Thaw Motility Research Articles

Seminal characteristics and efficacy of egg yolk free semen extender on cryopreservation of Murrah buffalo bull semen

The experiment was undertaken to study the efficacy of egg yolk free semen extender on cryopreservation of buffalo semen. A total of 24 semen ejaculates from Murrah bulls (5 to 6 years) maintained at Central Semen Station, Anjora, Durg, Chhattisgarh were divided into two aliquots and were diluted using Tris-fructose-egg yolk-glycerol (TFYG) and egg yolk free commercial extender (EYFCE, AndroMed). The overall average Murrah bull fresh semen volume (mL), sperm concentration (million/mL), initial progressive motility (%), live sperm (%), abnormal spermatozoa (%), intact acrosome (%) and HOS reactive sperm (%) were 4.90±0.49, 1286.92±100.23, 75.00±0.67, 81.45±0.61, 10.70±0.51, 80.16±0.43 and 68.83±0.40, respectively. There was significant difference between bulls in semen volume (P<0.01), individual motility (P<0.01) and total sperm abnormalities (P<0.05). The semen extended with TFYG and EYFC extenders were cryopreserved in french mini-straw (0.25 mL) using programmable bio-freezer. The mean values of post thaw motility (%), live sperm (%), abnormal sperm (%), intact acrosome (%), HOST (%) of Murrah bull semen extended with TFYG and EYFCE were 45.83±0.77 vs 49.16±1.33, 57.41±0.54 vs 58.62±0.73, 12.54±0.46 vs 12.45±0.63, 69.62±0.82 vs 71.45±0.64 and 56.45±0.52 vs 57.37±0.55, respectively. Post thaw motility, live spermatozoa and intact acrosome were significantly higher (P<0.01) in frozen thawed semen extended with as EYFCE compared to TFYG in Murrah bull semen. It is concluded that EYFCE results in improvement in post thaw semen characteristics viz. post thaw motility, live sperm percent and intact acrosome.

Reproductive performance of Jamunapari crossbred using frozen semen

Artificial insemination (AI) is modern techniques to introduce better and new genetic at the small holder farmer level by using frozen for livestock development. The aimed of the present study to popularization of goat’s artificial insemination to increase goat production with improved genetic merit by assessing the productive and reproductive traits using Jamunapari frozen semen. The study has been conducted in ten upazila namely Paba, Bagha, Nachal, Chapai sador under Rajshahi region and Birampur, Parbotipur, Fulbari, Kaliganj, Ulipur, Kawnia upazilas under the Rangpur regions. Artificial insemination has been performed in 260 numbers of does. The semen samples initially qualified for freezing was frozen as per the protocol standardized in BRAC bull station, stored in liquid nitrogen (LN2) having the post thaw motility of 50% and were later used for AI. The service per conception was 1.10 in the present study. Moreover, we found that overall conception rate was 67.1%. The average birth weight of male and female kids was 0.98 and 0.95 kg, respectively. In the present study kidding produced single, twins and triplets in 37.7, 50.65 and 11.69% of cases, respectively. The average post-partum heat period was 76.5 days. The results indicated frozen semen AI could successfully be used for preservation, conservation and propagation in Jamunapari goats. Bangladesh Journal of Animal Science 52 (3): 85-87.

A Comparative Study for Semen Quality Traits between Murrah and Nili Ravi Buffalo Breeding Bulls Maintained at an Organized Farm

Background: The aim of the study was to compare the effect of season on the semen quality traits of Murrah and Nili Ravi buffalo breeding bulls. Methods: For a period of 5 years, a total of 855 ejaculation records of 17 buffalo breeding bulls, including Murrah (11) and Nili Ravi (6) buffalo bulls maintained at an organised farm, were analysed for various semen traits using least squares analysis. Result: Breed had a significant effect (p less than 0.01) on semen volume, consistency, mass activity (MA), total sperm count (TSC) and post thaw motility (PTM) and a non-significant effect on colour and initial motility (IM). In Murrah bulls, except for semen volume, the other parameters viz. consistency, MA, TSC and PTM were found to be significantly higher than Nili Ravi bulls. Season of semen collection had a significant effect on consistency, TSC and PTM whereas a non-significant effect on semen volume, colour, MA and IM was observed. During the summer season, overall consistency and TSC were higher, but PTM was found to be comparatively lower. Season and breed interaction had a significant effect (p less than 0.01) on consistency, TSC and IM while a non-significant effect on semen volume, colour, MA and PTM was observed. In Murrah buffalo bulls, a non-significant effect of season on consistency was observed whereas, in Nili Ravi bulls, a significantly higher consistency during summer and spring and; a significantly lower consistency during the rainy season was observed. In Murrah bulls, the highest TSC were observed during the rainy and summer seasons; whereas, in Nili Ravi bulls only during the summer season. In Murrah bulls, semen consistency, MA and IM were lower during the spring season; semen volume was lower during the prewinter season; total sperm count was lower during the winter season. In Nili Ravi bulls, semen volume was lower during summer; consistency, MA and TSC were lower during the rainy season; and IM was lower during the spring season. The PTM was lower during the summer season in both the breeds.

