals underwent individual louse count and were divided into 2 homogenous groups according to louse count, age, number of lactations, days in milk, live BW, BCS, pregnancy status, and milk yield. Group A (n = 28) was treated by a pour-on formulation of ACYP, and Group S (n = 28) was treated by pour-on saline solution. Individual louse counts were performed weekly on 10 buffaloes in each group. Feed intake was recorded daily and the total mixed ration, individual ingredients, and orts were analyzed to calculate DM ingestion. Individual milk yield was recorded daily and milk samples were analyzed at the beginning of the trial, after 4 wk, and at the end of the trial to assess milk composition. Individual BCS was also evaluated simultaneously. Finally, the animals underwent synchronization of ovulation starting 4 wk after treatment and the pregnancy rate and the calving-conception interval were evaluated. Data were analyzed by the Mann-Whitney test and ANOVA for repeated measures. The infestation was constant in Group S, whereas no lice were present in Group A throughout the study. Daily DMI was similar in the 2 groups (16.7 ± 0.4 vs. 16.3 ± 0.3 kg/d in Group A vs. Group S, respectively), although buffaloes in Group A showed higher (P < 0.05) BCS score at the end of the trial (7.39 ± 0.1 vs. 7.14 ± 0.1 in Group A vs. Group S, respectively). The average milk yield/buffalo was higher (P < 0.05) in Group A compared to Group S (10.58 ± 0.1 vs. 10.39 ± 0.1 kg in Group A vs. Group S, respectively) and this was mainly due to the higher milk production recorded in buffaloes at less than 75 d in milk (11.81 ± 0

  11. In vitro evaluation of liquid-stored buffalo semen at 5°C diluted in soya lecithin based extender (Bioxcell®), tris-citric egg yolk, skim milk and egg yolk-citrate extenders.

    PubMed

    Akhter, S; Ansari, M S; Rakha, B A; Ullah, N; Andrabi, S M H; Khalid, M

    2011-02-01

    This study was designed to compare the quality of liquid-stored buffalo bull spermatozoa in soya lecithin based extender Bioxcell(®) (BIOX), milk (MILK), tris-citric egg yolk (TEY) and egg yolk-citrate (EYC) extender at 5°C. Semen was collected from five Nili-Ravi buffalo (Bubalus bubalis) bulls of 6-7 years of age with artificial vagina over a period of 3 weeks (two consecutive ejaculates once in a week). Semen ejaculates having more than 60% motility were pooled, split into four aliquots, diluted (37°C; 10 × 10(6) motile spermatozoa/ml), cooled from 37 to 5°C in 2 h (0.275°C/min) and stored for 5 days. Sperm motility, viability, plasma membrane integrity (PMI) and normal acrosomal ridge were studied at first, third and fifth day of storage. Higher values of progressive sperm motility (%), sperm viability (%), sperm PMI (%) and normal apical ridge (%) were observed in BIOX, MILK and TEY extenders at first, third and fifth day of storage than EYC extender. Progressive sperm motility, sperm viability and sperm PMI in BIOX(®) extender were not different from MILK and TEY extenders at 1st and third day storage period. However, at fifth day of storage, the values for these parameters remained significantly higher (p<0.05) in BIOX(®) compared with MILK, TEY and EYC extenders. At fifth day of storage, the semen quality parameters for Bioxcell(®) were comparable to those with MILK and TEY extenders at third day of storage. In conclusion, motility, viability and PMI of buffalo bull spermatozoa remained similar in Bioxcell(®) , milk and TEY extender at first and third days of storage at 5°C. Yet, the values for the aforementioned parameters in Bioxcell(®) were higher compared with milk, TEY and EYC extender at fifth day of storage at 5°C. © 2010 Blackwell Verlag GmbH.

  12. Influence of epidermal growth factor and insulin-like growth factor 1 on nuclear maturation and fertilization of buffalo cumulus oocyte complexes in serum free media and their subsequent development in vitro.

