Cattle Sperm Research Articles

Comprehensive Study of Sperm Proteins and Metabolites Potentially Associated with Higher Fertility of Zebu Cattle (Bos indicus) in Tropical Areas.

Crossbreeding of zebu cattle (Bos indicus) with European breeds (Bos taurus) producing crossbred cattle was performed to overcome the low growth rates and milk production of indigenous tropical cattle breeds. However, zebu cattle fertility is higher than those of crossbred cattle and European breeds under warm conditions. Combination study of proteomics and metabolomics toward Malaysian indigenous breed Kedah × Kelantan-KK (B. indicus) and crossbreed Mafriwal-M (B. taurus × B. indicus) to understand physiological reasons for higher thermotolerance and fertility in Zebu cattle sperm. 161 regulated metabolites and 96 regulated proteins in KK and M (p < 0.05) showed more efficient carbohydrate and energy metabolism, higher integrity of the DNA and plasma membrane, a lower level of reactive oxygen species, and higher levels of phospholipids, which confirmed higher sperm plasma membrane integrity in KK. A stronger antioxidant system and lower polyunsaturated fatty acids help KK sperm cope with oxidative stress under warm conditions. The higher abundance of flagella structural proteins in KK provides a stronger structure that supports sperm motility. Abnormality of flagella, plasma membrane disruption, and DNA fragmentation were higher in M. These findings provide selective molecular markers for developing high-producing and more thermotolerant cattle breeds in tropical areas (197 words).

THE EFFECTIVENESS OF VITAMIN C AND E ADDITION IN VARIOUS DILUENTS OF BRAHMAN CATTLE SPERMATOZOA AFTER FREEZING

This research aimed to determine the quality of Brahman cattle spermatozoa after freezing in various diluent ingredients (Egg Yolk Citrate, Egg Yolk Tris, Skim Milk) which added vitamin C and vitamin E. The research was carried out using 4 treatments with 3 different diluent ingredients, each of which was Vitamins C and E are added to each diluent ingredient at a dose adjusted to the type of diluent agent. The treatment given is P0: Diluent without Vitamin C and E; P1: Diluent + Vitamin C; P2: Diluent + Vitamin E; and P3: Diluent + Vitamin C and Vitamin E. Research data were analyzed using analysis of variance and followed by Duncan's test at the 5% and/or 1% level for different variables. The results showed that the addition of vitamin C and vitamin E had a significant effect (P0.05) on motility and viability in the egg yolk tris diluent and egg yolk citrate but had no significant effect (P0.05) in the skim milk diluent. The use of different diluents had no significant effect (P0.05) on motility, viability, and abnormalities. This research concludes that Vitamin C added to the diluent tris egg yolk, egg yolk citrate, and skim milk showed higher average motility, percentage of live spermatozoa, and abnormalities of post-freezing Brahman cow spermatozoa.

The effect of adding butterfly pea (Clitoria ternatea) flower extract to the coconut water-egg yolk diluent on the quality of Aceh cattle spermatozoa

Abstract This study aimed to determine the effect of adding butterfly pea flower (Clitoria ternatea) extract to the coconut water-egg yolk diluent on the quality of Aceh cattle spermatozoa stored at 4°C. The semen used is obtained from the ejaculate of Aceh cattle kept in the Saree artificial insemination center. The treatment in this research was the addition of butterfly pea flower extract to the coconut water-egg yolk diluent with concentrations of 0% (P1), 0.2% (P2), 0.4% (P3), 0.6% (P4), and 0.8% (P5). The spermatozoa quality variables measured were motility, viability, and abnormalities of spermatozoa after and before cooling. The results showed that adding butterfly pea flower extract to the diluent significantly affected spermatozoa motility and viability. Still, they did not affect the percentage of spermatozoa abnormalities before or after the cooling process. The quality of Aceh cattle spermatozoa before and after the cooling process was no different. Adding butterfly pea flower extract to a concentration of 0.6 g/100 ml in the diluent (P4) produced the best spermatozoa motility, viability, and abnormalities. The conclusion is that the addition of butterfly pea flower extract to the coconut water-egg yolk diluent can prevent a decrease in the quality of Aceh cattle spermatozoa stored at 4°C.

