Bull Semen Research Articles

Генетическое разнообразие сохраненной и предковой популяций чистопородного холмогорского скота Республики Коми

The genealogical information and genetic markers of different types were used to evaluate the genetic diversity of the endangered Kholmogory cattle breed in the Komi Republic. The main genealogical and gene pool of the ancestral population is available in the gene pool farms and the bank of cryopreserved semen of bulls. The effective number of the today’s population is sufficient to prevent the inbreeding from increasing above the acceptable limits for a short time. However, to maintain the genetic diversity for a long time, we need to attain at least a two-fold increase in the effective population number, primarily by increasing the number of seed donors. The use of different types of markers allows to obtain the exhaustive and objective information about the dynamics of the gene pool of the endangered breed.

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.

Correlation analysis of the PRM1, STK35 and IFT27 level of expression genes with the quality of native and deposited seed of holsting bulls

The research aim is a correlation analysis of the PRM1, STK35 and IFT27 genes mRNA expression with the native and decryoconserved sperm quality indicators of Holstein bulls to search for effective transcriptomic biomarkers of bull semen. In the study course, native and decryoconserved sperm of seven Holstein bulls were used. To solve the study problems, eight indicators of sperm quality were studied, the studied genes in native and decryoconserved spermatozoa expression was analyzed in real time PCR. Nonparametric probabilistic and statistical methods were used, the analysis of rank correlation was carried out using Spearman’s criterion. Higher expression of the studied genes was mainly noted in frozen--thawed sperm compared to native. The mRNA expression level of the protamine gene (PRM1) did not give a reliable correlation with sperm parameters: The level ITF27 gene mRNA expression was significantly positively correlated with the content of defective cells from frozen--thawed sperm (0.714, p=0.05) and dead cells (0.714, p=0.0545) from native sperm. A negative correlation was noted with the content of normal cells in frozen--thawed sperm (-0.750, p=0.038) and live cells (-0.714, p=0.0545) in native sperm. The ITF27 gene transcript (mRNA) showed a negative correlation (-0.703, p=0.0545) in terms of the acrosome defect of frozen--thawed spermatozoa. In terms of the content of reactive oxygen species (ROS), the mRNA of the ITF27 gene had a significant positive correlation (0.786, p=0.0251). The STK35 gene transcript (mRNA) was the only one of all the studied mRNAs that had an average negative correlation with sperm motility in native (-0.692, p=0.052) and frozen--thawed sperm (-0.876, p=0.035). These studies can be used to create a system of non-invasive transcriptional markers of bull sperm quality.

Correlation between in vitro sperm kinetic, oxidative stress assessments and field fertility of cryopreserved bull semen

This study assessed kinetic parameters and oxidative stress in bull sperm after post-thaw (PT) or after sperm selection by Percoll™ gradient, and thermo resistance test (SS + TRT) to identify useful indicators of field fertility. For the experiment, commercial doses of frozen semen were obtained from six Aberdeen Angus bulls. Three of the bulls were classified as high fertility and three as low fertility according to the IFert™ index provided by the international breeding company CRV Lagoa. Pooled semen samples were distributed between two treatment groups for analysis: post-thaw (PT) or sperm selection (SS) (Percoll™) and thermal resistance test (SS + TRT). The samples were evaluated using sperm kinetics (CASA) (motility %, progressive motility %, VCL µm/s, VSL µm/s, VAP µm/s, LIN %, STR % and WOB%), production of reactive oxygen species (ROS), lipid peroxidation, superoxide dismutase (SOD) enzyme activity and total antioxidant capacity. Data were analyzed using Two-Way ANOVA, considering the fertility index, the treatment used in the samples as effects, and the interaction between these factors. When a significant effect was observed, the values were compared using the Bonferroni test. A Pearson Correlation analysis was performed between the fertility indices and the sperm parameters analyzed in vitro, to evaluate the relationship between sperm quality and the fecundity rate obtained by the bulls. Sperm kinetic parameters, including total motility, progressive motile, and beat cross-frequency, were higher in low fertility compared to high fertility bulls (P < 0.05). However, curvilinear velocity was greater in high fertility bulls followed by SS + TRT. Straight-line velocity, average path velocity, linearity, and beat cross-frequency beat were higher in high fertility bulls after SS + TRT. Reactive oxygen species was correlated with fertility after SS. In addition, there was a decrease in lipid peroxidation was observed only in high fertility bulls. However, lipid peroxidation and high fertility were correlated after PT and SS + TRT. The combination of in vitro sperm kinetic parameters predicted in vivo fertility more accurately than individual kinetic parameters. The lipid peroxidation of sperm is an important indicator of fertility in bulls. High fertility bulls appeared to be more susceptible to lipid peroxidation, which was only reduced in high fertility bulls, suggesting that their sperm can repair the damage induced by oxidative stress.

