Addition of pomegranate juice ( Punica granatum ) in tris‐based extender improves post‐thaw quality, motion dynamics and in vivo fertility of Nili Ravi buffalo ( Bubalus bubalis ) bull spermatozoa (original) (raw)
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Cryopreservation of buffalo (Bubalus bubalis) semen-limitations and expectations
2017
Artificial insemination with cryopreserved semen is the most viable biotechnology for faster and increased genetic improvement in many species allowing for improved herd performance and productivity. Pakistan has 34.6 million buffalo which have major share in total milk produced in the country. Rapid increase in human population and demand for animal products motivated the researchers to increase per animal milk production using this biotechnology. In buffalo, natural breeding practice is common in the country compared to artificial insemination, low fertility rate with cryopreserved semen is main hindrance in its propagation. It is need of the hour to disseminate the knowledge of various factors and components contributing in buffalo semen cryopreservation to improve milk and fertility rate of this breed.
Journal of Advanced Biotechnology and Experimental Therapeutics, 2024
Abstract: Cryopreservation has been used extensively for cattle in Bangladesh, however no study was conducted on the cryopreservation of buffalo sperm in Bangladesh. Thus, the current study aimed to evaluate the suitability of different semen extenders for improving the post-thaw quality of Bangladeshi buffalo sperm under a manual cryopreservation protocol. The manual cryopreservation protocol was compared and optimized with a commercial biofreezer protocol. Then, the efficacy of different diluters was evaluated using the optimized manual cryopreservation protocol. Meanwhile, the post-thaw sperm quality in terms of motility and morphology was evaluated by computer assisted sperm analyzer during the optimization process. During manual cryopreservation, the first cooling from 37 °C to 5 °C was done in an equilibration chamber and the second cooling from 5 °C to-120 °C in a Styrofoam box using liquid nitrogen vapor from different distances (0.5, 1.5, 1.6, 2 and 3 inches). Simultaneously, another batch of sperm was cryopreserved using a programmable freezer. The highest number of motile sperms (62.67±1.12; P<0.01) and progressive motility (38.97±1.10; P< 0.001) was observed at 1.6 inches above liquid nitrogen, which were similar to the results obtained from automated biofreezer protocol (65.94±4.65 and 45.54 ± 3.64, respectively). To evaluate the semen extenders' efficacy, one locally developed Tris-fructose-egg yolk-based diluter and two commercial diluters (Andromed, and Triladyl) were used in the freezing of buffalo sperm. The highest recovery and conception rates were observed in sperm diluted with tris-fructoseegg yolk-based (TFE) (82.4% and 80%, respectively). Therefore, it is suggested that this manual cryopreservation protocol and the TFE diluter could be a suitable and inexpensive alternative for Bangladeshi buffalo sperm considering post-thaw sperm quality and fertility.
Buffalo Bulletin, 2022
In this study, efforts were made to investigate fresh semen parameters and to select a suitable extender for buffalo semen cryopreservation. Experiment I, fresh undiluted seminal parameters were determined while Experiment II, efficacy comparison of coconut water extender (CWE) with tris citric acid extender (TCAE) and skimmed milk extender (SME) was made. Four bulls single ejaculate was collected weekly for 5 and 10 weeks for Experiment I and II, respectively however osmotic pressure replicates were 16 and 6 for semen and seminal plasma, respectively. In Experiment I, each bull spermatozoa concentration, motility (%), semen volume and pooled semen percentage NAR, PMI, viability and MTT (3-(4, 5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide) reduction rate was checked. In Experiment II, pooled semen added to extenders and then equilibrated (4 o C) and filled to obtain 20×10 6 spermatozoa/0.5 ml straws before plunging in liquid nitrogen. Percentage thawed spermatozoa viability, normal acrosomal ridge (NAR), motility, DNA damage, plasma membrane integrity (PMI) and lipid peroxidation (nM) recorded. In Experiment I, seminal parameters as spermatozoa concentration (1226.43±71.48 million/mL), semen volume (2.84±0.14 mL), viability (90.05±0.71%), motility (77.13±0.71%), PMI (86.23±0.34%), NAR (94.67±0.30%) and MTT reduction rate (0.290±0.06) while osmotic pressure of seminal plasma (294.83±3.87 mOsm/ kg) and semen (290.87±2.58 mOsm/kg) was recorded. In Experiment II, TCAE higher (P<0.05) sperm motility noted compared to CWE and SME whereas percentage viability, NAR, PMI, DNA damage was non-significant. Lipid peroxidation compared to SME and TCAE was higher (P<0.05) in CWE. In conclusion, based on sperm motility and lipid per oxidation, TCAE was more efficient for cryopreservation of buffalo semen.
