Presence Of Sperm Research Articles

Male reproductive ageing: a tale of the whole ejaculate.

Ageing is nearly ubiquitous and encompasses all biological functions. We here focus on age-dependent changes in male reproductive capacity across a broad range of animal taxa. While there has been a long-standing focus on mating ability and overall reproductive success, we here highlight the underlying mechanisms that explain loss in fertilisation capacity in ageing males. Fertilisation is mediated by not only the presence of sperm, but also the cocktail of seminal fluid proteins that ensure sperm survival, capacitation and interaction with female physiology. Sperm ageing has received much attention in studies of male reproductive senescence; however, post-mating processes include a number of interlocked steps that together cumulate in successful fertilisation. As such we consider male ability to elicit female post mating responses such as uterine conformational changes, sperm storage and ovulation and the components within the ejaculate that mediate these post-mating processes. For the latter seminal fluid proteins are key and hence we reflect on age-dependent changes in quality of the entire ejaculate and its consequences for male reproductive capacity. While first studies accrue and highlight that changes in the non-sperm fraction can explain substantial variation in senescent male reproductive success and male ability to induce post-mating responses necessary for fertilisation many open questions still remain that warrant further investigations. One being what the potential age-dependent changes in composition are or whether there is a general decline and how this interacts with sperm to affect fertilisation success. Further, the impact females might have to ameliorate these changes will be an area of interest.

Infundibular sperm storage and uterine muscular twisting in the Amazonian lancehead, Bothrops atrox.

Although knowledge about the reproduction of snakes of the genus Bothrops has increased recently, many questions remain regarding sperm storage by these neotropical pitvipers. The relationship between two known sperm storage sites-the nonglandular uterus and the infundibulum-has yet to be detailed for Bothrops. In addition, the formation of uterine muscular twisting (UMT) and its function in sperm storage in the nonglandular uterus have been questioned. This article presents data on both sperm storage sites (posterior infundibulum and nonglandular uterus) in the Amazonian lancehead, Bothrops atrox, aiming to address the role of UMT in bothropoid reproduction considering female reproductive stages, timing of sperm storage, and morphology and histochemistry of sperm receptacles. The oviduct of B. atrox is composed of the infundibulum (anterior and posterior), the glandular uterus, the nonglandular uterus and the pouch. Sperm storage occurs in the posterior infundibulum and nonglandular uterus in vitellogenic females. In the posterior infundibulum sperm is stored in receptacles, while in the nonglandular uterus it is stored in crypts in the UMT of vitellogenic females. Both sperm storage sites possess granules testing positive for neutral mucopolysaccharides in the presence of spermatozoa, possibly favoring the conditions for spermatozoa surveillance during sperm storage in the reproductive tract.

Evaluation of sperm DNA fragmentation and chromatin structure in infertile men with immotile short‐tail sperm defect

Teratozoospermia is characterised by the presence of spermatozoa with abnormal morphology. One of the morphological disorders that lead to male infertility is immotile short-tail sperm (ISTS) defect. In this study, we evaluated the levels of chromatin packing and DNA fragmentation in patients with immotile short-tail sperm defect. Semen samples were obtained from 31 infertile men with ISTS as case group and 31 normozoospermic men as a control group. Protamine status was evaluated using chromomycin A3 (CMA3) staining and sperm DNA fragmentation assessed by sperm chromatin structure assay (SCSA) and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labelling (TUNEL). The percentage of positive CMA3 spermatozoa was significantly higher in patients' samples (22.6±6.9) compared with controls (16.3±4.2) (p<.05) and also mean (±SD) of sperm DNA fragmentation was significantly higher in patients compared with controls, as measured by TUNEL assay (10.45±4.60 vs. 7.03±2.86, p<.05) and SCSA (24.80±13.1 vs. 15.2±7.2, p<.05). According to our study, sperm DNA fragmentation and chromatin packing abnormality are significantly higher in the ISTS samples compared with normal samples. A possible explanation for this relationship is that sperm chromatin condensation and sperm flagellum formation occur during the same phase of spermatogenesis.

