Bull Semen Research Articles

Evaluation of a novel microfluidic chip-like device for purifying bovine frozen-thawed semen for in vitro fertilization

The aim of this study was to validate a novel sperm purification device, the VetCount™ Harvester, for use in bovine in vitro fertilization (IVF). The device's performance was compared to BoviPure™ gradient centrifugation, a commercially available and accepted routine technique. Semen quality parameters were assessed for frozen-thawed semen from six different bulls (n = 6) following sperm purification. For each bull two semen subsamples were purified utilizing BoviPure™ gradient centrifugation and the VetCount™ Harvester, including a third subsample as untreated control. Both treatments significantly increased the proportion of progressively motile sperm cells (84.4 ± 14.1% and 85.1 ± 7.8%, respectively) compared to the untreated semen (41.9 ± 18.8%). BoviPure™ gradient and VetCount™ Harvester selected predominantly viable acrosome intact (VAI) sperm cells with low membrane fluidity and low free intracellular calcium concentration [Ca2+]i (76.5 ± 4.4% and 78.6 ± 6.0%). Normalizing [Ca2+]i of VAI sperm cells (non-treated semen: [Ca2+]i = 1) VetCount™ Harvester purified spermatozoa (0.67 ± 0.10) showed significantly lower [Ca2+]i than BoviPure™ treated sperm (0.84 ± 0.14; P < 0.05). Subsequently, the fertilizing ability of the spermatozoa was evaluated performing a competitive fertilization assay. Sperm cells from both treatment groups were fluorescently labelled using different dyes and added in equal amounts to in vitro matured oocytes. After 18 h co-incubation, the origin of the fertilizing sperm cell was evaluated via fluorescence microscopy. In two bulls, VetCount™ Harvester selected sperm that fertilized significantly more oocytes then BoviPure™ treated sperm, in another bull it was the opposite. For three bulls no difference was observed.We conclude that the VetCount™ Harvester selects a high-quality, fertile sperm fraction from frozen-thawed bull semen. However, some considerations have to be kept in mind for the direct use of the isolated sperm fraction in IVF.

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&lt;0.005) to the semen volume, individual motility, and post- thawing motility except for semen volume and sperm abnormality(p&gt;0.05). However, initial motility, sperm concentration, and pre filling motility did not vary in months (p&gt;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&lt;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.

Association of A51703G and G51656T loci polymorphism in luteinizing hormone receptor (LHR) gene with sperm traits in Holstein bulls

Today, there is an increasing desire to use Candidate genes as molecular markers to improve reproductive traits. The luteinizing hormone receptor is expressed on the Leydig cells and plays a crucial role in gametogenesis initiation and maintenance. This study aimed to study the effects of luteinizing hormone receptor (LHR) gene polymorphisms (A51703G and G51656T) on bull sperm traits. Genomic DNA was extracted from the semen, blood, and samples of 150 AI Iranian Holstein bulls according to the established protocol [10] with minor modifications. Semen traits were routinely collected at bull semen collection centers from each animal between 1992 and 2015. The bull fertility traits include semen concentration (SC 108 per ml) sperm volume (SV ml), fresh sperm motility (FSM %), and post-tallow sperm motility (PTSM %). The mutation was detected with A PCR-RFLP method. The digestion of amplicons was also carried out with BfaI and HinfI restriction enzymes in A51703G and G51656T loci, respectively. The restriction profiles of PCR products indicated two alleles in each locus (A and G for A51703G and G and T for G51656T). The frequency of the A and T alleles were 0.6647 and 0.606, respectively. Analysis of variance showed that LHR loci had a significant effect on sperm characteristics in Iranian Holstein bulls (P < 0.05). Bulls with GG genotype surpassed those with GA and AA genotypes regarding SP, FSM, and PTSM in the A51703G locus. In the G51656T locus, the association result showed that bulls with AG genotype had higher than others in all traits (P < 0.05). In conclusion, these data suggest that LHR polymorphisms are an excellent genetic marker for breeding programs in male reproductive traits in the Iranian bull population.