Effect of Duck Egg Yolk on Cryopreservation and Fertility of Egyptian Buffalo Bull Semen.
Abstract: The objective of this study was to compare the effectiveness of different duck egg yolk (DEY) concentrations (10, 15 and 20% DEY) with chicken egg yolk (20% CHEY) on the cryopreservation of Egyptian buffalo spermatozoa following dilution, equilibration and freezing-thawing processes. For this purpose, one ejaculate of semen from each of three Egyptian buffalo bulls was collected twice each week for 4 weeks with artificial vagina (42ºC). Pooled ejaculates were divided into four parts and were diluted in Tric citric acid glycerol extender containing either 10 or 15 or 20% DEY or 20% CHEY at 37ºC. Extended semen was equilibrated for 4h at 5ºC and then was filled in 0.5 ml straws and frozen in liquid nitrogen. Thawing of semen was performed at 37ºC for 30s. Progressive sperm motility, live sperm and plasma membrane integrity (%) after different stages of cryopreservation were assayed. The results of postthaw sperm motility for all extenders containing DEY showed significantly (P < 0.05) higher values (52.1, 59.6 and 52.5% for 10, 15 and 20% DEY, respectively) compared to 47.9% for control extender (20% CHEY). Also, post equilibration and post-thaw live sperm and plasma membrane integrity were significantly (P < 0.05) higher in 15% DEY extender as compared with control extender (20% CHEY), the values being (78.4 vs. 70.7% and 85.8 vs. 79.8%) and (72.3 vs. 55.0 % and 80.7 vs. 69.1 %)respectively. Using a post-thawing semen containing 15% DEY yielded comparatively highest conception rate (65.8%) followed by 20% DEY (59.3%), 20% CHEY (58.6%) and 10% DEY (58.1%). In conclusion, DEY compared to chicken egg yolk in extender improves the frozen-thawed quality of Egyptian buffalo bull spermatozoa and fertility rate.
Effects of Thawing Methods (Techniques) on Freeze Thaw Buffalo Bull Sperm Quality
Egyptian Journal of Veterinary Sciences
B ACKGROUND: Semen cryopreservation involves preserving the sperms so they can be used later. This technique can prove to be a pathway for restoring the number of endangered species as their sperm can be preserved and used for fertilization later on. Objective: The propose of study is to find out the optimum thawing time, temperature and post thaw incubation period of cryopreserved buffalo sperm. Methods: Three buffalo bulls of the 1-2 year age were selected at Semen Production Unit, Qadirabad, Pakistan. Semen was collected by artificial vagina at 42 °C and 18 qualified ejaculates (two ejaculates/bull/week) for 3 weeks (replicate) were cryopreserved following a standard protocol. Frozen semen was divided in different groups, with each group being thawed at different temperature and time. Group I was subjected to slow thawing at 4-5°C, 2 nd group to moderate thawing at 35ºC-37 ºC , 3 rd group to rapid thawing while the 4 th one was subjected to air thawing. Moreover, optimum incubation period of cryopreserved sperm was evaluated by thawing and incubating 42 straws in a water bath at 37ºC in with 2 straws being assessed every hour up till 10 hours. Results: Thawed semen samples were assessed based on different semen quality parameters. Acrosome integrity, plasma membrane integrity and progressive motility of cryopreserved buffalo sperm were significantly higher (P<0.05) in Group II which was subjected to moderate thawing. However, sperm quality parameters remained same (P > 0.05) with different incubation time (from 1 hr to 11 hr at 37°C). Conclusion: In conclusion, moderate thawing is effective in protecting the sperm cell from freeze thaw damages during cryopreservation. Sperm functional quality parameters were very acceptable after even 5 h of incubation indicated the usefulness of these incubated semen straws for field insemination in areas where availability of liquid nitrogen containers is problematic. Digital thermos maintained at 37°C could be alternative for short distance travelling for taking semen doses at insemination site. Further research efforts are needed to verify the fertilizing ability of this straw incubated (37°C) semen.