Possible factors influencing post-ejaculatory changes of the osmolality of human semen in vitro.

The human ejaculate is made up of secretions from the different accessory sex glands that empty in sequence at ejaculation. However, the different secretions only mix completely in vitro when the entire ejaculate is collected in a container and handled in the laboratory. At ejaculation, proteins from the seminal vesicles form a gel in the ejaculate and semen cannot be properly analysed and processed until the gel is liquefied. During and after liquefaction, there is continuous enzymatic activities and an ongoing increase in osmolality. The aim of this study was to investigate possible factors that influence the increase in semen osmolality in vitro. Osmolality was measured by freezing-point depression. Prostatic secretion was measured as zinc concentration. The presence of spermatozoa neither influenced the actual measurement of semen osmolality, nor the increase in osmolality. Enzymatic inhibitors reduced the increase in osmolality, and semen dilution prevented any increase in semen osmolality. However, the increase in osmolality covaried with the seminal zinc concentration, indicating that the observed increase was related to factors of prostatic origin. A simple and convenient procedure to reduce the risk for osmotic challenges for spermatozoa during handling for assisted reproductive technologies might be early dilution of semen.

Comprehensive gene expression analysis by RNA sequencing of testicular tissue exosomes in azoospermic men: predicting the presence of sperm

Comprehensive gene expression analysis by RNA sequencing of testicular tissue exosomes in azoospermic men: predicting the presence of sperm

Host influence on the nutritional and reproductive status of Triatoma infestans (Klug) (Hemiptera: Reduviidae) peridomiciliary populations

Triatoma infestans is the main vector of Chagas disease in the southern cone of South America. This species is well adapted to living in rural houses and structures used for housing domestic animals (peridomestic habitats). In this study, we evaluated the relationship between the source of blood consumed by adults of T. infestans collected from different peridomestic habitats from two localities from Cruz del Eje department (Cordoba, Argentina) and their nutritional and reproductive status. In each individual, the ratio between body weight and total body length was used as an indicator of nutritional status (NS). The presence of sperm in spermathecae and the number of chorionated oocytes in ovaries and oviducts were considered indicators of reproductive status (RS) of females. The feeding source in the promesenteron of male and female insects was identified using anti-chicken, anti-goat, antihuman and anti-dog antisera. Chicken coops were the main peridomestic structure present in the study area as well as the peridomestic sites with the highest percentage of T. infestans . Insects collected from the different peridomestic structures showed a NS between 8 and 15 mg/mm. Of the evaluated females, 35.7% presented chorionated oocytes. Food profile analyses revealed that chicken was the main blood source. Independently of the blood source, the triatomines presented a NS between 8 and 15 mg/mm. No specimens feeding exclusively on human blood were found; nevertheless, of 31.48% of insects feeding on mixed blood sources, 59% included human blood. All T. infestans specimens that included human blood in the mixed blood source were collected from chicken coops and storerooms located in a 12-m area around domiciles. Human blood present in mixed blood meal of adult insects suggests that T. infestans moves from domiciles to peridomicilies and vice versa

Altered microRNA profiles of testicular biopsies from patients with nonobstructive azoospermia.

Many studies have shown that microRNAs (miRNAs) play vital roles during the spermatogenesis. However, little is known about the altered miRNA profiles of testicular tissues in nonobstructive azoospermia (NOA). Using microarray technology, the miRNA expression profiles of testicular biopsies from patients with NOA and of normal testicular tissues were determined. Bioinformatics analyses were conducted to predict the enriched biological processes and functions of identified miRNAs. The microarray data were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the results of which were then validated with a larger sample size. Correlations between the miRNA expression levels and clinical characteristics were analyzed. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic ability of miRNAs for azoospermia. Hierarchical clustering showed that 129 miRNAs were significantly differentially expressed between the NOA and control groups. Bioinformatics analysis indicated that the differentially expressed miRNAs were involved in spermatogenesis, cell cycle, and mitotic prometaphase. In the subsequent qRT-PCR assays, the selected miRNA expression levels were consistent with the microarray results, and similar validated results were obtained with a larger sample size. Some clinical characteristics were significantly associated with the expression of certain miRNAs. In particular, we identified a combination of two miRNAs (miR-10b-3p and miR-34b-5p) that could serve as a predictive biomarker of azoospermia. This study provides altered miRNA profiles of testicular biopsies from NOA patients and examines the roles of miRNAs in spermatogenesis. These profiles may be useful for predicting and diagnosing the presence of testicular sperm in individuals with azoospermia.