    PubMed

    Purohit, G N; Brady, M S; Sharma, S S

    2005-07-01

    The in vitro maturation, fertilization and development of Indian water buffalo (Bubalus sp.) cumulus oocyte complexes (COCs) to blastocysts were studied during culture, either in serum free tissue culture medium 199 (TCM 199) or Waymouth MB (WM). Based on different supplements added to these media, the experimental groups included: (a) no supplement (control); (b) hormones (FSH, LH and oestradiol) (c) Epidermal growth factor (EGF); (d) IGF-1; and (e) EGF + IGF-1. Experiments were conducted to note three end points: (1) nuclear maturation 24 h after culture (eight replicates); (2) fertilization 24 h after insemination (10 replicates); (3) development to blastocysts (nine replicates). The oocytes were cultured in groups of up to five per drop. Using a two-way (5 x 2) factorial model with interactions, the results were compared using generalized linear models with binomial errors and the logit link function. In experiment 1, the proportion of oocytes reaching metaphase II was higher for all the supplement treatments than the control treatment (t = 3.68, p < 0.0001). The proportion of oocytes reaching metaphase II was 74.7, 63.2, 64.7 and 81% with hormone (chi2 = 17.23, p < 0.0001), EGF (chi2 = 7.07, p = 0.007), IGF-1 (chi2 = 19.21, p = 0.002) and EGF + IGF-1 (chi2 = 33.04, p < 0.0001) supplementation, respectively, compared to 46.6% in the control (no supplement) group. Media did not have an effect on outcome. In experiment 2, the proportion of oocytes fertilized was significantly higher with hormones (31.0%, chi2 = 12.5, p = 0.0004), IGF-1 (35.7%, chi2 = 20.53, p < 0.0001), and the EGF + IGF-1 combination (49.7%, chi2 = 51.35, p < 0.0001) compared to control (16.2%). No significant effect of media was seen. In experiment 3, the proportion of oocytes that cleaved at 48 h after culturing was significantly higher for all supplement treatments compared to control. IGF-1 supplementation was the only treatment that did not produce a significantly higher rate of progression

  13. Sarcocystis rommeli, n. sp. (Apicomplexa: Sarcocystidae) from cattle (Bos taurus) and its differentiation from Sarcocystis hominis

    USDA-ARS?s Scientific Manuscript database

    Cattle (Bos taurus) are intermediate hosts for three named species of Sarcocystis, S. cruzi, S. hirsuta, and S. hominis. Recently, a fourth species was identified and named S. sinensis. However, S. sinensis originally named a species of Sarcocystis in water buffalo (Bubalus bubalis) in China. Based ...

  14. Hydrologic, Water-Quality, and Biological Data for Three Water Bodies, Texas Gulf Coast Plain, 2000-2002

    DTIC Science & Technology

    2003-01-01

    appreciable urban development near Dickinson has occurred since 1990, so it is likely that the percent- age of the basin that is urban is greater...Suckers Catostomidae Smallmouth buffalo Ictiobus bubalus 0 3 1 2 1 0 7 River carpsucker Carpiodes carpio 0 13 0 0 0 0 13 Bullhead catfish Ictaluridae

  15. Evaluation and Proposed Refinement of the Sampling Design for the Long Term Resource Monitoring Program’s Fish Component

    DTIC Science & Technology

    2002-10-01

    the collection of most age I+ channel catfish and smallmouth buffalo Ictiobus bubalus 3. Loss of fyke nets will eliminate population trend...Semotilus atromaculatus Catostomidae River carpsucker Carpiodes carpio Quillback C. cyprinus Highfin carpsucker C. velifer White sucker C. commersoni Blue

  16. Intraspecific and interspecific genetic variation of Gongylonema pulchrum and two rodent Gongylonema spp. (G. aegypti and G. neoplasticum), with the proposal of G. nepalensis n. sp. for the isolate in water buffaloes from Nepal.

