Relationship of motility and acrosome reaction on sexing sperm in Ongole Crossbred bull

Artificial insemination using sexing semen is expected to produce calves with the desired sex. One sexing sperm method is the percoll density gradient centrifugation method. This study aimed to determine the changes and the relationship between motility and acrosome reaction after sexing process using percoll density gradient centrifugation. The material used was semen of ±5 years old Ongole crossbred bull with a bodyweight of ±700 kg as many as three bulls with mass motility 2+ and individual motility 70%. The method used was to compare fresh semen with sexed semen after the cooling process. Parameters measured were motility characters using CASA analysis, which included motility parameters, progressive motility, capacitation, and no acrosome reaction. Statistical analysis used paired T-test to distinguish among fresh semen, after sexing and cooling process. In comparison, regression and correlation were used to analyze the relationship of capacitation and hyperactivation sperm with no acrosomal reaction with motility and progressive motility. The results showed that motility and progressive motility decreased after the sexing and cooling process. Meanwhile, the acrosomal reaction, capacitation, and hyperactivity increased.

USE OF CIDR AND ESTRADIOL CYPIONATE IN SYNCHRONIZATION PROTOCOLS ON ESTRUS PRESENTATION, PREGNANCY AND BIRTH RATE IN CREOLE SHEEP UNDER HIGH ALTITUDE CONDITIONS

The objective of this research was to evaluate the use of the intravaginal device (CIDR) and estradiol cypionate in synchronization protocols, on oestrus presentation, pregnancy and birth rate in Creole sheep under high altitude conditions. The study was carried out in a non-reproductive season, in the peruvian highlands, using 57 ewes. Four experimental groups were formed: group T1 (n=14) and T3 (n=14), CIDR progesterone device for 7 days and 12 days, respectively; group T2 (n=15) and T4 (n=14), CIDR progesterone device for 7 days and 12 days plus 1 mg of estradiol cypionate, 24 hours after removal of the device in both groups, respectively. Estrous presentation was observed from CIDR removal to 48 hours using vasectomized rams and IATF was performed with fresh semen 48 hours after device removal. The pregnancy rate was determined by ultrasonography at 46 and 90 days after FTAI and the birth rate was recorded. The chi-square test was used for statistical analysis. There was no difference (P>0.05) between groups, for estrous presentation, but there was difference (P<0.05) for pregnancy rate at 46 days between groups with: T1 (42.9%), T3 (38.5%), T4 (21.4%), with respect to T2 (0%) (The difference in results are shown in Table 2). There was a difference (P<0.05) for the pregnancy rate at 90 days: T1 (35.7%) and T3 (30.8%), with respect to T4 (7.1%) and T2 (0 %). The birth rate showed differences (P<0.05) for T1 (28.6%) and T3 (28.6%), with respect to T2 (0%) and T4 (0%). In conclusion, a high pregnancy rate was obtained with CIDR for 7 and 12 days, compared to the use of CIDR plus estradiol cypionate. However, no births were obtained with progesterone plus estradiol cypionate treatment.

Comparison of seminal characteristics of Aberdeen Angus, Holstein and Nelore bulls before and after cryopreservation

The aim of this study was to evaluate parameters indicative of sperm quality of fresh and post-thawed semen of Aberdeen Angus, Holstein and Nelore bulls. Thirty-nine bulls were used: Aberdeen Angus (n=13), Holstein (n=13) and Nelore (n=13). The ejaculate collects were performed twice a week using artificial vagina, totaling 792 semen collections, 307 for Aberdeen Angus, 225 for Holstein and 260 for Nelore bulls. After collection, fresh semen was evaluated and semen freezing was performed. After freezing, the batches were thawed and progressive motility was determined. The analysis of fresh semen showed that there was no difference (P = 0.053) between the Aberdeen Angus and Nelore breeds, while ejaculates from Holstein bulls showed a statistical difference (P = 0.024). As well, a difference (P<0.001) was identified in the sperm concentration of the three breeds. In the samples evaluated after thawing, a statistical difference was observed between Holstein and Nelore breeds (P<0.001), while the values of the Angus breed were similar to the other two breeds. The difference in motility of fresh and post-thawing semen showed that Nelore and Angus bulls showed greater variation in values between the analyzes (26.0±8.9% and 25.3±8.4%, respectively) showing a significant difference (P<0.001) in relation to Holstein bulls (20.6±9.3%) that obtained the smallest difference. The analysis of fresh and post-thawing semen did not show any significant difference (P=0.13) between breeds. In conclusion, the semen cryopreservation process causes a decrease in the physical parameters of the semen and these quality losses suffer interference according to the breeds.

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

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

Composition of Stallion Seminal Plasma and Its Impact on Oxidative Stress Markers and Spermatozoa Quality

The composition of seminal plasma of individual sires varies and so does the fertilizing ability. Micro and macro elements along with seminal enzymes, hormones, proteins, and lipids contained in seminal plasma are essential for the proper physiological function of spermatozoa. However, elevated levels against the normal physiological values, especially in the case of trace metals, result in the production of reactive oxygen species. The deficiency of antioxidants in the seminal plasma that could scavenge free radicals causes an impairment of spermatozoa quality. Ejaculates were obtained from 19 stallions. The fresh semen was analyzed to evaluate qualitative parameters of spermatozoa in terms of the motility, viability, and integrity of DNA. Separated seminal plasma underwent the assessment of the chemical and biochemical composition and RedOx markers. Based on the obtained concentrations of individual chemical elements, the correlation analysis suggested a negative impact of Cu in seminal plasma on the SOD, GPx, and LPO. Contrary, positive correlation was detected between FRAP and motility features. While Cu negatively correlated with sperm motion parameters, the adverse effect on viability was suggested for Cd. Our data suggest that seminal plasma has a potential due to its availability to become the potential biomarker of the reproductive health of farm animals.

