Semen Collection Method Research Articles

Sperm collection in the domestic cat: a comparison of two techniques

Semen collection in cats in the clinic setting can be difficult. However, semen analysis is vital when evaluating breeding soundness of a male. Electroejaculation (EEJ) is currently the most reliable semen collection method but requires specialized equipment and training of the operator. Chemical ejaculation followed by urethral catheterization (UC) is a technique that allows semen collection without special equipment: a catheter is placed into the urethra of a sedated tom and semen is collected passively into the catheter. Earlier studies used the sedative medetomidine at high doses for this procedure. However, medetomidine has been replaced with dexmedetomidine in some countries. This study sought to compare the results of EEJ and UC for semen collection in the domestic cat using dexmedetomidine, a potent α2-adrenoceptor agonist (α2A), as a substitute for medetomidine at the equivalent dose to that used in earlier studies. Twelve domestic cats were collected thrice at weekly intervals. All cats received intramuscular ketamine (5 mg/kg) and intramuscular dexmedetomidine (30 μg/kg) for initial cleanout via EEJ, then randomly underwent either EEJ or UC one week apart. The EEJ was performed under the same anesthetic protocol as the initial cleanout. The UC was performed using intramuscular dexmedetomidine at a dose of 60 μg/kg. Success of collection, total sperm number, sperm morphology, and motility characteristics were analyzed. Sperm was collected successfully from all 12 cats via EEJ and from 11/12 via UC. There were no significant differences in the percentage of total motile, progressively motile, or morphologically normal sperm between ejaculate types when averaged across all cats or individual cats. Although UC yielded a lower volume and higher concentration ejaculate, it consistently produced a lower total sperm number than ejaculates retrieved via EEJ (17.91 x 106 total sperm for UC versus 46.51 x 106 total sperm for EEJ). These results indicated that dexmedetomidine is a very effective sedative and performed satisfactorily in both procedures at the doses used in this study. It was also safe with no adverse effects on healthy toms. EEJ remained the most reliable in terms of assessing semen quality and retrieving semen with adequate number of sperm for breeding purposes. However, UC with dexmedetomidine at this dose demonstrated a 92% success rate, presenting itself as a remarkably consistent alternative.

Microbiota Analysis of Ejaculated Honey Bee Drone Semen and the Effect of Semen Collection Method on Bacterial Loads.

Artificial insemination in queen honey bees is the only tool that provides complete control over mating for research and breeding purposes, making it essential in genetic improvement and conservation programs in this species. The aims of this study were to characterize drone semen bacterial loads by culture-dependent and independent methods and to describe their variation depending on the method of semen collection, the colony and the apiary. In the first experiment, the bacterial loads of semen collected from the seminal vesicles or from ejaculates was studied using culture-dependent methods. The collection method had a significant influence on the overall bacterial count in semen. Out of the 42 semen samples analyzed, 26 (61.9%) tested positive for bacterial isolation. This encompassed the entirety of samples obtained from the seminal vesicles (21 of 21), whereas only 23.8% of those derived from ejaculates (5 out of 21) showed bacterial isolation. In the second experiment, next-generation sequencing techniques were used to describe the microbiome of ejaculated drone semen for the first time. The most abundant phyla were Proteobacteria, Firmicutes, Bacteroidota and Actinobacteriota, while the most abundant genera were Lactobacillus, Staphylococcus, Prevotella, Alloprevotella and Streptococcus. The results showed that the apiary had a significant effect on the community structure composition and abundance of the seminal microbiota, and significative differences in abundance were observed for the genera Sphingomonas, Methylobacterium-Methylorubrum, Bifidobacterium and Alloprevotella. Significant differences were also observed in the richness of the microbiota between apiaries and colonies.

