Comparative Study of Abnormalities of the Spermocytogram between Manual Techniques versus Automated SCA (Sperm Class Analyzer)

It is already well established that the percentage of normal sperm and specific sperm abnormalities has diagnostic value in vivo. The spermocytogram is an important part of the analysis of human semen, this kind of analysis, simple at first sight, brings real difficulties because the results from one laboratory to another are very relatively reliable. When analyzing abnormal sperm morphology, the observer has to estimate the respective size of the different spermatozoa or their components, the length of all the spermatozoa (too small or too big) and to identify the shape (sperm with a coiled flagellum, or no flagellum as an example). The objective of our work is first of all to make a comparative study of the results of the morphological evaluation of 50 semen samples between the manual technique versus the automated one (SCA) in order to find a possible correlation between these two techniques. According to the analytical stage of the manual and automatic results, the two latter led us to obtain significantly different percentages.

Oligogenic heterozygous inheritance of sperm abnormalities in mouse

Male infertility is an important health concern that is expected to have a major genetic etiology. Although high-throughput sequencing has linked gene defects to more than 50% of rare and severe sperm anomalies, less than 20% of common and moderate forms are explained. We hypothesized that this low success rate could at least be partly due to oligogenic defects – the accumulation of several rare heterozygous variants in distinct, but functionally connected, genes. Here, we compared fertility and sperm parameters in male mice harboring one to four heterozygous truncating mutations of genes linked to multiple morphological anomalies of the flagellum (MMAF) syndrome. Results indicated progressively deteriorating sperm morphology and motility with increasing numbers of heterozygous mutations. This first evidence of oligogenic inheritance in failed spermatogenesis strongly suggests that oligogenic heterozygosity could explain a significant proportion of asthenoteratozoospermia cases. The findings presented pave the way to further studies in mice and man.

Cytogenetic and testicular histological alterations induced by sulphur dioxide in dried apricot leather

Sulphur dioxide (SO2) is used as a preservative in food to prevent its discolouration, and to inhibit the growth of bacteria. Little data is available concerning its in vivo hazardous impact.The present study is therefore designed to examine the cyto-genotoxic potential and the testicular histological alterations in adult mice, induced by SO2 present in the dried apricot leather used to prepare the oriental drink Qamar Al-Deen. Two different forms of drinks were tested; cold and boiled drinks. Animals were placed into 4 groups. The first group received distilled water as a negative control.The second and third groups received orally the drink for 28 days in the form of a cold and a boiled drink, respectively. Animals of the fourth group received cyclophosphamide, they were used as a positive control for cyto-genotoxic tests. The chromosomal aberrations, as well as sperm abnormalities, were significantly elevated in animals that received the two different drink preparations. The mitotic index significantly decreased in comparison with negative and positive controls. Furthermore, histological examination showed different degrees of alterations in the testis. Our results suggest that the presence of SO2 inside the apricot leather might be responsible for these changes. Thus, these remarkable hazardous effects of SO2 on male albino mice could be used as a potential guide for the prediction of its human health impact. Furthermore, consumers could be advised to prevent excessive consumption of the drink (Qamar Al-Deen) prepared from dried apricot leather.

Correlation between age, testosterone and adiponectin concentrations, and sperm abnormalities in Simmental bulls

Background and Aim: Capacity for sperm production is affected by age, which is related to the morphology of sperm abnormalities and can affect fertility. The aim of this study was to evaluate the relationship between age and concentrations of testosterone and adiponectin with sperm abnormalities in Simmental bulls. Materials and Methods: The study used 11 bulls, separated into three groups. The first group consisted of five bulls aged 4-5 years, and the second and third groups each consisted of three bulls, aged 6-7 and 8-10 years, respectively. The average sperm motility of the animals ranged from 57.66±2.60% to 70.17±0.22%. Blood samples were obtained from the coccygeal region of the animals. Testosterone and adiponectin analysis was performed using the enzyme-linked immunosorbent assay method. Sperm morphology was evaluated using carbol fuchsin-eosin staining according to the Williams method. Finally, correlations between testosterone and adiponectin concentrations, age, and sperm abnormalities were analyzed using Pearson's correlation analysis. Results: The findings revealed a significant correlation (p<0.01) between the concentrations of testosterone and adiponectin (–0.538), age (–0.588), and abnormal sperm morphology (–0.912). Moreover, they revealed that the concentration of testosterone in the bulls aged 8-10 years was lower, at 21.89±4.56 ng/mL, compared to that in the bulls aged 4-5 years, at 36.15±1.29 ng/mL, and 6-7 years, at 35.16±5.39 ng/mL. The findings also revealed a positive correlation between adiponectin concentration and age (0.529) and sperm abnormalities (0.506). The increase in testosterone concentration was inversely related to the adiponectin concentration (–0.538). Moreover, the mean amount of abnormal sperm increased with increasing age: 3.82±0.33% in the group aged 4-5 years, and 4.40±0.72% and 10.20±1.97% in the groups aged 6-7 years and 8-10 years, respectively. Conclusion: The study data indicate that there is a decrease in testosterone concentration, a high adiponectin concentration, and an increase in abnormal sperm with increasing age in bulls.