Quantitative Characteristics of Crossing Bali Cows with Different Strain of Bulls in Waelata District Buru Regency

This study aims to determine the quantitative characteristics of the crossbreed Bali cattle with several male breeds such as the Ongole, Limousine, and Simmental breeds. The method used in this research is the observation method in the form of direct observation in the field and measurement of cattle. A total of 30 adult cattle aged 1.5-2 years, consisting of 10 heads each (5 males and 5 females) crosses through Artificial Insemination (AI) of Bali cattle using frozen semen of Ongole breed bulls (POBAL), Limousin (LIMBAL) and Simmental (SIMBAL). The results showed that POBAL cattle had an average body length of 114.4-116.0 cm, chest circumference 179.4-220.0 cm, shoulder height 126.2-143.8 cm, and body weight 407.15-586.84 kg . LIMBAL cattle have an average body length ranging from 129.4-151.2 cm, chest circumference 170.4-191.8 cm, shoulder height 123.0-151.6 cm, and body weight 370.184-457.122 kg. While the average body length for SIMBAL cattle ranges from 159.8-180.0 cm, chest circumference 166.6-219.8 cm, shoulder height 125.8-154.1 cm, and body weight 356.662-584.678 kg.

Proposed Marketing Strategy Development for Local Cattle Frozen Semen Sales (Case Study: Balai Inseminasi Buatan Lembang)

BIB Lembang was constructed in 1975 and became the first Artificial Insemination Center to be developed in Indonesia. BIB Lembang is also a Technical Implementation Unit which is under the Director General of Livestock and Animal Health and is responsible to the Director General of Livestock and Animal Health and is technically fostered by the Director of Livestock Breeding and Production with production duties. Marketing, testing, and monitoring of superior livestock semen quality, as well as preparation of artificial insemination methods. Meanwhile, based on the author’s initial research and problem-solving efforts, The research’s findings indicate that although BIB Lembang continues to have strong customer loyalty, a large number of its patrons still choose imported cattle frozen semen to local cattle frozen semen. The purpose of this study is to identify the factors that influence customer satisfaction in relation to service quality, as well as the impact of human resources capability and technological infrastructure support on the quality of local cattle frozen semen products. Additionally, a business strategy to enhance competitiveness in the local cattle semen market will be designed. Both quantitative and qualitative methods will be used in this study. STP methodology and customer analysis both employ quantitative methodologies. Internal study of “Current Marketing Capability” and the search for preferences in “secondary data” were used in the data collection process. Seven people were interviewed using the qualitative method, one of whom was the head of BIB Lembang. Based on the research findings, companies can implement business strategies such as service quality to boost sales of local frozen cow semen. These strategies should be complemented by superior human resource competence, adequate technological infrastructure, good online ordering facilities – Sijalu, and customer satisfaction. To develop a SWOT Analysis and TOWS matrix for BIB Lembang in order to obtain a competitive edge and boost sales of local cattle frozen semen, the internal and external aspects will be analyzed using PESTEL analysis, competitor analysis, current marketing capability, and competitor analysis.