Decoding the influence of semen collection processes on goat sperm quality from a perspective of seminal plasma proteomics

This study aims to assess the impact of semen collection methods on goat semen quality and seminal plasma (SP) proteomes. Semen was collected by artificial vagina (AV) or electro-ejaculator (EE) and semen parameters were evaluated. Tandem mass tag coupled with liquid chromatography–tandem mass spectrometry was used to screen SP differentially abundant proteins (DAPs) between EE and AV. PRM was used to confirm the reliability of the data. In contrast to EE, a lower volume, higher progressive motility and concentration were observed in AV. No differences were found in total motility, membrane integrity, acrosome integrity, and ROS production between EE and AV. In total, 1692 proteins were identified in SP, including 210 DAPs. Among them, 120 and 90 proteins were down-regulated and up-regulated in AV compared to EE, respectively. The GO annotation showed that DAPs are mainly localized in the membrane, involved in deference responses to bacterium, RNA processing, and related to oxidoreductase activity. KEGG demonstrated tight associations of DAPs with specific amino acids, carbon metabolism, citrate cycle, and propanoate metabolism. In conclusion, this study provides valuable insights into the effects of semen collection on goat semen quality and explores the potential action mechanism based on the modification of SP proteomes. Significance of the studyThe quality of fresh semen directly influences the results of artificial insemination and semen cryopreservation in livestock. This study represents the first attempt to evaluate the impact of semen collection methods including electroejaculation and artificial vagina on sperm quality and seminal plasma proteomes in goat. The results of this study demonstrated that semen collection methods directly impacted the quality of goat semen. Then, the proteomic strategy was used to explore the potential action mechanism of semen collection methods on sperm. Some differentially abundant proteins that potentially influence semen quality were identified. Furthermore, this study suggests the possibility of utilizing specific proteins as predictive markers for goat semen quality.

Evaluating sperm quality characteristics obtained through the female teaser method in native chicken breeds

<p class="MDPI17abstract"><strong>Objective: </strong>The quality and quantity of chicken sperm are critical factors influenced by the semen collection method<strong>. </strong>Therefore, this study aims to evaluate the sperm quality obtained from three native chicken breeds using the female teaser method.</p><p class="MDPI17abstract"><strong>Methods: </strong>These three native chicken breeds are Arabic, Birma, and Kampung Unggul Balitbangtan (KUB) chickens. Each breed comprised a group of three roosters, and the semen collection was performed ten times. Macroscopic and microscopic evaluations were conducted to assess the chicken sperm. The collected data were analyzed using Analysis of Variance (ANOVA) followed by Duncan's multiple range test (DMRT).<strong></strong></p><p class="MDPI17abstract"><strong>Result: </strong>The results of this study showed significant differences (P<0.05) in the sperm volume among the chicken breeds, with Birma (0.51 ml) and KUB chickens (0.60 ml) exhibiting distinct volumes compared to Arabic chicken (0.25 ml). Generally, the chicken sperm exhibited a milky white color and thick consistency. Microscopic evaluation yielded a mass motility rating of 3+. Meanwhile, individual motility and viability did not exhibit significant differences (P>0.05) among the three native chicken breeds. Abnormalities displayed significant variations (P<0.05), with Birma chicken showing the highest abnormality rate at 9.77%± 0.28. Sperm concentration did not exhibit significant differences (P>0.05), with the highest concentration observed in Arabic chicken at 4265.33±59.84 (x10<sup>6</sup> cells/ml). However, total sperm count and total motile sperm count exhibited significant variations (P<0.05), with the highest values recorded in KUB chicken at 2241.82±264.13 (x10<sup>6</sup> cells) and 1936.27±169.85 (x10<sup>6</sup> cells), respectively.<strong></strong></p><p class="MDPI17abstract"><strong>Conclusions: </strong>In conclusion, the semen collection method using the female teaser yielded native chicken sperm with high quality and quantity and meets the required standards for successful insemination. This method holds potential and is recommended for application in native chicken breeding programs.<strong></strong></p>

Semen Quality of Captive Fishing Cats (Prionailurus viverrinus) Obtained by Electroejaculation