Assess the quality of Murrah Buffalo (Bubalus bubalis) Bull Semen regarding seasonal and geographical variability in Nepal

We carried out this study to assess the quality of semen regarding the seasonal and geographical variability of Murrah buffalo breeding bull in Nepal. We aimed to access the quality of seme n of murrah buffalo bull considering the seasonal and geographical variability to help standardize quality semen producing protocol and concern. A total of eight Murrah buffalo bulls were used. Semen collection was done at once week and two ejaculations were considered at an interval of 30 minutes during the Summer (June to August), Autumn (September to November), Winter (December to February), and Spring (March to May) from 2017 to 2020 AD. Immediately after collection, initial motility, sperm concentration, pre-filling motility, post-thaw motility, live sperm, and abnormality of sperms were evaluated. Findings revealed that the season had affect (p<0.005) to the semen volume, individual motility, and post- thawing motility except for semen volume and sperm abnormality(p>0.05). However, initial motility, sperm concentration, and pre filling motility did not vary in months (p>0.05). It is thus well reflected that several traits related to quality and quantity of semen Murrah buffalo bulls may vary depending on the season of collection, location and age (p<0.05) with the semen traits of initial motility, sperm concentration, and pre-filling motility. Accordingly the semen collected during the autumn and spring season is revealed the percentage of average post thaw motility above the 52% that considered the success of artificial insemination is good for buffaloes.

Impact of Cysteine as a Semen Additive on Mitigating the Decline in Sperm Quality due to Summer Stress in Riverine Buffaloes (Bubalus bubalis)

The seasonal variations such as temperature, rain and humid environment greatly influences reproduction, and semen quality particularly in buffalo bulls, either positively or negatively. The heat stress coupled with freezing and thawing processes generates reactive oxygen species (ROS), which further reduce post-thaw spermatozoa quality. Application of cysteine as a semen additive have been reported to alleviate such damaging effect in canine, buck, bull, human, boar, and rooster. Therefore, objective of this study was to investigate ameliorating effect of cysteine on summer stress on buffalo spermatozoa. For the experiment, 24 ejaculates with individual progressive motility >70, volume >1mL and sperm concentration 1000 million/mL from four Murrah buffalo bulls were collected and divided into four groups as Control (Group-I), and Group-II, III, and IV (4, 8, and 12 mmol cysteine, respectively) containing 80 million spermatozoa per mL. Semen was filled (French mini straw), sealed and equilibrated at 40⸰C and frozen into LN2. The group containing L-cysteine at 4 mmol concentration had higher significant (p<0.05) difference in post thaw motility, viability, plasma membrane integrity and acrosome integrity, TAC, DNA integrity and lower significant (p<0.05) difference in spermatozoa abnormality, LPO and intracellular ROS. Moreover, cysteine concentration at 8 mmol has no beneficial effect compared to the Control, whereas Group-IV had greater adverse effect. This may be due to its toxicity at this concentration. It is evident from the study that the cysteine molecules at 4 mmol concentration produced more pronounced improvement in the freezability potential of buffalo bull spermatozoa.

