Buffalo Bulls Research Articles

Cloning and Expression of Leptospiral Immunoglobulin Like B Gene and Use of The Recombinant Antigen for The Diagnosis of Bovine and Caprine Leptospirosis

In the present study recombinant LigB protein is employed in latex agglutination test, which is a cross reacting lipoprotein able to detect acute infection caused by any pathogenic leptospiral serovars. It was employed for serodiagnosis of leptospirosis. The 46KDa 6X His tagged LigB protein, obtained by IPTG induction of recombinant E. coli M15 cells containing the N-terminal region of LigB gee in PQE30 expression vector, was purified by Ni-NTA affinity chromatography and adsorbed on latex bead surface for performing latex agglutination test against Leptospirosis suspected field sea. Western blot confirmed that rLigB is an immunodominant protein against which antibodies are produced during active infection. A total of 453 field sera, including 432 bovine sera, 18 caprine sera and three sera samples of buffalo bull collected post-mortem following death the animal from Indian Veterinary Research institute (IVRI) were tested using rLigB based LAT. The result showed that 300 sera were tested positive by rLigB based LAT, which were reconfirmed using microscopic agglutination test (MAT). The results from LAT were in concordance with MAT. In conclusion, under laboratory and field conditions, rLigB based LAT is a rapid, pen site, reliable diagnostic tool of high sensitivity and specificity for the detection of Leptospirosis. 
 Int. J. Appl. Sci. Biotechnol. Vol 8(2): 146-153

Nano-purification of raw semen minimises oxidative stress with improvement in post-thaw quality of buffalo spermatozoa.

The study consisted of application of anti-ubiquitin antibodies (Abs)-coated iron oxide-nanoparticles (IONPs) for minimisation of oxidative stress to contemporary live spermatozoa from the raw semen. Round-shaped IONPs (12.09±0.91nm) after two-stage functionalisation (silanisation and pegylation) were conjugated with Abs. Four aliquots from each of the 24 ejaculates (4 buffalo bulls) formed Control (Group I) and treatment (II, III and IV) groups; each containing 150±25 million dead/damaged spermatozoa. IONPs-Abs complex were added at ratio of 1:1 (0.5µg/ml), 1:2 (1.0µg/ml) and 1:4 (2.0µg/ml), respectively, in Groups II, III and IV. The semen quality parameters showed improvement at lag-stage (post-nano-purification before processing for cryopreservation). The mean post-thaw motility (%) in Group IV was found to be greater (p<.05) than Group I. Moreover, the overall DNA integrity (%) at post-thaw stage was improved in the nano-purified semen samples. The value of malondialdehyde was greater (p<.001) in Group I than Groups II, III and IV. The mean total antioxidant capacity and superoxide dismutase (U/mg protein) activity values in Group IV was greater (p<.05) than Group I. The study results show that IONPs conjugated with anti-ubiquitin Abs at 2.0µg/ml can be an effective dose for depletion of dead/damaged spermatozoa from buffalo ejaculates to minimise oxidative stress.

Assessment of semen characteristics and in vivo conception rate of preserved buffalo bull semen extended in tris enhanced with Diospyros kaki

BackgroundThe freeze-thaw process results in structural and functional damages caused by over accumulation of reactive oxygen species (ROS). Addition of exogenous antioxidants to semen extender is of a great importance to overcome the oxidative damage during the freezing process. The present study aimed to clarify the impact of Diospyros kaki on preserved buffalo semen quality. Tris extender enhanced with kaki juice (KEE) at concentrations 0% (control) and 1-10% (v/v). A twenty percent of freshly prepared egg yolk was added to the KEE extender (KEEY), then all tubes were centrifuged to eliminate any debris. Semen was added to the supernatants in other tubes. Semen was evaluated and the conception rate was implemented.ResultsSperm forward motility was significantly (P < 0.0001) kept high after 5 days of chilling for the concentrations 3, 4, and 5% with respect to the control and kept high (P < 0.01) at the other concentrations till the 4th day of chilling. Addition of KEE had significantly (P < 0.03) ameliorated post freezing sperm motility with all the concentrations of the extender except the concentration (10%). The highest spermatozoal motility was obtained with the concentration of 1% with respect to the control. Alive sperm%, abnormalities%, and % of intact spermatozoa membranes (HOST%) were similar to the control with all concentrations of kaki used. The conception rate was higher when 1-6% KEE were used.ConclusionsSome concentrations of Diospyros kaki improved buffalo bull semen quality post-chilling, post-freezing, the conception rate, and the concentration 1% gave the best results.