Fishing cat (Prionailurus viverrinus) is categorized in a group of small cats. The cat has brownish grey fur with dark brown spots and lines running across the length of body. It is wildlife conserved (IUCN) and protected as wildlife animal in Thailand. It is also in the CITES Appendix II. The aim of this study was to study semen quality of this cat by electroejaculation. Five male fishing cats aging approximately 13 years old and body weighed between 9-16 kg were employed in this study. Anesthetic by dexmedetomidine (0.02 mg/kg) and ketamine (4 mg/kg)) was used before semen collection. Stimulation patterns for electrical stimuli using 0.5-5 volts by autonomous program were applied into the rectum of each fishing cat using the rectal probe (r=1.27 cm). Semen was successfully collected from sixteen out of twenty attempts. Semen volume and sperm motility were much variable. Average semen volume was 43.13 µl (range between 5-200 µl). Average pH was 5.81 (range from 5-7). Average sperm motility was 28.25 % (range from 1-90 %). Average sperm concentration was 207.44 million/ml (range from 12-860 million/ml). The semen collection method developed in this study proved to be a useful technique for frozen semen preservation and planning of artificial insemination in order to propagate and conserve the threatened and endangered fishing cat species in the future.

Effect of Electroejaculation Protocols on Semen Quality and Concentrations of Testosterone, Cortisol, Malondialdehyde, and Creatine Kinase in Captive Bengal Tigers

The Bengal tiger (Panthera tigris tigris) is critically endangered, so assisted reproductive technologies, including artificial insemination, are important conservation tools. For wild and domestic felids, electroejaculation (EE) is the most common semen collection method, with protocols optimized to obtain sufficient amounts of viable sperm for artificial insemination. However, less attention has been paid to ensuring animal wellbeing during the process. This study examined the effects of three EE protocols (Low, 2–5 volts; Medium, 3–6 volts; High, 4–7 volts) on semen quality, testicular size, serum testosterone, creatine kinase (CK), and malondialdehyde (MDA) concentrations, and serum cortisol as a proxy for stress. Blood samples were collected before, during, and after each EE series. Seminal plasma pH, and sperm motility, viability, and morphology were evaluated after each procedure. Seminal plasma and sperm pellet MDA concentrations were also determined. Primary sperm abnormalities and seminal plasma MDA were higher in the Low compared to Medium and High voltage groups (p < 0.05). Serum CK in the High voltage group increased during the EE series (p < 0.05), suggesting the potential for muscle damage. However, no significant changes were observed for serum cortisol, testosterone, or MDA concentrations. Results suggest the Medium voltage protocol produced good quality samples at lower voltages than the High protocol with no negative effect on muscle function, which might be better for animal welfare.

Modified collection methods and quality of semen in Nigerian local turkeys (Meleagris gallopavo)

Semen collection in birds was initially achieved either via post-mortem drainage of the vas deferens or by cloacal retrieval after natural mating. Current techniques do not exclude stress and risk of injury during restraint or milking, especially in domestic turkeys. The present study was designed to develop more convenient methods of semen collection in turkeys. It involved five (n = 5) matured turkey toms weighing approximately 12 – 15 kg and two (n = 2) turkey hens. In the improved collection method, toms were stimulated by exposing them to hens, covered using a locally constructed wooden box. They were allowed to mount until there was evidence of ejaculatory response, demonstrated by increased abdominal pressure and cloacal contraction. After mounting, the tail feathers were lifted, and a collection vial was placed immediately below the cloaca to collect the ejaculates. In the modified abdominal massage, the turkey toms were restrained on sterna recumbency while the ejaculatory response was initiated by gently massaging the soft part of the abdomen and backward massage of the tail feathers. This is continued by rubbing either side of the cloaca till erection is achieved and semen is milked into a collection vial by gently squeezing the cloaca. Collections were made twice a week for four weeks using each method independently. The mean semen volume in the improved collection method was significantly (p < 0.05) higher than the modified abdominal massage. There was no significant (p > 0.05) difference in motility, abnormalities and concentration between both methods. The results from this study suggest that both collection methods were satisfactory for practical use and may offset some challenges associated with the most widely used abdominal massage technique of semen collection in turkeys.