Phytochemical Screening, Antioxidant Effect and Sperm Quality of the Bomba ceiba Stamen Extracts on Charolais Cattle Sperm Induced by Ferrous Sulfate.

Orange Bombax ceiba (B. ceiba) is an indigenous plant, and its stamen is an important ingredient in traditional Lanna food. There are limitations in scientific reports on the effects of the biological activities of B. ceiba stamens on the male reproductive system. This study aims to investigate the phytochemical compounds of the orange B. ceiba stamen and its potential effect on the antioxidant properties and quality of cattle sperm treated with Fe. The orange BUE had the highest total phenolics, total tannins, total monomeric anthocyanins, and maximal antioxidant potential. The orange BAE had the highest concentration of total flavonoids. LC-QTOF/MS showed that the orange BUE contained the highest number of phytochemical compounds related to male reproductive enhancement. The orange BUE enhanced sperm motility, and both the orange BUE and the BAE enhanced sperm viability and normal sperm morphology via free radical scavenging. It might be suggested that B. ceiba stamens have benefits for sperm preservation, sperm quality, and increasing the economic value of local plants, and that they may be developed and used to guard against oxidative stress from cryodamage induced by frozen semen technology.

Cold exposure impacts DNA methylation patterns in cattle sperm.

DNA methylation is influenced by various exogenous factors such as nutrition, temperature, toxicants, and stress. Bulls from the Pacific Northwest region of the United States and other northern areas are exposed to extreme cold temperatures during winter. However, the effects of cold exposure on the methylation patterns of bovine sperm remain unclear. To address, DNA methylation profiles of sperm collected during late spring and winter from the same bulls were analyzed using whole genome bisulfite sequencing (WGBS). Bismark (0.22.3) were used for mapping the WGBS reads and R Bioconductor package DSS was used for differential methylation analysis. Cold exposure induced 3,163 differentially methylated cytosines (DMCs) with methylation difference ≥10% and a q-value < 0.05. We identified 438 differentially methylated regions (DMRs) with q-value < 0.05, which overlapped with 186 unique genes. We also identified eight unique differentially methylated genes (DMGs) (Pax6, Macf1, Mest, Ubqln1, Smg9, Ctnnb1, Lsm4, and Peg10) involved in embryonic development, and nine unique DMGs (Prmt6, Nipal1, C21h15orf40, Slc37a3, Fam210a, Raly, Rgs3, Lmbr1, and Gan) involved in osteogenesis. Peg10 and Mest, two paternally expressed imprinted genes, exhibited >50% higher methylation. The differential methylation patterns of six distinct DMRs: Peg10, Smg9 and Mest related to embryonic development and Lmbr1, C21h15orf40 and Prtm6 related to osteogenesis, were assessed by methylation-specific PCR (MS-PCR), which confirmed the existence of variable methylation patterns in those locations across the two seasons. In summary, cold exposure induces differential DNA methylation patterns in genes that appear to affect embryonic development and osteogenesis in the offspring. Our findings suggest the importance of replicating the results of the current study with a larger sample size and exploring the potential of these changes in affecting offspring development.