Semen production and semen quality of indigenous buffalo breeds under hot semiarid climatic conditions in India

Semen data comprising of 97,023 ejaculates of 197 bulls from 6 buffalo breeds were analyzed. The traits considered were as follows: ejaculate volume, sperm concentration, mass activity, initial and post-thaw motility, total sperm, and total motile sperm before and after thawing as well as a composite trait equal to the theoretical number of doses which can be produced from each given ejaculate. The objective was to measure the semen production potential of indigenous buffalo bulls and identify factors affecting these traits. A linear mixed model was used, including a random bull effect along with other fixed factors: the order of the ejaculate on a particular day, the interval between collections, the time of collection, the breed, the age at collection, the semen collector, and the year and month of collection. The study showed breed wise variation for all traits. The first ejaculate of a bull on a particular day was superior to the second for nearly all the traits. Longer collection intervals are better than shorter intervals for all the parameters, although short collection interval of 2 to 4days produced higher yield in terms of total semen doses without hampering semen quality. The study also showed a slight decrease of semen quality with time of collection within a day. The Murrah breed showed comparatively consistent performance during their whole life compared with the other breeds. Repeatability estimates for semen traits were found to be low (0.09 for mass activity) to relatively large (> 0.4 for volume and concentration). A negative correlation was found between bull effects for semen volume and concentration while a high positive correlation was found between mass activity, initial motility, and post-thaw motility. Results of the study will help in suggesting suitable management and breeding plans for semen production traits.

Effects of Feeding and Maturation System on Qualitative Characteristics of Buffalo Meat (Bubalus bubalis)

Simple SummaryNowadays, consumers used to being properly informed when purchased animal products. They are influenced by some meat characteristics like colour and tenderness. Nutrition is an important strategy to influence these properties, together with post day ageing processes. In this regard, the inclusion of fresh forage during fattening period and a prolonged maturation time in a monitored refrigeration device as Maturmeat®, were assessed on 16 Italian Mediterranean buffalo bulls. Data regarding physical-chemical analyses, colour and texture on two muscles, the Semitendinosus and the Longissimus muscles, were recorded during two maturation times (30 and 60 days). The combined used of fresh forage inclusion and post dry ageing processes influenced in a different way the two considered muscles, suggesting that different muscles could have different properties. Moreover, meat colour changed in both muscles in animals fed green forage and maturated in Maturmeat® while no significant change was observed in the control group. In conclusion, the use of this monitored post dry ageing system could influence the saleable yield and productivity of buffalo meat and allow the creation of ready-to-eat food, produced starting from raw ground meat, that is very requested by the consumers.We aimed to evaluate the effects of post dry ageing (PDA) period on meat colour and rheological characteristics in 16 buffalo bulls fed two different diets: with (FRS) or without (CTL) rye grass. Animals were randomly divided into two feeding groups and slaughtered at 540 ± 4.7 and 533 ± 7.0 kg of live body weight, respectively, for the CTL and FRS group. After five days post-mortem ageing (T0), Semitendinosus muscle (ST) and Longissimus muscle (LD) underwent a prolonged maturation process in a controlled meat chamber for 30 days (ST) and until 60 days (LD). After 30 days (T1), significant changes (p < 0.01) in meat colour (ΔE) in both muscles of the FRS group was recorded, while no significant change was observed in CTL group. The FRS diet had a positive effect on textural properties of ST muscle compared to CTL diet, as well as hardness, chewiness and gumminess. All qualitative characteristics improved in the first period of PDA but, whereas LD showed to keep improving, extending the post-ageing period by further 30 days, the ST becomes un-processable at 60 days. In conclusion, a combined used of fresh feeding and PDA period could enhance both tenderness and colour in animal fed FSR.