Characterization of basal seminal traits and semen cryopreservation in Canada lynx (Lynx canadensis)

Little is known about the reproductive biology of the Canada lynx; virtually no data are available describing seminal parameters in this species and sperm cryopreservation studies have not been performed. Our aims were to 1) evaluate effectiveness of two semen collection methods: urethral catheterization (UC) and electroejaculation (EEJ); 2) characterize basal seminal traits throughout the breeding season; 3) compare effectiveness of semen cryopreservation using TEST egg yolk (TEY) or soy lecithin-based (SOY) media; and, 4) evaluate the ability of preserved Canada lynx sperm to fertilize domestic cat in vitro matured oocytes by heterologous in vitro fertilization. Testicular volume averaged 4.5±1.0 cm3. A significant relationship was found between animal body weight and testes volume (P<0.02), as well as for testicular volume and sperm counts per ejaculate (P<0.01). Fecal testosterone concentrations and sperm production were correlated (P<0.01). Electroejaculation was more reliable than UC for sperm collection (8.8 and 1.5 × 106 sperm, respectively; P<0.01). The percentage of sperm with normal morphology was greater in EEJ (29 ± 11 %) than in UC (18 ± 9 %) (P<0.05) samples and primary sperm structural abnormalities were more frequent in UC than EEJ samples (P<0.01). Overall, sperm quality was low in most males throughout the breeding season, but consistent with previous findings in the Lynx genus. Sperm production averaged 10.8 × 106 sperm/ejaculate with 45 % motility; 29 % normal morphology and 69 % acrosome integrity. Motility and acrosome integrity post-thaw declined over time, but were similar between spermatozoa cryopreserved in TEY and SOY media (P > 0.05). Cleavage rates did not differ between TEY (29 ± 17 %) and SOY (27 ± 14 %) and mean numbers of sperm bound to the zona pellucida was similar (20 sperm, TEY vs. 18 sperm, SOY), indicating that both extenders equally preserved the function of Canada lynx sperm for semen banking purposes. Our findings provide zoos and population managers with valuable information about normative reproductive traits in this species, establishing comparative benchmarks for assessing the reproductive potential of specific males recommended for breeding.

Alternative Opportunities to Collect Semen and Sperm Cells for Ex Situ In Vitro Gene Conservation in Sheep

The semen of domestic mammals is conventionally collected with an artificial vagina (AV) for artificial insemination (AI) or for short- or long-term storage. However, the procedure has certain drawbacks: animal training is not feasible in extensive animal care systems nor among wild species, as the trained animals sometimes fail to mount. Hence, there is a need for alternative semen collection methods. Electroejaculation (EEJ) and epididymal sperm recovery (ESR) are the two effective alternatives to AV. However, in recent years, animal welfare campaigners have called for the ban, in certain EU countries, of EEJ due to its inhumane nature. In this review, alternative methods of sperm collection (by EEJ and ESR, their qualities, and their freezing techniques) are highlighted, as well as the effects of EEJ on pre-freeze and post-thaw ram sperm quality parameters and the animal welfare progress made in EEJ between the 20th and 21st centuries. Additionally, the techniques for enhancing post-thaw sperm quality prior to freezing and for the freezing of EEJ and ESR spermatozoa are explored. ESR and EEJ are reliable alternatives to AV on certain occasions. EEJ is ideal for semen collection in wild or untrained animals, breeding soundness examinations, collection outside of the breeding season, and culling. At the same time, ESR is ideal in cases of castration, accidental death of elite sire, or postmortem for gene conservation purposes or assisted reproductive technologies (ARTs) studies.

Quercetin: Putative effects on the function of cryopreserved sperms in domestic animals.

Quercetin is one of the most used antioxidant flavonoids and largely exists in many fruits and vegetables because of its capability to scavenge the free reactive oxygen species (ROSs) by repressing lipid peroxy radical fusion, metal ion chelating through enzyme inhibition, and adopting the repair mechanisms. It also exhibits various biological actions, including antioxidative, anti-inflammatory and antimicrobial activities. Furthermore, it contributes well to sustaining the endogenous cellular antioxidant defence system. The process of cryopreservation is associated with increased oxidative stress, and some steps are potential sources of ROSs, including the method of semen collection, handling, cryopreservation culture media, and thawing, which result in impaired sperm function. Several antioxidants have been proposed to counteract the harmful impact of ROS during semen cryopreservation. The antioxidant capability of quercetin has been verified in different animal species for providing valuable defence to sperm during the cryopreservation process. The beneficial properties of quercetin on various parameters of fresh and post-thaw sperm in different species are clarified in this review. More in-depth investigations are required to clarify quercetin's mechanism of action in different animal species.