Effect of Supplemented Alcoholic Extract of Annatto (Bixa orellara) Seeds and α-Tocotrienol to Soy-Bean Lecithin Extender on Some Biomarkers of Cryopreserved Semen of Holstein Bulls

The purpose of this research was to examine the impact of Soy-bean lecithin extender combined with different quantities of alcoholic extract of annatto (Bixa orellara) and - tocotrienol on sperm abnormalities and certain biomarkers of Holstein bulls semen after being frozen for varying lengths of time. In the experiment, bull semen was pooled, diluted with Soy-bean lecithin extender, and distributed randomly among five groups: Diluted semen was put through the semen cryopreservation technique; the control treatment (C) consisted solely of soy-bean lecithin extender; T1 included 150mg annatto extract/25 ml; T2 contained 200mg annatto extract/25 ml; T3 contained 3mM tocotreinol; and T4 contained 4mM tocotreinol. After 48 hours, 3 months, and 6 months, both chilled and frozen semen were evaluated for abnormalities, Malondialdehyde (MDA), Total antioxidant capacity (TAC) concentration, and DNA fragmentation percentage. As shown by the data, there were no significant differences between the treatments in terms of total abnormalities in sperm after any of the cryopreservation times, but there were significant differences in terms of MDA and DNA fragmentation. Some of the biomarkers in Holstein bulls’ sperm were found to be significantly improved when a combination of alcoholic extract of annatto(Bixa orellara) and tocotrienol was added to Soy-bean lecithin.

Effect of Adding Kojic Acid and Sodium Alginate to Semen Extender of Buffalo Bulls on some Biological and Bacteriological Parameters

This experiment examined the effects of adding sodium alginate and Kojic acid as substitutes for conventional antibiotics to Skim milk extender on the characteristics of cryopreserved and frozen buffalo bull semen, as well as the evaluation of their additions as antibiotics that help lower the microbial load. Following the collection and dilution of the Skim milk extender, the experimental treatments were separated into five groups, as follows: T1: (control-) without adding any antibiotics; T2: (control+) adding the conventional antibiotics Gentamicin 0.4 IU and Tylosin 0.08 IU per 100 ml; T3: adding Kojic acid at (0.06 g/L) T4: adding sodium alginate at (0.6 mg/mL)T5: adding a combination of Kojic acid (0.06 g/liter) with sodium alginate (0.6 mg/ml) The diluted semen was cryopreserved in accordance with the recommended procedure, and the characteristics of the cryopreserved semen were then assessed. These characteristics included plasma membrane integrity and individual motility, as well as bacteriological tests that measured the total number of bacteria in the treatments, the total number of E. coli bacteria, and the total number of Staphylococcus bacteria for both cooled and frozen semen after 2 hours, 48 hours, 2 months, and 3 months post cryopreservation. The results of the study revealed There were significant enhancement differences (p 0.05) for the treatments T3, T4, and T5 in the percentages of individual motility and viability in comparison to the two control groups, as well as a significant reduction (p 0.05) in the total number of bacteria and the total number of E. coli and Staphylococcus bacteria for treatments T3, T4, and T5 after all periods of cooling and freezing in comparison with the two control groups. It is concluded from the current experiment that the addition of sodium alginate and kojic acid as an alternative substitute for commonly used antibiotics to the semen extender has a significant role in enhancing some characteristics of the buffalo bulls’ semen and helping to reduce the microbial load to a minimum.

Effect of Addition Different Levels of Kojic Acid and Sodium Alginate as a substitutes of conventional Antibiotic on the Characteristics of Cryopreserved Semen of Buffalo Bulls

This experiment examined the effects of adding sodium alginate and KOJIC acid as substitutes of Conventional antibiotics to soybean lecithin extender on the characteristics of cryopreserved and frozen buffalo bull semen, as well as evaluation of their additions as antibiotics that to help lowering the microbial load. Following the collection and dilution of in the soybean lecithin extender, the experimental treatments were separated into five groups, as follows: T1: (control-) without adding any antibiotics; T2: (control+) adding the conventional antibiotics Gentamicin 0.4 IU and Tylosin 0.08 IU per 100 ml; T3: adding Kojic acid at (0.06 g/L) T4: adding sodium alginate at (0.6 mg/mL)T5: adding a combination of Kojic acid (0.06 g/liter) with sodium alginate (0.6 mg/ml) The diluted semen was cryopreserved in accordance with the recommended procedure, and characteristics of the Cryopreserved semen were then assessed. These characteristics included sperm viability and individual motility, as well as bacteriological tests that measured the total number of bacteria in the treatments, the total number of E. coli bacteria, and total number of Staphylococcus bacteria for both cooled and frozen semen after 2 hours, 48 hours, 2 months, and 3 months post cryopreservation Results of the study revealed A significant enhancement differences (p 0.05) for the treatments T3, T4 and T5in the percentages of Individual motility and viability in comparison to the two control groups, as well as significant reduction( P 0.05) in the total number of bacteria and the total number of E. coli and Staphylococcus bacteria for treatments T3, T4 and T5after all periods at cooling and freezing in compared with the two control groups. it is concluded from the current experiment that addition of Sodium alginate and Kojic acid as an alternative substitute for commonly used antibiotics to the semen extender has a significant role in enhancing some characteristics of the buffalo bulls’ semen and helped to reduce the microbial load to a minimum.