The Success Rate of Artificial Insemination Using Post-Thawed Spotted Buffaloes Epididymal Sperm
Media Peternakan, 2013
Spotted buffalo, an exotic species that exists in Tana Toraja, South Sulawesi, Indonesia, is getting extinct due to high number of slaughtered during a funeral ceremony, called Rambu Solo', as well as special treatments that do not allow the male spotted buffaloes perform natural mating activity. According to that, the research was trying to start conservation program by collected the cauda epididymal sperm soon after slaughtered. Two egg yolk-based extenders with different buffers, tris hydroxyl amino methane (TEY20) and trisodium citrate dehydrate (CEY20), were used as comparison to evaluate the post-thawed epididymal sperm quality and fertilizing capacity in artificial insemination program. The results showed that the post-thawed progressive motility of epididymal sperm was 40% and 39.17%, while viability was 65.99% and 63.26% and membrane integrity was 65.43% and 63.03% in TEY20 and CEY20 extenders, respectively. The success rate of pregnancy was 46.67% using post-thawed epididymal sperm in TEY20 and 40% using the one in CEY20 extenders. In conclusion, tris-based and citrate-based extenders have similar ability to maintain the epididymal sperm quality and its fertilizing capacity.
Innovative strategies to improve fertility of buffalo semen
2015
The improvement of fertility is an important contribution that applied scientific research can provide to buffalo breeding, especially in productive areas like the Campania region. Many studies have been conducted to improve the management of the female reproductive system, while insufficient are the information about the effect of the male on the reproductive efficiency in this species. Buffalo is a photoperiodic animal that tends to increase the reproductive activity during periods when daylight hours decrease. In particular, in the male, it is manifested by a reduction of libido when daylight hours increase. This causes a reduction in the number of jumps and a worsening of the qualitative characteristics of the semen produced . Also, the cryopreservation process influences the quality of the semen causing premature sperm capacitation hence, reducing its longevity in the female reproductive tract. At present, for cryopreservation of semen, standard protocols are used equally for a...
Frozen-thawed quality of swamp buffalo spermatozoa cryopreserved with sugar palm juice extender
Buffalo Bulletin, 2020
The objective of this research was to examine the effectivity of sugar palm juice as an alternative extender to cryopreserve buffalo. Moreover, we expected to obtain the best composition of sugar palm juice that can be used widely as an alternative extender for buffalo semen cryopreservation. Semen was collected from swamp buffalo bull using artificial vagina. Fresh, diluted, equilibrated, and frozen-thawed was evaluated for general parameters, i.e spermatozoa progressive motility (M), viability (V) and membrane integrity (MI). Fresh semen that had good quality was diluted up to 1.108 sperm/ml in four different compositions of extenders, i.e lactose extender containing 20% egg yolk as control (C), 74% sugar palm juice + 20% egg yolk + 6% glycerol (SPG6), 73% sugar palm juice + 20% egg yolk + 7% glycerol (SPG7), and 72% sugar palm juice + 20% egg yolk + 8% glycerol (SPG8), respectively. Diluted-semen was equilibrated at 5oC for 4 h. It followed by frozen and stored in liquid nitrogen...