C-type natriuretic peptide treatment before in vitro maturation enhances maturation of yak oocytes and developmental competence of their crossbred embryos

ABSTRACT The effects of C-type natriuretic peptide (CNP) pre-treatment on the in vitro maturation (IVM) and developmental competence of yak (Bos grunniens) oocytes were evaluated. The results showed that 100 nM CNP could effectively maintain meiotic arrest of yak oocytes in vitro within 6 h. Yak oocytes matured in two-step IVM (pre-IVM with 100 nM CNP for 6 h followed by conventional IVM for 28 h) yielded a significantly (P < .05) increased cleavage rate, blastocyst rate after in vitro fertilization with cattle spermatozoa compared to the conventional one-step IVM method. The density of transzonal projections (TZPs) decreased after 6 h standard IVM (P < .05), whereas, COCs pre-IVM with 100 nM CNP for 6 h maintained the same density of TZPs compared to the fresh COCs. This pre-IVM resulted in lesser reactive oxygen species level (P < .05). Two-step IVM resulted in the increased glutathione levels, mRNA expression levels of BCL2, BMP15, and GDF9 (P < .05) in oocytes. In conclusion, these results indicate that CNP treatment (100 nM, 6 h) before IVM could enhance the maturation of yak oocytes and developmental competence of their crossbred embryos. This may have important applications in in vitro production of yak crossbred embryos.

Efek Penyimpanan Semen Beku Terhadap Kualitas Spermatozoa Sapi Simental Pada Suhu 5°C

This study aimed to determine the effect of frozen semen storage on sperm quality. quality of Simmental cattle spermatozoa at 5° C. This research was conducted laboratory at the UPTD Laboratory of the Livestock Breeding Center of the Jambi Provincial Food Crops Horticulture and Livestock of Jambi Province, from July 10 to August 22, 2023. August 22, 2023. Completely Randomized Design (CRD) was used in the study. with 5 treatments and 5 replications so there were 25 experimental units. Treatment PO (control) = direct thawing, P1 = storage for 15 minutes, P2 = storage for 30 minutes. storage for 30 minutes. P3 = storage for 45 minutes, P4 = storage for 60 minutes. storage for 60 minutes. The variables observed were spermatozoa motility, spermatozoa viability, spermatozoa abnormality. The results of analysis of variance showed that the storage of frozen semen of Simmental cattle at 5 ° C with different times had a significant effect (P &lt;0.05) on the motility of spermatozoa and spermatozoa viability and not significantly different (P&gt;0.05) on spermatozoa abnormality. Based on the research results obtained It is concluded that storage of frozen semen inside at 5 ° C can maintain the quality of frozen semen of Simental cattle with a storage time of 45 minutes.

The Effect of Additional Tomato Extract in Diluting Sodium Citrate and Egg Yellow in Kampung Chicken on The Quality of Semen Cow Limousine

Undiluted sperm will reduce fertility, therefore dilution is performed to maintain the quality of spermatozoa during storage. Tomatoes contain carbohydrates, vitamin A, vitamin C, protein, and lycopene as antioxidants. Carbohydrates and antioxidants act as a source of energy to potentially prevent free radicals that can damage cells. This study aims to determine the effect of adding tomato juice in sodium citrate diluent and free-range chicken egg yolks to the semen quality of limousin cattle. This study used a completely randomized design (CRD) with 4 treatments and 5 replications. The treatment consisted of a solution of sodium citrate and egg yolk (P0), sodium citrate and egg yolk + 10% tomato extract (P1), sodium citrate and egg yolk + 20% tomato extract (P2), and egg yolk sodium citrate + 30% tomato extract. % (P3). Parameters observed were motility, viability and abnormalities of semen spermatozoa of Limousin cattle. Observations were made at 0, 24, 48, and 72 hours after cooling. The results showed that the addition of 10% tomato extract (P1) had a significant effect on the highest percentage of motility, namely 42.00 ± 2.73c for the 48th hour, the highest viability 62.10 ± 4.51c for the 72nd hour, and lowest abnormality 16.60 ± 1.81a until the 72nd hour. The addition of 10% tomato extract (P1) in sodium citrate diluent and egg yolk resulted in the best motility, viability, and abnormality percentage of Limousin cattle spermatozoa.

