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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 10 6 total sperm for UC versus 46.51 x 10 6 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., Competing Interests: Declaration of competing interest There are no conflicts of interest to declare., (Copyright © 2024 Elsevier Inc. All rights reserved.)

As the number of cancer survivors increases, so does the demand for preserving male fertility after radiation. It is important for healthcare providers to understand the pathophysiology of radiation-induced testicular injury, the techniques of fertility preservation both before and during radiation, and their role in counseling patients on the risks to their fertility and the means of mitigating these risks. Impaired spermatogenesis is a known testicular toxicity of radiation in both the acute and the late settings, as rapidly dividing spermatogonial germ cells are exquisitely sensitive to irradiation. The threshold for spermatogonial injury and subsequent impairment in spermatogenesis is ~ 0.1 Gy and the severity of gonadal injury is highly dose-dependent. Total doses < 4 Gy may allow for recovery of spermatogenesis and fertility potential, but with larger doses, recovery may be protracted or impossible. All patients undergoing gonadotoxic radiation therapy should be counseled on the possibility of future infertility, offered the opportunity for semen cryopreservation, and offered referral to a fertility specialist. In addition to this, every effort should be made to shield the testes (if not expected to contain tumor) during therapy., Competing Interests: Competing interests: N/A. Conflict of interest: This research received no specific grant from any funding agency in the public, commercial or not-for-profit sectors. The authors declare no potential conflicts of interest, including financial interests, activities, relationships and affiliations., (© 2023 Greater Poland Cancer Centre.)

Semen collection methods vary greatly and rely on the practitioner's expertise, available materials, and the specific behavioral traits of the male animals involved. When it comes to domestic cats, wild felids, and canids, semen collection is particularly challenging. Thus, given the difficulty of semen collection in these species, pharmacological semen collection (PSC) stands out since it is a quick and straightforward method that does not require specific equipment. The PSC consists of administering α2-adrenergic receptor agonist drugs, mainly medetomidine, and dexmedetomidine, aiming semen release into the urethra with posterior urethral catheterization and sperm recovery. This technique was primarily described in domestic cats and wild felids, and despite the decreased seminal volume, the retrieved semen is highly concentrated and presents good quality. However, further studies are required to optimize semen collection in domestic dogs and wild canids. Therefore, the purpose of this review is to provide a comprehensive overview of the research developed on pharmacological semen collection (PSC) in the past few decades. The objective is to equip professionals with the essential knowledge required for the efficient application of this technique in both domestic and wild canids and felids and to make a valuable contribution to conservation efforts and the preservation of biodiversity, aligning with the principles of One Conservation., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare

Androgens and estrogens are steroid hormones that regulate reproductive processes in both males and females. Monitoring plasma levels of these steroids or their metabolites present in feces, offers diagnostic support for assessing the reproductive status of animals. Immunoassays are commonly used methods for quantifying these hormones, but their protocols require species-specific validation to ensure reliability. The objective of this study was to perform analytically and biologically validation of enzyme immunoassay (EIA) kits for measuring testosterone (T), estradiol (E 2 ), faecal androgen metabolites (fAM), and faecal estrogenic metabolites (fEM) in the tropical screech owl (Megascops choliba). Serum and fecal samples were collected from six adult females and six males both before and during breeding season, with males' gonadal activity assessed using electroejaculation (EE). The parallelism test confirmed the immunogenic similarity of the antigens in the estradiol and testosterone standards and the antigens in the serum samples and fecal extracts of M. choliba. Additionally, the EIA kits displayed nearly 100% recovery rates, and showed coefficients of variation ranging from 8% to 14% at the intra-assay level and from 10% to 16% at the inter-assay level, underscoring result reliability and consistency. In males, the highest serum T and fAM levels were recorded concurrently with the presence of spermatozoa in samples collected via EE. Although females did not exhibit oviposition events, significantly higher E 2 and fEM levels were observed in August compared to May, suggesting potential seasonal variations in estrogenic hormone production. Fecal androgen and estrogen levels were significantly different between sexes in August, with males having higher fAM and females having higher fEM levels. Overall, the immunoassays validated in this study were found to be efficient in diagnosing reproductive activity in owls., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

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.

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., (© 2022 Wiley-VCH GmbH.)

Transrectal ultrasonic-guided massage of the accessory sex glands (TUMASG) is a technique that allows collecting semen requiring few electrical stimuli or even no pulse. A long-acting analogue of oxytocin (carbetocin, 0.1 mg) was i.v. administered before TUMASG in 10 conscious bucks (Experiment 1) and 10 anaesthetized Iberian ibexes (Experiment 2) to shorten the time of semen collection, decrease the number of electrical stimuli and/or improve the semen quality. The ejaculated volume, concentration, quality parameters and kinetics variables of the sperm were determined in fresh semen. The time length of the procedures and the number of electric pulses applied were recorded. Furthermore, stress response indicators (number of vocalizations in Experiment 1; heart and respiratory rates, rectal temperature, cortisol levels, totals proteins and neutrophil-to-lymphocyte ratio in Experiment 2) were documented. In bucks, the administration of carbetocin tended to shorten the time needed for semen collection but no-showed differences in the fresh seminal quality. In the Iberian ibexes, there were no significant differences between groups in the time length of procedures or in the number of animals that ejaculated. Carbetocin administration only reduced the respiratory rate, did it modify fresh semen characteristics in ibexes. In conclusion, the administration of carbetocin did not appear as a useful tool to improve welfare during semen collection with TUMASG or semen quality in conscious bucks and anaesthetized ibexes, having only slight advantages related to the procedure., (© 2022 Wiley-VCH GmbH.)

Oxytocin is a peptide hormone that mainly functions to control the contractility of smooth muscles and sex-related steroidogenesis in male reproductive tracts. However, specific information concerning this hormone in controlling the reproductive organs of cats is limited. This study aimed to investigate the expression of oxytocin receptors (OTRs) and their signal mediator via prostacyclin synthase (PTGIS) in reproductive structures following oxytocin assisted electroejaculation. In Experiment 1, the testis, cauda epididymis and vas deferens from five cats were examined by immunohistochemistry and quantitative polymerase chain reaction in order to study the responses of OTR and PTGIS mRNA to oxytocin injection. Experiment 2 examined the effect of oxytocin administration prior to electroejaculation on ejaculate characteristics and sperm quality in terms of motility, viability and fertilizing ability. Immunohistochemistry revealed the expression of OTRs in Leydig's, peritubular myoid cells and some spermatogenic cells. The expression was found in the epithelium and smooth muscle of the epididymis and vas deferens. After oxytocin administration, the OTR mRNA was upregulated in the epididymis (p > .05) and vas deferens (p = .01). The expression level of PTGIS mRNA increased in the response to oxytocin treatment only for the vas deferens (p > .05). Oxytocin treatment before electroejaculation resulted in an approximately twofold increase in sperm concentration and total sperm output/ejaculate, while this intervention did not significantly affect ejaculate volume, sperm quality or fertilizing ability. This study concluded that the oxytocin cascade is locally present in the reproductive structures and plays a role in promoting sperm delivery during electroejaculation in cats., (© 2022 Wiley-VCH GmbH.)