Analysis of meiotic segregation by triple-color fish on both total and motile sperm fractions in a t(1p;18) river buffalo bull.

Chromosomal aberrations are relatively frequent pathologies in both humans and animals. Among them, translocations present a specific meiotic segregation pattern able to give a higher percentage of unbalanced gametes that can induce fertility problems. In this study, the meiotic segregation patterns of 1p, 1q and 18 Bubalus bubalis chromosomes were analyzed in both total sperm fraction and motile sperm fraction of a t(1p;18) carrier and a control bulls by triple-color FISH analysis with a pool of specific BAC probes. The frequencies of each total sperm fraction products in the carrier resulting from alternate, adjacent I, adjacent II and 3:1 segregation were 39%, 20%, 1% and 38%, respectively. On the other hand, the frequencies of each motile sperm fraction products in the carrier resulting from alternate, adjacent I, adjacent II and 3:1 segregation were 93%, 5%, 0% and 2%, respectively. The frequencies of normal sperms in the carrier were 27% and 69% in total sperm fraction and motile sperm fraction, respectively. The frequencies detected in motile sperm fraction were also validated by comparison with bull’s progeny. To our knowledge, this is the first report on the meiotic segregation patterns in motile sperm fractions of B. bubalis bull carrying a chromosomal translocation. These data suggest that translocation has a very limited effect on aneuploidy in the gametes, and therefore, on the reproductive abilities of the bull.

Effect of partial deoxygenated extender on different post thaw incubation periods of physico-morphological and functional attributes during cryopreservation of buffalo spermatozoa

The study was designed to explore the effect of partial deoxygenated extender of different post thaw incubation periods on physico-morphological attributes during cryopreservation of buffalo semen. Semen samples with mass motility of ≥4+ and individual progressive motility ≥ 80 collected from murrah buffalo bulls were utilized. Immediately after collection, a portion of semen was evaluated for progressive motility, hypo-osmotic swelling response (HOS response) and acrosomal intactness and other portion was divided into two equal groups. Group I was kept as control and diluted with Egg yolk-Tris-Glycerol (EYTG) with a maximum of 60×106 sperm/mL. Group II was diluted with partially deoxygenated EYTG extender. Both the groups were subjected to equilibration and freezing. Both the group of semen samples were evaluated at pre-freeze and post-thaw stage of progressive motility, viability, HOS response and acrosomal intactness. Post-thaw motility and viability was studied at 0, 15, 30, 45 and 90 minutes of incubation. At pre-freeze and post thaw stage, significantly (p

The addition of resveratrol in tris citric acid extender ameliorates post-thaw quality parameters, antioxidant enzymes levels, and fertilizing capability of buffalo (Bubalus bubalis) bull spermatozoa

Resveratrol is a natural grape-derived polyphenol with potent antioxidant properties that protect spermatozoa against lipid peroxidation (LPO) by eradicating free radicals. The objectives of this study were to 1) appraise the effects of resveratrol in extender on post-thaw quality parameters, antioxidant enzymes, adenosine triphosphate (ATP), DNA fragmentation, LPO and 2) fertilizing capability of buffalo bull spermatozoa. Semen was collected from four fertility proven bulls with artificial vagina thrice, evaluated initially, and diluted in five different extenders containing resveratrol (T4 = 100 μM, T3 = 50 μM, T2 = 20 μM, T1 = 10 μM), and control (no resveratrol), and evaluated after post-dilution and post-thawing stage of cryopreservation. Analysis of variance revealed that, there was no difference (P > 0.05) in any parameters due to treatments at post-dilution. However, at post-thawing, sperm progressive motility (%), plasma membrane integrity (%), mitochondrial membrane potential (%) and ATP levels (nmol/106) were found higher in semen samples cryopreserved in T3 and 4 than control. Sperm supravital plasma membrane integrity (%) and viable/acrosome integrity were found higher in semen samples cryopreserved in T4 than control and T1. Furthermore, sperm catalase (U/mL), glutathione peroxidase (μM) and superoxide dismutase (U/mL) concentrations were found significantly higher in resveratrol treated groups as compared to the control. Conversely, DNA fragmentation (%) and LPO (μM/mL) were significantly (P > 0.05) decreased in semen samples cryopreserved in T4 in comparison to the control. Fertilizing capability was found higher in semen samples cryopreserved in T4 as compared to the control (%, 77.33 vs. 57.41, P < 0.05). It is concluded that the addition of resveratrol in extender ameliorates quality parameters, antioxidant enzymes levels and fertilizing capability, and alleviate DNA fragmentation and LPO in buffalo spermatozoa during cryopreservation.