Effects of cysteamine supplementation on cryopreserved buffalo bull semen quality parameters

This work aimed to determine the effect of cysteamine (25, 50, 100 and 200 μM) incorporated during dilution on frozen thawed buffalo semen quality. Semen was collected twice weekly for 7 consecutive weeks from three Egyptian buffalo bulls using an artificial vagina. Semen samples were pooled and extended with a Tris-based extender, cooled, equilibrated and finally frozen in liquid nitrogen. The diluted semen was evaluated for motility, viability, morphology, plasma membrane and DNA integrity, in addition to oxidative stress and in vitro fertilizing capability. The post thaw motility and velocity parameters noticeably increased with different concentrations of cysteamine (mainly 100 μM) during different incubation periods. The post thaw sperm viability and normality significantly (p < 0.05) improved with concentrations of 50 and 100 μM. Plasma membrane integrity substantially increased at all concentrations of cysteamine. Cysteamine reduced alanine aminotransferase (at all concentrations), aspartate aminotransferase (at 25–100 μM), and creatine kinase (at 100 and 200 μM). Cysteamine at a concentration of 100 μM noticeably enhanced the total antioxidant capacity and glutathione peroxidase and decreased nitric oxide production. Cysteamine, at concentrations of 100 and 200 μM, increased the DNA intensity in the comet head (%) and decreased the DNA % in the comet tail. The comet tail length and moment substantially decreased at concentrations of 50–200 μM. Cysteamine did not affect the in vitro fertilizing capability of sperm. In conclusion, cysteamine incorporation (mainly at a concentration of 100 μM) in buffalo semen extender showed varying protective effects on different sperm parameters against cryo-damage; however, it did not affect the in vitro fertilizing capacity of sperm.

PREDICTION OF FROZEN SEMEN DOSES PRODUCTION IN DAIRY STUDS USING MACHINE LEARNING ALGORITHM

The ability to predict the frozen semen doses produced per ejaculate would be of considerable benefit for the management of skill, human resource, capital and time. The new computing paradigm called machine learning involves in predicting dependent variable by learning complex and non-linear relationship among independent variables. The purpose of this study is to develop prediction model using one of the conventional and machine learning modelling techniques called Multiple Linear Regression (MLR) and Artificial Neural Network (ANN) respectively. A Total of 1,57,532 ejaculates data were used for modelling. The modelling involved prediction of frozen semen doses produced per ejaculate using independent variables namely volume of ejaculate, ejaculate number, sperm concentration, initial motility and post thaw motility. Various combinations of architectural parameters were employed to explore optimum configuration for each model. The ANN (R2=90.66) modelling was observed to be efficient over MLR (R2=73.52). The root mean squared error (RMSE) value was found to be lower in ANN (33.89) when compared to MLR (57.31). Hence, the ANN modelling approach is efficient to predict frozen semen doses that could be produced per ejaculate.

Cryoprotectant With A Mitochondrial Derived Peptide, Humanin, Improves Post-Thaw Quality Of Buffalo Spermatozoa

BACKGROUND:Semen cryopreservation results in deleterious effects on spermatozoa, including lipid peroxidation and a reduction in the total antioxidant components of seminal plasma. The ultimate outcome of these changes is a reduction in post-thaw semen quality. A mitochondrial derived peptide, humanin, a potent cytoprotective and antioxidant agent was used in the present study.OBJECTIVE:To evaluate the efficacy of a mitochondrial-derived peptide, humanin to improve the post-thaw quality of buffalo spermatozoa.MATERIALS AND METHODS:A total of 18 ejaculates from three Murrah buffalo bulls (n=6 each) were collected. Each ejaculate was divided into four aliquots. The first aliquot was diluted with standard EYTG dilutor (Group I, control), whereas the other three aliquots were diluted with EYTG supplemented with 2 μM (Group II), 5 μM (Group III) and 10 μM humanin (Group IV), respectively. Semen was evaluated for physico-morphological and functional attributes such as progressive motility, viability, abnormality, acrosome integrity, plasmamembrane integrity of fresh samples, pre-freeze and post-thaw stages. Oxidative stress parameters [lipid peroxidation (LPO) and total antioxidant capacity (TAC)] were also measured at the pre-freeze and post-thaw stages.RESULTS:Humanin s upplementation resulted in significantly higher (p≤0.05) post- thaw motility in all treatment groups and, higher (p≤0.05) viability in Groups III and IV in comparison to the control at the post-thaw stage. Spermatozoa with intact acrosome and plasma membran e were higher (p≤0.05) in Groups III and IV as compared to Group s I and II. The LPO levels at the post- thaw stage were found to be lower (p≤0.05) in all treatment groups versus the control group, whereas, higher (p≤0.05) TAC value s were recorded in Groups III and IV in comparison to the control and Group II.CONCLUSION:Humanin supplementation in the extender improved the freezabilty of buffalo spermatozoa.