A novel, patented method for semen collection in dromedary camel (Camelus dromedarius).

In the current article, a developed, patented method denoted the 'Camel Semen Collection Kit-CSCK', was designed to solve the problem of semen collection in dromedary camels. CSCK is composed of three main parts: (1) Semen collection sac: made from supersensitive flexible low-density polyethylene- (LDPE); (2) Metal stainless steel applicator: designed to introduce the collection sac intravaginally and fixate it to the vaginal wall of a female camel through air insufflation; (3) Fixation sticker: a cushion sheet sticker is used to secure the outer portion of the collection sac to the female's perineal area. Semen was collected twice a week from eight dromedary bulls by using electroejaculation (EJ), artificial vagina (AV) and CSCK. Successful semen collections were 81.3%, 84.4% and 43.8% using EJ, CSCK and AV techniques respectively. Semen obtained by EJ technique showed lower semen volume, gross activity, sperm concentration, total sperm motility and percentage of live sperm cells compared to the other two techniques. Semen collected by CSCK showed a longer collection period and higher volume, gross activity, sperm motility and percentage of live spermatozoa and a lower rate of visible contamination compared to AV technique. The advantages and disadvantages of the three techniques were compared and discussed. In conclusion, CSCK represents a practical and easy method to reliably collect high-quality semen from any untrained male dromedary camel and may facilitate the widespread application of assisted reproductive technologies (ARTs) on a large scale in this species.

Male Fertility Preservation: A boon for young cancer survivors

Diagnosis of any ailment especially cancer is found to be pivotal to evaluating the type of treatment that needs to be administered to man. It aids in subsequent prognosis and timely recovery in patients. When concerned with male cancer survivors, the emphasis on their fertility health is always an issue. As the numbers of survivors are increasing day by day due to the advanced medical and technological approaches, man could look with confidence to a life of ease from cancer. To review and compile all the feasible as well as relevant information about the preservation of male fertility from published resources. Reputed databases were searched for content based on specific keywords like “fertility preservation after cancer treatment", "methods of male gamete preservation", "methods of semen collection for preservation", "fertility preservation", "erectile dysfunction" and "testicular cancer and fertility". The year of publication for articles under study was restricted from 2016-2021 in most of the databases. It was found that oncologists generally recommended preservation of the male fertility before the commencement of the cancer treatment procedures. Preservation of fertility among young men should be considered in all patients before initiating any kind of prognosis related to the disease.

An easy, safe, and practical method for semen collection in dromedary camels

In the current study, we have developed an easy, practical, and reliable method to collect semen from camel bulls. Simply, intromission ofa super-sensitive artificial insemination glove intravaginally with an outer fixation in the perineal region from lateral and dorsal sides of thevulva, mating, glove withdrawal and semen collection were the successive steps. Bull behaviour before and during the mating process wasobserved. The collected semen was evaluated in aspects of semen colour, volume and sperm cell concentration. Also, individual spermmotility and semen fertility by applying in-vitro fertilization were investigated after semen dilution in a new developed extender for camelsemen (100 mmol/l Tris, 65 mmol/l lactose, 50 mmol/l glucose, 40 mmol/l galactose, 125 mg/100ml BSA, 50 IU/ml catalase enzyme,1000 IU/ml penicillin G and 500µg/ml streptomycin: pH 7.0). Presence of the intravaginal glove and its fixation by the external adhesivetape did not affect the bull willingness and did not interrupt the mating process. Semen collection was successful in 112 out of total128 trials (87.5%) and the lengthy copulation was (10.3 ± 6.2 min - Mean ± SD). Mean parameters of the collected ejaculates were:volume of 8.3 ± 3.6 ml (Mean ± SD); sperm cell concentration of 437.6 ± 62.4 X 106 (Mean ± SD) sperm/ml; and sperm individualmotility after liquefaction for 45 min was 68.2 ± 11.4 % (Mean ± SD). Semen was grayish white to creamy white in colour dependingon sperm cell concentration. Finally, using of the collected diluted semen in IVF technique showed cleaved, morula and blastocyst rates66.1%, 43.5% and 25.8%, respectively. In conclusion, using of intravaginal super-sensitive AI gloves is a practical technique for semencollection and will facilitate the application of the new reproductive technologies on a large scale in dromedary camels.Keywords: Semen collection, Camel, In-vitro fertilization