Age-Dependent Changes in Fresh Semen Quality of Swamp Buffalo (Bubalus bubalis)

The purpose of this study was to evaluate the fresh semen quality of swamp buffalo raised in Lembang Artificial Insemination Center with respect to different age. The semen was collected from one swamp buffalo bull at three different stages (5, 6, and 7 years old). The evaluation of semen quality was conducted directly upon collection. Semen color, consistency, and mass sperm motility were analyzed descriptively, while semen volume, pH, sperm concentration and individual sperm motility were analyzed using analysis of variance followed by least significant difference test. The results showed that the dominant physical characteristics of swamp buffalo’s semen were milky white color with moderate consistency and fast mass sperm motility (++). There was a highly significant effect (P<0.01) of age on semen volume, sperm concentration, and individual sperm motility, whereas semen pH was not affected (P>0.05) by age. Swamp buffalo had higher (P<0.01) semen volume and sperm concentration at 7 years old, but with lower (P<0.01) individual sperm motility as compared to 5 and 6 years old. In brief, this study reveals that semen production of swamp buffalo is dependent of age with 7 years old become the most superior age.

Correlations Amongst Functional and Morphological Attributes and Oxidative Markers of Fresh and Cryopreserved Semen of Gir and Murrah Bulls

This study was undertaken during the winter season on healthy mature Gir cattle and Murrah buffalo bulls (n=3 each). The semen samples (6 ejaculates/bull, total 36 ejaculates) collected in the morning using artificial vagina were evaluated for routine seminal attributes, including acrosomal and plasma membrane integrity. The samples were then diluted @ 100 million sperm/ml with tris fructose yolk glycerol extender without and with sericin @ 0.1, 0.25, 0.5 and 1.0% (w/v), filled in French mini-straws, and frozen in LN2 using biofreezer as per standard freezing protocol. Straws were thawed in water bath at 37°C for 30 sec and evaluated for post-thaw quality, viz., motility, viability, morphology, acrosome integrity and plasma membrane integrity (HOST). Lipid peroxidation (malondialdehyde - MDA production) and activities of enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were assessed as oxidative markers in seminal plasma of freshly diluted and frozen-thawed semen samples. Sericn at 0.5% level significantly (p less than 0.01) improved the post-thaw sperm quality with reduced oxidative stress in both the species. The breed-wise correlation coefficients (r) among sperm quality attributes and oxidative markers were studied in fresh and frozen-thawed semen of each species, and also for fresh with frozen-thawed semen. The findings revealed significant interrelationships amongst most of the attributes of fresh as well as post-thawed semen and also of fresh semen attributes with those of cryopreserved semen including oxidative markers in both the species. Sperm motility estimation in fresh, pre-freeze and post-thawed semen was a legitimately good indicator of quality of spermatozoa at various steps of semen processing/freezing, and its fertilizing potential. Thus, the sperm motility, HOS test and either MDA or SOD/GPx activity alone may be used as valuable and practical tools for routine assessment of bovine semen quality considering significant correlations found between them.