The Cryoprotective Potential of Propolis Supplemented in Frozen-Thawed Bull Semen: Biochemical and Physiological Findings

In this study, the cryoprotective effect of different doses of propolis (P) on bull semen, which has strong pharmacological properties thanks to its rich phenolic components, was investigated biochemically and physiologically. Semen samples were collected from Simmental breed bulls via the artificial vagina and pooled. After dividing into five groups, control (C: no additive), and four different dose P (200, 100, 50, and 25 μg/mL) groups, the final concentration was diluted to 16×106 per straw. Semen samples were equilibrated at 4°C for approximately 4 hours, then placed in French straws and frozen. After thawing, sperm motility and kinetic parameters, DNA integrity by single-cell gel electrophoresis, sperm abnormalities by liquid fixation, and lipid peroxidation levels by the colorimetric method was analyzed by Computer-Assisted Semen Analyzer. P added to the diluent showed no effect on motility and kinetic parameters at P25 and P50 (p &gt;0.05), while P100 and P200 had a negative effect (p &lt;0.001). The addition of P (25 and 50) had a treatment effect on tail abnormality compared to C (p &lt;0.05). Especially P50 had a positive effect on tail length, tail DNA, and tail moment, while P100 and P200 caused DNA damage (p &lt;0.001). MDA levels increased in all P dose groups compared to C (p &lt;0.001). This study has clearly demonstrated that P25 and P50 supplements could be used therapeutically in the treatment of sperm abnormalities and prevention of DNA damage in post-thawed bull sperm.

P–049 The presence of double heads is associated with increased frequency of the other sperm abnormalities

Study question Is there an association between the presence of spermatozoa with double heads and the other sperm abnormalities in human semen? Summary answer Patients with double-headed spermatozoa had a significantly increased percentage of morphological abnormalities (head, midpiece and tail defects). What is known already The morphological evaluation of spermatozoa has a prognostic value for successful IVF procedure. It has been proven that certain morphological defects have a negative impact on fertilization, embryo quality, and pregnancy outcome in in-vitro fertilization cycles. Sperm abnormalities, such as double head, double tail and thin midpiece are rarely observed. However, their effect on the other sperm defects has not been well studied yet. The aim of this study was to examine the effect of the presence of double-headed spermatozoa on the frequency of occurrence of the other sperm defects. Study design, size, duration This retrospective study includes 2140 men aged between 18 and 73 years, with a mean of 36 years. It was conducted at Nadezhda Women’s Health Hospital, Bulgaria between October 2015 and August 2020. A comparative analysis was performed between semen samples with and without double-headed spermatozoa and the other sperm abnormalities, as well as the percentage of morphologically normal forms. Participants/materials, setting, methods Morphological analysis was performed according to the Kruger’s strict criteria. Totally 23 types of abnormalities were determined: head defects (small, large, amorphous, elongated, round, pear-shaped, double, acephalic, detached head, small and large acrosomal areas and spermatozoa without acrosome), midpiece defects (thick, bent, asymmetric, thin midpiece and cytoplasmic droplets), tail defects (stumped, coiled and double tail), acrosomal vacuoles, nuclear vacuoles and multiple defects. Statistics: Mann-Whitney U-test and T-test; P ≤ 0.05. Main results and the role of chance Presence of double-headed spermatozoa was observed in 12.62% (270/2140) of the studied samples. In these patients the frequency of occurrence of double-headed spermatozoa ranged between 1% and 29% with a mean of 0.41%±1.71%. Men with double-headed spermatozoa had significantly higher percentage of spermatozoa with small heads (24.51%±22.65%, P = 0.04), round heads (11.69%±10.13%, P &lt; 0.01), nuclear vacuoles (10.64%±5.25%, P &lt; 0.01), sperm without acrosome (9.76%±8.61%, P = 0.05), asymmetric midpiece (4.73%±3.96%, P &lt; 0.05), bent midpiece (8.9%±7.22%, P &lt; 0.01), thin midpiece (2.13%±4.44%, P &lt; 0.01), double tail (1.78%±0.8%, P &lt; 0.01), detached head (1.98%±1.42%, P &lt; 0.01), stumped tail (6.03%±5.19%, P = 0.02), and cytoplasmic droplets (8.86%±5.02%, P &lt; 0.01) compared to the patients without double-headed spermatozoa. Moreover, the percentage of sperm with multiple defects in the double-headed group was significantly higher (35.53%±29.91%, P &lt; 0.01), while the percentage of normal forms was significantly lower (2.93%±3.64%, P &lt; 0.01) compared to the patients without double heads. Limitations, reasons for caution In this study unequal sample sized groups were compared. We also need to investigate whether the obtained results will be confirmed in patients with certain pathological states, such as oligozoospermia, teratozoospermia, and asthenozoospermia. Wider implications of the findings: The present study revealed that the presence of double-headed spermatozoa in the ejaculate is related to an increased frequency of the other semen abnormalities. The double-headed spermatozoa could be used as an indicator for the total morphological quality of human spermatozoa Trial registration number Not applicable