Detection of SARS-CoV-2 in semen samples: a scoping review

Background: many doubts about the infection of SARS-CoV-2 were raised, such as sexual transmission, sterility, and changes in fertility procedures; however, information is not clearly stated and organized. Purpose: to review and summarize scientific evidence on detection of SARS-CoV-2 in semen samples of Covid-19 patients. Methods: literature search in PubMed, Web of Science, Scopus, Medline and Embase databases, and followed Scoping Review protocol defined by Joanna Briggs Institute (JBI) after the guiding question “Is it possible to detect SARS-CoV-2 in the semen of adult patients with a confirmed diagnosis of Covid-19?” Results: 287 studies were identified, and, after discerning analysis, 9 studies published in the English language were selected. Three researchers analyzed the studies for SARS-CoV-2 presence in the seminal fluid, patients’ severity, days since the onset of disease, diagnosis confirmation, semen collection method, viral analysis method, and sample numbers. Conclusions: it was not possible to find strong evidence to confirm the presence or absence of Covid-19 in the semen of adult patients. New studies on the subject should be better designed, taking into account the possible anatomical and functional conditions and changes of the male reproductive system during and after the infection by SARS-CoV-2.

21 Impact of Breed and Method of Semen Collection on Viability and Membrane Integrity of Goat Spermatozoa

Abstract Goats have potential to be genetically improved for meat and milk. Artificial insemination is an important technique used to enhance genetic gain through dissemination of selected male traits. One reason for huge variability in fertility rates in goats is the semen quality, which is affected by breed, collection method, and freezing procedures. Here we investigated effect of breeds (Saanen versus Alpine) and semen collection methods (artificial vagina; AV versus electroejaculation; EE) on the quality of goat spermatozoa. Semen collected from Saanen and Alpine bucks by either AV or EE was evaluated for its color, consistency, and volume. Besides, sperm motility, concentrations, viability, abnormalities, and membrane integrity were assessed. At least three ejaculates were collected from each breed and data were analyzed by student t-test. Results showed that colors of samples collected from the two breeds were white to yellow and the consistency ranged from watery to creamy. Irrespective to the collection method, average volume of ejaculates was higher for Alpine than Saanen (1350 vs. 880 µl for AV and 1066 vs. 987.5 µl for EE). Sperm motility and concentrations were higher (P ≤ 0.05) in Alpine than Saanen semen for both methods. Within the same breed, these parameters were higher (P ≤ 0.05) for AV than EE. Percentage of viable spermatozoa was higher for Alpine than Saanen bucks (80.0% vs. 64.5% in AV) and (52.42% vs. 41.35% in EE). However, sperm abnormalities were lower (P ≤ 0.05) in Alpine semen. Sperm membrane integrity tended to be higher in Alpine semen. In conclusion, semen collected from Alpine bucks had higher quality than those obtained from Saanen ones. Furthermore, collection of semen by AV improved sperm motility and viability than EE in both breeds. These findings suggest that both breeds and method of semen collection could influence the viability and fertility of goat sperm.

Practical Method for Freezing Buck Semen.