The aims of this study were to investigate the proteome of koala spermatozoa and that of the prostatic bodies with which they interact during ejaculation. For this purpose, spermatozoa and prostatic bodies were fractionated from the semen of four male koalas and analysed by HPLC MS/MS. This strategy identified 744 sperm and 1297 prostatic body proteins, which were subsequently attributed to 482 and 776 unique gene products, respectively. Gene ontology curation of the sperm proteome revealed an abundance of proteins mapping to the canonical sirtuin and 14-3-3 signalling pathways. By contrast, protein ubiquitination and unfolded protein response pathways dominated the equivalent analysis of proteins uniquely identified in prostatic bodies. Koala sperm proteins featured an enrichment of those mapping to the functional categories of cellular compromise/inflammatory response, whilst those of the prostatic body revealed an over-representation of molecular chaperone and stress-related proteins. Cross-species comparisons demonstrated that the koala sperm proteome displays greater conservation with that of eutherians (human; 93%) as opposed to reptile (crocodile; 39%) and avian (rooster; 27%) spermatozoa. Together, this work contributes to our overall understanding of the core sperm proteome and has identified biomarkers that may contribute to the exceptional longevity of koala spermatozoa during ex vivo storage., (© 2021 Wiley-VCH GmbH.)

The objective of the present study was to evaluate the effect of 24 h' fasting prior to semen collection by electroejaculation on behavioral responses, volume of rectal fecal content, bladder size, penis protrusion, erection, ejaculation stimuli, and ejaculate parameters in young Simmental bulls. Twenty-two Simmental beef bulls with an age of 13.2 ± 1.2 mo (mean ± SD) were used in a prospective randomized blinded controlled cross-over design with two pens fasted for 24 h (n = 9; FAS group), and the other three pens were non-fasted (n = 13; CON group). The bulls were maintained under confined conditions without access to pasture. One week later, the pen treatments were inverted, and semen was collected again under the same conditions and by the same team. The behavioral responses, volume of fecal rectal content, bladder size, as well as the number of stimuli required to obtain penis protrusion, erection, and ejaculation to electroejaculation were measured. The following ejaculate parameters were measured: volume, concentration, spermatozoa motility, and morphology. The behavioral response of the bulls to electroejaculation was not different between the CON group and the FAS group (3.2 ± 0.5 and 3.0 ± 0.7, respectively; P = 0.36). Bladder size was significantly reduced in the FAS group compared with the CON group (2.3 ± 0.8 vs. 2.8 ± 0.9, respectively; P = 0.02). The volume of feces in the rectum was not different between the two groups (CON was 2.3 ± 1.7 and FAS was 3.0 ± 1.8; P = 0.23). Compared with the CON group, the FAS group showed a higher proportion of penis protrusion (100% versus 81.8%, P = 0.10), erection (100% versus 81.8%; P = 0.10), and ejaculation (100% versus 90.9%; P = 0.49). The combined efficiency of penis protrusion, erection, and ejaculation (CE-PPEE) in the FAS group was superior to that of the CON group (P = 0.001). The number of stimuli necessary for penis protrusion, erection, and ejaculation for the CON group were 13.5 ± 3.7, 14.9 ± 3.7, and 20.8 ± 5.8, and they were 15.0 ± 4.2, 16.6 ± 4.2, and 20.2 ± 8.1 for the FAS group. The number of stimuli for penis protrusion (P = 0.09), erection (P = 0.08), and ejaculation (P = 0.77) were no different between the two groups. Ejaculate volume was 4.0 ± 2.6 ml and 4.1 ± 2.3 ml for the CON and FAS groups, respectively (P = 0.90). The motility was 1.4 ± 0.7 and 1.4 ± 0.8 for the CON and FAS groups, respectively (P = 0.72). The concentration of spermatozoa was 336.2 ± 273.1 million and 421.1 ± 300.6 million for the CON and FAS groups, respectively (P = 0.31). The percentage of normal spermatozoa was 50.9 ± 18.8 and 45.6 ± 14.3 for the CON and FAS groups, respectively (P = 0.16). It was concluded that fasting for 24 h prior to semen collection by electroejaculation reduced the bladder size and increased the proportion of bulls with penis protrusion, erection, and ejaculation without any difference detected in behavioral responses, volume of rectal fecal content, and ejaculate parameters., Competing Interests: Declaration of competing interest All authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Objective: To demonstrate the proper technique to perform electroejacuation (EEJ) in men with spinal cord injury (SCI) for the purpose of inducing ejaculation., Design: A video demonstration of the proper technique to perform EEJ in men with SCI using the Seager model 14 electroejaculation machine., Setting: Major university medical center., Patient(s): Men with SCI; institutional review board approval was obtained, and all subjects signed an informed consent form., Intervention(s): Spinal cord injury occurs mostly in young men where the majority suffer from ejaculatory dysfunction. The method of choice to induce ejaculation in penile vibratory stimulation (PVS). PVS is successful in 86% of men with SCI whose level of injury is T10 or rostral. If PVS fails or the level is Caudal to T10, the patient is referred for EEJ. This video will demonstrate the proper technique for successful ejaculation using EEJ. Patients with history of autonomic dysreflexia or their level of injury is T6 or rostral are pretreated with 10-20 mg of nifedipine sublingually 10 minutes before stimulation. The patient is then placed in the lateral decubitus position. The bladder is emptied, and a buffer is instilled. An anoscopy is performed, and a rectal probe is placed. A current is delivered until an antegrade ejaculation is retrieved. A retrograde specimen is collected and examined for sperm identification. Patients with complete SCI (no sensory or motor function is preserved in sacral segments S4-S5) can undergo EEJ without anesthesia. Patients with incomplete SCI (significant nerve sparing or normal sensations) will experience pain during stimulation, and general anesthesia is recommended without the use of muscle relaxing agents., Main Outcome Measure(s): Successful ejaculation after performing EEJ in men with SCI., Result(s): Electroejacuation is successful in 95% of men with SCI and in nearly 100% if general anesthesia is used. Outcomes of in vitro fertilization or intracytoplasmic sperm injection after EEJ showed 37.5% pregnancy rate per cycle, 50.0% pregnancy rate per couple, 33.3% live birth rate per cycle, and 43.8% live birth rate per couple. No complications due to EEJ were observed in 953 trials, and none occurred in the patients presented in this video demonstration., Conclusion(s): Electroejacuation is a safe and reliable method for induction of ejaculation in men with SCI who fail a trial of PVS., (Copyright © 2021 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

The giant anteater (Myrmecophaga tridactyla) is being threatened by natural habitat destruction and fragmentation, illegal hunting and road kills. In this context, the generation of basic information on the reproductive parameters of this species is vital, aiming to improve reproductive management via, amongst others, assisted reproductive technologies. This study aimed to describe the morphological and functional features of semen collected from captive giant anteaters. Electroejaculation was performed in 13 animals housed in zoos located in São Paulo state, Brazil. Semen samples were collected from 13 animals in 16 procedures. Samples were evaluated for volume, motility, vigor, pH, concentration, sperm morphology, and functional tests. The following mean values were obtained: volume 1.28 ± 0.27 mL; motility 28.3 ± 6.2%; vigor 2.4 ± 0.25; concentration 129.4 ± 36.1 × 10 6 sperm/mL; pH 7.4 ± 0.2. Total acrosome, head, midpiece, and tail sperm abnormalities were 3.2 ± 0.8%, 25.4 ± 3.6%, 20.7 ± 3.2%, and 14.7 ± 2.6%, respectively. Intact acrosome was found in 83.7 ± 3.1% and intact membrane in 81.1 ± 4.0% of all samples collected. Mitochondrial activity was 66.4 ± 6.0% (Class I), 18.7 ± 2.9% (Class II), 8.0 ± 2.0% (Class III), 3.9 ± 1.0% (Class IV), and 3.0 ± 0.9% (Class V). Sperm DNA fragmentation rate was 13.2 ± 3.7%. These results indicated that electroejaculation is a feasible method for semen collection in giant anteaters, allowing a more detailed description of the semen in this species., (© 2021 Wiley Periodicals LLC.)