Options for Fertility Treatments for Trans Women in Germany.

Fertility preservation in trans women is a crucial but thus far neglected component in the gender confirming treatment in Germany. It is difficult for trans women to access reproductive health care because centers offering treatment, psychological guidance, gender confirming surgery, as well as reproductive health services are scarce in Germany. Legal, social, or financial issues as well as individual patient comorbidities prevent trans women from receiving appropriate counselling. This review provides an overview on options of fertility preservation in trans women. We consider recent publications on testicular regression at the time of gender confirming surgery demonstrating presence of sperm or at least spermatogonia in the majority of tissues. This may open options for cryopreservation of sperm or testicular stem cells in trans women even at the final stage of transition. Hence, standardized urological procedures (i.e., sperm cryopreservation after masturbation or sperm extraction from the testicular tissue) and experimental approaches (cryopreservation of testicular tissue with undifferentiated spermatogonia) can be offered best at the initiation but also during the gender confirming process. However, counselling early in the gender confirming process increases the chances of fertility preservation because gender confirming hormone therapy has an impact on spermatogenesis.

Nasal administration of gonadotropin releasing hormone (GnRH) elicits sperm production in Fowler\u2019s toads (Anaxyrus fowleri)

BackgroundDeclining amphibian populations around the world necessitate the establishment of captive assurance colonies as a hedge against extinction. For species that are difficult to breed in captivity, assisted reproductive techniques, such as treatment with exogenous hormones, are necessary for successful reproduction. The purpose of this study was to determine whether intranasal administration of a gonadotropin releasing hormone analog (GnRHa) elicits sperm production in anurans.MethodsMale Anaxyrus fowleri (n = 15/trt) were nasally administered GnRHa (1, 5, 10, and 20 μg) in phosphate buffered saline (PBS) using a pipette. Spermic urine was collected from animals over an 8 h period. Samples were assessed for sperm presence, motility, and concentration.ResultsTreatment of male toads with a PBS control and 1 μg of GnRHa did not elicit sperm production. Nasal administration of GnRHa resulted in sperm production from 60, 93, and 80% of males in the 5, 10, and 20 μg treatment groups, respectively. Sperm motilities averaged 70, 63, and 52% within the 5, 10, and 20 μg treatment groups, respectively, with the highest (p < 0.05) sperm motility observed using 5 μg of GnRHa. Significantly higher sperm concentrations were observed in males treated with 10 μg of GnRHa compared with 5 or 20 μg of GnRHa.ConclusionNasal administration of GnRHa was successful in eliciting spermiation from male Anaxyrus fowleri, which typically began less than 3 h after treatment. Nasal administration of GnRHa may provide a novel non-invasive method of hormone delivery for at-risk amphibian species that have low reproductive output.

Toxicologic Pathology Forum*: Opinion on Sexual Maturity and Fertility Assessment in Long-tailed Macaques ( Macaca fascicularis) in Nonclinical Safety Studies.