Effects of Adding Glutathione to AndroMed Diluent on Intact Plasma and Acrosome Membranes, and Progressive Motility of Cattle Spermatozoa During Freezing Processes

Adding endogenous antioxidants to the diluent is significantly associated with semen quality during the freezing process. This study aimed to investigate the effects of adding glutathione to AndroMed diluent on the preservation of crucial sperm attributes, namely, intact plasma membrane (IPM), intact acrosome membrane (IAM), and progressive motility of Bali cattle spermatozoa. A completely randomized design was used, and spermatozoa samples were obtained from a Bali cattle and divided into two diluent treatment groups (36 diluent samples in each group with six replications), namely pure AndroMed as the control and a group with the addition of glutathione (1 mmol) to AndroMed. Each treatment was replicated six times and evaluated at three freezing stages, including post-dilution, post-equilibration, and post-thawing, for crucial sperm properties. The results indicated that fresh Bali cattle spermatozoa had progressive motility, IAM, and IPM of 75%, 89%, and 88%, respectively. During the freezing process, there was a significant decrease in semen quality, including progressive motility, IAM, and IPM of spermatozoa after dilution to post-equilibration and post-equilibration to post-thawing in both treatment groups. Meanwhile, the addition of 1 mmol of glutathione to AndroMed diluent had a significant difference in increasing progressive motility, IAM, and IPM of Bali cattle spermatozoa at each stage of semen freezing, including post-dilution, post-equilibration, and post thawing when compared with controls. Based on the results, it can be concluded that adding 1 mmol of glutathione to the AndroMed diluent enhanced the quality and integrity of Bali cattle semen, including progressive motility, IAM, and IPM during the freezing process.

Ameliorative effect of crocin on post-thaw quality, fertility-associated gene expression and fertilization potential of buffalo (Bubalus bubalis) bull sperm.

Buffalo bull sperm suffer more cryoinjuries due to lipid peroxidation of high structural polyunsaturated fatty acid contents than cattle sperm. Consequently, the post-thaw fertilization potential of buffalo bull sperm is compromised. Crocin is a carotenoid known for its antioxidant potential through scavenging reactive oxygen species. Objectives of the current study were to investigate the effect of crocin addition in the semen extender on post-thaw quality, fertility-associated gene expression and fertilization potential of buffalo bull sperm. Semen samples (n = 32) from four Nili-Ravi buffalo bulls were extended with tris-citric acid extender containing different concentrations of crocin (0 mM; control, 0.5, 1, 1.5 and 2 mM). The extended semen was packed in 0.5 mL French straws (25 × 106 sperm/straw) and cryopreserved in liquid nitrogen. Computer-assisted semen analysis, hypo-osmotic swelling test, normal apical ridge assay, Rhodamine 123, acridine orange, propidium iodide staining, and thiobarbituric acid reactive substances assay were performed to assess sperm motility parameters, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential, DNA integrity, viability, and lipid peroxidation, respectively. Expression levels of sperm acrosome-associated SPACA3, DNA condensation-related PRM1, anti-apoptotic BCL2, pro-apoptotic BAX, and oxidative stress-associated ROMO1 genes were evaluated through qPCR. The fertility of semen doses containing the most potent concentration of crocin (based on optimum post-thaw semen quality) was compared with control during the breeding season. Buffaloes (n = 400; 200/group) were inseminated 24 h after the onset of oestrus and transrectally palpated for pregnancy at least 60 days post-insemination. Results revealed that 0.5 and 1 mM crocin improved sperm post-thaw total motility, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential and viability, and 1 and 1.5 mM crocin enhanced catalase activity and reduced lipid peroxidation compared to control (p < .05). Moreover, 1 mM crocin improved sperm post-thaw progressive motility, kinematics, and DNA integrity, and 1.5 mM crocin enhanced plasma membrane integrity than control (p < .05). Expression levels of SPACA3, PRM1 and BCL2 genes were higher (p < .05) with 1 mM crocin compared to other groups. In contrast, no difference (p > .05) was noticed in expressions of BAX and ROMO1 genes among all groups. The fertility rate of semen doses containing the most potent concentration (1 mM) of crocin was higher (p = .0465) compared to control (56 ± 0.03% vs. 46 ± 0.04%, respectively). In conclusion, 1 mM crocin in the semen extender improves post-thaw quality, fertility-associated gene expression and fertilization potential of buffalo bull sperm.