The domestic ferret (Mustela putorius furo) provides a good model for developing new reproductive technologies for use with threatened related species. Such technologies could also be used in the reproductive management of this pet species. The present work reports an improved freezing protocol for ferret sperm. Semen was collected by electroejaculation plus rectal massage (in an attempt to reduce the electrical stimulation necessary) from five adult male ferrets, and then subjected to one of two freezing protocols: (a) from 5 to -35°C at 40°C/min, then from -35 to -65°C at 17°C/min, and finally from -65 to -85°C at 3°C/min-a decelerating freezing rate; and (b) from 5 to - 10°C at 5°C/min, and then from -10 to -130°C at 60°C/min-an accelerating freezing rate. After thawing, the viability and acrosomal integrity of the sperm frozen via the two-step accelerating method were better than those frozen via the three-step decelerating method (43.3 ± 3.5% and 71.2 ± 3.4% compared with 29.7 ± 3.7% and 58.8 ± 3.4% respectively; p < .05). No differences were seen between the methods with respect to sperm motility variables; most sperm (>90%) remained static with both freezing methods. In conclusion, although the method with accelerating freezing rate was associated with better post-thaw sperm viability and acrosome integrity values, neither of the two freezing methods tested provided adequate motility results after thawing. Combining rectal massage with electrical stimuli seemed to reduce the number of the latter required for successful sperm collection., (© 2020 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

We aim to evaluate our experience, comparing intracytoplasmic sperm injection (ICSI) outcomes of cycle using fresh versus thawed electroejaculated spermatozoa. All consecutive couples undergoing ICSI cycles using electroejaculated spermatozoa, during a 16-year period, were evaluated. Embryological/laboratory variables of the ICSI cycles were assessed and compared between those utilising fresh (fresh group) versus thawed (thawed group) electroejaculated spermatozoa. Fifty-seven couples were evaluated, 30 used a fresh electroejaculated spermatozoa in 55 ICSI cycles, while 27 used a thawed sperm sample in 41 ICSI cycles. There were no in-between group differences in the mean numbers of oocytes retrieved per oocyte retrieval nor the percentage of MII oocytes. The fresh group demonstrated significantly higher fertilisation (71.5% vs. 64.1%, respectively, p < .05), top-quality embryos (66.5% vs. 54.9%, respectively, p < .02), clinical pregnancy per transfer (41.3% and 21.2%, respectively, p < .05) and cumulative clinical pregnancy (58.2% vs. 26.8%, respectively, p < .001) rates, as compared to the thawed group. Independent of the source of spermatozoa used, no pregnancy was achieved following ICSI utilising immotile spermatozoa. In conclusion, ICSI cycles using ejaculated spermatozoa of patients suffering from neurologic or psychogenic anejaculation are reassuring. The use of fresh ejaculated spermatozoa retrieved on the day of the female spouse oocyte retrieval might improve outcome. Whenever a thawed electroejaculated spermatozoa yield no motile spermatozoa, emergency electroejaculation is mandatory., (© 2020 Blackwell Verlag GmbH.)

The endangered black-footed ferret (BFF; Mustela nigripes ) is an important example of the benefits of assisted reproduction in species conservation with both semen evaluation and artificial insemination using fresh and frozen sperm being successfully incorporated into the breeding program. Currently, electroejaculation (EE) is routinely utilized for semen collection in BFFs, a technique that requires custom equipment and experienced operators, and does not consistently yield viable samples in this species. In this case study, we evaluated the feasibility of urethral catheterization (UC) for semen collection, a method predominately tested in domestic and non-domestic felids, on four occasions (three BFF males). After general anesthesia with a combination of ketamine, midazolam and α2-agonist dexmedetomidine (thought to promote semen release into the urethra), a lightly lubricated, flexible feeding tube was passed into the urethral opening and advanced ~7-8 cm into the urethra. A syringe attached to the feeding tube was used to apply mild negative pressure to collect sperm. Semen samples were successfully collected on all four attempts. Sperm characteristics ranged as follows: 10.5-26.0 × 10 6 sperm/ml concentration, 50-90% motility and 36-61% normal sperm morphology. This is the first report of the use of UC as a potential alternative to EE in the BFF, a more field-friendly technique that is less invasive and more consistent for obtaining samples free of urine contamination., (© The Author(s) 2020. Published by Oxford University Press and the Society for Experimental Biology.)

The pampas deer is an endangered species, from which reproductive biology little is known. We aimed to describe and compare the reproductive seasonal patterns of adult and yearling pampas deer stags throughout the year, including morphological traits, testosterone concentration, sperm morphology and cryoresistance pattern changes. Six adult (AS) and five yearling (YS) stags were captured with anesthetic darts once in winter, spring, summer and autumn to study morphological variables, serum testosterone and semen. Adult males were heavier, their neck girth tended to be greater and their testosterone concentration was higher than in YS. Animals were heavier in summer and autumn. Neck girth and testosterone concentration were greater in autumn. Scrotal circumference, testicular volume and gonado-somatic index varied with seasons, decreasing from winter to spring, increasing in summer and remaining in greater values in autumn. Sperm quality had maximum values from summer to winter. However, the cryoresistance ratio of motility score was greater in spring. In conclusion, in the captivity conditions, pampas deer stags seems to present a light seasonal reproductive pattern, with maximum testis size, testosterone secretion and fresh semen quality in autumn. Nevertheless, sperm cryoresistance ratio seemed to remain stable along the year. Although YS were still growing, they achieved similar semen quality than AS., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

The guinea pig, as a livestock species, is still developing and growing throughout Peru and neighboring countries, as reflected by its increasing export since 2000. However, the selection of proven fertile males is tedious due to the absence of seminal parameter standards and the lack of safe semen collection techniques. Thus, pregnancy detection or live births are required for males' selection. The purpose of this study was to describe the qualitative and quantitative semen parameters of fertile guinea pig males, to set reference values, and to validate a novel electroejaculation technique for the species. Semen was collected at weekly intervals from sixteen fertile males. Four transrectal electroejaculations were performed per male with 95% successful collections, yielding 39 viable semen samples. Seminal characteristics were as follows: pH 7.0 ± 0.13; ejaculate volume 0.67 ± 0.55 mL; sperm motility 90.81 ± 6.64%; sperm concentration 36.7 ± 28.41 × 106 sperm/mL; sperm count 20.09 ± 17.56 × 106 sperm/ejaculate; percentage of abnormal morphology 18.26 ± 8.52%; and percentage ubiquitinated spermatozoa 5.57 ± 6.28%. These values will serve as a reference to detect best breeding and infertile males rapidly. The described techniques are reproducible by commercial producers.