If nonhuman primates represent the only relevant species for nonclinical safety evaluation of biotechnology-derived products, male and female fertility effects can be assessed in repeat dose toxicity studies given that sexually mature monkeys are used. This opinion piece provides recommendations for determining sexual maturity and when/how fertility assessments should be conducted in the cynomolgus monkey. Male sexual maturity should be proven by presence of sperm in a semen sample, female sexual maturity by at least two consecutive menstrual bleedings. As per regulatory guidance, default parameters for an indirect assessment of fertility in both sexes are reproductive organ weight and histopathology. Beyond default parameters, daily vaginal swabs are recommended for females, and for males, it is recommended to include blood collections (for potential analysis of reproductive hormones), testis volume sonography, and collection of frozen testis samples at necropsy. Only if there is a cause for concern, blood collection for potential reproductive hormone analysis should be conducted in females and semen analysis in males. In principle, adverse reproductive effects can be detected within 4 weeks of test article administration, depending on study design and reproductive end point chosen. Therefore, there are options for addressing reproductive toxicity aspects with studies of less than 3 months dosing duration. *This is an opinion article submitted to the Toxicologic Pathology Forum. It represents the views of the authors. It does not constitute an official position of the Society of Toxicologic Pathology, British Society of Toxicological Pathology, or European Society of Toxicologic Pathology, and the views expressed might not reflect the best practices recommended by these Societies. This article should not be construed to represent the policies, positions, or opinions of their respective organizations, employers, or regulatory agencies.

Surrogate production of Salmo salar oocytes and sperm in triploid Oncorhynchus mykiss by germ cell transplantation technology

Surrogate broodstock technology is emerging as a promising biotechnology for xenogeneic production of gametes of highly valuable fish species. In this study, we produced gametes of landlocked Atlantic salmon using triploid rainbow trout as recipient through germ cell transplantation technique in embryos. Spermatogonial cells were obtained from Atlantic salmon juveniles and transplantation conducted into the coelomic cavity of 150 mixed-sex triploid embryos of rainbow trout. Colonization efficiency assessed in larvae four weeks post-transplantation showed presence of PKH-26-labeled cells in 61.1% of individuals. Analysis in sexually mature adults revealed presence of donor-derived sperm in four transplanted males (10%), whereby one of them reached sexual maturity in the first year. Transplanted females became mature after the second year and donor-derived oocytes were detected in four individuals (12.1%). Milt produced by transplanted triploid males were used for artificial insemination. Genetic analysis of hatchlings revealed that progeny was composed by pure Atlantic salmon and therefore were donor-derived. In conclusion, this study shows that surrogate broodstock can be used to produce oocytes and spermatozoa using species from different genus. The production of surrogate gametes using species with a shorter life cycle represents an efficient approach to streamline the development of improved strains in aquaculture industry.

The obstructive interval predicts pregnancy rates in post-vasectomy patients undergoing ICSI with surgical sperm retrieval.

Are the outcomes of (i) surgical sperm retrieval (SSR) and (ii) intracytoplasmic sperm injection (ICSI) influenced by the obstructive interval (time elapsed since vasectomy)? Medical records from 148 patients (194 cycles) with secondary azoospermia due to vasectomy, who presented for percutaneous epididymal sperm aspiration (PESA) and ICSI in a private university-affiliated IVF centre, from January 2012 to February 2017, were analysed in this historical cohort study. The obstructive interval was recorded for each couple, and its influences on the outcomes of SSR and ICSI treatment were investigated using general mixed models with adjustment for potential confounders. Clinical pregnancy rate was the main outcome measure. The obstructive interval was negatively correlated with the presence of spermatozoa (β = -0.032, P = 0.009) and motile spermatozoa (β = -0.031, P = 0.010) during PESA. The need to convert to testicular sperm aspiration was significantly influenced by the obstructive interval (β = 0.012, P = 0.003). The blastocyst development rate on day 5 was inversely correlated with the obstructive interval (β = -0.011, P = 0.014). Implantation and clinical pregnancy rates were negatively influenced by the obstructive interval (β = -1.107, P = 0.039 and β = -0.016, P = 0.031, respectively). The receiver operating characteristic curve analysis demonstrated that the obstructive interval has a predictive value on the achievement of clinical pregnancy (area under the curve = 0.667, P = 0.001, Youden index 0.3385, associated criterion >17 years). Men undertaking vasectomy should be made aware of the long-term effects and their implications for future reproductive treatment.

What can we deduce about the reproductive condition of spinifex hopping mice (Notomys alexis) from external examination?