The Effect of Adding Sugar Cane (Saccharum officinarum L.) Water in Skim Milk and Egg Yolks on The Quality of Simmental Cow

Sugarcane is a type of grass plant that is widely cultivated by the community and has a fairly high nutritional content. Sugarcane juice contains sucrose which consists of glucose and fructose. Sucrose contained in sugarcane juice serves as a source of energy for spermatozoa. This study aims to determine the effect of adding sugarcane juice to skim milk diluent and egg yolk on motility, viability and semen abnormalities of Simmental cattle. This research was conducted at the UPT Artificial Insemination Department of Food Security and Animal Husbandry of North Sumatra Province. The method used in this study was a completely randomized design with 5 treatments and 4 replications consisting of P0: Skimmed milk and egg yolk diluent without sugarcane juice, P1: Skim milk and egg yolk diluent + 25 ml of sugarcane juice, P2: Milk diluent skim and egg yolk + 35 ml of sugarcane juice, P3: Diluent of skim milk and egg yolk + 45 ml of sugarcane juice. Parameters measured were motility, viability and sperm abnormalities of Simmental cattle semen. The results of this study indicate that the P1 treatment with 25 ml of sugarcane juice was able to maintain motility during 72 hours of storage by 40%, the highest mean value of spermatozoa viability during 144 hours of storage (60.90% ± 1.08), and the lowest spermatozoa abnormality value during storage 144 hours (17.10% ± 1.08). The conclusion of this study was that 25 ml of sugarcane juice in skimmed milk and egg yolk diluent could maintain the quality of Simmental cattle spermatozoa.

Review Jurnal: Efektivitas Penggunaan Kuning Telur Berbagai Jenis Unggas Sebagai Pengencer Semen pada Ternak

A method is needed that can be used to maintain sperm quality in order to produce superior seeds. The use of poultry egg yolk as a diluent is of course widely used because poultry eggs can maintain the good quality of cattle sperm. This research aims to determine the level of effectiveness of egg yolks of various types of poultry as a sperm diluent on the motility and viability of cattle sperm. The method used in this research is the literature review method and the descriptive method. The results showed that the use of free-range chicken egg yolk and quail egg yolk as sperm diluent showed the highest motility values. The highest percentage of viability was in purebred chickens, while the lowest abnormality was in dilution with goose yolk eggs.

Comparative RNA isolation methods from fresh ejaculated spermatozoa in Sahiwal cattle (Bos indicus) and Murrah buffalo (Bubalus bubalis) bulls for high quality and enhanced RNA yield

Sperm mRNA transcriptional profiling can be used to evaluate the fertility of breeding bulls. The aim of the study was to compare the modified RNA isolation methods for higher RNA yield and quality from freshly ejaculated sperm of cattle and buffalo bulls. Ten fresh ejaculates from each Sahiwal (n = 10 bulls × 10 ejaculates) and Murrah bulls (n = 10 bulls x 10 ejaculates) were used for RNA isolation. From the recovered live sperm, total sperm RNA was isolated by conventional methods (TRIzol, Double TRIzol), membrane-based methods combined with TRIzol (RNeasy + TRIzol) with the addition of β-mercaptoethanol (BME) and Kit (RNeasy mini) methods in fresh semen. Among different isolation methods; the membrane-based modified methods combined with TRIzol (RNeasy + TRIzol) with the addition of β-mercaptoethanol (BME) resulted significantly (p < .05) higher total RNA quantity (300–340 ng/µL) and better purity in different concentrations of spermatozoa viz., 30–40 million, 70–80 million and 300–400 million sperm. The study concluded that the inclusion of BME to the combined membrane-based methods with somatic cell lysis buffer solution was best for constant increased yield and purity of RNA isolation from Sahiwal cattle and Murrah buffalo bull sperm.