    PubMed

    Setsuda, Aogu; Da, Nengtai; Hasegawa, Hideo; Behnke, Jerzy M; Rana, Hari Bahadur; Dhakal, Ishwari Prasad; Sato, Hiroshi

    2016-02-01

    The gullet worm (Gongylonema pulchrum) has been recorded from a variety of mammals worldwide. In an earlier study, we demonstrated two separate transmission cycles in cattle (Bos taurus) and wild mammals in Japan based on nucleotide sequences of the ribosomal RNA gene (rDNA) and cytochrome c oxidase subunit I (cox-1) region of mitochondrial DNA of multiple isolates of different origins. Our earlier study additionally demonstrated two major cox-1 haplotypes of G. pulchrum prevalent in cattle in Japan. In the present study, we collected G. pulchrum from cattle and goats (Capra hircus) in Alashan League, Inner Mongolia, China; Gongylonema aegypti from spiny mice (Acomys dimidiatus) in the Sinai Peninsula, Egypt; and Gongylonema neoplasticum from a black rat (Rattus rattus) in Okinawa Island, Japan, to analyze their genetic relationships with G. pulchrum in Japan. The gullet worms from Alashan League had almost identical rDNA nucleotide sequences and two cox-1 haplotypes as seen in G. pulchrum from the cattle in Japan. The two rodent Gongylonema spp. had distinct rDNA nucleotide sequences compared with those of G. pulchrum; only the 18S and 5.8S rDNA sequences showed high identities at 97.2-98.7%, while the remaining sequences were less than 75% identical. The 18S, 5.8S, and 28S rDNA sequences of the two rodent Gongylonema spp. showed nucleotide identities of 99.8% (1811/1814), 100% (158/158), and 98.9% (3550/3590), respectively. The cox-1 regions showed 91.6% (338/369)-92.1% (340/369) identities, with completely identical amino acid sequences. The genetic diversities of three distinct Gongylonema spp. and their possible intraspecific genetic variation may allow us to resolve the taxonomic position of Gongylonema spp. which display few obvious morphological differences from their congeners. Consequently, the Gongylonema isolate from water buffaloes (Bubalus bubalis) in Nepal reported in our previous study is concluded to be a new species, and Gongylonema nepalensis n

  17. Heat stress and antioxidant enzyme activity in bubaline ( Bubalus bubalis) oocytes during in vitro maturation

    NASA Astrophysics Data System (ADS)

    Waiz, Syma Ashraf; Raies-ul-Haq, Mohammad; Dhanda, Suman; Kumar, Anil; Goud, T. Sridhar; Chauhan, M. S.; Upadhyay, R. C.

    2016-09-01

    In vitro environments like heat stress usually increase the production of reactive oxygen species in bubaline oocytes which have been implicated as one of the major causes for reduced developmental competence. Oocytes during meiotic maturation are sensitive to oxidative stress, and heat stress accelerates cellular metabolism, resulting in the higher production of free radicals. Therefore, the aim of present work was to assess the impact of heat stress during meiotic maturation on bubaline cumulus-oocyte complexes (COC), denuded oocytes (DO), and cumulus cell mass in terms of their oxidative status. Accordingly, for control group, COC were matured at 38.5 °C for complete 24 h of meiotic maturation and heat stress of 40.5 and 41.5 °C was applied to COC during the first 12 h of maturation and then moved to 38.5 °C for rest of the 12 h. In another group, COC after maturation were denuded from the surrounding cumulus cells by manual pipetting. Results indicated that the production of reactive oxygen species (ROS), lipid peroxides, and nitric oxide (NO) was significantly ( P < 0.05) higher in the oocytes subjected to heat stress (40.5 and 41.5 °C) during meiotic maturation compared to the oocytes matured under standard in vitro culture conditions (38.5 °C). Also, the antioxidant enzymatic activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase were significantly ( P < 0.05) increased in all the treatment groups compared to the control group. Therefore, the present study clearly establishes that heat stress ensues oxidative stress in bubaline oocytes which triggers the induction of antioxidant enzymatic defense system for scavenging the ROS.

