Quality of chilled indigenous ram semen using skim milk-based extenders

 Abstract : This research aimed to know the potential of ethanol extract of roselle fruits in improving the quantity and quality of sperm in mice induced nicotine. The method is implemented with the experimental method that uses a completely randomized design 30 male mice. This study consists of five treatments and six replications. Treatment for the provision of nicotine with a dose of 0,07mg / 10 g body weight and roselle fruit ethanol extract at a dose consisting of 0 mg / 10 g BB, 3 mg / 10 g BB, 6 mg / 10 g BB, and 9 mg / 10 g BB. Roselle fruit extract given by gavage and nicotine via subcutaneous injection for 35 days. Parameters measured were the number, viability, normal morphology, motility, and sperm plasma membrane integrity. The data were analyzed by analysis of variance and real distance difference test (BJND). The average number of sperm in mice P1, P2, P3, and P4 is 103.17 x 106 / ml, 153 x 106 / ml, 157 x 106 / ml, 146 x 106 / ml. The percentage of sperm viability in mice P1, P2, P3, and P4 is 47%, 60.67%, 70%, 68.67%. The percentage of normal sperm morphology P1, P2, P3, and P4 is 42.5%, 59.5%, 73.67%, 62.83%. The percentage of sperm motility in mice P1, P2, P3, and P4 is 41.3%, 61%, 63.7%, 59%. The percentage of plasma membrane integrity mice P1, P2, P3, and P4 is 42.4%, 70.4%, 71%, 62.9%. The results showed that ethanol extract of roselle fruits at a dose of 3 mg / 10 g BB maximal increase the number, the percentage of normal morphology, the percentage motility, sperm plasma membrane integrity mice and 6 mg / 10 g BB increased the percentage of sperm viability nicotine-induced mice. Results of this study are expected to be additional material on learning Biology Semester II Class X Basic Competencies 3.3 Describe the characteristics Divisio and its Role in the World of Plants for Survival on Earth. Keywords : quantity and quality of mice sperm, roselle, nicotine

The objective of this study was to assess the association of exposure to metal mixtures with semen quality and sperm DNA integrity of coke oven workers (n = 96). Urinary six metals (cadmium, lead, arsenic, zinc, selenium, and copper) were quantified using inductively coupled-mass spectrometry. Semen quality parameters included sperm concentration, sperm concentration, sperm motility, sperm morphology, and sperm viability. Sperm DNA fragmentation and 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodGuo) adducts served as biomarkers for assessing sperm DNA integrity. Bayesian kernel machine regression with the hierarchical variable selection process was used for analyzing both individual and joint effects of the metal mixture on the outcomes of semen samples, while adjusting for age, smoking, alcohol consumption, job length, and body mass index. The metal mixture was associated with reduced sperm concentration, motility, viability, and normal morphology. It was novel that a dose-response relationship was observed between exposure of the metal mixture and semen quality. Among the metals tested, cadmium had a reverse relationship with sperm motility, viability, and normal morphology, and a non-linear relationship with sperm viability and sperm motility. The metal mixture and individual metals were not associated with sperm DNA fragmentation and 8-oxodGuo. In conclusion, exposure to metal mixtures and cadmium may exert an association with semen quality and had no association with sperm DNA breakages.

A cross-sectional study was conducted to determine the semen parameters (i.e. volume, concentration, motility, viability and normal morphology) of proven fertile males in Singapore and compare it with the World Health Organization (WHO) recommended normal values and to examine some factors that may affect spermatogenesis. A total of 243 men, whose wives were pregnant at the time of collection of semen, provided a semen sample each after sexual abstinence for 3 days. A questionnaire was used to elicit occupational exposure, alcoholic consumption, smoking history and past significant medical history. Most subjects had normal sperm volume (56.4%), concentration (79.8%), motility (69.5%) and viability (53.5%) based on WHO criteria. However, fertile men had a low mean percentage of normal sperm morphology (20.0%), although they were normally distributed. Cigarette smoking was associated with significantly lower semen volumes even after adjusting for alcohol consumption. The sperm parameters (i.e. volume, density, motility, viability and normal morphology) were not significantly associated with ethnic differences. The WHO criterion for normal sperm morphology is too stringent, and should be adopted with caution. Normal sperm morphology is but one of many parameters for assessment of fertility. Social alcohol consumption, cigarette smoking, and 'recent fever' did not appear to affect sperm quality in this group of fertile men.

Should a semen analysis be ordered in a man with history of previous fertility?