Genetic and non-genetic factors affecting conception rate of frozen semen in small holder dairy farmer system of rural India.

Fertility traits are as important as production traits in crossbred bovine population. Assessment of the fertility of bull in any frozen semen production program and evaluation of the conception rate under actual field conditions provide valuable information. The objective of this retrospective study was to estimate the effect of genetic and non-genetic influence on conception rate of frozen semen bulls maintained at Bharatiya Agro-Industries Foundation (BAIF) Pune based on the conception rate in cattle population of small holder dairy farmer system. The data comprising of 1,08,238 insemination records pertaining to 83 Holstein Friesian pure and crossbred bulls available at BAIF Pune were used to analyze conception rate. The fixed effect solutions and covariance components were estimated by linear mixed model using the restricted maximum likelihood method in WOMBAT software. The genetic correlations were estimated using bivariate analysis between post thaw motility and conception rate. The study was based on fertility related information from cows maintained in different villages of India and thus reflects the actual fertility of frozen semen used. The study was suggestive of influence of very small fraction of genetic effect and higher impact of management effect on conception rate. Fertility-related information available from this study is an invaluable asset in decision making process of breeding policies.

Kualitas semen sapi pejantan berdasarkan umur, suhu, dan kelembaban di Taman Ternak Pendidikan Universitas Airlangga

This research was conducted at the Teaching Farm of the Faculty of Veterinary Medicine, Airlangga University, to know the effect of age, temperature, and humidity on the quality of semen of bulls which are turned into frozen semen. This research uses fresh semen samples of bulls of various ages grouped into 3 groups, young, middle age, and old groups and measures temperature and humidity during the storage process. Fresh semen is collected using an artificial vagina and then examined macroscopic and microscopic tests which is then checked for post thawing motility. Data analysis uses Analysis of Variants (ANOVA) followed by Duncan. The results showed that the percentage of spermatozoa motility of fresh semen and post thawing differed and age affected the quality of semen produced. There was no effect of temperature and humidity when processing of collecting semen. Bull’s semen at Teaching Farm Faculty of Veterinary Medicine Airlangga University has good quality and is suitable for Artificial Insemination.

Semen quality and production potential of Indian buffalo breeds

The objective of the study was to evaluate non-genetic factors on semen traits. The overall means for the semen quality traits: ejaculate volume, sperm concentration, total sperm, mass activity, initial and post thaw motility, hypoosmotic swelling test, acrosome integrity of fresh and frozen test were 4.39±0.01 ml, 1.29±0.001 billion, 5.48±0.01 billion, 2.55±0.003, 73.41±0.0004%, 55.9±0.0002%, 59.01±0.001%, 84.98±0.002% and 71.13±0.002%, respectively. Breeds were found to differ in all the semen production and quality traits studied except acrosome integrity of fresh semen. Summer for semen production, and winter and monsoon for semen quality traits were comparatively better season. The quality traits improved with age and declined slightly in the later life.

Effect of cervical insemination with frozen semen on fertility of Indian goat breed

Artificial Insemination (AI) has great potential to multiply superior quality of goat with faster rate in spite of lower conception rate. This technique will spread elite genetic material throughout a population and is also important for breed conservation and has paved the way for other reproductive biotechnologies. Ejaculates from bucks aged between 2–4 years old from Jamunapari, Barbari, Sirohi and Jakhrana were collected using artificial vagina, twice a week. Immediately after collection, the volume, colour, consistency, and mass motility of ejaculate were assessed and were extended with Tris -Egg yolk- Fructose diluent having 10% (v/v) egg yolk and glycerol 6% (v/v). Sperm concentrations were adjusted to 1×108/ml and diluted semen was equilibrated at 5°C for 4 h before being frozen. The post thaw motility in Jamunapari and Barbari was significantly higher then Jakhrana and Sirohi. In two major breeding seasons (May–June and October–November), 181 goats of different breeds (Barbari, Jakhrana, Jamunapari and Sirohi) including 35 goats in villages were intra-cervical inseminated with frozen semen in natural estrous condition. The kidding percentage in Barbari, Jamunapari, Jakhrana and Sirohi were 53.12±2.40%, 34.61±1.96%, 26.53±2.12% and 28.57±2.32% respectively. Overall, a success rate of 37.57% was recorded on the basis of actual kidding rate irrespective of goat breed maintained at this institute under semi-intensive management system. The best post thaw quality of buck semen and comparative deeper cervical insemination in Barbari and Jamunapri breeds results in higher conception and kidding percent compared to other breeds (Sirohi and Jakhrana).