Oxford nanopore sequencing as an alternative to reduced representation bisulphite sequencing for the identification of CpGs of interest in livestock populations

Epigenetic marks could potentially explain a portion of the phenotypic variance related to traits of interest in animal breeding models. DNA methylation is the most studied epigenetic mark, involving the addition of a methyl group to the fifth carbon of a cytosine, which transforms it into 5-methylcytosine. This mark is commonly associated with inhibiting gene expression without altering the nucleotide sequence, particularly when located within promoter regions. While bisulphite sequencing is the gold standard for detecting methylation marks, new techniques have emerged to address bisulphite's limitations. Some of these limitations include the confounding effects of bisulphite treatment on DNA methylation and C to T polymorphisms, as well as the inability to distinguish between DNA methylation and hydroxylation. In this context, we propose employing Nanopore sequencing to identify methylation marks. We conducted sequencing on six bull semen samples using both Nanopore and reduced representation bisulphite sequencing. CpGs were filtered based on two coverage thresholds (4x and 7x). Our results indicated that Nanopore methylation data exhibited a correlation exceeding 0.95 with bisulphite sequencing data. The correlation decreased to 0.67 when analysing only CpGs with methylation frequencies ranging from 0.1 to 0.9, due to the lack of enough coverage for ONT. We also examined differentially methylated cytosines identified by each sequencing method. The overlap between the two sets of DMCs and the associated genes was limited, as RRBS library preparation predominantly amplifies gene promoter regions, while ONT covers more intergenic regions. Interestingly, both methods highlighted differentially methylated genes and positions linked to quantitative trait loci of significant economic interest in animal breeding, suggesting promising applications for ONT in the field of agri-genomics.

Characteristics and Kinematics of Bali Bull Sperms after Thawing Using Tris Soy Lecithin

Soy functions as an extracellular cryoprotectant, which can maintain the integrity of spermatozoa cell membranes with its main content is lecithin. Lecithin from soybeans protects sperm cells from cold stress and reduces the effects of oxidative stress during cryopreservation. This study aimed to analyse the effect of various levels of lecithin diluent on the quality of Bali bull semen during cryopreservation. Semen collection of Bali bull was carried out once a week during four times consecutively using an artificial vagina. The semen was then diluted using the essential ingredient Tris Aminomethan with the addition of powdered soy lecithin; P1 (1 %), P2 (3 %) and P3 (5 %), respectively. Andromed® (K1) as a positive control and Tris without soy lecithin (K2) as a negative control. The parameters observed were motility, progressive motility, kinematics, viability, membrane integrity, and acrosome integrity. The results of this study showed that the dilution of semen with soy lecithin before and after thawing the semen quality was not significantly different (P< 0.05) in motility, viability, plasma membrane integrity, and acrosome integrity. Meanwhile, the kinematics of VAP, VCL, VSL, DAP, DCL, and DSL showed that the average quality increased at P3 compared to K1, K2, P1, and P2, which decreased after thawing or were significantly different (P< 0.05). It can be concluded that Bali bull semen diluted with 3% and 5% of tris soy lecithin produces good characteristics and kinematics, can protect spermatozoa from cold shock.