Colloidal centrifugation with Androcoll-E™ prolongs stallion sperm motility, viability and chromatin integrity

BackgroundThe pathogenesis of teratozoospermia (<4% morphologically normal sperm cells) and the relationship between sperm morphological abnormalities and abnormal sperm nuclear DNA fragmentation, which are considered indicators of male fertility, have not been elucidated. Our research was designed to determine the prevalence of different sperm DNA fragmentation (SDF) levels in men with teratozoospermia and to establish a discriminating threshold value for SDF in assessing sperm morphology.MethodsBasic semen characteristics and detailed sperm morphological analysis (head, neck, midpiece, and tail defects and excess residual cytoplasm) (WHO, 2010), and the nuclear sperm DNA dispersion test were performed on semen samples obtained from 523 men with teratozoospermia (n=296) and those without teratozoospermia (n=227).ResultsSubjects with abnormal sperm morphology had not only lower results for standard sperm characteristics, including detailed sperm morphological abnormalities, but also a higher proportion of sperm cells with SDF vs. men with normal sperm morphology. Moreover, significantly fewer subjects with low SDF levels (≤15%), more subjects with high SDF levels (>30%) and a higher odds ratio (OR) for having high SDF levels were found in the group of men with teratozoospermia vs. men without teratozoospermia. However, the receiving operating characteristic (ROC) curve analysis indicated that a SDF >18% was a significant negative predictive value to distinguish between men with normal sperm morphology or men with abnormal sperm morphology. The optimal area under the ROC curve (AUC) was 0.746. In the group of men with teratozoospermia, a higher incidence of men with >18% SDF and a higher OR for having >18% SDF were observed. SDF negatively correlated with sperm number, morphologically normal sperm cells, sperm motility and sperm vitality but positively correlated with the teratozoospermia index (TZI) and detailed sperm morphological abnormalities.ConclusionsThe obtained findings demonstrated that: (I) detailed sperm structural defects coexist with abnormal nuclear sperm DNA dispersion, (II) men with teratozoospermia may have a higher risk for sperm DNA damage, (III) the calculated optimal SDF value of 18% measured by the DNA sperm dispersion test is the best criterion to predict normal and abnormal sperm morphology.

The effects of adding mixed chicken and Japanese quail egg yolks (EYs) to the cryodiluent on the quality of ram semen before freezing and post-thawing were evaluated. Additionally, the composition of chicken and quail egg EYs and their mixture were analyzed for results explanation. The semen was collected from rams (n = 5) and extended with cryodiluent containing the EY of chicken, quail or their mixture (1:1). The extended semen was chilled slowly to 5 °C within 2 h and equilibrated for 2 h, before frozen on the liquid nitrogen vapor and cryopreserved at −196 °C. The straws were evaluated before freezing and post-thawing for sperm motility, vitality and abnormality besides plasma-membrane and DNA integrities. The moisture, ash, protein, and fatty acid (FA) contents of chicken EY, quail EY and their mixture were analyzed. Sperm vitality, plasma membrane integrity and DNA integrity before freezing were significantly (P < 0.05) higher in quail EY than chicken EY and mixed EYs cryodiluent. The chicken EY extender significantly improved the vitality, plasma membrane and DNA integrities of post-thawed ram semen in comparison with quail EY or mixed EYs extenders. While, the post-thawing sperm abnormalities was lower (P ≤ 0.05) in quail EY than chicken EY and mixed EYs cryodiluent. The post-thawing sperm motion kinetics parameters were higher in quail EY than chicken EY and mixed EYs cryodiluent. The highest percentages of moisture, ash, saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs) were detected in quail EY had. While, the highest percentages of fat, protein and polyunsaturated fatty acids (PUFAs) were detected in chicken EY. In conclusion, using of chicken EY can improve total motility, vitality, plasma membrane integrity and DNA integrity of cryopreserved ram semen. While, using of quail EY can improve sperm abnormalities and kinetic motion parameters of cryopreserved ram semen. Mixing chicken and quail EYs added no value for post-thawing ram semen parameters.