Seasonal influence on expression of heat shock proteins (HSP70 and HSP90) vis-à-vis functional competence of Gir bull semen

The success of assisted reproduction relies on functional competence of frozen-thawed semen. Heat stress affects protein folding leading to aggregation of mis-folded proteins. Hence, a total of 384 (32 ejaculates/bull/season) ejaculates from six matured Gir bulls were used to evaluate physico-morphological parameters, the expression of HSPs (70 and 90) and fertility of frozen-thawed semen. The mean percent individual motility, viability and membrane integrity were significantly (p < 0.01) higher in winter compared to summer. Out of 1200 Gir cows inseminated, 626 confirmed pregnant and the mean conception rate of winter (55.04 ± 0.35) was significantly (p < 0.001) higher than summer (49.33 ± 0.32). A significant (p < 0.01) difference in concentration of HSP70 (ng/mg of protein) but not HSP90was observed between the two seasons. The HSP70 expression in pre-freeze semen of Gir bulls had significant positive correlation with motility (p < 0.01, r = 0.463), viability (p < 0.01, r = 0.565), acrosome integrity (p < 0.05, r = 0.330) and conception rate (p < 0.01, r = 0.431). In conclusion, the season influences physico-morphological parameters and expression of HSP70 but not HSP90 in Gir bull semen. The HSP70 expression is positively correlated with motility, viability, acrosome integrity and fertility of semen. The semen expression of HSP70 may be utilized as biomarker for thermo-tolerance, semen quality and fertilizing capacity of Gir bull semen.

Bulls fed a high gain diet decrease blastocyst formation after in vitro fertilization.

Bulls used in cattle production are often overfed to induce rapid growth, early puberty and increase sale price. While the negative consequences of undernutrition on bull sperm quality are known, it is unclear how a high gain diet influences embryo development. We hypothesized that semen collected from bulls fed a high gain diet would have a reduced capacity to produce blastocysts following in vitro fertilization. Eight mature bulls were stratified by body weight and fed the same diet for 67 d at either a maintenance level (0.5% body weight per day; n = 4) or a high gain rate (1.25% body weight per day; n = 4). Semen was collected by electroejaculation at the end of the feeding regimen and subjected to sperm analysis, frozen, and used for in vitro fertilization. The high gain diet increased body weight, average daily gain, and subcutaneous fat thickness compared to the maintenance diet. Sperm of high gain bulls tended to have increased early necrosis and had increased post-thaw acrosome damage compared with maintenance bulls, but diet did not affect sperm motility or morphology. Semen of high gain bulls reduced the percentage of cleaved oocytes that developed to blastocyst stage embryos. Paternal diet had no effect on the number of total or CDX2 positive cells of blastocysts, or blastocysts gene expression for markers associated with developmental capacity. Feeding bulls a high gain diet did not affect sperm morphology or motility, but increased adiposity and reduced the ability of sperm to generate blastocyst stage embryos.

When the plan goes awry: how to negotiate estrus synchronization errors in beef cattle

Estrus synchronization protocols vary widely, and errors during implementation are not uncommon. When an error is made, recommendations for resolution must consider the initial purpose of implementing the synchronization protocol in the herd. The ideal solution is to convert the protocol to a different published protocol. When this is not possible, the safest solution for maintaining an acceptable pregnancy rate is often either restarting a synchronization program or potentially foregoing artificial insemination for the season. Knowledge of estrous cycle physiology and how hormone treatments manipulate the cycle are foundational when attempting to modify a protocol based on the specific error that is made. Errors made early in a synchronization protocol tend to be more manageable than those made toward the end of the protocol. The use of less valuable semen and immediate introduction of a clean-up bull should be considered to maximize pregnancy outcomes when attempting to correct an error during estrus synchronization.

