Morphometric characterization and classification of vicuña sperm (Vicugna vicugna) using ISAS® CASA-Morph system

The aim of the study was to determine the morphological and morphometric characteristics of the vicuña sperm (Vicugna vicugna), looking for possible subpopulations in its structure. Semen was collected by electroejaculation method from three adult male vicuñas weighing 50.33 ± 2.52 kg, once per animal. The volume, color and concentration, sperm morphology and morphometry were determined. The samples for the analysis of sperm morphology and morphometry were stained with Hemacolor®, and analyzed using the CASA-Morph, Integrated Semen Analysis System (ISAS®v1). Five forms of the vicuña sperm head were distinguished: normal, piriform, long, short and rounded. The morphometric parameters determined were the length, width, area, perimeter, ellipticity, elongation, regularity and rugosity of the vicuña sperm head, percentage of acrosome, head gray level, as well as the width, area, distance and angle of insertion of midpiece of the sperm. In relation to morphology, significant differences (P <0.05) were found in the percentage distribution of head shapes, with the normal shape (55.7%) being the majority and different from the other shapes. Significant differences (P <0.05) were found between animals in the morphometric variables of head, percentage of acrosome, ellipticity, rugosity, elongation and gray level; while the variables of the midpiece and regularity did not show differences (P> 0.05). The morphometric variables were distributed in four main components (PCA) called elongation, area, circularity and midpiece width, which explained 84.59% of the total variance. The cluster analysis determined five subpopulations (SP): SP1 grouped small cells of low length, width and area (18.8%); SP2 of sperm of large size, both in area and width of the head (17.38%); SP3 of rounded cells with high values of percentage of acrosome and head gray level (24.04%); SP4 of spermatozoa of intermediate size and elongated, with greater ellipticities and elongation (23.61%); and SP5 of cells of intermediate size and short, with low values of area and length (16.71%).

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.

Establishment of semen collection technique using electroejaculator and semen cryopreservation of Javan leopard (Panthera pardus melas Cuvier, 1809)

Background and Aim: The Javan leopard (Panthera pardus melas Cuvier, 1809) is a subspecies of Panthera pardus spp., spread across the African and Asian regions. Information on reproductive aspects is crucial for wild animals, including the Javan leopard. In this study, we aimed to develop electroejaculator (EE) techniques and evaluate cryopreservation success in Javan leopard semen. Materials and Methods: The semen of four adult Javan leopards was collected once a week using EE. Placement of the EE probe in the rectum was performed after ultrasound imaging (ultrasonography) to determine the prostate body location. The semen obtained was then evaluated macroscopically and microscopically. Three Javan leopards were used for cryopreservation. The ejaculate was divided into two parts [i.e., one part diluted with AndroMed® (Minitüb, Tiefenbach, Germany) and the other part with Steridyl® (Minitüb, Tiefenbach, Germany)] at a 1:1 ratio immediately after collection and evaluation. The semen was then packed in a 0.25 mL MiniStraw® (Minitüb, Tiefenbach, Germany) then equilibrated at 4°C for 2 h. After equilibration, the straw was then frozen in liquid nitrogen vapor. Frozen semen was then stored in containers until further evaluation. Results: The results showed that ejaculation response occurred at all levels of stimulation, while erections did not always occur. The fastest ejaculation and erection occurred at the fourth voltage. The macroscopic evaluation showed that the semen volume was 0.80±0.26 mL, cloudy white, pH 7.44±0.14, and with watery semen consistency. The microscopic evaluation showed that the sperm motility was 66.98±0.39%, with sperm viability of 75.6±1.79%. Sperm concentration was 62.17±46.95×106 mL–1 with a total concentration of 42.14±23.51×106 cells. Normal sperm morphology is only 40.72±6.26%. Conclusion: This study concluded that the development of a semen collection technique using an EE preceded by imaging of the EE probe location using ultrasound was effective for the ejaculation of Javan leopards. The characteristics of the semen of the Javan leopard showed moderate semen volume, sperm motility, and viability. Javan leopard showed low sperm concentration and normal sperm morphology.

METRONIDAZOLE INDUCES SIGNIFICANT PATHOLOGICAL ALTERATIONS IN THE MALE REPRODUCTIVE SYSTEM OF MICE

The present study was aimed to investigate the possible effects caused by the antibiotic, metronidazole, on the histology of the testis and the morphology of the sperms of mice. To achieve this, adult male albino Swiss Mice were orally administrated with 0.1 ml (100 mg\ kg ) of the commercially used metronidazole (Flagyl) for 60 consecutive days. The results showed significant decreases (p<0.05) in the mean weight of body and testis in the treated mice as compared to the distilled water-treated control group. The treatment also caused several histopathological changes in the testis which included necrosis, congestion, hemorrhage, edema, germ cell downfall and rupture in the seminiferous tubules. The tests of the sperm morphology revealed a number of deformations as a result of the treatment, such as the appearance of headless, bend, broken-tailed, hook-tailed, and two-headed sperms, as well as sperms with cytoplasmic droplets. We conclude that from these results that orally administered Metronidazole exerts negative effects on the male reproductive system of mice .