Field studies often use external examination of the vagina or the swelling of the scrotum to make deductions about the reproductive condition of rodents. In this study we sought to determine what information on reproductive condition could be gained from external examination of the reproductive anatomy of spinifex hopping mice (Notomys alexis) using individuals from a captive colony. Female N. alexis with perforate vaginae had a significantly larger mean uterine mass and larger mean maximum ovarian follicle diameter than non-perforate females. Corpora lutea were recorded in two perforate animals but were not present in females with a ‘pinhole’-size perforate or non-perforate vagina. In male hopping mice scrotal bulge size was unrelated to testes mass, ventral prostate mass or presence/absence of sperm in the cauda epididymides. Males with dark scrotal pigmentation had a significantly smaller mean testes mass than males with light or no pigmentation. However, there was no relationship between scrotal pigmentation and ventral prostate mass or sperm presence in the cauda epididymides. Our study suggests that vaginal perforation is an acceptable indicator of sexual maturity in female hopping mice, but scrotal bulge size and scrotal pigmentation should not be used to predict reproductive condition of males.

The annual cycle of spermatozoa content in the vas deferens and some reproductive parameters in the narrow-clawed crayfish Pontastacus leptodactylus (Eschscholtz, 1823)

Understanding the annual dynamic of gametes in crayfish is useful for successful management of crayfish population. The annual variations in spermatozoa content of vas deferens, gonado-somatic index (GSI), hepatosomatic index (HSI), vas deferens index (VDI), and testicular index (TI) were investigated in male Pontastacus leptodactylus. The spermatozoa content, GSI, HSI, VDI, and TI of P. leptodactylus significantly (P < 0.05) changed during different seasons of the year. For example, significantly higher spermatozoa number, GSI, VDI, and TI were recorded in January (P < 0.05). On the other hand, the HSI reached the peak in August (P < 0.05). The spermatophore degeneration started in April. Although the vas deferens lacks spermatophore structure during the summer, the presence of spermatozoa was observed throughout the year. This study represents a step forward in understanding the seasonal spermatozoa content and the reproductive status of male P. leptodactylus.

Potential roles of neutrophils in maintaining the health and productivity of dairy cows during various physiological and physiopathological conditions: a review.

Neutrophils represent the first line of innate immunity and are the most prominent line of cellular defence against invading microorganisms. On stimulation, they can quickly move through the walls of veins and into the tissues of the body to immediately attack or monitor the foreign antigens. Neutrophils are highly versatile and sophisticated cells which are endowed with highly sensitive receptor-based perception systems. They were traditionally classified as short-lived phagocytes actively involved during infection and inflammation, but recently, it has been seen that neutrophils are capable of detecting the presence of sperms during insemination as well as an implanting embryo in the female reproductive tract. These specialised phagocytes play a major role in tissue remodelling and wound healing, and maintain homeostasis during parturition, expulsion of placenta, folliculogenesis, corpus luteum formation and luteolysis. Here, we review the role played by neutrophils in maintaining homeostasis during normal and inflammatory conditions of dairy cattle. We have summarised the alteration in the expression of some cell adhesion molecules and cytokines on bovine neutrophils during different physiological and physiopathological conditions. Some emerging issues in the field of neutrophil biology and the possible strategies to strengthen their activity during the period of immunosuppression have also been discussed.

Prospective analysis of factors predicting feasibility & success of longitudinal intussusception vasoepididymostomy in men with idiopathic obstructive azoospermia.

