EFFECT OF BRADYKININ ON FREEZABILITY OF ABATTOIR DERIVED MURRAH BULL EPIDIDYMAL SPERMATOZOA

Abnormal sperm count and motility on semen analysis are not sufficiently predictive of abnormal Kruger morphology

Effect of graphene oxide as cryoprotectant on post-thaw sperm functional and kinetic parameters of cross bred (HF X Sahiwal) and Murrah buffalo (Bubalus bubalis) bulls

Background and Aim:Various factors can reduce the quality of semen used for artificial insemination and have an impact on fertility decline, such as poor handling during frozen semen distribution. This study was aimed at assessing the quality of frozen-thawed semen after distribution in the field and its importance in maintaining fertility.Materials and Methods:The Brahman Cross (BX) breeding program of PT Lembu Jantan Perkasa, Indonesia, was used. This program was preferred due to its adherence to guidelines that limit the effects of extraneous factors that may affect semen quality. Frozen-thawed semen samples from eight bulls with the same production code were analyzed and compared between the production site (artificial insemination [AI] center) and the field (BX breeding program). Total and progressive motility (PM) of sperm were determined using computer-assisted semen analysis. Plasma membrane integrity (PMI) was assessed using hypoosmotic swelling test, sperm viability using Eosin-Nigrosin staining, acrosome integrity using trypan blue-Giemsa staining, morphological abnormalities using William staining, and DNA fragmentation using toluidine blue staining. The fertility rate was determined using the conception rate (%) derived from AI data based on 502 AI services and 478 cows in the BX breeding program. A t-test was used to compare the quality of frozen-thawed semen before and after distribution. The relationship between the qualities of frozen semen after distribution in the field with fertility was analyzed using Pearson correlation.Results:There was no significant difference (p>0.05) in the quality of frozen-thawed semen (sperm motility, PMI, viability, acrosome integrity, abnormalities, and DNA fragmentation) between the production site (AI center) and after distribution in the field (BX breeding program). The semen met the minimum standards for AI programs. Total motility (r=0.986), PM (r=0.961), sperm viability (r=0.971), PMI (r=0.986), and acrosome integrity (r=0.992) were all positively correlated (p<0.05) with fertility rate; while sperm abnormalities (r=−0.996) and sperm DNA fragmentation (r=0.975) were negatively correlated (p<0.05) with fertility rate.Conclusion:The study showed that to achieve the maximal and optimal fertility rate in bulls in an AI program, the overall quality of frozen-thawed semen in all aspects is critical. This can be achieved if the handling during distribution and storage, as well as the various factors that may affect the quality of semen in the field, can be controlled properly.

Buffalo semen collected from Murrah bull were cryopreserved and evaluated for different motility parameter, kinematics and plasma membrane integrity. Buffalo bulls were maintained uniform standard management and nutritional practices. Semen was collected regularly twice a week semen collection schedule from four (04) Murrah bull. Collected semen was immediately transported to laboratory and evaluated for different macroscopic parameter (color, volume and thickness). Fresh semen was then diluted with saline solution and evaluated for sperm concentration, motility, sperm kinematics and morphology. Semen samples that fill all the standard were selected for freezing and diluted with Tris-egg yolk citrate diluter. Diluted semen was equilibrated, cryopreserved and finally evaluated for post thaw sperm quality. Different motility parameter (total, progressive, static and slow motility) varied significantly (p&lt;0.01) irrespective of different freezing stages. Significantly higher progressive sperm motility and viability of buffalo spermatozoa were observed at fresh semen whereas lower progressive sperm motility and viability was found at post thaw stage. Total and progressive motility reduced by 2.5 and 2.12% following equilibration, whereas following cryopreservation, total and progressive motility reduced by 35.7 and 28.51% and static motility increases accordingly (35.4%). Significantly higher plasma membrane integrity of sperm was observed at fresh semen followed by pre freeze and post thaw semen. Following freezing, integrity of plasma membrane reduces at the rate of 10.81% and 26.7% at pre freezing and post thaw stages. Significantly higher average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were found for fresh semen followed by pre-freeze and post-thaw semen. Frozen buffalo semen with higher progressive motility and motion characteristics may be produced if motility losses can be reduced during freezing stage as this stage results higher motility losses. Asian Australas. J. Biosci. Biotechnol. 2022, 7(2), 75-81