Background Data on the effect of obesity on seminal fluid and men fertility are inconsistent. The aim of this study was to evaluate the impact of obesity on semen characteristics in infertile men. Patients and methods A cross-sectional study was conducted on seventy-four infertile men who met inclusion criteria. Semen sample were collected and sperm concentration, progressive motility, total motility and normal sperm morphology were assessed in accordance with WHO 2010 criteria. For each patient weight and height were measure and patients were divided by BMI into normal weight (BMI: 18.5–24.9 kg/m2, n = 30), overweight (BMI: 25–29.9 kg/m2, n = 30) and obese (BMI: ≥30 kg/m2, n = 14). Seminal fluid parameters were compared among the three groups. Results Sperm concentration was lower in obese men but it did not differ significantly from those of normal weight and overweight infertile men (25.71 ± 22.16, 34.33 ± 31.11, 36.07 ± 31.24 and million/ml respectively, P > 0.05). Sperm progressive motility, total motility and normal sperm morphology also were not significantly different among the three groups. Conclusion Our findings suggest that obesity may have no influence on sperm concentration, motility and normal morphology in infertile men.

Data on the effect of obesity on seminal fluid and men fertility are inconsistent. The aim of this study was to evaluate the impact of body mass index (BMI) on semen characteristics in infertile men.A cross-sectional study was conducted on seventy-four infertile men who met inclusion criteria.Semen sample were collected and sperm concentration,progressive motility,total motility and normal sperm morphology were assessed in accordance with WHO 2010 criteria. For each patient weight and height were measure and patients were divided by BMI into normal weight (BMI: 18.5–24.9 kg/m2,n=30),overweight (BMI: 25–29.9 kg/m2,n=30) and obese (BMI:≥30 kg/m2, n=14). Seminal fluid parameters were compared among the three groups.Sperm concentration was lower in obese men but it did not differ significantly from those of normal weight and overweight infertile men (25.71±22.16,34.33±31.11,36.07±31.24 and million/ml respectively,P&gt;0.05). Sperm progressive motility,total motility and normal sperm morphology also were not significantly different among normal weight,overweight and obese infertile men.Our findings suggest that BMI may have no influence on sperm concentration,motility and normal morphology in infertile men.

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

Evaluation of Tris–citric acid, skim milk and sodium citrate extenders for liquid storage of Punjab Urial (Ovis vignei punjabiensis) spermatozoa

2033 GENOME-WIDE ASSOCIATION STUDY IDENTIFIES CANDIDATE GENES FOR ABNORMAL SPERM MORPHOLOGY IN TWO COHORTS OF MEN

Sperm banking and AI could benefit conservation of endangered African wild dogs (AWD). However, it is not clear whether their strict dominance hierarchy causes subfertility in subdominant males that typically do not breed. Our study investigated the effect of dominance on male reproductive parameters, including faecal glucocorticoids (fGCM) and androgens (fAM), testis and prostate volume, preputial gland size, semen collection success, and the number, motility, morphology, viability, acrosome integrity (PSA-FITC), and DNA integrity (TUNEL) of spermatozoa collected by electroejaculation. Samples were obtained from n = 12 captive AWD (4 US packs) in the pre-breeding season and n = 28 captive AWD (n = 11 from 4 US packs; n = 17 from 3 Namibian packs) in the breeding season. Male hierarchy was clearly determined by behavioural observations in all but 1 Namibian pack. Data were grouped by dominance status and means were compared by ANOVA or t-test; P ≤ 0.05 was significant. In the pre-breeding season, there was no significant difference in body weight, fGCM, fAM, or prostate and testis volume between dominance groups. Semen was successfully collected from all alphas but only half the subdominants; urine contamination was negatively associated with dominance. Sperm quality was low (17.3 ± 10.2% total motility, 12.8 ± 8.5% progressive motility, 27.4 ± 11.5 × 106 ejaculated spermatozoa, 40.6 ± 9.8% normal morphology, 63.1 ± 5.1% viability, 72.6 ± 5.2% acrosome integrity) with no difference observed in any parameter except progressive motility and normal sperm morphology, which were significantly lower in subdominants (27.7 ± 16.8% v. 0.0 ± 0.0% and 59.8 ± 13.0% v. 21.4 ± 5.7%). From pre-breeding to breeding season, testis and prostate volume increased significantly, particularly in beta and gamma males respectively. Prostate volume was higher in alpha than beta males (16.0 ± 6.4 cm3 v. 5.7 ± 1.4 cm3), but testis volume, body weight, fAM, and fGCM did not differ between dominance groups (12.0 ± 0.9 cm3, 28.5 ± 0.8 kg, 0.51 ± 0.07 µg g−1, and 30.6 ± 2.3 ng/g of dry weight). Semen was successfully collected from 75% of males with reduced urine contamination. Collection success, urine contamination, and preputial gland size were not associated with dominance. Sperm quality improved with significantly greater number, viability, and total motility. However, sperm quality did not differ between dominance groups (47.4 ± 6.7% total motility, 30.5 ± 5.8% progressive motility, 32.3 ± 9.2 × 106 ejaculated spermatozoa, 50.9 ± 5.2% normal morphology, 74.4 ± 4.2% viability, 85.6 ± 3.0% acrosome integrity, and 99.7 ± 0.1% DNA integrity). In conclusion, subdominant males are at higher risk of urine contamination and have lower sperm motility and normal morphology when semen is collected in the pre-breeding season. However, their semen is of similar quality to dominant males in the breeding season, indicating that reproductive suppression of subdominant males is only behavioural. Thus, AWD males of all social ranks in the breeding season are suitable candidates for sperm banking.