Background & objectives:Microsurgical reconstruction for idiopathic obstructive azoospermia is a challenging procedure, and selection of appropriate patients is important for successful outcomes. This prospective study was done to evaluate the ability of scrotal ultrasound measurements to predict the surgical feasibility and determine factors that could predict a patent anastomosis following vaso-epididymal anastomosis (VE) in men with idiopathic obstructive azoospermia.Methods:In this prospective study, men diagnosed with idiopathic obstructive azoospermia, scheduled for a longitudinal intussusception VE, underwent a scrotal ultrasound measurement of testicular and epididymal dimensions. During surgery, site and type of anastomosis, presence of sperms in the epididymal fluid and technical satisfaction with the anastomosis were recorded. All men where VE could be performed were followed up for appearance of sperms in the ejaculate. Ultrasound parameters were compared between men who had a VE versus those with negative exploration. Predictive factors were compared between men with or without a patent anastomosis.Results:Thirty four patients were included in the study conducted between September 2014 and August 2016 and a VE was possible in only 19 (55%) patients. Of these 19 patients, six had a patent anastomosis with one pregnancy. Preoperative ultrasound measurements could not identify patients where a VE could not be performed. Motile sperm in the epididymal fluid was the only significant predictor of a successful anastomosis.Interpretation & conclusion:Forty five per cent of men planned for a VE for idiopathic obstructive azoospermia could not undergo a reconstruction. Ultrasound assessment of testicular and epididymal dimensions could not predict the feasibility of performing a VE. The presence of motile sperms in the epididymal fluid was the only significant predictor of a patent VE in our study.

The reproductive pattern of the Gerbilliscus cf. leucogaster (Rodentia: Muridae) from Namibia

Very little is known about the reproductive biology of the Gerbilliscus cf. leucogaster (Peters, 1852) despite its wide distribution throughout the southern African subregion. Body mass, reproductive tract morphometrics, and gonadal histology were studied over 12 months in wild caught Gerbilliscus cf. leucogaster from the central part of Namibia to gain insights into the reproductive pattern of this species. The number of Graafian follicles and corpora lutea in 93 females increased at the end of the dry period (September) and throughout the wet months of the year (October–May) relative to that of the dry season (June–August). Pregnant and lactating females were recorded during the wet months of the year, with a peak observed during February. Testicular mass relative to body mass, testicular volume, and seminiferous tubule diameter in 64% of males increased significantly during the rainfall period (October–June). In addition, 8% of males exhibited little spermatogenesis and 28% showed no spermatogenesis or presence of sperm in the epididymis during the dry period (June–August). These findings suggest that the Gerbilliscus cf. leucogaster breeds predominantly during the rainfall period in Namibia when the food resources are more abundant.

Mating status of brinjal shoot and fruit borer Leucinodes orbonalis Guenee (Crambidae: Lepidoptera) male adults caught in sex pheromone traps

Brinjal Shoot and Fruit Borer, Leucinodes orbonalis , is a major constraint in brinjal production. Investigation was taken up to understand the mating status of L. orbonalis male adults caught in sex pheromone trap. The genetalia of the male moths were examined for the presence of sperm cells and sperm bundles. The results indicated that 55.38 per cent of males caught in the trap were unmated whereas 26.15 per cent of them are partially mated and 18.40 per cent males are spent ones. The study indicated presence of L orbonalis male moths of different mating status in the pheromone catches.

Сравнительная оценка эффективности иммунохроматографических методов обнаружения спермы при судебно-медицинской экспертизе половых преступлений

The article presents a review of the literature of native and foreign authors on the application of confirmatory methods for determining the presence of sperm on material evidences. The article compares the main immunochromatographic methods. It provides analysis of the acts of forensic medical reports on the examination of sexual crimes.

INFECTED FRESHWATER SNAILS, BIOMPHALARIA ALEXANDRINA, THE INTERMEDIATE HOST OF SCHISTOSOMA MANSONI

The objective was to study the chromosomes of Biomphalaria alexandrina snails (class Gastropoda) in control and infected state which could be helpful in understanding how host-parasite relationships in feasible and effective control measures. The chromosomal changes were studied using the air-drying method. The results showed that B. alexandrina had a diploid chromosome number, 2n = 36. Also, the meiotic stages were detected as early- leptotene and late leptotene, zygotene, diplotene, metaphase I. The result also revealed the presence of a primitive sperm, with a conical head and a very long, uni-flagellate tail. A comparative meiotic chromosome analysis between the control and infected ones showed some significant differences, as pachytene and diplotene were more condensed.