This study was conducted to determine the best combination between two collection method and two extenders in the cryopreservation of semen from creole bulls adapted to highlands of the Ecuadorian Andes. Sixty ejaculates from three adult Creole bulls were evaluated after collection by artificial vagina (AV) and electroejaculation (EE). Semen samples were split into two aliquots and diluted with a soy lecithin extender (Andromed®; A) or an egg yolk-containing extender (Triladyl®; T) and packed in straws of 0.25 ml with 20 × 10 6 sperms. Optical microscopy and computer-assisted semen analysis system (CASA) were used to evaluate semen quality characteristics. The effects of collection methods and extender type as well as its interaction were evaluated by a factorial ANOVA and Bonferroni's test. Semen samples collected with EE and frozen with T (EE-T) and A (EE-A) had greater proportion of spermatozoa with optical assessed individual progressive motility (IPM), plasmatic membrane intact (HosT), and lower tail abnormalities than those obtained with AV and frozen with the same extenders (AV-T and AV-A); however, differences were significant only between EE-A and AV-T. CASA assessment indicated that the total mobility (TM) was greater (P < 0.05) in semen samples diluted with T, although these samples had a greater proportion (P < 0.05) of sperms with local motility (LM) and fewer immobile sperms (IS), than those extended with A. Generally, semen samples obtained with EE or AV and diluted with T seems to be the best option to ciopreserve gametes of Creole bulls raised in highlands of Ecuadorian Andes.

The aim of this study was to evaluate home-made and commercial extenders for the cryopreservation of Rusa deer semen. After collection by electroejaculation, six ejaculates were diluted and frozen in TES-based, Tris-based and Triladyl ® extenders. Subjective motility, viability, morphology, acrosome integrity and membrane functionality were assessed post-thawing and after 1-hr incubation at 37°C (Thermal stress test). Total and progressive motility, and kinematic parameters were also assessed through CASA system. Post-thawing sperm progressive motility (PM), velocity according to the straight path (VSL) and linearity (LIN) showed significant differences, and higher values were detected for spermatozoa diluted with Triladyl ® and TES (p < 0.05) as compared with Tris (PM of Triladyl ® 14.7% vs. 3.2% TES and 2.5% Tris; VSL 56 for Triladyl ® , 59.2 for TES and 41.7 for Tris; LIN 45.6 for Triladyl ® , 52 for TES and 36.5 for Tris). Triladyl ® and TES extender led to better post-thawing sperm parameters, but these preliminary results need to be verified through artificial insemination trials., (© 2018 Blackwell Verlag GmbH.)

This study was conducted on red foxes to determine the appropriate voltage in electroejaculation for semen collection from stud males, and to confirm whether frozen semen with bovine semen extender can be used for artificial insemination. The proper load voltage for electroejaculation was 3-4 V based on semen collection rates and concentrations of spermatozoa. Frozen semen was prepared according to the known procedure for cows. In frozen-thawed semen, a relatively high conception rate (81.3%) was obtained in vixens, in which the optimum insemination time was detected by vaginal electrical resistance. These findings demonstrate that the restricted condition for semen collection by electroejaculation with cryopreservation of semen using bovine semen extender can be applied to artificial insemination of red foxes.

The aim of our study was to compare the quality parameters of fresh feline ejaculates collected by three different techniques-urethral catheterization after medetomidine administration (CT), electroejaculation (EE) and epididymal slicing after orchiectomy (EP). A total of 34 adult male cats (Felis catus) were included in the study. In all male cats, the sperm collection was performed under general anaesthesia by three collection methods in the following order: urethral catheterization, electroejaculation and epididymal slicing. The sperm parameters evaluated were as follows: volume, motility, viability, sperm concentration, total sperm count and morphological examination. The highest quality semen parameters were achieved using EE. The comparison of results of the evaluated sperm quality parameters from EE and EP showed significant differences only in one case-the percentage of head abnormalities and lower percentage of head abnormalities were achieved using EE compared to EP: 8.5% (3.0%-21.0%) versus 10.0% (4.0%-22.0%). Semen collected by CT rendered the lowest quality samples when compared to sperm samples collected by EE and EP, especially with respect to the motility and total sperm count which were significantly lower (p < 0.001). Our study showed that sperm samples collected by EE and EP result in better quality of feline ejaculates compared to collection by CT from sperm samples collected from the same male cats. These results demonstrate the necessity of further research of urethral catheterization as a novel technique of semen collection in male cats., (© 2018 Blackwell Verlag GmbH.)

Background: This report presents a case where electroejaculation (EEJ) was used for semen cryopreservation (SCP), prior to gonadotoxic anti-cancer treatment in a 14-year old boy diagnosed with Hodgkins disease., Method: Two sessions of EEJ were performed with an interval of 48 hours., Results: No complications were seen and the procedures resulted in nine frozen straws of motile spermatozoa., Conclusion: EEJ is a safe and feasible procedure for SCP in an adolescent cancer patient who is unable to masturbate or use penile vibratory stimulation (PVS).