Low concentrations of 3-O-methylglucose improve post thaw recovery in cryopreserved bovine spermatozoa

A number of studies have explored the use of membrane permeable (usually metabolizable) and membrane impermeable saccharides to protect cells in general, and sperm in particular during cryopreservation. Critical concentrations for protective levels of sugars frequently range between 50 mmol/L and 500 mmol/L, where efficacy is attributed to the sugar's membrane stabilizing and glass forming attributes and colligative effects that reduce intra- and extracellular salt concentrations during freezing. Many studies on bull sperm have demonstrated that both permeating and non-permeating sugars have negligible positive effects on post-thaw viability. Recently, however, a non-metabolizable sugar, 3-O-Methylglucose (3-OMG), was shown to protect hepatocytes during liver cryopreservation at 0.1–0.3 mol/L. Because glucose is readily transported into sperm, we hypothesized that 3-OMG could be a new class of cryoprotectant to explore in bull sperm. Here we present positive results demonstrating that 3-OMG improves post thaw viability in bull sperm, and that this effect is not likely due to improved glass forming capabilities. In particular, in experiment 1, 3-OMG was added to the Tris-egg yolk-glycerol base media at levels from 0 mmol/L to 200 mmol/L. Semen from four bulls was collected and diluted with one of the cryopreservation media, cooled, and frozen following industry standard practices. Motility and mitochondrial activity were negatively impacted when concentration of 3-OMG was more than 25 mmol/L. Therefore, we explored lower concentrations in experiment 2, where semen from eight bulls was used to evaluate concentrations 5 mmol/L, 15 mmol/L and 25 mmol/L of 3-OMG compared with control. Motility and progressive motility in 5 mmol/L 3-OMG and in the control were significantly higher than 15 mmol/L and 25 mmol/L 3-OMG, whereas mitochondrial activity and acrosome integrity in 5 mmol/L 3-OMG were significantly better than the control freezing medium. In experiment 3, to evaluate whether the improved effects of 3-OMG are due to its non-metabolizing property, or due to colligative effects, we compared post-thaw viability in semen from four bulls cryopreserved with 5 mmol/L glucose, sucrose, or 3-OMG. Motility and progressive motility was significantly improved in 3-OMG compared to glucose or sucrose groups which were comparable to the EY control. In conclusion, 3-OMG at a concentration of 5 mmol/L in Tris-egg yolk-glycerol medium improves the post thaw motility, progressive motility and viability of bull sperm. The mechanism of action is not understood but because the efficacy is maximal at low concentrations, it is not likely due to improved intra- or extracellular glass forming abilities and may demonstrate a different protective mechanism than was shown in hepatocytes.

PENGARUH LAMA WAKTU PENGHITUNGAN STRAW DAN POSISI JARAK STRAW DIATAS PERMUKAAN NITROGEN CAIR TERHADAP MOTILITAS DAN VIABILITAS SEMEN BEKU SAPI MADURA POST THAWING

Aims of this research was to determine the effect of the duration of straw count and the position of straw from liquid nitrogen surface to the motility and viability of post thawing frozen Madura cattle. This study used a sample derived from Madura cattle (Pasean OO 1220) from BBIB singosari as much as 54 straw divided into 9 treatments. P1 (1cm 10 sec), P2 (1cm 20 sec), P3 (1cm 40 sec), P4 (2cm 10 sec), P5 (2cm 20 sec), P6 (2cm 40 sec), P7 (3cm 10 sec), P8 (3cm 20 sec), P9 (3cm 40 sec) and repeated as much as 6 times then observed for motility and viability of post thawing spermatozoa. The data obtained were analyzed using SPSS 16.0. The results showed that the duration of straw count and the position of straw from liquid nitrogen significant (P&lt;0,05) on post thawing motility. The position of straw from liquid nitrogen signifant (P&lt;0,05) on post thawing viability. The conclusion of this research showed that 1 cm 40 sec give the higher post thawing motility.