PSV-9 Cold Exposure Impacts Dna Methylation Patterns of Genes in Cattle Sperm Involved in Early Embryonic and Bone Development

Abstract DNA methylation and other epigenetic modifications are indispensable for maintaining sperm quality, fertilization capacity, and embryonic and postnatal development. In mice, environmental factors, such as stress, nutrition, or exposure to the cold condition, have been demonstrated as factors that alter methylation marks of sperm that are passed to the subsequent generation through transgenerational inheritance and genomic imprinting. Breeding bulls from northern regions of the USA are exposed to extreme cold for 2 to 3 months during the winter; however, no single study has analyzed the consequences of this cold exposure on the methylation pattern of bull sperm, which constitutes a significant knowledge gap. Therefore, this study aimed to explore the effects of cold exposure on the overall and gene-specific methylation status in sperm. We collected semen from 5 bulls during winter seasons after cold exposure and at normal temperatures during late spring. Whole Genome Bisulfite Sequencing (WGBS) was conducted to obtain the DNA methylation profile of these semen samples and to identify unique genes that have differentially methylated regions due to cold exposure (Figure 1). Cold exposure did not change the overall methylation level between the two groups but induced 438 Differentially methylated regions (DMRs) that overlapped with promoters, introns, exons, intergenic regions, shores, and shelves of CpG island (CGI) in 186 unique genes. We also identified nine unique differentially methylated genes (DMGs) (Pax6, Macf1, Mest, Ubqln1, Smg9, Trappc9, Ctnnb1, Lsm4, Peg10) involved in embryonic development and nine unique DMGs (Prmt6, Nipal1, C21h15orf40, Slc37a3, Fam210a, Raly, Rgs3, Lmbr1, Gan) involved in osteogenesis (Table 1). DMRs were located in the promoter regions and introns of these genes which preferentially involve gene silencing and alternative splicing. Among the DMGs involved in embryonic development, Mest and Peg10 are two paternally imprinted genes where only the paternal allele expresses. Peg10, required for optimal placental growth and trophoblast proliferation, overlapped with a hypermethylated DMR in the promoter region. Mest, another paternally imprinted gene, the downregulation of which results in embryonic growth retardation, had hypermethylation in introns. Moreover, Methylation-specific PCR (MS-PCR) verified the methylation changes in the identified regions. It was the first study to investigate the effect of cold exposure on DNA methylation of cattle sperm and suggest its association with altered sperm DNA methylation profiles. Differential methylation appears to alter gene expression and affect early embryonic development, osteogenic activity, and overall offspring growth performance through the imprinting effects.

Identifikasi Morfometri Spermatozoa Sapi Simental dan Sapi Ongole

The aim of the research was to determine the morphometry comparison of spermatozoa of simental cattle and ongole cattle such as head length, head width, head area, middle piece length, tail length and overall body length. This research was carried out at the UPTD Laboratory of the Livestock Breeding Center, Horticulture and Animal Husbandry Food Crops Service, Jambi Province. This methodology uses exploration and observation of spermatozoa, using a microscope with a magnification of 400 times. The samples measured were 480 spermatozoa, taken from frozen semen randomly at the Department of Horticultural Food Crops and Livestock, Jambi Province, then smear preparations were made with eosin dye. The variables observed were head length (PK), head width (LK), head area (LKP), middle piece length (PMD), tail length (PE) and overall body length (PTK). The research results showed that the morphometry of simental cattle spermatozoa PK (9.46 µm), LK (5.11 µm), LKP (41.96 µm), MD (13.77 µm), PE (47.68 µm) and PTK ( 70.92 µm). Meanwhile, the size of the spermatozoa of Ongole cattle PK (9.48 µm), LK (5.17 µm), LKP (42.57 µm), MD (13.83 µm), PE (47.96 µm) and PTK (71.28 µm); The statistical results obtained were not significantly different in the morphology of the two cow spermatozoa. The relationship between PTK parameters of simental cattle spermatozoa and PE was very significant (r=0.870) and likewise for Ongole Cow Spermatozoa, the relationship between PTK and PE was very significant (r=0.953). This situation shows that there is a very close relationship between body length and tail length. The percentage of X chromosome simental cattle spermatozoa was (44.38%) and Y chromosome spermatozoa was (55.62%), while the percentage of X chromosome Ongole cattle spermatozoa was (45.83) and Y chromosome carrying spermatozoa was (54.17).