  18. Heat stress and antioxidant enzyme activity in bubaline (Bubalus bubalis) oocytes during in vitro maturation.

    PubMed

    Waiz, Syma Ashraf; Raies-Ul-Haq, Mohammad; Dhanda, Suman; Kumar, Anil; Goud, T Sridhar; Chauhan, M S; Upadhyay, R C

    2016-09-01

    In vitro environments like heat stress usually increase the production of reactive oxygen species in bubaline oocytes which have been implicated as one of the major causes for reduced developmental competence. Oocytes during meiotic maturation are sensitive to oxidative stress, and heat stress accelerates cellular metabolism, resulting in the higher production of free radicals. Therefore, the aim of present work was to assess the impact of heat stress during meiotic maturation on bubaline cumulus-oocyte complexes (COC), denuded oocytes (DO), and cumulus cell mass in terms of their oxidative status. Accordingly, for control group, COC were matured at 38.5 °C for complete 24 h of meiotic maturation and heat stress of 40.5 and 41.5 °C was applied to COC during the first 12 h of maturation and then moved to 38.5 °C for rest of the 12 h. In another group, COC after maturation were denuded from the surrounding cumulus cells by manual pipetting. Results indicated that the production of reactive oxygen species (ROS), lipid peroxides, and nitric oxide (NO) was significantly (P < 0.05) higher in the oocytes subjected to heat stress (40.5 and 41.5 °C) during meiotic maturation compared to the oocytes matured under standard in vitro culture conditions (38.5 °C). Also, the antioxidant enzymatic activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase were significantly (P < 0.05) increased in all the treatment groups compared to the control group. Therefore, the present study clearly establishes that heat stress ensues oxidative stress in bubaline oocytes which triggers the induction of antioxidant enzymatic defense system for scavenging the ROS.

  19. The Olduvai buffalo Pelorovis and the origin of Bos

    NASA Astrophysics Data System (ADS)

    Martínez-Navarro, Bienvenido; Antonio Pérez-Claros, Juan; Palombo, Maria Rita; Rook, Lorenzo; Palmqvist, Paul

    2007-09-01

    The origin of the genus Bos is a debated issue. From ˜ 0.5 Ma until historic times, the genus is well known in the Eurasian large mammal assemblages, where it is represented by Bos primigenius. This species has a highly derived cranial anatomy that shows important morphological differences from other Plio-Pleistocene Eurasian genera of the tribe Bovini such as Leptobos, Bison, Proamphibos-Hemibos, and Bubalus. The oldest clear evidence of Bos is the skull fragment ASB-198-1 from the middle Pleistocene (˜ 0.6-0.8 Ma) site of Asbole (Lower Awash Valley, Ethiopia). The first appearance of Bos in Europe is at the site of Venosa-Notarchirico, Italy (˜ 0.5-0.6 Ma). Although the origin of Bos has traditionally been connected with Leptobos and Bison, after a detailed anatomical and morphometric study we propose here a different origin, connecting the middle Pleistocene Eurasian forms of B. primigenius with the African Late Pliocene and early Pleistocene large size member of the tribe Bovini Pelorovis sensu stricto. The dispersal of the Bos lineage in Western Europe during middle Pleistocene times seems to coincide with the arrival of the Acheulean tool technology in this continent.

  20. Investigation of the Fisheries of Africa, Buck, and Judd Lakes, Louisiana.

    DTIC Science & Technology

    1987-10-01

    0.3048 metres pounds (mass) 0.4535924 kilograms * To obtain Celsius (C) temperature readings from Fahrenheit (F) readings , use the following formula...C - (5/9) (F - 32). To obtain Kelvin (K) readings , use K - (5/9) (F - 32) + 273.15. 3 INVESTIGATION OF THE FISHERIES OF AFRICA, BUCK, AND JUDD LAKES...carps Common Carp (Cyprinus carpio) x x 2,4 Catostomidae - suckers Smallmouth buffalo (Ictiobus bubalus) x x x 2,4 Bigmouth buffalo (Ictiobus

  1. A Buffalo Story.

    ERIC Educational Resources Information Center

    Reville, Eugene T.

    The Buffalo Public Schools have developed magnet schools to aid desegregation plans and to provide for different student needs and learning styles. The Academic Challenge Center is designed to meet concerns about declining reading and math scores. The Buffalo Traditional School emphasizes traditional learning styles and puts a high priority on…

  2. The commercial fishery in Lake Oahe, North and South Dakota, 1964-70

    USGS Publications Warehouse

    Higham, Joseph H.