Cryopreserving genetic resources is becoming increasingly important for species management. In the zoo-based red wolf (Canis rufus) population, inbreeding continues to increase in the absence of new founders. Through banking sperm, we preserve genetic diversity and create the ability to decrease inbreeding accumulation in the future. The quality and quantity of banked sperm can be affected by cryopreservation media and semen collection methods. This study's objectives were to further optimize semen extender used for red wolf sperm cryopreservation, investigate effects of post-thaw holding temperature, and to determine if urethral catheterization is an effective method for semen collection in this species. Semen collection via electroejaculation (EE) was performed on 39 adult red wolf males (ages 1 to 11) from 15 institutions. Urethral catheterization (UC) was attempted on a subset (n = 14) of those males, prior to EE. Thirteen different semen extenders were used for cryopreservation, which varied in osmolarity (HI or NORM), sugar source (glucose, fructose, or a combination), and cryoprotectant (glycerol or DMSO). Significant decreases in percent motility, forward progressive status (FPS), and acrosomal integrity were observed over time across all extenders (P < 0.0001). Among the extender components examined, post-thaw sperm motility and FPS were lower in DMSO versus glycerol based treatments (P < 0.005). Therefore, DMSO should be considered unsuitable as a cryoprotectant when freezing red wolf sperm. Effects of osmolarity and sugar source were minimal and temporally variable, however notably, a higher percentage of morphologically normal sperm were observed in the fructose-based extenders compared to glucose-based extenders post-thaw (P < 0.05). Additionally, post-thaw sperm motility and FPS declined more rapidly in samples maintained at 37 °C compared to samples held at room temperature (P < 0.05). Greater volumes of semen were collected using EE compared to UC (P = 0.041), and sperm samples collected using EE also had greater motility and FPS (P < 0.05). Additionally, though no gross morphological differences were observed, there were fewer sperm with intact acrosomes in the samples collected via UC (P = 0.0443). Thus, UC should not be considered sufficient for semen collection in red wolves when the desired fate of sperm is cryopreservation and/or AI. However, UC does provide an opportunity for a basic reproductive evaluation of a red wolf male., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Baseline information about the reproductive physiology of an endangered species is vital to captive breeding programs. This study analyzed reproductive parameters from eleven captive Chacoan peccaries (Catagonus wagneri) (mean age: 10.5 ± 1.2 years old) in Paraguay. After immobilization, testis length, width and firmness (hard to flaccid, 1-3) were assessed, followed by electroejaculation and analysis of semen. Samples were examined for percentage and progressive motility, total cell count, percentage live spermatozoa and morphology. Mean (±SEM) testis volume and firmness were 24.7 ± 1.8 cm 3 and 2.1 ± 0.1, respectively. Mean ejaculate volume was 2.9 ± 0.7 ml with a pH of 7.7 ± 0.3. Few male peccaries had motile spermatozoa (n = 5/9) with a mean percentage and progressive motility of 18.3 ± 8.5% and 0.6 ± 0.3, respectively. The mean percentage of live spermatozoa was 25.1 ± 5.6%. Male peccaries had a low percentage of normal spermatozoa (12.4 ± 2.5%). The mean total count of spermatozoa per ejaculate was also quite low at 1.58 ± 1.01 million total spermatozoa per ejaculate. Spermatozoa defects were predominantly primary (77.7%) with the most common spermatozoa defects being tapered head (19.0 ± 7.4%), diadem/crater (17.7 ± 2.8%), and excess residual cytoplasm (9.6 ± 2.5%). Male age was not correlated with semen parameters (percent live: r=-0.19; motility percentage: r = 0.01; percent normal spermatozoa: r = 0.38; total count: r = 0.29; p>0.05). Evaluation of additional males from this population as well as other captive populations at various time points during the year is warranted., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study was designed to evaluate the efficiency of medetomidine anesthesia for semen collection through urethral catheter in wild and captive jaguars. Six captive and five wild jaguars were chemically restrained with a combination of medetomidine (0.08-0.1 mg/kg) and ketamine (5 mg/kg). After medetomidine administration the urethra was catheterized using a urinary tomcat catheter (1 mm diameter × 130 mm length) to collect semen from all animals. By using this technique, we could collect an average of 347.2 μl of semen containing 2,635.2 sperm/ml. Forward progressive motility, sperm progressive motility, and sperm morphology analysis demonstrated that the methodology did not affect sperm quality. Thus, urethral catheterization after medetomidine administration is a practical and efficient method to collect high-quality semen from wild and captive jaguars; this will enable the development of reproductive assisted technologies for jaguars., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The demand for feline semen collection, evaluation, and subsequent use is growing as a way to preserve important genetic materials. This article describes semen collection methods using a variety of techniques that can be applicable in almost any setting. Also discussed are cryopreservation methods that optimize sperm survival., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: It is generally accepted that non-human primates (NHP) represent the model of choice for integrative studies of testicular function and endocrine control. However, there are many species-specific differences that necessitate identification prior to the selection of an appropriate model for these studies. In an NHP breeding facility, this opportunity of selection is usually presented during breeding periods when it is crucial to determine which individuals should be maintained as breeders. With reference to adult males and their use in breeding programs and reproductive studies, it is therefore imperative to document the normal semen and sperm values, expected seasonal changes and the variabilities found within samples and among individuals. The comparison of closely related species that differ by breeding seasonality will, therefore, highlight their value in reproductive research., Methods: Semen samples were obtained by rectal probe electroejaculation (RPE). The seminal and sperm characteristics of captive-bred Vervet monkeys (Chlorocebus aethiops) (n = 10) and Rhesus monkeys (Macaca mulatta) (n = 10) were evaluated and compared. Parameters such as semen volume, pH, sperm concentration, and sperm motility were analyzed by means of a computer-aided sperm analysis (CASA) system., Results: Large variations in semen and sperm parameter values indicated differences between species and samples. Monthly variations were observed for the Vervet regardless of breeding and conceptions that occurred throughout the year. In contrast, Rhesus seminal characteristics indicated a clear seasonal trend., Conclusion: Non-human primates have long provided as research models for studying complexities of human reproductive biology. The baseline values reported from this study can be applied as guidelines during the selection of male individuals for reproductive studies. Of further interest is the comparative data on semen and sperm parameters between two congeneric species that differ by seasonal versus aseasonal breeding., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Most men with spinal cord injury (SCI) are infertile due to a combination of erectile dysfunction, ejaculatory dysfunction, and abnormal semen quality. This article addresses issues that should be considered when managing the reproductive health of men with SCI. The authors present recommendations based on their decades of experience in managing the reproductive health of more than 1,000 men with SCI. Men with SCI face obstacles when pursuing sexual activity and/or biologic fatherhood. Hypogonadism and premature symptoms of aging may interfere with sexual function. Erectile dysfunction is prevalent in the SCI population, and treatments for erectile dysfunction in the general population are also effective in the SCI population. Most men with SCI cannot ejaculate with sexual intercourse. The procedures of penile vibratory stimulation (PVS) and/or electroejaculation (EEJ) are effective in obtaining an ejaculate from 97% of men with SCI. The ejaculate often contains sufficient total motile sperm to consider the assisted conception procedures of intrauterine insemination or even intravaginal insemination at home. If PVS and/or EEJ fail, sperm may be retrieved surgically from the testis or epididymis. Surgical sperm retrieval typically yields enough motile sperm only for in vitro fertilization with intracytoplasmic sperm injection. The majority of new cases of SCI occur in young men at the peak of their reproductive health. With proper medical management, these men can expect to experience active sexual lives and biologic fatherhood, if these are their goals. Numerous tools are available to physicians for helping these patients reach their goals.

The preservation of spermatozoa is an important tool used in conservation programs to increase the genetic diversity of threatened and endangered species. Although routinely used to manage conservation programs for higher vertebrates, there have been limited attempts to establish reproductive assistance programs for tortoises. The purpose of this study was to develop a model for collecting and characterizing semen in Testudinidae. Semen was collected from 13/16 (81.2%, 95% CI: 62-100) adult male leopard tortoises ( Stigmochelys pardalis ) via electroejaculation under propofol anesthesia. Semen samples were collected most frequently after the second series of electrostimulations (6/13, 46.1%), with fewer animals producing semen after the first (5/13, 38.5%) or third (2/13, 15.4%) electrostimulations. The average volume of a semen sample in the tortoises was 0.26 ml (standard deviation: 0.16, minimum-maximum: 0.1-0.6), the average spermatozoal concentration was 101.62 × 10 6 /ml, and the average motility at time of collection was 57.3%. A rapid decrease in motility was observed in refrigerated samples over 24 h resulting in a median motility of 0% at 24 h post-collection. The results of this study suggest that electroejaculation is a safe and efficient method for collecting semen from leopard tortoises.

Aim: The objective of this research is to report parameters for breeding soundness evaluation, semen extension, and cryopreservation in Rusa timorensis ., Materials and Methods: Seven healthy stags were chosen for semen collection using an electroejaculator. The collections were performed twice in a breeding season between February and June 2016. Samples were collected between 2 and 3 weeks interval, collected twice for each animal. Semen was evaluated, extended, and cryopreserved using four different extenders; Andromed ® , BioXcell ® , Triladyl ® , and a modified Tris-egg yolk combined with Eurycoma longifolia Jack., Results: R. timorensis semen characteristics according to volume (ml), color, sperm concentration (10 6 /ml), general motility (%), progressive motility (%), and % morphology of normal spermatozoa are 0.86±0.18 ml, thin milky to milky, 1194.2±346.1 10 6 /ml, 82.9±2.8%, 76.1±4.8%, and 83.9±4.8%, respectively., Conclusion: Semen characteristics of R. timorensis collected by electroejaculation is good allowing for cryopreservation and future artificial insemination work. The most suitable extender for Rusa deer semen is Andromed ® .

Objectives: To evaluate treatment results of electroejaculation in patients with spinal cord injuries and the additional value of repeated electroejaculation., Methods: We carried out a retrospective chart analysis of all spinal cord injury patients treated with electroejaculation at University Medical Center Utrecht, Utrecht, the Netherlands, from January 1994 to July 2015. Data were collected on the patients' demographics and medical history. We evaluated sperm quality according to World Health Organization standards, pregnancy and delivery rates., Results: A total of 230 electroejaculation procedures were carried out in 47 patients. In 227 of 230 electroejaculations (98.7%), an ejaculate could be obtained. In 169 of 230 (73.5%) electroejaculation procedures, it was possible to yield semen containing progressively motile spermatozoa. In 18 of 47 (38.3%) patients, no semen of sufficient quality could be yielded during the first electroejaculation. Repeated electroejaculation resulted in ejaculates containing progressively motile spermatozoa in seven of 18 (38.9%) of these men. Procreation was attempted through in vitro fertilization/intracytoplasmic sperm injection in 17 couples; of these, 14 of 17 (82.4%) couples achieved pregnancy., Conclusions: In the majority of spinal cord injury patients treated with electroejaculation, it is possible to obtain semen that can be used for assisted reproductive technologies. Repeated electroejaculation should be considered when the first procedure fails., (© 2016 The Japanese Urological Association.)