The Difference in Subpopulation Structures in Sex-Sorted and Non-Sorted Semen by Discriminant Analysis of Bull Sperm Motility Data Collected by a Computer-Assisted Sperm Analysis System

We examined the motility characteristics of sperm from sex-sorted bull semen as a model for low-quality (expected low conception rate) sperm compared to that of sperm from non-sorted semen (expected average conception rate) using cluster analysis followed by discriminant analysis. The results indicated that sex-sorted semen contained sperm with hyperactivation-like motility as the main subpopulation immediately after thawing, and this subpopulation decreased after 2 h of incubation. The main subpopulation in the non-sorted semen had progressive motility that was maintained during incubation. A conventional comparison of the mean values of kinematic parameters could not distinguish the samples. In conclusion, discriminant analysis using data from a cluster analysis of motile sperm may accurately describe differences in the structures of sperm motility subpopulations between low and normal fertility semen.

Candidate genes and proteins regulating bull semen quality: a review

Candidate genes and proteins regulating bull semen quality: a review

The influence of L-proline and fulvic acid on oxidative stress and semen quality of buffalo bull semen following cryopreservation.

This study investigates the effects of cryopreservation and supplementation of Azeri water buffalo's semen with proline (Lp) and fulvic acid (FA). Therefore, this study aimed to assess motility parameters, sperm viability, oxidative stress parameters, and DNA damage to detect the optimum concentrations of Lp and FA for buffalo semen cryopreservation. Thirty semen samples of three buffalo bulls were diluted in Tris-egg yolk extender and divided into 12 equal groups including control (C), Lp-10, Lp-20, Lp-40, Lp-60, Lp-80 (containing 10, 20, 40, 60, 80mM L-proline, respectively), FA-0.2, FA-0.5, FA-0.8, FA-1.1, FA-1.4 and FA-1.7 (containing 0.2%, 0.5%, 0.8%, 1.1%, 1.4% and 1.7% fulvic acid, respectively). The velocity parameters, TM and PM were improved by FA-1.7, FA-1.4, Lp-40 and Lp-60 groups compared to the C group but no significant difference was found regarding the amplitude of lateral head displacement and straightness compared to the control groups. The percentage of sperm viability and PMF were increased by FA-1.7, FA-1.4, FA-1.1, Lp-40 and Lp-60 groups compared to C group, while in terms of sperm DNA damage FA-1.7, FA-1.4, FA-1.1, Lp-10, Lp-20, Lp-40 and Lp-60 groups showed better results compared to C group. The results also showed that FA-1.7, FA-1.4, FA-1.1, Lp-20, Lp-40 and Lp-60 groups could improve TAC, SOD, GSH and decrease MDA levels. Also, FA-1.7, FA-1.4, Lp-20 and Lp-40 groups could improve GPx levels but just FA-1.7, and Lp-40 groups could improve CAT levels compared to C group. Thus, it can be concluded that L-proline and fulvic acid supplementations can improve the quality parameters of post-thawed buffalo bull semen.

Evaluation Deoxyribonucleic acid (DNA) fragmentation of local Indonesian cattle frozen sperm using Halomax method