Fresh and Post-Thaw Seminal Attributes of Jafarabadi Buffalo Bulls

The study was conducted on four Jafarabadi breeding bulls, 5-6 years old to know the fresh and post-thawed seminal characteristics based on total of 192 semen ejaculates evaluated and cryopreserved over one year period. The mean values of fresh neat seminal characteristics of Jafarabadi bulls, viz., ejaculate volume (ml), colour/density (score), sperm concentration (million/ml), mass activity (score), initial motility (%), live sperm (%), abnormal sperm (%), HOS reactive sperm (%) and acrosomal integrity (%) were 5.19±0.18, 2.38±0.10, 1253.36±24.75, 3.73±0.05, 80.31±0.05, 86.20±0.64, 5.00±0.40, 85.75±0.43 and 93.56±0.56, respectively, whereas the mean post-thawed sperm characteristics, viz., progressive sperm motility, live sperm (%), abnormal sperm (%), HOS reactive sperm (%), acrosomal integrity (%) and first insemination conception rate (%) observed were 57.60±0.36, 66.34±0.53, 8.85±0.33, 56.97±0.46, 75.26±0.17 and 44.63±0.14, respectively. The semen quality of fresh and post-thawed samples observed was within normal limit for use in breeding program with satisfactory first insemination conception rate.

Comparative efficacy of conventional stains for evaluation of plasmalemma and acrosome integrity of buffalo spermatozoa

The aim of the present study was to compare the efficiency of various conventional single and dual staining protocols for assessment of spermatozoa attributes. The efficiency of single stains such as Eosin-Nigrosin (EN) and Trypan blue (TB) for viability and Giemsa (G) for acrosome integrity vis-a-vis dual stains such as Trypan blue-Giemsa (TB-G) and Eosin Nigrosin-Giemsa (EN-G) were compared. Ejaculates (12) from 3 buffalo bulls were used for evaluation of livability and acrosome integrity using said stains. The smears stained with EN-G were difficult to visualize distinctly and provided obscure results. A nonsignificant difference was found between EN, TB and TB-G for spermatozoa viability. However, at recommended concentration, TB yielded faint images, thus making the observation unnecessarily tedious. However, staining with TB-G was found advantageous for simultaneous determination of viability and acrosome status. Moreover, the results between G and TB-G deferred nonsignificantly indicating similar effectiveness of both the stains. Overall, TB-G can be a reliable, quick and convenient method for determining viability and acrosome integrity simultaneously for routine semen evaluation.

Utility of first lactation fat energy corrected milk yield as a trait for genetic evaluation of Mehsana buffalo bulls using various sire evaluation methods

India being a vegetarian country, milk is the major source of dietary bio-energy, but majority of animals are routinely being evaluated on the basis of their milk producing ability. The present study was aimed to come up with a sire evaluation methodology based on first lactation Fat Energy Corrected Milk Yield (FBE) in order to obtain an accurate and unbiased estimate of breeding value of Mehsana buffalo bulls and ranking them on the basis of their daughter's performance for future herd improvement. Data for the present study included 7825 she buffaloes in their first lactations, extended over a period of 25 years (1989 to 2013), from field progeny testing programme of Dudhsagar Research and Development Association (DURDA), Dudhsagar Dairy, Mehsana, Gujarat. The data were classified into different subclasses based on period, season, cluster and age at first calving group. The average breeding values of Mehsana buffalo bulls evaluated for FBE by least squares method (LSM), best linear unbiased prediction sire model (BLUP-SM) and best linear unbiased prediction animal model (BLUP-AM) methods were 1215.89, 1185.7 and 1185.7 kcal, respectively. BLUP-AM method had lowest error variance as compared to LSM and BLUP-SM methods of sire evaluation. This indicated that BLUP-AM was most efficient method.

Individual Variation in Semen Characteristics of Murrah Buffalo Bulls

This study evaluated the variation in semen characteristics between two individual Murrah buffalo bulls. Two hundred and ninety semen samples were involved in this study. The results showed that most of the semen had a milky white color with moderate consistency. The Bull B1 tended to have higher proportion of creamy color and moderate consistency than the Bull B2. The ejaculatory volume is varied (P&lt;0.01) between Bull B1 and Bull B2. However, semen pH and sperm concentration of Bull B1 and B2 were equal (P&gt;0.05). The individual bull had a highly significant effect (P&lt;0.01) on sperm motility of fresh semen. The individual variation (P&lt;0.05) also found on sperm motility of before freezing. Whereas, sperm motility of post-thawing semen and recovery rate value were not significantly influenced (P&gt;0.05). Moreover, it was found that the frozen semen production differed (P&lt;0.01) between two individual bulls, with bull B1 had higher production than the bull B2. This study clearly demonstrates that the frozen semen production of Murrah buffalo is highly dependent on the individual bull. However, the variation in frozen semen production between two bulls studied here is still qualified to be used for artificial insemination.