An evaluation of a large database of red wolf fresh ejaculate characteristics (n = 427 ejaculates from 64 wolves) was undertaken to increase knowledge of seminal characteristics in the red wolf and evaluate possible relationships between inbreeding, age, and seminal quality. Phase microscopy analysis of electroejaculates collected over 14 natural breeding seasons was compared with animal ages and inbreeding coefficients. Ejaculate volume increased and sperm concentration and total count decreased as wolves aged (P < 0.01, 0.001, and 0.05, respectively), and the proportion of sperm cell morphological abnormalities was greater in animals with higher coefficients of inbreeding (P < 0.001), particularly for older animals (P < 0.001). Moreover, the mean coefficient of inbreeding of animals that had failed to reproduce given at least one opportunity during their lifetimes was significantly greater than that of wolves with proven fertility, and wolves of proven fertility exhibited higher sperm concentrations and total counts than nonproven wolves. Thus, as the captive red wolf population becomes more inbred, the maximum age of reproduction is likely to decrease; an important finding to consider when projecting population dynamics and determining pairing recommendations., (Copyright © 2016 Elsevier Inc. All rights reserved.)

To address the need to preserve current genetic diversity before it is lost forever; further studies to adapt assisted reproductive technologies to various endangered species are needed, among other things. Roe deer (Capreolus capreolus), an over abundant wild deer, can serve as model species to develop or improve sperm cryopreservation of threatened or endangered deer species. The aim of this study was to compare the ability of three diluents (Berliner Cryomedium [BC]; Tris, citric acid, glucose [TCG]; TES, Tris, glucose) to support chilling, cryopreservation (with 5% glycerol; G) and postthaw incubation (at 22 °C and 37 °C) of roe deer spermatozoa collected by electroejaculation. Berliner Cryomedium was the diluent that better preserved roe deer spermatozoa during refrigeration, able to maintain motility for at least 14 days, longer than the other extenders. BC + G was the extender of choice for cryopreservation, showing higher viability compared with TCG + G (66.7 ± 3.4 vs. 54.5 ± 6.5; P < 0.05) and higher level of acrosome integrity compared with TES, Tris, glucose + G (79.4 ± 3.4 vs. 67.9 ± 5.0; P < 0.05). Maintaining the samples at 22 °C after thawing presented higher values in various parameters compared with 37 °C. The knowledge gained through this study can potentially act as a preliminary step toward development of new protocols to help increase the reproductive success of biologically similar, yet endangered, wild species., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Differential phenotypic characteristics for taxonomic diagnosis purposes are well determined in the genus Saimiri (squirrel monkey). However, data on its reproductive characteristics are lacking. Our aim was to determine testicular biometry and correlate with seminal analysis in captive (Saimiri collinsi) and free living (Saimiri vanzolinii, Saimiri cassiquiarensis, and Saimiri macrodon) squirrel monkeys. Testicular length, width, height, circumference, and volume were measured. Testicular biometry showed no differences between right and left testicles within the same species, as well as among species. Semen collected by electroejaculation was constituted of a liquid and coagulated fraction, or only one of them. No significant difference was observed between mean volumes of liquid (49.2 ± 68.9 μL: S. collinsi; 28.3 ± 59.8 μL: S. vanzolinii; 5 ± 7.1 μL: S. cassiquiarensis; and 0 μL: S. macrodon) and coagulated (65.4 ± 142.1 μL: S. collinsi; 125.8 ± 142.5 μL: S. vanzolinii; 175 ± 176.8 μL: S. cassiquiarensis; and 500 μL: S. macrodon) fractions within species or when each fraction was compared among the studied species. No correlation between testicular volume and seminal volume was observed when liquid (R = 0.31, S. collinsi; R = -0.69, S. vanzolinii) and coagulated (R = 0.32, S. collinsi; R = -0.37, S. vanzolinii) fractions were evaluated. No sufficient data were obtained for the other two species. Seminal quality was similar among species, and the most common defect was coiled tail. The method of electroejaculation yielded satisfactory results on these species, under field conditions., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Great apes are mammals close to humans in their genetic, behavioral, social and evolutionary characteristics and new genomic information is revolutionizing our understanding of evolution in primates. However, all these species are endangered. While there are many global programs to protect these species, the International Union for Conservation of Nature (IUCN) projects that in a near future the wild populations will decrease significantly. Nowadays, the relevance of captive populations of great apes is becoming critical for research and understanding of pathophysiology of diseases. In this report, the evaluation of infertility in a group of captive chimpanzees maintained at Leon's Zoological Park using a human infertility protocol is described. Our results suggested that infertility in this group was due to low hormonal levels and sperm alterations in the male characterized by hormonal assessment and a sperm sample obtained by electroejaculation and cryopreserved using human protocols. In the females, it was demonstrated that it is possible to follow the follicular cycle using non-invasive methods based on morphological changes in genitalia, detection of blood in urine and measurement of hormones in saliva samples; concluding that fertility in females was normal. Also, we demonstrate that human artificial insemination procedures may be applied. Our human approach was successful in finding the infertility cause in this group of captive chimpanzees. In countries with limited resources, collaboration of zoos with human infertility clinics can be beneficial for research and management of reproductive aspects of great apes.