Abstract. Prabowo TA, Bintara S, Yusiati LM, Sitaresmi PI, Widayati. 2023. Evaluation Deoxyribonucleic acid (DNA) fragmentation of local Indonesian cattle frozen sperm using Halomax method. Biodiversitas 24: 2225-2230. Deoxyribonucleic acid (DNA) is a molecular chain containing genetic material that determines the quality of each individual produced. DNA damage in cattle can affect fertility and miscarriage of embryos. There is no simple research on DNA damage in local Indonesian cattle (Bos indicus Linnaeus 1758); therefore, miscarriage and infertility cases in Indonesia are still high, one of which can be attributed to sperm DNA damage. This study aimed to evaluate DNA damage in the frozen semen of local Indonesian cattle and suggested SDF (Sperm DNA Fragmentation) inclusion screening as the main requirement for frozen semen. The materials used in this study were 54 straws of frozen semen from nine males from three different types of local cattle, including Bali, Madura, and Aceh. Each type consisted of three bulls, each with six replicates. Sperm-Bos-Halomax was used to examine sperm DNA damage. Data were analyzed using one-way analysis of variance (ANOVA). Aceh cattle had the highest post average compared to Bali and Madura at 63.33±2.43%. Abnormality of Balinese cattle had the lowest average compared to Aceh and Madura at 8.83±1.04%. Evaluation of sperm DNA damage in local cattle using the Sperm-Bos-Halomax® method showed that the highest average DNA damage was 1.72±0.62% in Aceh, and the lowest was 1.67±0.66% in Madura cattle. The sperm DNA damage in Bali cattle was 1.67±0.70%. The sperm DNA quality in the frozen semen of the local cattle in this study was good, but it is still necessary to determine SDF as the main standard in frozen semen.

Reproductive Performance of Bali Cows through Artificial Insemination Program in Polewali Mandar Regency, West Sulawesi Province

Intensification of artificial insemination (AI) in Bali cattle is a government program to increase the population and productivity of local cattle. The objective of this study was to examine the reproductive performance of Bali cows through the AI program in Polewali Mandar Regency, West Sulawesi. The study was conducted in two different locations, i.e. in the low lands (&lt;300mASL) including Wonomulyo and Campalagian Districs, and high lands (&gt;300mASL) including Limboro and Bulo Districts. Data collection was carried out on farmers and inseminators profiles, and cow’s reproductive parameters including service per conception (S/C), conception rate (CR), calving rate (CvR), calving interval (CI) and Calf Mortality (CM). These data were calculated and analyzed descriptively. The cows in this study were raised by farmers in small-scale farm, mostly under 5 heads. The cattle breeding through AI program was carried out by inseminators with over 10 years of experience. In the lowlands, S/C values were 1.26-1.17 and CI were 360 to 366.31 days, respectively. Bali cows that were inseminated using Bali bull semen had a CvR of 75.69% which were higher than those using Simmental bull semen (CvR 49.69%;). While in the highlands, Bali cows that were inseminated with Bali bull semen tended to have higher S/C (1.30) and CI (356.95 days) values but had CvR of 61.00% and CR 77.00% which was also higher than that of Simmental bull semen (S/C 1,50; CI 412,86 days; CvR 50.00%; CR 66.67%). The high S/C in the highlands (1.50) is due to location constraints or the distance between the livestock and the inseminator's residence which is quite far, so it takes time to carry out AI services and is one of the factors that can cause the provision of AI services for female cows in heat is not optimal. In the highlands, most of the cattle are grazed in the grazing areas, making it difficult for breeders to detect estrus. In general, Bali cattle produced by AI in the low lands of Polewali Mandar district tended to have better reproductive performance than those in the high lands. The S/C in the low lands (1.26 and 1.17) is lower than that in the high lands (1.3 and 1.5). Keywords: AI, Bali cows, lower land, high land reproductive performance

Bull Semen Obtained on Beef Farms by Electroejaculation: Sperm Quality in the First Two Hours of Storing with Different Extenders and Holding Temperatures.

Sperm quality decreases over time, so bull semen may need to be preserved after field collection. However, the effect of handling such semen samples from commercial farms and placing them in very short-term storage has not been elucidated. Therefore, ejaculate from 25 bulls from 1 dairy and 14 beef cattle farms were collected under farm conditions and evaluated for semen quality during the first two hours after collection. Two commercial extenders (AndroMed® and BIOXcell®) and two different storage temperatures (5 °C and room temperature) were used to evaluate the influence on semen quality and sperm kinetics in ejaculates grouped into three evaluation times, based on time since collection (Time 1: <75 min, n = 7; Time 2: 75-105 min, n = 11; and Time 3: 105-120 min, n = 7). Classical semen parameters, sperm motion kinetics by CASA and colony-forming units were assessed. The differences between both extenders in curvilinear and straight-line velocities (VCL and VSL) for the different time groups (Time 2 and Time 3) were statistically significant for p < 0.05. AndroMed® showed lower VSL, straightness and linearity in sperm compared to BIOXcell® (p < 0.05). In conclusion, AndroMed® induced more curvilinear movement, while BIOXcell® stimulated straighter motility.