Factors Affecting Fresh Semen Quality in Pasundan Cattle at UPTD BPPIBTSP Ciamis

Background: Pasundan cattle are local cattle native to Indonesia. One way to conserve beef cattle genetics is to use Artificial Insemination technology. The success of Artificial Insemination can be influenced by the quality of semen. Purpose: To determine factors affecting fresh semen quality in Pasundan cattle at UPTD BPPIBTSP Ciamis. Methods: The data were obtained through observations on seven Pasundan bulls in March 2021 towards fresh semen quality and some factors influencing it. The Pasundan bulls observed were seven productive males. Results: The fresh semen quality of Pasundan cattle, such as volume, color, and pH, showed good result,s but the average consistency and concentration of spermatozoa were still below the standard. The factors that can affect the fresh semen quality are the breed of beef cattle, age, body weight, feed, season, exercise, and frequency of semen storage. Conclusion: The determining factors that can cause the consistency and concentration of Pasundan cattle’s spermatozoa at UPTD BPPIBTSP Ciamis are feed and season.

Total Aseptization of Boar Semen, to Increase the Biosecurity of Reproduction in Swine

The aim of the study was to establish the complete microbiological profile of boar semen (Sus scrofa domesticus) and to choose the most effective antiseptic measures in order to control and optimize AI reproduction in pig farms. One hundred and one semen samples were collected and analyzed from several pig farms. The microbiological profile of ejaculates was determined by evaluating the degree of contamination of fresh semen and after dilution with specific extenders. The bacterial and fungal load of fresh boar semen recorded an average value of 82.41/0.149 × 103 CFU/mL, while after diluting the ejaculates the contamination value was 0.354/0.140 × 103 CFU/mL. Twenty-four germs (15 bacterial and 9 fungal species) were isolated, the most common being Candida parapsilosis/sake (92%) and Escherichia coli (81.2%). Modification of the sperm collection protocol (HPBC) reduced contamination in raw sperm by 49.85% in bacteria (significant (p < 0.00001) and by 9.67% in fungi (non-significant (p < 0.111491). The load in bacteria and filamentous fungi can be controllable, but not in levuras fungi. Some fluconazole-added extenders (12.5 mg%), ensure fungal aseptization, and even an increase in sperm progressivity (8.39%) for at least a 12 h shelf life after dilution. Validation of sperm aseptization was done by maintaining sow fecundity unchanged after AI (insignificant p > 0.05).

Search for Associations of FSHR, INHA, INHAB, PRL, TNP2 and SPEF2 Genes Polymorphisms with Semen Quality in Russian Holstein Bulls (Pilot Study)

The aim of the study was to search for new mutations in the previously studied gene loci of follicle-stimulating hormone receptor (FSHR), inhibin α (INHA), inhibin β A (INHAB), prolactin (PRL), transition protein 2 (TNP2), and sperm flagella 2 (SPEF2) by sequencing, as well as the search for associations of previously identified mutations at these loci with fresh semen quality in Russian Holstein bulls. Phenotypic data from 189 bulls was collected. Data was analyzed for most bulls for three years of semen collection. The maximum value of each semen quality indicator (doublet ejaculate volume, sperm concentration, progressive motility and total number of spermatozoa) were selected. SNPs were identified in the FSHR, INHA, INHAB, TNP2, SPEF2 genes. The PRL gene did not have polymorphism. Significant (p < 0.05) associations of polymorphisms in the FSHR gene with double ejaculate volume, concentration and total number of spermatozoa were identified. Polymorphism in the INHA gene was significantly associated (p < 0.05) with sperm concentration. Polymorphism in the INHAB gene was significantly associated (p < 0.05) with doublet ejaculate volume and total number of spermatozoa. Polymorphisms in the TNP2 and SPEF2 genes did not have significant associations with semen quality. The SNPs studied in our pilot work may be considered as candidate genetic markers in the selection of bulls.

Effects of Three Semen Extenders, Breeding Season Month and Freezing–Thawing Cycle on Spermatozoa Preservation of Portuguese Merino Sheep

This study aimed to evaluate and compare the effect of three semen extenders (S-EXT) on 22 spermatozoa (SPZ) parameters (subjective and computer-assisted sperm analysis evaluations), before and after semen cryopreservation throughout different months of the breeding season in the Portuguese Merino breed. According to the multivariable model, the SPZ viability (alive %), kinetics subjective individual motility, total motility, total progressive motility and its subpopulations, and beat cross frequency) were higher in the egg yolk-based S-EXT improved by Estação Zootécnica National (Portugal) than in Ovixcell® or Andromed® extenders. All the differences were only observed in thawed semen, except for total motility and total progressive motility, in which Ovixcell® also showed the poorest results on fresh semen. An interaction effect between S-EXT and semen processing was observed on 72.3% (17/22) of the evaluated parameters, evidencing a variable cryoprotective action between S-EXT. The SPZ viability was poorer in the onset of the breeding season (end of April/early May) than in the previous middle breeding season (November/early December), suggesting the influence of a short anoestrous season on ejaculate quality, even though the volume and SPZ concentration of the ejaculates remained stable throughout the experiment. Additionally, S-EXT x semen processing x month interaction effect on 59.1% (13/22) of the evaluated parameters evidenced the importance of SPZ time collection in a natural environment to cryopreserve ram’s semen. We concluded that, overall, the egg yolk-based S-EXT provided a greater value to the cryopreservation of Merino rams´ semen. Nevertheless, the causes of the interaction effect between S-EXT, semen processing and/or month on several SPZ parameters should be addressed, including SPZ molecular research in new studies, in order to improve egg yolk-based as well as in egg yolk-free-based S-EXT.