    1974-01-01

    Ten species were commercially harvested in Lake Oahe, among which bigmouth buffalo (Ictiobus cyprinellus) predominated (63.8% of the total weight), smallmouth buffalo (Ictiobus bubalus) and goldeye (Hiodon alosoides) ranked second, and third. Variations in the seasonal and annual production of buffalo were governed by market conditions and availability. Production of minor species depended on the amount of fishing effort directed toward the harvest of buffalo. Gill and hoop nets were the principal fishing gears. The fishing season usually extended from April through December; most fishing was in May through August.

  3. 1. Photocopy of photograph (from the Buffalo and Erie County ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    1. Photocopy of photograph (from the Buffalo and Erie County Historical Society) Photographer unknown 1859 GENERAL VIEW FROM THE SOUTHEAST - Buffalo Lighthouse, Buffalo Harbor, Buffalo River & Lake Erie, Buffalo, Erie County, NY

  4. 2. Photocopy of photograph (from Buffalo and Erie County Historical ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    2. Photocopy of photograph (from Buffalo and Erie County Historical Society) Photographer unknown 1928 GENERAL VIEW FROM THE SOUTHWEST - Buffalo Lighthouse, Buffalo Harbor, Buffalo River & Lake Erie, Buffalo, Erie County, NY

  5. Purification, characterization, structural analysis and protein chemistry of a buffalo heart galectin-1.

    PubMed

    Ashraf, Ghulam Md; Rizvi, Sabika; Naqvi, Shoa; Suhail, Nida; Bilal, Nayeem; Hasan, Shirin; Tabish, Mohammad; Banu, Naheed

    2010-11-01

    A soluble β-galactoside-binding lectin was purified by gel filtration chromatography from Bubalus bubalis heart. Its metal-independent nature, molecular weight of 14.5 kDa, preferential affinity for β-D-lactose, and 87-92% identity with carbohydrate recognition domain of previously reported galectin-1 confirmed its inclusion in galectin-1 subfamily. Stokes radii determination using gel filtration under reducing and non-reducing conditions revealed its homo-dimeric nature, further confirming its Gal-1 nomenclature. The purified lectin was found to be the most stable mammalian heart galectin purified till date, suggesting its preferential use in various recognition studies. Treatment of the purified lectin with oxidizing agent, thiol blocking reagents, denaturants, and detergents resulted in significant changes in UV-VIS, fluorescence, CD and FTIR spectra, which strongly emphasized the important aspect of regular secondary structure of galectins for the maintenance of their active conformation. Reduction of the activity of the purified lectin after oxidation by H2O2, with remarkable fluorescence quenching, may suggest potential role for galectin-1 in free radical-induced, oxidative stress-mediated cardiovascular disorders. The predictions of bioinformatics studies were found to be in accordance with the results obtained in wet lab.

  6. 76 FR 20530 - Safety Zone; Boom Days, Buffalo Outer Harbor, Buffalo, NY

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-13

    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone; Boom Days, Buffalo Outer Harbor, Buffalo... temporary safety zone in the Buffalo Outer Harbor, Buffalo, NY for the Boom Days Fireworks. This zone is intended to restrict vessels from Doug's Dive, the NFTA small boat harbor and a portion of the Buffalo...

  7. Underway view from port side in the Buffalo River with ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Underway view from port side in the Buffalo River with water cannons spraying. City of Buffalo in background. TC - Fireboat EDWARD M. COTTER, Moored on the Buffalo River at 155 Ohio Street, Buffalo, Erie County, NY

  8. UPPER BUFFALO WILDERNESS AND BUFFALO ADDITION ROADLESS AREA, ARKANSAS.

    USGS Publications Warehouse

    Miller, Mary H.; Armstrong, Michelle K.

    1984-01-01

    The Upper Buffalo Wilderness and Buffalo Addition Roadless Area covers about 19 sq mi in the Ozark National Forest, Newton County, Arkansas. No metal-bearing minerals were observed during geologic mapping, and analyses for zinc and lead contents in surface rock and sediment samples from the study area are not anomalous. Exploratory drilling into the Boone Formation and (or) the Everton Formation will be necessary to evaluate the mineral-resource potential of zinc and lead in the study area.