Reducing oxidative stress by κ-carrageenan and C60HyFn: The post-thaw quality and antioxidant status of Azari water buffalo bull semen

Azeri water buffalo is a species of great interest due to the high quality of its products such as milk. Due to the decreasing trend of its number and risk of extinction in the future, our attention is directed towards ensuring the preservation of its genetic reserves by keeping its sperm. Using antioxidants in semen extender is one of the ways to reduce the detrimental effects of freezing process on post-thawed quality of spermatozoa. This study was conducted to determine the effect of κ-carrageenan (k-CRG) and C60HyFn supplemented semen extender on the quality of post-thawed Azari water buffalo spermatozoa. A total of 30 semen samples were obtained from three buffaloes using an artificial vagina (twice a week for five weeks = 10 replicates). The samples (n = 3) from each replicate were pooled and divided into equal aliquots to prepare 14 extender groups, including control (C), k-0.2, K-0.4, K-0.6, K-0.8 (containing 0.2, 0.4, 0.8 mg K-CRG/mL, respectively), C-0.1, C-0.2, C-0.4, C-0.8, C-1, C-5, C-10, C-20, and C-40 (containing 0.1, 0.2, 0.4, 0.6, 0.8, 1, 5, 10, 20, 40 μM C60HyFn, respectively), and then frozen. After thawing, motility and velocity parameters, plasma membrane integrity (PMI) and functionality (PMF), DNA damage, Hypo-osmotic swelling (HOS) test, malondialdehyde (MDA), total antioxidant capacity (TAC), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase glutathione activities and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging were evaluated. In vivo fertility was compared between k-0.6, C-1 and control groups. 60 buffalo were inseminated 24 h after the onset of estrus. The diagnosis of pregnancy was performed rectally at least 60 days after fertilization. Total and progressive motility and velocity parameters were improved by k-0.4, k-0.6, k-0.8, C-0.4, C-0.8, C-1, C-5, and C-10 groups) compared to the other groups. Plasma membranes integrity and PMF were improved by k-0.4, k-0.6, C-0.4, C-0.8, C-1, C-5, and C-10 groups compared to other groups, while in terms of sperm DNA damage K-0.4, K-0.6, K-0.8, C-0.2, C-0.4, C-0.8, C-1, C-5, and C-10 groups showed better results compared to the control group. The evidence also showed that k- 0.4, k-0.6, k-0.8, C-0.4, C-0.8, C-1, C-5, and C-10 groups could improve TAC, and decrease MDA levels. Also, k-0.4, k-0.6, k-0.8, C-0.2, C-0.4, C-0.8, C-1, C-5, and C-10 groups could improve GPx, CAT, and GSH levels, but no significant difference was found regarding SOD compared to the other groups. DPPH scavengers were tested by K-0.6, K-0.8 and C-1, C-5, C-10, C-0.8, C-0.4 and C-0.2 groups and compared to other groups improved. The fertility rate [70% (14/20)] was higher in C-1 than other groups. To conclude that k-CRG and C60HyFn supplementation can increase the quality parameters of cryopreserved buffalo semen after thawing and that 1 M C60HyFn can increase in vivo fertility of buffalo semen.