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

Simple SummaryThis study investigates how carvacrol and thymol affect the quality of semen from Beni Arouss bucks stored in skim milk at 4 °C. Semen from eight bucks was collected weekly for 11 weeks, pooled, and divided into three groups: one diluted in skim milk, and the others diluted in skim milk supplemented with 200 µM of carvacrol and thymol. Sperm motility, viability, abnormalities, membrane integrity, lipid damage, and bacterial growth were assessed during 48 h of storage at 4 °C. After 48 h, carvacrol improved sperm motility, viability, and reduced bacterial growth and lipid damage. Thymol showed similar benefits but did not enhance progressive motility. These beneficial effects are due to the antimicrobial properties of these two compounds, offering potential benefits for livestock breeding.This study aims to investigate the impact of carvacrol and thymol on the quality of Beni Arouss buck semen stored in skim milk at 4 °C. Ejaculates were collected from eight Beni Arouss bucks weekly for 11 weeks, pooled, and then divided into three equal parts. Samples were diluted to 400 × 106 sperm/mL in skim milk (control) and skim milk supplemented with a single dose of 200 µM carvacrol and thymol each. Evaluations of sperm motility, viability, abnormalities, membrane integrity, lipid peroxidation, and bacterial growth were conducted at 0, 6, 24, and 48 h of liquid storage at 4 °C. After 48 h of storage, the results indicate that the addition of carvacrol positively influences total and progressive motility and viability. However, it also leads to a decrease in lipid peroxidation and bacterial growth compared to the control group (p < 0.05). Thymol showed similar results to carvacrol, except for progressive motility (p > 0.05). Bacterial growth was negatively correlated with total and progressive motility and viability (p < 0.05), while no correlation between lipid peroxidation and these parameters was observed (p > 0.05). In conclusion, the addition of carvacrol and thymol to skim milk extender moderately improves the quality of Beni Arouss buck semen after 48 h storage at 4 °C due to its antimicrobial activity.

Testicular temperature gradient and vascular perfusion as predictors of semen quality in summer-stressed Murrah bulls.

The aim of this study was to investigate the relationship between the levels of insulin-like growth factor-1 (IGF-1) in serum and seminal plasma and the characteristics of semen in Beetal bucks (Capra hircus). A total of 12 adult Beetal bucks were involved in the study, with each buck providing six ejaculates collected using a standard artificial vagina (n = 72 total). Only qualified semen samples (volume of 0.7 mL, a mass motility rating of 3+ or higher on a 0-+ scale, and individual progressive motility of 80% or more) divided into three fractions were processed for estimation of IGF-1 and other seminal parameters like motility, viability, acrosome integrity, sperm abnormality and superoxide dismutase (SOD) activity. The first and second fraction were diluted and extended with Optixcell extender (1:15 ratio). The first ejaculate fraction was processed for studying fresh semen parameters and the second fraction was cryopreserved for evaluating frozen semen parameters. French mini straws (0.25 mL) were used for semen filling, and polyvinyl alcohol powder of different colours was used for sealing the extended semen. The third fraction of each ejaculate was centrifuged at room temperature (1100 × g for 7 min) to separate the seminal plasma. Additionally, blood samples were taken from each buck on the same day as semen collection, resulting in a total of 36 blood samples. The results revealed a significant positive correlation (r = .4243; p < .05) between the concentration of IGF-1 in both serum and seminal plasma of the Beetal bucks. Furthermore, the concentration of IGF-1 in serum showed significant positive correlations with sperm viability (r = .554; p < .05), acrosome integrity (r = .527; p < .05), post-thaw sperm motility (r = .407; p < .01), post-thaw sperm viability (r = .426; p < .01) and post-thaw acrosome integrity (r = .333; p < .05). However, it had a significant negative correlation with SOD activity in fresh semen (r = -0.458; p < .01). Moreover, the concentration of IGF-1 in seminal plasma demonstrated significant positive correlations with individual progressive motility (r = .341; p < .05), sperm viability (r = .527; p < .05), acrosome integrity (r = .539; p < .05), sperm plasma membrane integrity (r = .464; p < .05), post-thaw sperm motility (r = .644; p < .01), post-thaw sperm viability (r = .643; p < .01), post-thaw acrosome integrity (r = .487; p < .01) and post-thaw sperm plasma membrane integrity (r = .521; p < .01). Additionally, it showed a significant negative correlation with SOD activity in both fresh semen (r = -0.714; p < .01) and frozen semen (r = -0.558; p < .01) of Beetal bucks. Based on these findings, IGF-1 in seminal plasma can be considered as a potential biomarker for the selection of bucks for breeding purposes.

Semen preparation methods and sperm apoptosis: swim-up versus gradient-density centrifugation technique

Aim: The present study was carried out to examine the relationships among frozen-thawed semen fertility, physical parameters, seminal quality, and testosterone concentration in Murrah buffalo bulls. Materials and Methods: A total of 30 breeding Murrah buffalo bulls (either progeny tested or under progeny testing program) were randomly selected from two government bull farms in Punjab. None of the bulls selected for this study had any preceding physical abnormality. A field fertility trial was conducted to determine the first service conception rate (FSCR). The number of females inseminated per bull semen was 10. All the bulls were inspected for structural soundness, measurement of scrotal circumference, testicular biometry, and internal pelvic area (IPA). Frozen-thawed semen was evaluated for total motility, progressive motility, viability, concentration, abnormality, and hypo-osmotic swelling test (HOST). Testosterone was estimated in blood plasma, seminal plasma as well as frozen-thawed semen extracts for establishing relationship. Results: The FSCR was 48% in the bulls having a scrotal circumference of ≥44 cm, although, there was no significant correlation between FSCR and scrotal circumference. Similarly, no consistent relationship existed between sperm concentration and scrotal circumference. A positive correlation was observed between IPA and FSCR (r=0.294). Of the six post-thaw seminal components (total motility, progressive motility, viability, HOST (%), total abnormality and concentration) only total motility had a high significant (p<0.01) correlation with FSCR (r=0.694). Varied correlations existed between other seminal parameters and fertility. Using a simple regression analysis, the post-thaw motility, IPA, prepuce length and testosterone (independent variables) combined to explain approximately 62% of the variation in the FSCR (dependent variable). Conclusion: The present study indicated that despite low to high correlations between seminal characteristics, physical parameters, fertility, and testosterone; the observations support the importance of these components and their function in maintaining semen quality and subsequent fertility.