  9. 33 CFR 117.773 - Buffalo River.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 1 2011-07-01 2011-07-01 false Buffalo River. 117.773 Section... DRAWBRIDGE OPERATION REGULATIONS Specific Requirements New York § 117.773 Buffalo River. (a) The draw of the Michigan Avenue bridge, mile 1.3, at Buffalo, shall operate as follows: (1) From March 22 through...

  10. 33 CFR 117.773 - Buffalo River.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 1 2012-07-01 2012-07-01 false Buffalo River. 117.773 Section... DRAWBRIDGE OPERATION REGULATIONS Specific Requirements New York § 117.773 Buffalo River. (a) The draw of the Michigan Avenue bridge, mile 1.3, at Buffalo, shall operate as follows: (1) From March 22 through...

  11. 33 CFR 117.773 - Buffalo River.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 1 2013-07-01 2013-07-01 false Buffalo River. 117.773 Section... DRAWBRIDGE OPERATION REGULATIONS Specific Requirements New York § 117.773 Buffalo River. (a) The draw of the Michigan Avenue bridge, mile 1.3, at Buffalo, shall operate as follows: (1) From March 22 through...

  12. 33 CFR 117.773 - Buffalo River.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Buffalo River. 117.773 Section... DRAWBRIDGE OPERATION REGULATIONS Specific Requirements New York § 117.773 Buffalo River. (a) The draw of the Michigan Avenue bridge, mile 1.3, at Buffalo, shall operate as follows: (1) From March 22 through...

  13. 33 CFR 117.773 - Buffalo River.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 1 2014-07-01 2014-07-01 false Buffalo River. 117.773 Section... DRAWBRIDGE OPERATION REGULATIONS Specific Requirements New York § 117.773 Buffalo River. (a) The draw of the Michigan Avenue bridge, mile 1.3, at Buffalo, shall operate as follows: (1) From March 22 through...

  14. Underway view from starboard side in the Buffalo River with ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Underway view from starboard side in the Buffalo River with grain elevators and lift bridge in background. TC - Fireboat EDWARD M. COTTER, Moored on the Buffalo River at 155 Ohio Street, Buffalo, Erie County, NY

  15. 77 FR 39408 - Safety Zone; Buffalo July 4th Fireworks, Lake Erie, Buffalo, NY

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-03

    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone; Buffalo July 4th Fireworks, Lake Erie, Buffalo, NY AGENCY: Coast Guard, DHS. ACTION: Temporary final rule. SUMMARY: The Coast Guard is establishing a temporary safety zone on Lake Erie, Buffalo, NY. This safety zone is intended to...

  16. Lake Effect Snow Covers Buffalo

    NASA Technical Reports Server (NTRS)

    2002-01-01

    An average of one foot of snow per day has fallen on Buffalo, New York, since Christmas Eve, resulting in a total of up to 5 feet from December 24-28. The snow fell very heavily, with accumulations of up to 3 inches per hour. Cold winds blowing along the surface of Lake Erie pick up warmth and moisture, which falls as snow as the warm air rises. This image was acquired by the Geostationary Operational Environmental Satellite (GOES), operated by NOAA, on December 27, 2001, at 12:32 p.m. EST. The scene shows thick bands of clouds extending from the eastern tip of Lake Erie and over Buffalo. The arrows show the wind direction, which is blowing down the length of the lake. Image and animation by Robert Simmon, based on data from the NASA GOES Project Science Office.

  17. Lake Effect Snow Covers Buffalo

    NASA Technical Reports Server (NTRS)

    2002-01-01

    An average of one foot of snow per day has fallen on Buffalo, New York, since Christmas Eve, resulting in a total of up to 5 feet from December 24-28. The snow fell very heavily, with accumulations of up to 3 inches per hour. Cold winds blowing along the surface of Lake Erie pick up warmth and moisture, which falls as snow as the warm air rises. This image was acquired by the Geostationary Operational Environmental Satellite (GOES), operated by NOAA, on December 27, 2001, at 12:32 p.m. EST. The scene shows thick bands of clouds extending from the eastern tip of Lake Erie and over Buffalo. The arrows show the wind direction, which is blowing down the length of the lake. Image and animation by Robert Simmon, based on data from the NASA GOES Project Science Office.