Sperm parameters and mitochondrial DNA sequence variants among patients at a fertility clinic in Ghana

Purpose The present study sought to investigate the common abnormalities and mtDNA mutations in the sperm of Ghanaian men attending the fertility Clinic at the Korle-Bu Teaching Hospital (KBTH). The study therefore provides a baseline data mtDNA mutations in a cross-section of Ghanaian men on referral to the fertility clinic at the KBTH. Materials and methods The semen of 55 men attending the fertility clinic were collected from the Urology and the Obstetrics and Gynaecology Departments of the KBTH. Demographic and clinical data were also collected using questionnaires. Semen analyses were performed and were followed by amplification and purification of mtDNA from total DNA extracted from the semen. Sequencing of the mtDNA amplicons was performed using the next generation sequencer (Illumina-MiSeq). Results Asthenozoospermia, oligospermia and oligoasthenoteratozoospermia were observed in 1.79%, 5.36% and 28.57%, respectively, of the study participants. There was no association between drinking and/or smoking and history of gonorrhea infection on sperm status/morphology. A total of 785 point mutations were detected in the non-coding control regions, rRNA genes, tRNA genes and the coding regions of the mtDNA samples from the participants. Amongst these mutations, 16 transition mutations were predominantly detected in the mtDNA samples. Missense mutations that were present in only specific sperm abnormalities were identified and they may contribute to infertility in the study population. Conclusion The present study has identified various abnormal sperm phenotypes that are prevalent in the study population and provided a baseline data on mtDNA mutations in the spermatozoa of the patients. A wide range of sperm abnormalities were detected in the study population with no association with life style or history of gonorrhea infection. The mtDNA point mutations detected in the selected genes that were analysed were mostly transition mutations. These transition mutations might be critical for the development of abnormal sperm phenotypes underlying male infertility in the Ghanaian population.

The The Effect of Age and Breed on The Quality of Bull Semen in The Regional Artificial Insemination Centre

The success of beef cattle reproduction development in Indonesia very much depends on the quality of the semen produced by the Indonesian Institute for Artificial Insemination. Good quality bull will benefit farmers, frozen semen producers, and the country in large. This study aimed to determine effect of age and breed on the libido and quality of bull semen collected in the Regional Artificial Insemination Center (RAIC). This study used PO, Simmental, and Brahman Bull in RAIC Yogyakarta, with a total amount of 15, aged 4-7 years old. Semen was collected using an artificial vagina, semen evaluation used a spectrophotometer, and eosin-nigrosin staining method used for sperm viability and abnormalities examination. The result of this study indicated that bull breed did not affect the libido value and the semen quality of bulls (P>0,05). Volume, concentration, motility, and viability in various age groups did not show significant differences (P>0,05), but age groups had a significant effect on sperm abnormalities (P<0,05). The group of bulls aged 5-6 years showed the best quality compared to other groups, with the volume 4,58 ± 1,37 ml, concentration 1,91 ± 0,36 x109/ml, motility 3,80 ± 1,30, viability 89.83 ± 10.69%, and sperm abnormalities 2.51 ± 0.81%. It can be concluded that breed did not affect the bull libido and semen quality, while age had an effect on bull sperm abnormalities.

Assessment of Semen Characteristics Among Three Phenotypes of Chicken Raised in Akko, Gombe State of Nigeria

In poultry breeding program the success of artificial insemination (AI) is highly influenced by the quality of semen. The aim of the present research was to evaluate the variations in semen characteristics of three phenotypes of chicken of the same specie raised in Akko, Nigeria. A total of 9 cocks from threedifferent phenotypes; Red feathered (n = 3), White feathered (n = 3) and Black feathered (n =3). The semen was evaluated for macroscopic (i.e. semen volume and colour) and microscopic (i.e. sperm concentration, motility and morphology) criteria after being collected by abdominal massage method. There were phenotypes variation (P > 0.05)effects on semen motility, number of live/dead sperm and sperm abnormalities. No differences(P>0.05) were observed on volume, colour, concentration, mass motility and pH of semen. The observed sperm progressive motility was ranged from 90.5 ±1.21% to 95.09±0.82%. The White feathered strain had the highest sperm progressive motility (P<0.05)and highest value for live and normal sperm (93.5±0.63% and 87.90±0.25%), while the black feathered had the least and (92.4±0.73% and 85.5±0.50%) respectively. The Red feathered strain had the highest value for both dead and sperm head defect (9.2±0.33%and 7.0±0.52%). Whilst, the Black feathered strain scored the highest percentage of both sperm tail and neck defects with values ranged from 11.4±0.43%to 14.2±1.08% and 19.4±0.53% to 23.6±0.69%, respectively. This study suggests that there are large variations present in semen characteristics of different phenotypes of cocks; White feathered strain is likely have better semen characteristics compared to Red and Black feathered strains. Therefore, White feathered strain can potentially be used in artificial insemination (AI) for chicken production and improvement.