Seasonal variations in histomorphology and histochemistry of the prostate gland of buffalo bulls

The prostate glands of buffalo bulls were collected from an abattoir immediately after slaughter and were fixed in 10% neutral buffered formalin for paraffin block preparation by an acetone-benzene schedule. Paraffin sections of 3-5 μm were obtained on glass slides with a rotary microtome and were stained with various histomorphological and histochemical stains. The distributions of glandular acini and ducts were variable throughout the length of the pelvic urethra. The disseminate part of the prostate gland was derived from the epithelium of the pelvic urethra and was a compound tubulo-alveolar type of gland. The glandular acini comprised three types of cells, i.e. principal (serous cells), mucous, and narrow dark cells. In winter, the lumen of the acini became wider than in summer. The amount of glandular tissue decreased from winter to summer, whereas a reverse pattern was observed in the stroma. The prostatic concretions were larger and centrally placed during winter but became peripheral in summer. The average epithelial height of prostatic acini varied significantly between seasons. The activity of neutral mucopolysaccharides, basic proteins, lipids, and phospholipids was higher in winter than summer, whereas that of acid mucopolysaccharides was higher in summer than winter.

Relationship between angioarchitecture of the testicular artery and spermiogram parameters in Egyptian buffalo bulls (bubalus bubalis).

The current study aimed to investigate the effect of testicular artery angioarchitecture on the spermiogram parameters in Egyptian Buffalo bulls. Eight adult buffalo bulls aged between 2 and 8years were used for semen evaluation. For anatomical studies, the masculine gonads were collected after slaughtering 30 adult bulls and prepared for injection by different masses (Urographine® , Latex and Epoxy) through the testicular artery. The mass activity of the ejaculate was assessed immediately after collection. The sperm motility in fresh bull ejaculate was more than 80%. The overall mean percentage of sperm abnormalities was <18%. The recorded sperm abnormalities were mostly secondary one including distal protoplasmic droplet, fragmented tail, detached head, detached galea capitis and bent tail. The highest percentage of sperm viability was recorded just after sperm collection (alive>85%). The results revealed that testicular artery can be divided into three parts (abdominal, funicular and marginal parts) along its course. The coils of the funicular part forming a cone-like structure with its base fixed to the head of the testis. Two epididymal branches to the head and tail of epididymis emanate from the funicular part which continues as pars marginalis on the lateral surface of testis before its division into the lateral and medial testicular arteries on approaching the tail extremity of the testis. The increase in the length of the testicular artery with increase in the size of the testes played a great role in the degree of complexity of the architectural vascular patterns. The degree of complexity is affected by the number of coils formed by the vessel. The increase in the convolutions of the vessel will reduce the speed of blood flow to the gonads. Thus in turn will enabling the thermoregulatory mechanism to work more efficiently and will affect the semen value.

In vitro Supplementation of Linoleic Acid Improves Quality of Cryopreserved Buffalo Semen

Present study was designed to evaluate the effect of linoleic acid (LA) supplementation in extender on post thaw quality of cryopreserved buffalo semen. Semen was collected from three adult Nili Ravi buffalo bulls of same age with artificial vagina (42°C) for five weeks (replicates; N=30). Qualified semen ejaculates (&gt;1mL volume, &gt;60% motility, &gt;0.5 billion/mL concentration) were diluted in tris-citric acid extender containing 0.0 (control), 5.0, 10.0 and 20.0ng mL-1 of LA and were cryopreserved using standard procedures. Sperm motility and plasma membrane integrity were improved plessthan0.05) in extender containing 10.0 ng mL-1 of LA compared to other treatments and control while number of acrosome intact live sperm, chromatin integrity and number of morphologically normal sperms remained the same. In conclusion, LA supplementation in extender at 10.0 ng mL-1 was found to be beneficial to improve post thaw quality of cryopreserved buffalo semen.