The cryoprotective effect of Guar gum co-supplemented with ethylene glycol and glycerol in Simmental bull semen

Sperm cryopreservation often reduces sperm quality by forming of intra- and extracellular ice crystals. Various compounds widely used to counteract this effect. The guar gum was considered as an extracellular cryoprotective substance. The present study evaluated the impact of the co-supplementation of guar gum with ethylene glycol or glycerol in the cryopreservation of bull sperm. Four ejaculates from 4 bulls were pooled and divided into ten groups consisting of 4 controls (glycerol 6%, ethylene glycol 6%, glycerol 3.5%, and ethylene glycol 3.5%, and six treatment groups including guar gum in 0.001% and 0.002% alone and or co-supplemented either with 3.5% glycerol or 3.5% ethylene glycol and frozen in liquid nitrogen. The sperm motility, viability, plasma membrane and DNA integrity, apoptotic-like changes, antioxidant capacity (TAC), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activities evaluated. The groups contained 3.5% glycerol + 0.001% guar gum, 3.5% ethylene glycol + 0.001% guar gum, and 0.001% guar gum alone showed higher values for live sperm, antioxidant enzymes, membrane integrity, mitochondrial membrane potential (MMP), fertilization, cleavage, and blastocyst rates; and lower values for apoptotic-like changes, H2O2 level, and DNA damage than the control groups. In conclusion, adding guar gum to the bull sperm diluent either alone or combined with glycerol or ethylene glycol ameliorated sperm viability and kinematic parameters and antioxidant capacity while reducing DNA damage and apoptotic-like changes. Guar gum also has improved embryo development. Due to its cost-effectiveness and physicochemical properties, guar gum is a promising supplement for bull sperm cryopreservation.

Laboratory testing and on-site storage are successful at mitigating the risk of release of foot-and-mouth disease virus via production of bull semen in the USA.

Thousands of frozen bovine semen doses are produced daily in the US for domestic use. An incursion of foot-and-mouth disease (FMD) in the country would pose strong challenges to the movements of animals and animal products between premises. Secure supply plans require an estimation of the risk associated with target commodities and the effectiveness of mitigation measures. This study presents the results of a quantitative assessment of the risk of release of FMD virus from five of the largest commercial bull studs in the US via contaminated frozen processed semen. The methodology from a previous study was adapted to better fit the US production system and includes more recent data. Two models were combined, a deterministic compartmental model of FMD transmission and a stochastic risk assessment model. The compartmental model simulated an FMD outbreak within a collection facility, following the introduction of a latent-infected bull. The risk of release was defined as the annual likelihood of releasing at least one frozen semen batch, defined as the total amount of semen collected from a single bull on a given collection day, containing viable FMD virus. A scenario tree was built using nine steps leading from the collection to the release of a contaminated batch from a given facility. The first step, the annual probability of an FMD outbreak in a given facility, was modeled using an empirical distribution fitted to incidence data predicted by five models published between 2012 and 2022. An extra step was added to the previously published risk pathway, to account for routine serological or virological surveillance within facilities. The results showed that the mitigation measures included in the assessment were effective at reducing the risk of release. The median annual risk of release from the five facilities was estimated at less than 2 in 10 billion (1.5 x 10-10) in the scenario including a 30-day storage, routine genome detection assays performed every two weeks and RT-PCR testing of the semen. In this scenario, there was a 95% chance that the risk of release would be lower than 0.00041. This work provides strong support to the industry for improving their response plans to an incursion of FMD virus in the US.

Cryopreservation of Simmental cattle semen with egg yolk from different avian species and level glycerol of different in tris diluent

Cryopreservation of Simmental cattle semen with egg yolk from different avian species and level glycerol of different in tris diluent

Cryopreservation of Simmental cattle semen with egg yolk from different avian species and level glycerol of different in tris diluent

Cryopreservation of Simmental cattle semen with egg yolk from different avian species and level glycerol of different in tris diluent

Effects of hyaluronic acid on sperm parameters, mitochondrial functionand apoptosis of spermatozoa in Simmental bulls with good and poor freezing ability.