PENGARUH JARAK DAN LAMA WAKTU PROSES PENGHITUNGAN STRAW SEBELUM DISTRIBUSI TERHADAP KUALITAS SEMEN BEKU SAPI SIMENTAL POST THAWING

Aims of this research was to determine the effect of distance and the duration of straw count above liquid nitrogen to the Quality of Frozen Simental Cattle. This study used a sample derived from simmental cattle from BBIB singosari as much as 48 straw divided into 8 treatments. P1(0,5cm 15 sec), P2(0,5cm 30 sec), P3( 0,5cm 45 sec), P4(0,5cm 60 sec), P5(1,5cm 15 sec), P6(1,5cm 30 sec), P7(1,5cm 45 sec), P8(1,5cm 60 sec) and repeated as much as 6 times then observed for motility and viability of post thawing spermatozoa. The data obtained were analyzed using SPSS 16.0. The data obtained were analyzed using SPSS 16.0. The results showed that the duration of straw count and the position of straw from liquid nitrogen not significant (P&gt;0,05) on post thawing motility. The position of straw from liquid nitrogen not signifant (P&gt;0,05) on post thawing viability. The conclusion of this research showed that 0,5 cm 15 sec give the higher post thawing motility.

Effect of Age and Season on the Seminal Traits of Pure HF Bulls Under Tropical Condition

The study was conducted to assess the effect of age and season on seminal traits of pure HF bulls under tropical condition. A total of 30 HF bulls of age group up to 3, 4-5, 6-7, 8 and above 8 years were selected and divided into 3 seasons viz rainy, summer and winter. The breeding bulls were kept under intensive system of rearing as per MSP guidelines of India. Semen was collected, evaluated and data analyzed by using standard procedures. The overall production and all the seminal traits viz volume, initial motility and post thaw motility was found to be better in summer season as compare to rainy and winter season. However, there was no significant change in concentration due to season. Among all the age groups, 8 years and above were found to produce highest volume, post thaw motility and eventually more production. Highest percentage of initial motility was recorded among the youngest group of bulls and concentration was significantly (plessthan0.01) high in the age group 6-7years followed by 8 and above 8 years of age. The study revealed that age and season have highly significant (plessthan0.01) effect on all the seminal traits and production.

Comparative Study between Slow Conventional Freezing and Cryoprotectant-Free Vitrification of Human Spermatozoa in Large Volume

Background: Cryopreservation is the collection, freezing, and long term storage of sperm, and is a highly effective method of protecting male fertility. Cryopreservation of semen has been widely used as a vital method for fertility preservation of male patients before undergoing chemotherapy, radiotherapy, and/or surgery that may lead to testicular failure or ejaculatory dysfunction. Objective: The aim of this study was to compare the aseptic technology of cryoprotectant-free vitrification of human spermatozoa in large volume, to conventional freezing protocol as regards post thawing motility, vitality and sperm DNA fragmentation. Patients and methods: This study included a total of 20 male patients seeking seminal fluid analysis, attending at the andrology laboratory of a specialized IVF center (ADAM International Hospital for Fertility and Sterility, Giza, Egypt. Patients were presented with the diagnoses of normozoospermia, oligozoospermia (either isolated or combined with asthenozoospermia or teratozoospermia). Results: Motility of (in a large volume (300 µl) in the absence of permeable cryoprotectants displayed significant statistically lower levels as compared to conventional Sperm Freezing. It was shown in different groups at different times (post thawing and 1-hour and 24-hour) of assessment that motility of vitrified spermatozoa decreases in comparison with slow conventional freezing as we go from the baseline. DNA fragmentation of vitrified spermatozoa showed higher levels as compared to conventional slow freezing but there is no significant statistical difference between vitrification and conventional slow freezing in DNA fragmentation. Conclusion: It could be concluded that vitrification technique was quite far away from comparison with slow conventional freezing protocol, and still need for further modifications and wide scale of study to achieve the good results.