022 Metabolomics markers associated with fertility in bull semen

022 Metabolomics markers associated with fertility in bull semen

Plasma membrane integrity and acrosomal integrity of fresh and frozen Bali bull semen based on different ejaculate volume

Semen volume is one of the measuring points used to determine the quality of a bull’s semen in one ejaculate. Plasma Membrane Integrity (PMI) and Acrosomal Integrity (AI) are more detailed microscopic examinations and are used to determine the ability of spermatozoa to fertilize. The aim of this study was to evaluate the PMI and AI of fresh and frozen semen of Bali bull based on different ejaculate volumes. This research was conducted in March-April at Artificial Insemination Centre (AIC) in Pucak, South Sulawesi. The method used in this study was an experimental method with 4 treatments of ejaculate volume 1-3 ml; 3.1-5 ml; 5.1-7 ml; and 7.1-9 ml for 3 replicates each. The observed variables were PMI and AI. The results showed that the volume variation had no significant effect (P>0.05) on the PMI and AI values of fresh Bali cattle semen, as well as post-thawing frozen semen. The average percentage of PMI in fresh semen was 63.49±1.46% and post thawing was 37.66±2.82%, while the average AI percentage for fresh semen was 73.17±2.06% and post thawing was 50.92±2.64%. The conclusion of this study is the volume of ejaculate does not affect the plasma membrane integrity and acrosomal integrity of Bali cattle semen, both fresh and frozen.

The quality of fresh and frozen semen Bali bull assessed from different ejaculate volume

Semen volume is the amount of ejaculate released by male Bali cattle during one ejaculation, which consists of spermatozoa and seminal plasma. The purpose of this study was to determine the effect of Bali cattle ejaculation volume on the quality of fresh and frozen semen. Semen collection was carried out using an artificial vaginal collection method. The collected semen was then evaluated with microscopic parameters: motility, viability, and abnormalities. Semen came from 7 male Bali bulls. The data obtained were analyzed by one-way ANOVA. The results showed that there was no significant difference (P>0.05) between the volume of Bali cattle ejaculation on the quality of fresh semen and also post thawing based on the parameters tested. The sperm concentration of Bali bulls has average 935.75 ± 80.25 x 106. The average percentage of fresh semen motility was 70.79±1.26 while that for frozen was 47.83±1.89. The average viability of fresh spermatozoa was 80.04±3.68 while frozen 52.87±0.67. The average abnormality value of fresh spermatozoa was 8.54±1.88 while frozen was 23.83±4.22. Based on these results, it was concluded that the different volume of ejaculate of Bali cattle did not affect the quality of fresh semen of Bali cattle, both fresh and frozen based on the parameters of motility, viability, and abnormalities.

Artificial Insemination of Cattle: Current and Future Trends

Artificial Insemination of Cattle: Current and Future Trends

Genome-wide association study reveals candidate markers related to field fertility and semen quality traits in Holstein-Friesian bulls

In vitro assessment of bull semen quality is routinely used in bull semen processing centres in order to ensure that semen destined to be used in the field has passed minimum standards. Despite these stringent quality control checks, individual bulls that pass the quality control checks can still vary in field fertility by up to 25%. A genome-wide association study was undertaken to determine genetic markers associated with prefreeze and post-thaw bull sperm quality traits as well as field fertility. Genome-wide association analysis was performed using a single nucleotide polymorphism (SNP) regression mixed linear model in WOMBAT. Genes within a 250 Kb span of a suggestive (P ≤ 1 × 10−5) SNP were considered as candidate genes. One SNP was associated with adjusted pregnancy rate, and 21 SNPs were associated across the seven semen quality traits (P ≤ 1 × 10−5). Functional candidate genes include SIPA1L2 which was associated with adjusted pregnancy rate. This encodes a Rap GTPase-activating protein involved in Rap1 signalling pathway and was previously found to play a role in the process of sperm differentiation. Gene ontology (GO) analysis also identified significantly enriched biological processes involved protein tyrosine kinase activity including genes such as DYRK1A, TEC and TXK that were associated with sperm motility prior to freezing. Another candidate gene associated with post-thaw sperm motility was FHDC1 which coordinates actin filament and microtubule dynamics. The induced 11 GO terms in the ejaculates rejected after freezing trait were related to ATPase, phosphatase and hydrolase activity. These results reveal novel specific genomic regions and candidate genes associated with economically important phenotypes such as field fertility and semen quality traits.