Preservation of Simmental bull sperm at 0°C in Tris dilution: effect of dilution ratio and long-distance transport.

This study aimed to assess the impact of the dilution ratio of Tris diluent, storage at 0°C, and long-distance transportation on the spermatozoa of Simmental cattle. It also validated the feasibility of the regional distribution of fresh semen. In experiment 1, semen was diluted at four dilution ratios (1:6, 1:9, 1:12, and 1:15) to determine the optimal dilution ratio of Tris diluent. In experiment 2, we assessed sperm viability, progressive motility (objectively assessed by computer-assisted sperm analyzer), and acrosome intactness in Tris dilutions kept at constant 0°C for 1, 3, 6, 9, and 12 days. We compared them to Tianshan livestock dilutions (Commercial diluent). In experiment 3, semen was diluted using Tris diluent, and sperm quality was measured before and after long-distance transport. Artificial insemination of 177 Simmental heifers compared to 156 using Tianshan Livestock dilution. The outcomes demonstrated that 1:9 was the ideal Tris diluent dilution ratio. The sperm viability, Progressive Motility, and acrosome integrity of both Tris and Tianshan dilutions preserved at 0°C gradually decreased over time. sperm viability was above 50% for both dilutions on d 9, with a flat rate of decline. The decrease in acrosome integrity rate was faster for Tianshan livestock dilutions than for Tris dilutions when stored at 0°C for 1 to 6 days. There was no significant difference (p>0.05) in sperm viability between semen preserved in Tris diluent after long-distance transportation and semen preserved in resting condition. The conception rates for Tris dilution and Tianshan livestock dilution were 49.15% and 46.15% respectively, with no significant difference (p>0.05). This shows that Tris diluent is a good long-term protectant. It has been observed that fresh semen can be successfully preserved for long-distance transport when stored under 0°C conditions. Additionally, it is feasible to distribute semen regionally.

Effect of Green Tea Extract on Spermatozoa Quality of Peranakan Ongole Bull On Frozen Storage

Low-temperature storage can often result in spermatozoa damage. It occurs due to Reactive Oxygen Species, leading to changes in the lipid composition of the membrane. Antioxidant compounds are needed to prevent ROS. Green tea extract (Camelia sinensis) can be used as an antioxidant agent. This study aimed to determine the effect of green tea extract in tris egg yolk on spermatozoa quality of Ongole Peranakan (PO) cattle. This study used a completely randomized design (CRD) with five extract treatments (0%, 1%, 2%, 3%,4%) and four repetitions. The parameters used were motility, viability, and membrane integrity. The data were analyzed with ANOVA and Duncan's test. Based on the results of the ANOVA test, the addition of green tea extract has a significant effect on spermatozoa quality of PO cattle in frozen storage (P0.005). The best results were obtained by the addition of 3% extract that was able to maintain the quality of post-freezing spermatozoa with average motility, viability, and membrane integrity of 47.44±0.166, 55.14±0.449, and 53.72±0.599. The addition of 3% green tea extract into tris egg yolk can maintain motility, viability, and membrane integrity in spermatozoa of PO cattle inside frozen storage, by adding green tea extract to diluent, spermatozoa can be stored for a long time at low temperatures making it easier for people to carry out artificial insemination.