Comparative Study between Slow Conventional Freezing and Cryoprotectant-Free Vitrification of Human Spermatozoa in Large Volume

Background: Cryopreservation is the collection, freezing, and long term storage of sperm, and is a highly effective method of protecting male fertility. Cryopreservation of semen has been widely used as a vital method for fertility preservation of male patients before undergoing chemotherapy, radiotherapy, and/or surgery that may lead to testicular failure or ejaculatory dysfunction. Objective: The aim of this study was to compare the aseptic technology of cryoprotectant-free vitrification of human spermatozoa in large volume, to conventional freezing protocol as regards post thawing motility, vitality and sperm DNA fragmentation. Patients and methods: This study included a total of 20 male patients seeking seminal fluid analysis, attending at the andrology laboratory of a specialized IVF center (ADAM International Hospital for Fertility and Sterility, Giza, Egypt. Patients were presented with the diagnoses of normozoospermia, oligozoospermia (either isolated or combined with asthenozoospermia or teratozoospermia). Results: Motility of (in a large volume (300 µl) in the absence of permeable cryoprotectants displayed significant statistically lower levels as compared to conventional Sperm Freezing. It was shown in different groups at different times (post thawing and 1-hour and 24-hour) of assessment that motility of vitrified spermatozoa decreases in comparison with slow conventional freezing as we go from the baseline. DNA fragmentation of vitrified spermatozoa showed higher levels as compared to conventional slow freezing but there is no significant statistical difference between vitrification and conventional slow freezing in DNA fragmentation. Conclusion: It could be concluded that vitrification technique was quite far away from comparison with slow conventional freezing protocol, and still need for further modifications and wide scale of study to achieve the good results.

Effect of azolla supplementation in feed on semen freezability in bucks

The present study was conducted to evaluate the effect of azolla supplementation on semen freezability in Barbari bucks. Ten adult Barbari bucks (2–4 years old) were selected, and divided into control and treatment group (5 bucks in each group) as per completely randomized design. Bucks of control group were fed with 400 g concentrate pellet/day along with 6–7 h grazing while bucks of treatment group were supplemented with 100 g fresh azolla along with 400 g concentrate pellet and 6–7 h grazing. Semen ejaculates (8) were collected from each buck of control and treatment groups after 60 days of azolla supplementation. Immediately after collection, semen samples were evaluated for colour, volume, density, mass motility, progressive motility, live sperm count, acrosomal integrity and hypo-osmotic swelling positive spermatozoa. Then semen samples were diluted with tris–citric acid fructose diluents having 6% (v/v) glycerol as cryoprotective agent and 10% (v/v) egg yolk. After equilibration, semen samples were filled in straws and vapour frozen for 10 min 4 cm above the liquid nitrogen and finally stored into liquid nitrogen container. Post thaw motility, live sperm count, abnormalities, acrosomal integrity and hypo-osmotic swelling test were conducted to check the effect of azolla feeding on the freezability. Post thaw motility, live sperm count, acrosomal integrity and hypo-osmotic swelling positive spermatozoa were significantly higher in treatment group. It could be concluded that dietary azolla supplementation to breeding bucks improved the quality and freezability of semen favouring the use in artificial insemination program.

Effect of Age and Frequency of Collection on Quality of Jersey Bulls Semen at National Livestock Breeding Center (NLBC), Nepal

Objective: To determine the effect of age and frequency of collection on quality parameters of Jersey bull semen at National Livestock Breeding Center (NLBC), Pokhara.&#x0D; Materials and Methodology: Nine Jersey bulls were selected randomly from NLBC. To obtain the effect of age, six bulls of three different age interval (3-4, 5-7 and 8-9 years) were selected randomly. Remaining three bulls were used to determine the effect of frequency of collection on semen quality. Studies were conducted for three months (Nov/Dec to Dec/Jan, 2016). Semen quality was analyzed using CASA system and SMILE software. ANOVA and paired t- tests were applied to find out level of significance. All the statistical analyses were done using SPSS 17.0&#x0D; Results: Sperm concentration, post- thaw motility, live sperm, and abnormal sperm were significantly affected by the age of the bulls. Semen quality was found superior in the bulls belonging to 5-7 years of age intervals. Significantly higher value for semen volume, sperm concentration and post- thaw motility was obtained on second collection (P&lt;0.05). However, volume initial motility, pre-filling motility, live sperm and abnormal sperm percentage did not differ significantly among different age intervals and between the frequencies of collection.&#x0D; Conclusion: Different quality parameters of semen like volume, concentration, initial motility, pre-filling motility, post thaw motility, live and normal spermatozoa percentages were higher in 5-7 years old bulls and in second collection of semen.&#x0D; Int. J. Appl. Sci. Biotechnol. Vol 7(1): 88-95