  18. Phylogeography and Domestication of Chinese Swamp Buffalo

    PubMed Central

    Xie, Wen-Mei; Xu, Ping; Chang, Ti-Cheng; Liu, Li; Cheng, Feng; Zhang, Run-Feng; Lan, Xian-Yong; Chen, Hong; Lei, Chu-Zhao

    2013-01-01

    To further probe into whether swamp buffaloes were domesticated once or multiple times in China, this survey examined the mitochondrial DNA (mtDNA) Control Region (D-loop) diversity of 471 individuals representing 22 populations of 455 Chinese swamp buffaloes and 16 river buffaloes. Phylogenetic analysis revealed that Chinese swamp buffaloes could be divided into two distinct lineages, A and B, which were defined previously. Of the two lineages, lineage A was predominant across all populations. For predominant lineage A, Southwestern buffalo populations possess the highest genetic diversity among the three hypothesized domestication centers (Southeastern, Central, and Southwestern China), suggesting Southwestern China as the most likely location for the domestication of lineage A. However, a complex pattern of diversity is detected for the lineage B, preventing the unambiguous pinpointing of the exact place of domestication center and suggesting the presence of a long-term, strong gene flow among swamp buffalo populations caused by extensive migrations of buffaloes and frequent human movements along the Yangtze River throughout history. Our current study suggests that Southwestern China is the most likely domestication center for lineage A, and may have been a primary center of swamp buffalo domestication. More archaeological and genetic evidence is needed to show the process of domestication. PMID:23437167

  19. In Buffalo, Opening Doors for the Overlooked

    ERIC Educational Resources Information Center

    Honawar, Vaishali

    2007-01-01

    This article describes the Buffalo Prep program. Housed at University of Buffalo, the program identifies disadvantaged but talented minority children, places them in academic-enrichment classes, and then finds them spots at private schools and a more selective public high school in the area to complete their precollegiate careers. In addition to…

  20. Phylogeography and domestication of Chinese swamp buffalo.

    PubMed

    Yue, Xiang-Peng; Li, Ran; Xie, Wen-Mei; Xu, Ping; Chang, Ti-Cheng; Liu, Li; Cheng, Feng; Zhang, Run-Feng; Lan, Xian-Yong; Chen, Hong; Lei, Chu-Zhao

    2013-01-01

    To further probe into whether swamp buffaloes were domesticated once or multiple times in China, this survey examined the mitochondrial DNA (mtDNA) Control Region (D-loop) diversity of 471 individuals representing 22 populations of 455 Chinese swamp buffaloes and 16 river buffaloes. Phylogenetic analysis revealed that Chinese swamp buffaloes could be divided into two distinct lineages, A and B, which were defined previously. Of the two lineages, lineage A was predominant across all populations. For predominant lineage A, Southwestern buffalo populations possess the highest genetic diversity among the three hypothesized domestication centers (Southeastern, Central, and Southwestern China), suggesting Southwestern China as the most likely location for the domestication of lineage A. However, a complex pattern of diversity is detected for the lineage B, preventing the unambiguous pinpointing of the exact place of domestication center and suggesting the presence of a long-term, strong gene flow among swamp buffalo populations caused by extensive migrations of buffaloes and frequent human movements along the Yangtze River throughout history. Our current study suggests that Southwestern China is the most likely domestication center for lineage A, and may have been a primary center of swamp buffalo domestication. More archaeological and genetic evidence is needed to show the process of domestication.

  1. 33 CFR 117.955 - Buffalo Bayou.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 1 2013-07-01 2013-07-01 false Buffalo Bayou. 117.955 Section 117.955 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY BRIDGES DRAWBRIDGE OPERATION REGULATIONS Specific Requirements Texas § 117.955 Buffalo Bayou. (a) The draw of...

  2. 33 CFR 117.955 - Buffalo Bayou.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Buffalo Bayou. 117.955 Section 117.955 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY BRIDGES DRAWBRIDGE OPERATION REGULATIONS Specific Requirements Texas § 117.955 Buffalo Bayou. (a) The draw of...