Ability of Melon (Cucumis melo. L) Fruit Juices Based Tris Diluent on The Quality of Frozen Spermatozoa of Bali Cattle After Thawing

The high potassium content in melon plays a role in maintaining the normal function of the muscles, heart, nervous system, and potassium is also the main regulator of blood pressure. The aim of this study was to evaluate the effect of using melon juice (Cucumis melo L.) as a diluent on the quality of spermatozoa and semen of Bali cattle both before and after freezing. The Banyumulek Artificial Insemination Center produces sperm used in Bali cattle breeding. The technique used was a completely randomized design (CRD) with 4 treatments and 15 replications. The first treatment was 100% egg yolk tris buffer without the addition of melon juice (P0), the second treatment was 80% buffer without the addition of 20% melon juice (P1), the third treatment was 60% buffer with the addition of 40% melon juice (P2). The fourth treatment was 40% buffer with the addition of 60% melon juice (P3). The outcomes showed that the worth of flawless plasma film and mortality upsides of ox-like semen after weakening and subsequent to defrosting were not essentially unique. Be that as it may, not altogether disparate in that frame of mind of motility, level of reasonability, level of ordinariness and post-defrosting anomalies. According to the findings of this study, melon juice can be used as an alternative extender for fresh Bali cattle sperm and preserve the quality of spermatozoa.

Effect of Addition of Green Tea Extract (Camellia sinensis) in Egg Yolk Tris Diluter on Spermatozoa Quality in Bali Cattle (Bos sondaicus) After Freezing

This study aimed to determine the effect of adding 96% ethanol extract of green tea (Camellia sinensis) in egg yolk tris diluter on the quality of spermatozoa of Bali cattle (Bos sondaicus) after freezing. The samples were divided into four different treatment groups, which contained semen sample + 13% glycerol + each egg yolk tris diluter + green tea extract (P0) 0 mg/100 ml, (P1) 0,05 mg/100 ml, ( P2) 0,10 mg/100 ml, and (P3) 0,15 mg/100 ml. The variables observed included motility, viability, and spermatozoa abnormalities. Data were analyzed using OneWay ANOVA and continued with Duncan's test (p&lt;0,05). The results showed that the P3 group had motility of 49,00 ± 2,15, viability of 61,92 ± 1,70, and abnormality of 4,24 ± 2,16 significantly compared to the other groups. In conclusion, the P3 group with the addition of 0,15 mg/100 ml of green tea could increase the motility, viability, and abnormal spermatozoa of Bali cattle.

Detection of Plasma Membrane Alpha Enolase (ENO1) and Its Relationship with Sperm Quality of Bali Cattle

Bali cattle are the indigenous cattle well known attributed to their high fertility. Various proteomes in sperm are substances that can determine sperm quality and fertility. One of which is alpha enolase (ENO1). This study aims to assess the presence of ENO1 in the plasma membrane of sperm and its relationship with the semen quality of Bali cattle. The study used 30 ejaculates from 5 bulls for sperm quality assessment and detection of ENO1. Sperm quality is indicated by total motility, progressive motility, kinematics motility, viability, plasma membrane integrity, and ENO1 quantity. Sperm motility and kinematics motility were measured using CASA, while viability was assessed by eosin-nigrosine differential staining. The HOS test was used to determine plasma membrane integrity, and ENO1 quantity was measured by the ELISA method. Data were analyzed using ANOVA (randomized group design), linear regression, Pearson’s correction, and the t-Student test. The results showed that ENO1 was detected in the plasma membrane at 1.27 ng/106 sperms. The amount of alpha-enolase in the plasma membrane of Bali cattle sperm is affected by sperm concentration (p<0.01) and not involved in sperm motility. There was no correlation between plasma membrane ENO1 quantity and semen quality. The results of this experiment indicated that alpha-enolase in the plasma membrane of Bali cattle sperm is affected by sperm concentration but not related to sperm quality.