Semen collection for cryopreservation is a key step for small ruminant conservation programs. While in these species semen is mainly collected via artificial vagina (AV), electroejaculation (EE) provides a viable alternative for untrained males. Herein we investigated the effect of semen collection method on post-thaw sperm quality by comparing two small ruminant species, sheep and goats. Semen from Blanca-Celtibérica bucks and Manchega rams was collected by AV and EE on the same day and cryopreserved using a standard protocol. At thawing, sperm motion parameters were evaluated by CASA, whereas membrane stability (YO-PRO-1), sperm viability (propidium iodide, PI) and mitochondrial activity (Mitotracker Deep Red) were analyzed using flow cytometry. The semen collection method negatively affected post-thaw sperm quality in bucks but not in rams. Thus, in bucks, post-thaw sperm motility was higher for samples collected by AV as compared to those obtained via EE. Similarly, post-thaw sperm parameters evaluated by flow cytometry were worse for buck samples collected by EE than those collected by AV in the same species, or than ram samples regardless of collection method. These results suggest that ovine and caprine spermatozoa have a different response to the cryopreservation process depending upon the semen collection method used. We hypothesize that the EE procedure may lead to changes in the composition of the ejaculate in bucks that would make spermatozoa more susceptible to the cryopreservation process, whereas this procedure would have had no effect on ram spermatozoa. This assumption requires further investigation., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The giant anteater (Myrmecophaga tridactyla) is considered vulnerable to extinction. Scientific data on the reproductive parameters of this species are scarce. Semen from eight free-ranging giant anteaters was collected to establish its characteristics and the effects of cooling and storage at 5 °C after dilution with the BotuCrio extender without cryoprotectant. The ejaculate presented two distinct sequential fractions, including a whitish fraction, which was milky and rich in sperm cells, and a gel fraction, which was colorless, viscous, and azoospermic. The mean ± standard error of the mean values of the seminal characteristics were as follows: volume of the first fraction, 0.75 ± 0.1 mL; motility, 75 ± 2.9%; vigor, 3.2 ± 0.3; sperm motility index, 68.8 ± 4.3; concentration, 108.5 ± 13.4 × 10(6)/mL; plasma membrane integrity index, 71 ± 4.0%; spermatic defects detected using modified Karras staining, 35.5 ± 3.3%; and spermatic alterations identified by differential interference contrast microscopy, 48.3 ± 6.8%. During refrigeration, the semen presented decreasing motility from 0 to 18 hours, sperm motility index decreased from 0 to 24 hours, and vigor did not change in the first 6 hours and then decreased to 18 hours., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The aim of the current study was to compare sperm quality characteristics of the collared peccary (Pecari tajacu) following freezing in extenders supplemented with whole egg yolk and different concentrations of low-density lipoproteins (LDL). Semen from 11 adult males was obtained by electroejaculation and evaluated for sperm motility, vigor, morphology as well as membrane integrity analyzed by the hypo-osmotic swelling (HOS) test and a fluorescent staining. Moreover, the semen was diluted in a Tris-based extender containing 20% egg yolk (control group) or 5, 10 or 20% LDL (treatment groups). The semen samples were frozen in liquid nitrogen and thawed in a water bath for 60s at 37°C. The treatments did not affect (p>0.05) sperm vigor, morphology or membrane integrity analyzed by the HOS test. However, post-thaw sperm motility was significantly higher (p<0.05) in the extender supplemented with 20% LDL (36.4 ± 5.3%) compared with the egg yolk extender and extender supplemented with 10% LDL. Furthermore, the percentage of membrane-intact frozen-thawed spermatozoa analyzed by the fluorescent staining was significantly higher (p<0.05) in the extender supplemented with 20% LDL (27.4 ± 6.5%) than in the other groups. In conclusion, 20% LDL can be used to substitute the whole egg yolk as a cryoprotective additive for freezing semen of the collared peccary., (Copyright © 2015 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.)

Alpha2-adrenergic agonists are used to immobilize many veterinary species, but use has been infrequently linked to urine contamination of semen collected via electroejaculation. The objective of the study was to compare the α2-agonists medetomidine and dexmedetomidine on urine contamination of semen in anesthetized cheetahs (Acinonyx jubatus) during electroejaculation procedures. From 2009-2012, a retrospective medical record review revealed 21 anesthesia events in 12 adult male cheetahs. Animals were immobilized with combinations of Telazol® (2.33±0.43 mg/kg) and ketamine (2.38±1 mg/kg); Telazol (1.17±0.14 mg/kg), ketamine (1.17±0.14 mg/kg), and medetomidine (0.012±0.0017 mg/kg); or Telazol (1.59±0.1 mg/kg), ketamine (1.59±0.1 mg/kg) and dexmedetomidine (0.01±0.001 mg/kg). Semen was successfully collected in all animals; four animals anesthetized with medetomidine had urine contamination (P=0.037). Medetomidine may contribute to urine contamination; however, further investigation is needed to determine significance in cheetahs.

Scientific interests and conservation needs currently stress the necessity to better understanding bat reproductive biology. In this study, we present the first, safe, inexpensive, and reliable method to obtain sperm from a microbat species (Carollia perspicillata) by electroejaculation. This method revealed to be highly efficient (100% success rate). We obtained ejaculates composed of two characteristically different fractions. We compared three buffers and recommend using an Earle's balanced salt solution as a semen extender. Earle's balanced salt solution provided significant repeatable measure of swimming ability (intraclass correlation coefficient: 0.74, P < 0.01) and proportion of motile sperms (intraclass correlation coefficient: 0.08, P = 0.01) and allowed sperm to maintain optimal swimming capacity over time. None of the buffers could dissolve all the coagulated sperm. Although the trypsin buffer freed a larger fraction of spermatozoa in the ejaculate, it impaired swimming ability without improving motility, viability, and stamina. We thus argue that the sperm population analyzed with Earle's balanced salt solution is a representative of the ejaculate. Finally, we found that the mean sperm velocity of C perspicillata (78.8 μm/s) is lower than that predicted by regressing sperm velocity on relative testes mass, a proxy of sperm competition. The question as to whether C perspicillata is an outsider for sperm velocity, or whether bats evolved yet another unique mechanism to cope with sperm competition deserves more investigations., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Here we show the effects of repeated electroejaculation (EE) on mean values of motility, mitochondrial functionality, and expression of active caspases on goat sperm obtained by EE. Evaluations were done using CASA and flow cytometry. A strategy for identification of kinematic sperm subpopulations, when individual data of sperm are not provided by the CASA system, is provided. Fifty semen samples, five of each of ten adult creole goats, were obtained by electroejaculation. Mean values of total motility, progressive motility and flow cytometry evaluations were compared among EEs. Relationships among mean values of variables were investigated using Spearman correlation and principal component analysis (PCA). For identification of kinematic sperm subpopulations, PCA followed by hierarchical clustering was applied on data of the intervals provided automatically by the CASA system. Total motility does no change after repeated EE. Mean values of motility parameters and molecular markers were unrelated in multivariate space, but bivariate correlations were found. Values in upper and lower intervals defined clearly the sperm subpopulations, which had motility parameters changing over time. Taken together, our results show that repeated EE does not affect mean values of total motility, that molecular markers are not related with motility parameters, and that it is possible to identify kinematic sperm subpopulations when individual data, of motility parameters, are not provided by the CASA system., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)

Many aspects of fertility rely on intact neurologic function and thus neurologic diseases can result in infertility. While research into general female fertility and alterations in male semen quality is limited, we have an abundance of knowledge regarding ejaculatory dysfunction following nerve injury. Normal ejaculation is the result of coordinated reflex activity involving both the sympathetic and somatic nervous systems. Nerve injury can result in retrograde ejaculation, and anejaculation. With retrograde ejaculation, the ejaculate is propelled into the bladder instead of out through the urethra. In mild cases this condition can be reversed by sympathomimetic medications and, in more severe cases, sperm cells can be extracted from the bladder following ejaculation. With anejaculation, the ejaculatory reflex is not activated by normal sexual stimulation. In such cases, the first choice of treatment is assisted ejaculation, preferably by penile vibratory stimulation. If vibratory stimulation is unsuccessful, then ejaculation can almost always be induced by electroejaculation. In cases where assisted ejaculation fails, sperm can be retrieved surgically from either the epididymis or from the testis. Once viable sperm cells have been obtained, these are used in assisted reproductive techniques, including intravaginal insemination, intrauterine insemination, and in vitro fertilization/intracytoplasmic sperm injection., (© 2015 Elsevier B.V. All rights reserved.)

Pampas deer (Ozotoceros bezoarticus) is a native endangered species. Knowledge of the basic spermiogram characteristics and the morphometric descriptors is necessary to effectively develop sperm cryopreservation. In other species, sperm sub-population is related to sperm cryo-resistance. The objective was to provide a general description of the sperm, including sperm head morphometric descriptors, its repeatability, and the existence of sperm sub-populations. Sperm were obtained from adult males by electroejaculation during the breeding season. The motility score was 3.4 ± 0.2 (mean ± SEM) and progressive motility was 59.4 ± 3.7%. Ejaculated volume was 413.9 ± 51.0 μl, the total number of sperm ejaculated was 321.2 ± 55.4 × 10(6). Also, 63.3 ± 3.1% of the sperm were morphologically abnormal and 23.7 ± 2.3% had acrosome damage. The sperm head length was 7.6 ± 0.01 μm, width 4.4 ± 0.01 μm, area 28.1 ± 0.07 μm(2) and the perimeter was 21.9 ± 0.04 μm. There was a positive relationship among morphometric descriptors and the motility score, overall motility and progressive motility. Also length (P=0.011), width (P=0.003), area (P=0.006) and perimeter (P=0.009) of sperm head obtained in two different collections were positively related. Overall, the low concentration, volume, overall quality and abnormal morphology, and wide variation of these variables may be a limitation for the development of sperm cryopreserved banks. There were three sperm sub-populations with different morphometric characteristics. The morphometric descriptors are maintained similarly among different collections., (Copyright © 2014 Elsevier B.V. All rights reserved.)