Effect of animal age, postmortem chilling rate, and aging time on meat quality attributes of water buffalo and humped cattle bulls.

The study was aimed to investigate the influence of animal age, post-slaughter chilling rate, and aging time on meat quality of M. longissimus dorsi (LD) of water buffalo (Bubalus bubalis) and humped cattle (Bos taurus indicus) bulls. After slaughtering, one side of carcasses was subjected to rapid chilling (RC) (0±2°C) and other side was hanged in controlled room temperature (25±2°C) for 3hr, then allowed to the chiller (0±2°C). The meat quality traits were analyzed at 1, 7, and 14days of storage. It was noted that rapidly chilled carcasses from the younger animals of both species missed the ideal pH/temperature window, which affects the toughness of the meat. Buffalo meat presented higher shear force, color L* values, and lower b* value as compared to the cattle meat. Moreover, meatshear force values decreased while all color coordinates and cooking loss values increased with lengthening the storage time in both age groups of cattle and buffalo. In conclusion, the tenderness of cattle meat was superior to that of buffalo and RC adversely affect the shear force values of young cattle and both age groups of buffalo bulls.

Impact of in vitro fertilization by refrigerated versus frozen buffalo semen on developmental competence of buffalo embryos.

The objective of this study was to evaluate the fertility of buffalo semen for in vitro embryo production (IVEP) by comparing the effectiveness of refrigerated versus frozen semen. Three OPU sessions were held at 30-day intervals. For oocyte fertilization three buffalo bulls were used, one per session. At each OPU-IVEP session, one ejaculate was collected and divided into two equal aliquots. Each aliquot was either refrigerated at 5ºC/24 hours or frozen. A TRIS extender containing 10% low density lipoproteins, 0.5% lecithin and 10 mM acetylcysteine was used adding 7% glycerol for freezing. Sperm motility/kinetic was evaluated by CASA and sperm membrane integrity by the hypoosmotic swelling test. The evaluations were performed at 0 h (post final dilution at 37ºC), at 4 and 24 hs post-incubation at 5ºC and post-thaw. At 24 hs incubation and immediately post thaw sperm cells were used for in vitro fertilization of buffalo oocytes equally distributed between both groups. Cleavage rates and embryo development were followed. The embryo/matured and embryo/cultured rates were 25.4 x 14.0% and 29.4 x 18.5% (P<0.05), for chilled and frozen semen, respectively. It is concluded that cooled semen can be used for in vitro embryo production in buffalo and that a better efficiency may be expected for cooled compared to frozen semen.

Effect of Supplementation of Polyvinylpyrrolidone in Extender on Buffalo Semen Parameters

The current study was planned to evaluate the supplementation of Polyvinylpyrrolidone in extender on cryopreservation of Nili-Ravi buffalo bull semen. The semen samples were collected from Nili-Ravi buffalo (Bubalus bubalis) bull kept at SPU Qadirabad, District Sahiwal, Pakistan. Qualifying semen ejaculates having motility &gt;60%, volume &gt;5-6ml and concentration &gt;0.5 billion/ ml were diluted 50 × 106 motile sperm ml approximately at 37°C in Tris-citric acid extender supplemented with different concentrations of PVP (0.01, 0.05, 0.1mM). The extender without PVP was kept as control. Semen was stored at 4°C for a period of 2 h and kept at 4°C for 4h. Semen was filled in 0.5 ml French straws using suction pump at 4°C, plunged and stored in liquid nitrogen (-196°C). Semen straws were rewarmed at 37°C for 30 seconds and assessed for sperm motility, plasma membrane integrity (PMI), dead sperm percentage and the live sperm percentage. The data on the role of PVP on different parameters of semen quality were analyzed by using ANOVA and RCBD. Higher percentage (P&lt; 0.05) of sperm motility (66.1±7.51 and 59.4±10.72) and PMI (72.9±5.39 and 75.7±6.5) was observed in extenders having 0.05 mM and 0.1mM PVP compared to extenders having 1.5mM PVP and control. The percentage acrosomal integrity was observed greater (P&lt; 0.05) in extended semen containing 0.1mM (68.2±0.50) PVP compared to extenders having 0.01 and control.