  3. 33 CFR 117.955 - Buffalo Bayou.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 1 2012-07-01 2012-07-01 false Buffalo Bayou. 117.955 Section 117.955 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY BRIDGES DRAWBRIDGE OPERATION REGULATIONS Specific Requirements Texas § 117.955 Buffalo Bayou. (a) The draw of...

  4. 33 CFR 117.955 - Buffalo Bayou.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 1 2011-07-01 2011-07-01 false Buffalo Bayou. 117.955 Section 117.955 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY BRIDGES DRAWBRIDGE OPERATION REGULATIONS Specific Requirements Texas § 117.955 Buffalo Bayou. (a) The draw of...

  5. 33 CFR 117.955 - Buffalo Bayou.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 1 2014-07-01 2014-07-01 false Buffalo Bayou. 117.955 Section 117.955 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY BRIDGES DRAWBRIDGE OPERATION REGULATIONS Specific Requirements Texas § 117.955 Buffalo Bayou. (a) The draw of...

  6. Monitoring the Freezing Point of Buffalo Milk.

    PubMed

    Pesce, Antonella; Salzano, Caterina; De Felice, Anna; Garofalo, Francesca; Liguori, Salvatore; De Santo, Annunziata; Palermo, Pierpaolo; Guarino, Achille

    2016-04-19

    The aim of this study was to evaluate the basic freezing point of buffalo milk. Bulk milk samples were collected from buffalo and cattle farms in Caserta area from 2008 to 2014. The analysis involved a total of 1886 buffalo milk samples and 1711 bovine milk samples. These were also tested for fat, protein and lactose contents by means of infrared spectrometry. The freezing point was determined by means of a thermistor cryoscope. Data underwent statistical analysis. Our research showed an average freezing point of -0.528°C for buffalo milk and -0.522°C for bovine milk. Given the lack of data on the freezing point of buffalo milk, our study provides the first indication of a basic freezing point of the milk of this species in Italy.

  7. Papillomatosis in buffaloes: a less-known disease.

    PubMed

    Somvanshi, R

    2011-08-01

    Scant information is available on papillomatosis in buffaloes, and it is an almost unknown disease. It has been described from India, Italy and Turkey. Buffalo papillomatosis occurs in cutaneous and mucosal forms. Cutaneous papillomatosis is manifested as cutaneous wart (CW) and teat papilloma types. The condition is known to be caused by bovine papillomaviruses (BPV)-1 and 2 and their mixed infections. Buffalo CWs are experimentally transmissible to hamsters, cattle as well as buffaloes. Once BPV establishes infection in buffaloes, infection spreads from buffalo to buffalo, without cattle intermediary. Histologically, CWs are mostly diagnosed as fibropapillomas. The mucosal form occurs as urinary bladder tumours similar to enzootic bovine haematuria which is also associated with bracken fern infested areas. BPVs are yet to be demonstrated in teat papillomas and urinary bladder tumours of buffalo cases. Papillomatosis in buffaloes is a little-known disease, but it is a separate infectious ailment of buffaloes and deserves more attention by researchers.

  8. Arboviruses recovered from sentinel livestock in northern Australia.

    PubMed

    Gard, G P; Shorthose, J E; Weir, R P; Walsh, S J; Melville, L F

    1988-10-01

    Over 700 arboviruses were recovered between 1981 and 1987 from the blood of sentinel livestock near Darwin. Twenty-three isolates were made from sheep, goats, swamp buffalo (Bubalus bubalis) and horses, and the remainder were from cattle. The isolates have been typed as 27 separate viruses belonging to the bluetongue, epizootic haemorrhagic disease, Palyam, Simbu, bovine ephemeral fever, Tibrogargan and alphavirus groups. Ten of these viruses have not been isolated elsewhere in Australia and four have been isolated only in Darwin. Considerable annual variations in virus activity and in the durations of detectable viraemia were observed.

  9. Domestic livestock resources of Turkey: water buffalo.

    PubMed

    Yilmaz, Orhan; Ertugrul, Mehmet; Wilson, Richard Trevor

    2012-04-01

    Water buffalo are an ancient component of Turkey's domestic livestock resources. Commonly referred to as the Anat