Simple SummaryGoat semen was previously considered to be problematic to freeze because of reactions between the semen and the components of the freezing media that were available at the time. However, there have been reports of several successful attempts to freeze goat semen in recent decades using various protocols, resulting in usable post-thaw sperm samples. In the present study, we adapted some of these methods to suit the particular conditions under which we had to work. We were able to produce thawed samples with acceptable sperm quality which were sent to a sperm bank for long-term storage.Although several protocols for cryopreserving buck semen are described in the literature, they differ widely in factors such as season and method of semen collection, extender and sperm concentration. Therefore, choosing a protocol that is suitable for a particular on-farm situation can be problematic. In the present study, semen was collected by artificial vagina from seven bucks on a farm located approximately 90 minutes’ drive away from the laboratory, about 6 weeks before the start of the goat breeding season. The semen was immediately extended in warm semen extender containing soy lecithin and was placed in an insulated box with a cold pack for up to 4 h, during semen collection from the remaining bucks and subsequent transport to the laboratory. Following centrifugation at 4 °C and resuspension in the soy lecithin extender to a sperm concentration of 800 × 106 spermatozoa/mL, 0.25 mL plastic straws were filled and frozen in racks 4 cm above the surface of liquid nitrogen. This simple protocol resulted in an acceptable post-thaw quality for all seven bucks, with a mean post-thaw motility of 55 ± 21% and mean fragmented chromatin of 3.27 ± 1.39%. Normal sperm morphology was >90% in all ejaculates. The semen was sent to a gamete bank for long-term storage.

Is Male Infertility a Concern of Time? Comparison of Semen Parameters over a Five Year Period among Sub-fertile Men Visiting a Fertility Centre in Chennai, Tamil Nadu, India

Introduction: Semen parameters, especially sperm count, concentration, progressive motility and volume have been said to be diminishing over time among men in developing countries like India. Aim: To compare the semen parameters (semen concentration, normal forms, progressive motility and defects) of sub-fertile males visiting ARC fertility Centre in Chennai. Materials and Methods: This cross-sectional study was conducted in Advanced Reproductive Care, Inc. (ARC) International Fertility and Research Centre located in Chennai, Tamil Nadu, India, from the period of January 2015 to December 2019. Total 50 semen samples were selected randomly each year bringing the total to 250 samples which were included in the study. Masturbation was the major method of semen collection. The semen was analysed for the various semen parameters like semen concentration, normal forms, progressive motility and defects in the laboratory under microscope on wet preparation of slides as per the standard guidelines. The various semen parameters along with the age of the participants were entered in Microsoft excel and analysed by Statistical Package for the Social Sciences (SPSS) version 22.0. Results: Total 250 semen reports were included in the study. They belong to age group of persons between 20 to 60 years. More than half of them were 30 to 40 year for all the five years. It was found that there was a statistically significant mean difference among progressive motility (25.14±16.37 in 2015 to 17.30±11.41 in 2019; p-value=0.009), semen concentration (78.1±61.15 in 2015 to 65.96±57.85 in 2019; p-value=0.001) among the semen samples over the years. There was a decreasing trend with the semen parameters being lower than optimal over the years. Conclusion: The study findings suggested that there is a decrease in semen parameters over the years and further studies have to be carried out to find the various environmental and lifestyle factors which could be responsible for the same.

Comparative ejaculatory response, fresh and frozen semen quality and fertility to artificial vagina vs electroejaculation method of semen collection in mithun (Bos frontalis) bulls.

The study aimed to describe a comparative ejaculatory response, fresh and frozen-thawed semen quality and fertility of semen collected by artificial vagina and electroejaculation in mithun. Experimental bulls were divided into two groups, G-I: young bulls (n=4; 4-5years of age) and G-II: older bulls (n=4; 8-10years of age). Sixteen ejaculates were collected from each group G-I (AV1 ) and G-II (AV2 ) by artificial vagina method (control). Thirty-six ejaculates were collected from the same bulls from each group G-I (EE1 ) and G-II (EE2 ) by electroejaculation method (treatment). The study did not reveal any significant (p>0.05) difference in the ejaculatory responses between EE1 and EE2 . Mann-Whitney U test indicated that salivation discomfort sign score was significantly (p<0.05) higher in EE1 . Fresh and frozen-thawed semen quality parameters, and motility and velocity profiles recorded by computer-assisted sperm analyser were significantly (p<0.05) lower in electroejaculation than the artificial vagina. The conception rates (AV1 vs EE1 & AV2 vs EE2 ) at day 35-45 post insemination were nonsignificantly higher (p>0.05) in the artificial vagina group. It concluded that, although artificial vagina method has better semen quality, nevertheless, electroejaculation has the potential for semen collection from free-range mithun bulls to incorporate in assisted reproductive technology procedures.