This study was carried out to evaluate the effect of electroejaculation (EEJ) on myocardial function, acid-base balance, and hematobiochemical profiles in male dromedary camels. Twenty sexually mature, apparently healthy male camels were assigned to EEJ. Parallel, eight naturally mated male camels were enrolled as a control group. Three blood samples were collected from each camel: just before (T0), directly after (T1), and 24 hours after (T2) EEJ or natural mating. The serum concentrations of the cardiac biomarker troponin I (cTnI), blood gas parameters, and hematobiochemical profiles were determined. Nineteen camels were ejaculated by the end of the second circuit and one by the end of the first circuit. In both groups, the mean heart and respiratory rates had increased significantly immediately after the procedure, but returned to normal values 24 hours after the procedure. The mean serum concentration of cTnI had increased significantly in all camels after EEJ, but not in controls. However, at 24 hours post-EEJ, the serum concentration of cTnI did not differ significantly compared with baseline values. The blood pH and base excess had decreased, and the PCO2 and lactic acid had increased after EEJ. The EEJ provoked decreases in hematocrit and mean corpuscular volume. In the control group, the base excess, HCO3(-), TCO2, anion gap, and lactic acid increased slightly after mating but did not reach a significant level compared with premating values. It is concluded that EEJ in camels results in a reversible myocardial injury, changes in the acid-base status, and increase the lactic acid concentration., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objective: To evaluate the feasibility of electroejaculation to perform semen cryopreservation in pubertal boys before gonadotoxic therapy and to review the literature on this topic., Design: Retrospective cohort study and review of the literature., Setting: Academic children's hospital., Patient(s): Boys diagnosed with cancer to whom sperm cryopreservation was offered before the start of gonadotoxic therapy., Intervention(s): We studied the outcome of electroejaculation, including patient characteristics, hormone levels, and pretreatment semen parameters., Main Outcome Measure(s): Semen cryopreservation., Result(s): Pretreatment semen samples were obtained by masturbation in 106/114 boys with cancer, of which 78/106 were adequate for preservation. Electroejaculation was offered to 11 boys, of which three of 11 samples appeared adequate for preservation. Reviewing all reported electroejaculation cases in children with cancer in the literature, 13/29 (45%) cases were successful. Testosterone levels were higher in patients with successful sperm yield obtained by electroejaculation (median, 8.3 nmol/L [5.2-42.4] in successful harvests, vs. median 1.7 nmol/L [0.01-17.9] in unsuccessful harvests)., Conclusion(s): Semen cryopreservation should be offered to all pubertal boys diagnosed with cancer. If masturbation fails, electroejaculation can be considered as a useful option for semen cryopreservation and leads to adequate material for cryopreservation in about half of the cases., (Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Objective: To evaluate the results of treatment with electroejaculation (EEJ), intrauterine insemination (IUI), and IVF/intracytoplasmic sperm injection (IVF/ICSI) in patients with psychogenic anejaculation (PAE)., Design: Retrospective clinical study., Setting: Academic tertiary referral fertility center., Patient(s): Eleven male patients diagnosed with psychogenic anejaculation (PAE) were included. Median age at the time of first treatment with EEJ was 33.0 (interquartile range, 29.0-36.0) years., Intervention(s): Electroejaculation, IUI, and IVF/ICSI., Main Outcome Measure(s): Semen analysis, fertilization rate, implantation rate, pregnancy rate, and delivery rate., Result(s): A total of 60 EEJs were performed in 11 patients. Mean VCM (volume [mL] × concentration [sperm cells/mL] × percentage progressive motile cells) of the retrieved sperm of all EEJs was 17.5 × 10(6) (SD 16.5 × 10(6)). Yielded semen was used in a total of 26 ICSI procedures in seven couples. The fertilization rate was 65.6% (80 of 122). The ICSI cycles resulted in five pregnancies; of these, one resulted in a spontaneous abortion in the first trimester. Three couples were treated with 34 IUI cycles, which resulted in live births in four pregnancies., Conclusion(s): Electroejaculation is a suitable and effective treatment that can be used in men with psychogenic anejaculation. The retrieved semen can be used successfully in assisted reproductive technology treatment. In this study EEJ resulted in pregnancies and the birth of eight healthy children., (Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Objective: To describe the outcomes of electroejaculation (EEJ) and testicular sperm extraction (TESE) performed for fertility preservation among male patients who are unable to ejaculate or have nonobstructive azoospermia/severe oligospermia before definitive cancer therapy., Design: Retrospective cohort study., Setting: Tertiary cancer referral center., Patient(s): Forty-nine patients seeking fertility preservation before definitive cancer therapy, with anejaculation, religious or cultural objections to masturbation, azoospermia, or severe oligospermia requiring either EEJ or TESE., Intervention(s): EEJ and TESE., Main Outcome Measure(s): Sperm retrieval rates., Result(s): Fifty-nine percent of patients overall and 60% of adolescents/young adults had sperm retrieved for cryopreservation. EEJ was successful in retrieving sperm in 60% of adolescents. Of all adolescents and young adults undergoing TESE, 33% had sperm retrieved for cryopreservation. No complications were reported. Chemotherapy was commenced without delay in all patients requiring it, frequently on the same day as the sperm retrieval., Conclusion(s): EEJ and TESE can be safely and successfully used for fertility preservation before cancer therapy among boys and young adult men who are unable to provide a semen specimen or have nonobstructive azoospermia, and they should be considered in all men meeting this patient profile., (Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Background: The increasing prevalence of cancer survivors who are infertile due to gonadal failure highlights the importance of fertility preservation prior to gonadotoxic treatments. Adolescent cancer patients may not be mature enough to produce sperm by masturbation, leading to the use of alternative methods for obtaining sperm for cryopreservation. The aim of the current study was to evaluate the safety and efficacy of electroejaculation (EEJ) for cryopreservation among adolescent cancer patients., Procedure: This retrospective cohort study included 45 adolescent (12-18 years old) cancer patients who underwent EEJ during 2002-2012 in an academic tertiary referral fertility center. Sperm cryopreservation, ejaculate parameters, and procedure complications were evaluated., Results: EEJ was performed without documented complications. Sperm was successfully obtained in 30 (66.7%) patients. Retrieval failures included ejaculates without sperm in 8 patients (17.8%) and no ejaculate in 7 patients (15.5%). Cryopreserved ejaculates were characterized by severe asthenospermia, normal sperm concentration, and low volume. Retrieved sperm group was further divided to 19 motile sperm ejaculates with significantly higher volume, sperm concentration, and total count compared to 10 non-motile sperm patients., Conclusions: EEJ is a safe and feasible procedure for cryopreservation in adolescent cancer patients who are unable to masturbate. The wide diversity of EEJ outcome and ejaculate parameters may represent a continuum of pubertal changes in that population., (© 2013 Wiley Periodicals, Inc.)