Tinospora cordifolia modulates the seminal parameters, leakage of intracellular enzymes and seminal antioxidants in equilibrated and cryopreserved semen of Sahiwal bulls.

The present study was undertaken to assess the effects of stem extract of Tinospora cordifolia (Giloy or Guduchi) in the semen extender on seminal parameters, leakage of intracellular enzymes and antioxidants in semen of Sahiwal bull. A total of 48 ejaculates from four bulls were selected for the study. Spermatozoa of 25×106 were incubated in 100, 300 and 500 μg of stem extract of Guduchi as Gr II, III and IV, respectively and pre-freeze and post-thaw semen samples were analysed for seminal parameters (motility, viability, total sperm abnormality; TSA, plasma membrane integrity; PMI, and acrosomal integrity; AcI), intracellular enzymes (aspartate aminotransferase; AST and lactate dehydrogenase; LDH) and seminal antioxidants (superoxide dismutase; SOD and catalase) in comparison with an untreated control group (Gr I). The results revealed that stem extract treated semen had significantly (p<0.05) higher motility, viability, PMI, AcI, SOD and catalase and had significantly (p<0.05) lower TSA, AST and LDH compared to those in untreated control group at pre-freeze and post-thaw stages. Semen treated with 100 μg stem extract /25×106 spermatozoa had significantly (p<0.05) higher motility, viability, PMI, AcI, SOD and catalase and had significantly (p<0.05) lower TSA, AST and LDH compared to those in control, 300, and 500 μg treated groups at pre-freeze and post-thaw stages. Further, these seminal parameters and antioxidants were showing decreasing trend and TSA and leakage of intra-cellular enzymes were showing increasing trend from Gr II to Gr IV at pre-freeze and post-thaw stages. Thus, 100 μg/25×106 spermatozoa were optimum or suitable dose for cryopreservation of Sahiwal bull semen. The study concluded that T. cordifolia stem extract 100 μg/25×106 spermatozoa in the semen extender can be effectively utilized to reduce the oxidative stress and improve the pre-freeze and post-thaw seminal parameters in Sahiwal bull. However, further studies on effects of different concentrations of stem extract on in-vitro or in-vivo fertility trials are to be conducted to assess the impact of the stem extract supplementation in the semen extender on field pregnancy outcome in bovine species.

Efek Tipe Kelahiran Terhadap Estrus Post-Partum Induk Sapi Bali Akseptor Inseminasi Buatan

Cases of dystocia is impact on delaying uterine involution, post-partum estrus and the next mating. Research has been carried out about type of birth process and its effect on post-partum estrus post-artificial insemination (AI) of Bali cows in Pujut District, Central Lombok Regency. The aim was to determine the effect of the type of birth process on post-partum estrus of cows. The research used descriptive method with the material were 40 Bali cows in 4 (four) villages, namely Pengengat, Teruwai, Mertak and Bangket Parak. The research variables were the type of birth process and post-partum estrus, as well as supporting variables such as the age of cow, body condition score (BCS), body weight, type of bull semen and days open. The data were statistically analyzed including the mean, standard deviation and percentage, followed by t-test. The results showed that 52.25% of Bali cows in Pujut District gave birth normally and 47.5% suffer light dystocia with birth assistance. The mean age of cows with dystocia was 29.53 ± 5.72 months by the BCS of 2.53, compared to 31.62 ± 5.61 months by the BCS of 2.81 in normal birth. The mean weight was 319.00 ± 29.72 kg compared to 342.81 ± 49.36 kg. The mean days open was 199.13 days compared to 195.67 days and post-partum estrus was 122.00 days compared to 105.00 days. In conclusion, the onset of post-partum estrus and days open in Bali cows that suffer light dystocia with birth assistance is longer than normal birth.