Bulls with varying freezability exhibit substantial variation in semen characteristics after cryopreservation. Sperm freezability is positively correlated with membrane cholesterol content, membrane integrity, mitochondrial activityand antioxidant content. The purpose of this study was to determine the optimal concentration of hyaluronic acid (HA) in bull sperm with different cryotolerances. Simmental bulls (n = 10) semen samples were taken and categorized based on their progressive motility (PM) after freeze-thawing: Group I, consisting of bulls (n = 5) with progressive sperm motility ≥45%, was considered good freezability ejaculates (GF), and Group II, including bulls (n = 5) with progressive sperm motility ≤30%, was considered poor freezability ejaculates (PF) bulls. Semen samples were diluted with a Tris-egg-yolk-glycerol (TEYG) extender containing various concentrations of HA: without HA (control), 1 mM HA, 2 mM HAand 4 mM HA. After the freeze-thaw process, sperm kinematics, plasma membrane and acrosome integrity, mitochondrial activityand apoptotic status were evaluated. The addition of 1 mM HA to the diluent of bulls with GF increased PM and linearity (LIN) compared to the control group (p < 0.05). Normal morphology was improved after thawing in the samples treated with 1and 2 mM HA in the GF and PF bullsrespectively. The membrane and acrosome integrity of GF bulls treated with 1 mM HA was significantly (p < 0.05) greater than that of the control groups. Adding 1 mM HA to the extender of bulls with GF and PF improved the proportion of viable cells compared with the highest concentration (4 mM) of HA. The mitochondrial activity of PF bulls treated with 1and 2 mM HA was significantly (p < 0.05) greater than that of the controls and 4 mM HA. Finally, it can be concluded that adding low doses of HA (1 mM) to the TEYG extender of GF and PF bulls ameliorated the post-thaw semen quality.

Influence of the coincubation of post-thawed bull semen with elevated β-hydroxybutyrate concentrations on sperm characteristics.

In this study, the relationships between post-thaw bull sperm characteristics and hyperketonemic conditions after coincubation with cow plasma or media were determined to investigate if such a condition could affect bull sperm characteristics. Two experiments were conducted. In experiment 1, blood samples were collected from 31 cows to prepare plasma. Cows were independently categorized into two groups according to plasma β-hydroxybutyrate (BHB) concentrations (above or below 1.2 mM). Thawed bull semen was diluted and incubated with diluted plasma; motility parameters were evaluated using Computer Assisted Semen Analysis (CASA). In experiment 2, a pooled sample of thawed semen was diluted and divided into three aliquots: without BHB (control) and treated with either 1.2 mM (1.2) or 3 mM (3) BHB. In addition to motility, flow cytometric analyses were carried out. In experiment 1, the overall motility decreased significantly in plasma containing high (≥1.2 mM) BHB compared to plasma containing low (<1.2 mM) BHB. In experiment 2, the overall motility tended to be lower in BHB (3 mM)-supplemented samples. The supplementation of 3 mM BHB increased the proportion of live superoxide-positive sperm and sperm with high mitochondrial potential, while the DNA fragmentation index decreased.

Abnormal centriolar biomarker ratios correlate with unexplained bull artificial insemination subfertility: a pilot study

The mechanisms underlying male infertility are poorly understood. Most mammalian spermatozoa have two centrioles: the typical barrel-shaped proximal centriole (PC) and the atypical fan-like distal centriole (DC) connected to the axoneme (Ax). These structures are essential for fertility. However, the relationship between centriole quality and subfertility (reduced fertility) is not well established. Here, we tested the hypothesis that assessing sperm centriole quality can identify cattle subfertility. By comparing sperm from 25 fertile and 6 subfertile bulls, all with normal semen analyses, we found that unexplained subfertility and lower sire conception rates (pregnancy rate from artificial insemination in cattle) correlate with abnormal centriolar biomarker distribution. Fluorescence-based Ratiometric Analysis of Sperm Centrioles (FRAC) found only four fertile bulls (4/25, 16%) had positive FRAC tests (having one or more mean FRAC ratios outside of the distribution range in a group’s high-quality sperm population), whereas all of the subfertile bulls (6/6, 100%) had positive FRAC tests (P = 0.00008). The most sensitive biomarker was acetylated tubulin, which had a novel labeling pattern between the DC and Ax. These data suggest that FRAC and acetylated tubulin labeling can identify bull subfertility that remains undetected by current methods and may provide insight into a novel mechanism of subfertility.