This study aimed to determine the quality of Ettawah crossbred buck semen diluted in skim milk, egg yolk-citrate extenders, or their combination, supplemented with epigallocatechin-3-gallate (EGCG). Semen was collected from Ettawah crossbred bucks using an artificial vagina. Six ejaculates were used for replication and divided into three extender treatments: T1, skim milk; T2, egg yolk-citrate; and T3, a combination of both, each supplemented with 1.5 µg/mL EGCG. The extended semen was stored in a refrigerator at 5 °C. Diluted samples were evaluated every 24 hours until spermatozoa motility declined to 30%, the minimum requirement for artificial insemination. The results showed that spermatozoa motility, viability, plasma membrane integrity, and morphological abnormality decreased during six days of storage at 5 °C. Spermatozoa motility remained above 30% for five days in T1 (36.50±1.22%) and T2 (41.67±2.06%), and for six days in T3 (43.33±1.03%). On day six, semen in the combined extender supplemented with EGCG (T3) showed significantly higher motility, viability, and membrane integrity, and lower morphological abnormalities (p &lt;0.05) than the other treatments. In T2, all spermatozoa quality parameters were higher (p &lt;0.05) than in T1. It can be concluded that the combined skim milk-egg yolk-citrate extender containing 1.5 µg/mL EGCG best maintained the spermatozoa quality of Ettawah crossbred bucks during six days of storage at 5 °C. Based on spermatozoa motility, the semen remained suitable for artificial insemination.

Oxidative stress has been identified as a major cause of low seminal fertility. Among the components of stallion seminal plasma, some enzymatic and non-enzymatic antioxidants have been identified, which protect sperm from injurious effects of reactive oxygen species (ROS); however, the characterisation of these components is still in preliminary stages, as well as their relationship with freezability. The aim of this study was to evaluate the effect of some components of seminal plasma (SP) on stallion semen freezability. Semen of 30 Colombian Creole horses, and a total of 60 ejaculates, were collected. Semen was centrifuged to recover the SP. It was lyophilized and some components were assayed: total protein concentration (TP) by Bradford assay, CRISP3 protein concentration by ELISA, vitamin C (CVIT), vitamin E (EVIT) and vitamin A (AVIT), by HPLC; content of iron (Fe), copper (Cu), magnesium (Mg) and Zinc (Zn) by atomic absorption spectroscopy flame. Semen was supplemented with 10% stallion lyophilized SP and cryopreservation was performed. Post-thaw, total motility (TM), progressive motility (PM), straight line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF), were assessed using a computer-assisted sperm analysis (SCA®, Microptic SL, Barcelona, Spain). Sperm viability (SV) was determined by the Live/Dead kit (Molecular Probes Inc., Eugene, OR, USA). Normal sperm morphology (NM) was performed by the supravital technique and plasmatic membrane integrity (MI) was evaluated by the hypo-osmotic test. For statistical analysis, completely randomised mixed models were fitted. Levels according to the concentration of components of SP (high, medium, and low) were established. Comparisons of the means between levels were done with Tukey’s test. The significance level used for all assessments was P &lt; 0.05. Means for TP of 0.35 mg BSA/g, CRISP3 of 55.22 ng/mg, CVIT of 2.66 mg/g, EVIT of 72.36 µg/g, AVIT of 37.37 µg/g, Fe of 17.37 mg/kg, Cu of 33.64 mg/kg, Mg of 109.08 mg/kg, and Zn of 0.49 g/100 g of SP were found. We found that a high level of CRISP3, AVIT, Cu, and Fe had higher results for post-thaw TM, PM, NM; medium levels of TP and Mg showed higher post-thaw TM, PM, NM, and MI; and lower levels of Zn had better results for post-thaw TM, PM, VCL, and VAP. In contrast, high and medium levels of CVIT had a deleterious effect on post-thaw TM, PM, SV, NM, and MI. We concluded that there is a relationship between concentrations of seminal plasma components and stallion semen freezability.