The effect of N-acetyl cysteine consumption on men with abnormal sperm parameters due to positive history of COVID-19 in the last three months

Male infertility is an important factor accounting for 40-50% of infertility cases that may be due to disturbance in one of the parameters as concentration, motility and morphology observed in one or two semen analysis with an interval of 1 and 4 weeks. COVID-19 may affect male fertility through virus division, cytotoxic effects on testicular tissue and immunopathological effect. N-acetyl cysteine (NAC) improved sperm concentration and acrosome reaction while reducing reactive oxygen species (ROS) and oxidation of sperm DNA. This interventional study was conducted on 200 men who were referred to private infertility clinics for female factor (their previous semen analysis was normal) and got COVID-19 infection in the last 3 months showing an impairment of the latest semen analysis due to COVID. Men were placed in two groups of control (n = 100) and intervention (NAC consumption). Subjects who got COVID-19 infection had a significant impairment of sperm quality (sperm concentration, sperm motility, and normal sperm morphology) compared to their semen analysis evaluated before the COVID-19 infection. NAC consumption significantly improved sperm total motility, sperm morphology and sperm concentration. COVID-19 infection has a negative effect on sperm parameters. NAC supplementation may have positive effect on sperm parameters.

Sheng Jing Decoction Can Promote Spermatogenesis and Increase Sperm Motility of the Oligozoospermia Mouse Model

Sheng Jing Decoction (SJD), as a traditional Chinese medicine prescription, is mainly be used to treat male infertility. However, the pharmacological functions and molecular mechanisms of SJD are poorly understood. In this study, we investigated the functions of SJD on spermatogenesis and sperm motility and explored the potential mechanisms involved. Here, we demonstrated that high, medium, and low doses of SJD are effective in restoring the impairments of the whole body and testicular tissue by cyclophosphamide inducing and to rescue the damage of testicular tissue cells including Sertoli cells and germ cells. SJD can partly restore the decrease in sperm concentration, sperm vitality, sperm motility, and normal sperm morphology rate in oligozoospermic mouse models. Ki67 staining analyses confirm SJD can promote testicular tissue cell proliferation. Real-time RT-PCR analyses also reveal that SJD can upregulate the expression of proliferation-associated gene Lin28a and differentiation-associated genes Kit, Sohlh2, and Stra8. SJD can also reduce the impairment of mitochondrial membrane potential (MMP) and sperm plasma membrane integrity by cyclophosphamide inducing. Our results reveal that SJD is effective in improving both sperm quantity and quality by increasing the sperm concentration, sperm vitality, sperm motility, and normal sperm morphology rate. SJD can promote spermatogenesis by upregulating the expression of the proliferation-associated gene Lin28a and the differentiation-associated genes (Kit, Sohlh2, and Stra8). SJD can sustain MMP and sperm plasma membrane integrity to increase sperm motility.

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.

A Comprehensive Understanding of Shukrala Karma in Male Infertility

Male reproductive health depends upon the normal structures and functions of Shukrava srotas. Shukra is composed of both sperm and semen and considered one of the important factors for fertilization. It is stated as Phalavat shukra. Preceptors of Ayurveda have described eight types of Shukra dosha and their treatments. Low sperm count, decrease sperm motility, abnormal sperm morphology, ejaculatory problems, sexual dysfunctions, environmental exposures (radiation, pollution, and stress etc), lifestyle habits (smoking, alcohol, recreational drugs etc), varicocele, hormonal imbalances, DNA damage, and reactive oxygen species (ROS) etc are causative factors for Male infertility. Thousands of years back, ancient system of medicine has mentioned the word Purusha vandya (male infertility) and different pharmacological activities, medicinal formulas, therapies to treat sperm disorders, semen impairments, and sexual dysfunctions. Vajeekarana is one of the special branches of Astanga Ayurveda which maintains the fertility and management of male infertility. Shukrala, Shukra janana, Shukra shodhana, Shukra rechaka, Shukra pravataka and Shukra sthambhaka etc., are the pharmacological activities mentioned to treat Shukradoshas. Shukrala karma mainly composed of Shukra vrudhikara and Shukra srutikara means which enhances the Shukra (semen and sperm) quantitatively and qualitatively and facilitates its ejaculation. Therefore, an attempt has been made to establish the concept of Shukrala karma and its therapeutic applicability in the management of male infertility.