Investigation of the Effect of Disinfectant and Antioxidant Intrauterine "Anionic Oxygen" Application on Fertility after Artificial Insemination in Dairy Cows

In the study, a total of 30 clinically healthy fertile Holstein cows (aged 3-8 years old; with 500-700 kg average live weight and 28 litres average daily milk) were used in a private dairy farm in Milas district of Muğla province. Randomly selected cows were divided into two groups (control and treatment) (n=15 cows/group) with almost similar body condition scores (BCS) (as all having approximately BCS of 2.75±0.25 units, 1-5 scale). Cows were monitored for natural oestrus, especially during the 2 months postpartum. Control cows (n=15) showing oestrus were inseminated (throughout the 3 consecutive occasions at maximum) by routine AI via recto-vaginal intrauterine route using frozen bull semen (0.25 ml dose; 7.5 x 106 total motile sperm) and thawing at 37 °C for 30 seconds. Inseminations were carried out for 7 months (from December 2021 to June 2022). For the treatment group, the cows were given the antimicrobial agent (UB20 O8®) intrauterine 30 minutes after each AI. The disinfectant dose, given to the corpus uteri region after the AI, was used as a total of 50 ml mixture (40 ml warm saline and 10 ml disinfectant). Throughout the study, animals were routinely given a farm-specific "dairy cow" diet (with 23% protein/concentrated feed). In addition, cows were provided regularly with clean drinking water, shade, and free movement in paddocks. Total pregnancy rates of cows and the 'pregnancy index' (the number of inseminations per pregnancy) of 35-60-day post-insemination as checked via ultrasound examination were analysed statistically by using Chi-Square and Wilcoxon tests. The possible difference between the groups based on pregnancy rate and pregnancy index data was considered significant at the P&amp;lt;0.05 level. According to the results obtained, numerical differences observed due to local treatment after all insemination periods were not statistically significant (P&amp;gt;0.05). Pregnancy rates following the 1st, 2nd, and 3rd inseminations in the treatment and control groups were 60.00% vs. 46.66%, 86.66% vs. 66.66%, and 93.33% vs. 80.00%, respectively. The total pregnancy indexes in the groups were 1.57 (14 pregnancies/22 inseminations; treatment) and 2.25 (12 pregnancies/27 inseminations; control). Briefly, it was observed that the pregnancy rate and pregnancy index in the treatment group were consistently but only numerically higher than those in the control group, indicating that anionic oxygen could be an effective alternative for intrauterine treatment leading to high fertility early postpartum.

The Quality of Cryopreserved Dairy Bulls Semen along Supply-Chain Used for Artificial Insemination in Tanzania

Several factors are said to influence semen quality in breeding bulls. This research was carried out to investigate the effect of season and cold value chains on semen quality parameters. A total of 1652 semen production records from 2018 to 2021 from 10 dairy bulls preserved at the National Artificial Insemination Centre (NAIC), USA River, Arusha, Tanzania, and 540 cryopreserved semen examined were analyzed using Analysis of Variance (ANOVA) and the results were expressed as mean ± standard error of the mean using statistical analysis system (SAS, 2004), to determine whether there were significant differences among variables tested. All semen traits (VOL, SPC, PM, TM, PM, TM, VER, MP, and HP) were significantly (P &lt;0.05) affected by season and cold value chain. During the cold season, the highest values of SPC, PM, TM, PM, TM, VER, MP, and HP were observed as compared to that of the warm season. Furthermore, semen quality decreased gradually from pre-state (before freezing) to post-state (after freezing for 48 hours) with minor affection for semen motility. There was a higher decrease (P &lt;0.05) in quality from cryo-stored semen straw to those of the same batch retrieved back from the field. The semen quality parameters were satisfactory in the first three stages when compared to the semen straw from the field which was observed to have poor motility which could be due to poor handling, leakage of storage and transportation containers, and sometimes the delay of refilling the LN2. Finally, during the cold season and in all stages of semen production the semen is qualitatively and quantitatively good.