The development of biometric techniques in domestic animals has greatly advanced scientific practices in wildlife research. The association between seminal characteristics and body and testicular biometry enables the selection of suitable breeders, though appropriate measurement techniques are required. The present study assessed differences among conventional methods and formulas for estimating testicular parameters. Testicular length, width, and thickness were measured using three methods in 13 adult male domestic cats. Testicular area, volume, and weight were estimated, from which the gonadosomatic index (GSI) was calculated. Sperm were collected using an alpha-2 adrenergic agonist and urethral catheterization, and characterized in terms of volume, vigor, total motility, progressive motility, concentration, plasma membrane integrity, and morphology. The three methods were consistent in terms of testicular area, volume, weight, and GSI. Moderate positive correlations were observed for testicular weight (r = 0.61, p < 0.05) and GSI (r = 0.58, p < 0.05). Testicular parameters showed strong positive correlations among each other (r > 0.80, p < 0.05). We observed a moderate positive correlation between head length and progressive motility (r = 0.65, p < 0.05). In conclusion, all testicular measurement and estimation techniques showed comparable performance. Therefore, testicular biometry is useful for selecting breeding males in feline conservation programs, wherein larger body biometrics are related to improved seminal and reproductive parameters.

The clinical content of preconception care: preconception care for men

Recovery of normal testicular temperature after scrotal heat stress in rams assessed by infrared thermography and its effects on seminal characteristics and testosterone blood serum concentration

This study investigated the impact of equilibration time on the quality of frozen-thawed semen in Pesisir bulls, focusing on post-thaw motility, viability, abnormalities, and intact plasma membrane.Semen samples were equilibrated for 2, 4, 6, 8, and 10 hours before freezing.Fresh semen had an average volume of 3.82mL, a sperm concentration of 762.510 6 /mL, and motility of 73.75%.Post-thaw results showed that motility decreased with longer equilibration times, from 68.17% at 2 hours to 58.96% at 10 hours.Viability remained relatively stable, decreasing slightly from 73.38% at 2 hours to 68.88% at 10 hours.Abnormality rates improved, decreasing from 23.17% at 2 hours to 19.75% at 10 hours.The percentage of intact plasma membranes varied, peaking at 8 hours (54.38%) and declining to 46.29% at 10 hours.Correlation analysis revealed the following relationships: Post-Thawing Motility: =64.25+2.501X-0.304X 2 (R=0.987);Post-Thawing Viability: =74.13+0.036X-0.057X 2 (R=0.767);Post-Thawing Abnormality: =26.00-1.486X+0.084X 2 (R=0.963);Post-Thawing Intact Plasma Membrane: =47.03+1.487.X-0.140X 2 (R=0.135).These findings suggested that while longer equilibration times improved certain parameters, such as abnormalities, they adversely affected motility and membrane integrity.Identifying the optimal equilibration time was crucial for balancing these effects and enhancing the overall quality of frozen-thawed Pesisir bull semen for artificial insemination programs.

&#x0D; &#x0D; &#x0D; &#x0D; Purpose: To investigate the in vitro effects of methanol leaf extract of E. hirta (MLEEH) on the motility, viability and morphology of caprine spermatozoa.&#x0D; Methods: The effect of MLEEH treatment (1.25, 2.5, 5, 10 and 20 mg per mL) on caprine sperm percentage total and progressive motility, viability and total abnormalities were evaluated at 1, 5 and 10 min post-treatment. Sperm revival test was used to evaluate the reversibility of sperm incapacitation following MLEEH treatment.&#x0D; Results: There were significant interactions (p &lt; 0.001) between the effects of MLEEH concentration and the duration of treatment on sperm total motility, progressive motility and viability. Increase in MLEEH concentration and the duration of treatment caused significant decreases (p &lt; 0.05) in sperm total motility, progressive motility and viability, whereas sperm morphology was not altered. Washing and supplementation of MLEEH-treated sperm failed to revive sperm motility.&#x0D; Conclusion: E. hirta treatment causes concentration-dependent and time-dependent decreases in total and progressive sperm motility and sperm viability, as well as irreversible immobilization of spermatozoa. These findings suggest possible adverse effects of E. hirta on the fertility of males. Thus, the extract can be potentially developed as an antifertility or contraceptive agent.&#x0D; &#x0D; &#x0D; &#x0D;