Pharmacologically Induced Ex Copula Ejaculation in Horses and Donkeys.

Pharmacologically induced ex copula ejaculation is a method used for collection of semen when the traditional methods of semen collection are not feasible. Common indications for this method include health issues that either preclude the physical act of mating or result in impaired erection and ejaculation. The method also offers an alternative when there is a lack of equipment and facilities required for semen collection using the conventional artificial vagina method. A variety of pharmacological protocols for ex copula ejaculation in stallions have been evaluated in both research and clinical settings with no serious side-effects reported. In general, these protocols included tricyclic antidepressants, alpha-2 adrenergic receptor agonists, and smooth muscle stimulators, either individually or in combination. Although there is a lot of variation in the ejaculatory rates among studies, a combination of imipramine and xylazine appears to be an effective option for inducing ejaculation in stallions. In cases where this protocol is not successful, collections should be reattempted using a combination of imipramine, detomidine, and oxytocin. Regardless of the protocol used, a quiet environment with minimal disturbance is associated with a better outcome. In contrast to the body of literature available on pharmacologically induced ex copula ejaculation in horses, only a few studies have been published so far on this topic in donkeys. Further studies are warranted to investigate whether pharmacologically induced ex copula ejaculation is an effective method of semen collection in jacks and to compare different pharmacological protocols for inducing ejaculation in jacks.

From the Semen Collection Method to the Hatchlings: The Use of Cryopreserved Sperm from Pheasants Fed an Antioxidant-Enriched Diet.

Simple SummaryAlthough cryopreservation techniques for use in bird species have advanced greatly over recent decades, especially in relation to domestic species, major gaps in our knowledge and technical capacities remain due to the complexity of the process and the unique particularities of sperm from different species. The hatchability of chicks is the decisive parameter that demonstrates the quality of a frozen–thawed sperm. Since very little information has been published about the common pheasant, a total of six artificial inseminations (AIs) were performed at 3–4-day intervals with doses of 35 × 106 of normal live thawed sperm on a total of 40 females. The inseminated sperm were collected from pheasants fed either a basal diet or an antioxidant-enriched diet and were then processed using a pellet freezing–thawing method, in which dimethylacetamide was used as a cryoprotectant. Regardless of the male birds’ dietary group, the resulting fertility rate from frozen–thawed sperm was approx. 30%, with 8–9 chicks hatching for every 100 eggs incubated.A widely used approach to preserving genetic diversity in birds involves the cryopreservation of semen. In this process, cells are subjected to physical and chemical stresses, but not all cell species respond equally. Many studies have been published on the freezing–thawing of sperm cells from a wide variety of domestic and wild species, on issues ranging from the sperm quality to different protocols, fertilisation success rates, etc. Nevertheless, very little information is available on the common pheasant. To fill this gap, the aim of this study was to describe the pheasant semen collection method, evaluate some qualitative parameters of sperm from males fed an antioxidant-enriched diet, and to test the in vivo fertilising capacity of the cryo-preserved semen. The freezing protocol employed involved pellets thawed by the hotplate method. Dimethylacetamide was used as a cryoprotectant at a final concentration of 6%. A total of six AIs were performed at 3-4-day intervals on a total of 40 females with doses of 35 × 106 of normal live thawed sperm. Males receiving the enriched diet produce more abundant and concentrated ejaculates. Freeze–thawed sperm lost 85% of their initial mobility, and diet influenced neither sperm mobility nor viability. The enriched diet did improve the number of normal freeze–thawed cells and was associated with a lower sperm fracture incidence. Regardless of the dietary group, frozen–thawed sperm resulted in a fertility rate of 30%, with 8-9 chicks hatching for every 100 eggs incubated.