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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 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. • 50 mcg/kg dexmedetomidine produces a representative sperm sample in cats. • Chemical and electroejaculation yield comparable ejaculate concentration in cats. • Chemical and electroejaculation motilities and morphology are comparable in cats. • Electroejaculation yields higher volume and sperm number than chemical ejaculation. • Nucleocounter and hemocytometer concentration measurements are equivalent. [ABSTRACT FROM AUTHOR]

Aiming at solving the problem of crablet production for mud crab commercial farming, an artificial insemination (AI) method in the mud crab, Scylla olivacea, was described. Placing a pair of cathodes to the inner coxa of both sides of the swimming paddles and an anode to the inner abdominal flab upon the hind gut of sexually mature S. olivacea males and applying 5–6 V of DC current for 3–5 s induced sperm mass expulsion. The sperm mass, comprising several spermatophores containing > 108 sperm/mL, was inserted into the vagina of the newly molted S. olivacea female. Out of 50 female trials, 10 of them developed ovarian maturation within 45 days, and 3 of these 10 crabs spawned. At full maturation of the ovary, needle aspiration of the ovarian tissue examined under light microscopy revealed approximately 50% mature oocytes with elevated fertilization membrane, indicating successful fertilization. Histological and ultrastructural studies of the seminal receptacle revealed numerous sperm with acrosome reaction (AR) within its lumen, revealing fertilization potential of the artificially inseminated sperm. Histologically, the mature ovary at 45 days following AI was composed of mature oocytes surrounded by numerous sperm with AR, suggesting sperm migration from the seminal receptacle into the ovarian tissue. This study reveals a high possibility of using AI for S. olivacea that could lead to the commercial supply of the crablets and a new insight into the internal fertilization of this economic species. [ABSTRACT FROM AUTHOR]

Zoo managed cheetahs provide an insurance population for wild cheetahs that are under threat of extinction from habitat loss, lack of prey, competition, pet trade and poaching for skin and bones. Assisted reproductive techniques including artificial insemination, in vitro fertilization, and embryo transfer augment natural breeding programs but rely on good quality semen for best results. It is understood that anesthesia can affect semen characteristics such as ejaculate volume, total sperm count, sperm motility, and incidence of urine contamination. Thus, the aim of this study was to conduct a retrospective analysis of 23 years of data to investigate sperm parameters of semen collected under anesthesia using medetomidine in combination with butorphanol and midazolam or Telazol® alone. Electroejaculation records (Medetomidine, Butorphanol, and Midazolam anesthetized n = 59 ejaculates, from 30 cheetahs, Telazol® anesthetized, n= 169 ejaculates, from 72 cheetahs) were evaluated for incidence of urine contamination. Electroejaculation records (Medetomidine, Butorphanol, and Midazolam anesthetized n = 21 ejaculates, from 17 cheetahs, Telazol® anesthetized, n = 143 ejaculates, from 63 cheetahs) were evaluated for total sperm count, total motility, ejaculate volume, and testicle size. Telazol® treated cheetahs had a numerically higher total sperm count (Median ± SD: 42.58 ± 77.8 × 106 spermatozoa) compared to those treated with medetomidine (Median ± SD: 31.2 ±44.58 × 106 spermatozoa), and a significantly (p < 0.05) higher sperm motility (Median ± SD: 70.0 ± 9.71%) compared to medetomidine (Median ± SD: 53.0 ± 16.41%) treated cheetahs. The findings of this study indicate that medetomidine anesthesia results in significantly lower sperm motility and Telazol® anesthesia results in a higher total sperm count and motility, thus resulting in higher quality ejaculate. This information can aid in the veterinary management of the species when involved in genome resource banking and assisted reproductive technologies. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Assisted reproductive technologies are playing an ever-increasing role in the management of ex situ rhinoceros populations. Traditionally, sperm collection in rhinos is conducted via electroejaculation (EEJ) or gamete rescue. Sperm collection via urethral catheterization (UC) is gaining popularity in wildlife reproductive sciences as it requires less specialized equipment, is faster, and less stimulatory than EEJ. This study investigated the use of UC to collect sperm from Southern white (Ceratotherium simum simum; n = 3) and Eastern black (Diceros bicornis michaeli; n = 6) rhinoceros as an alternative or complementary to collection via EEJ. UC samples (n = 14) exhibited high sperm concentration (1.6 ± 0.49 ×109 sperm/mL) and low volume (0.98 ± 0.26 mL). Samples obtained via EEJ (n = 11) were of moderate sperm concentration (0.14 ± 0.05 ×109 sperm/mL) and high volume (30.6 ± 7.2 mL). UC samples were processed for cryopreservation in OptiXcell extender. Motility of UC sperm decreased post-thaw (56 ± 2.4%) over pre-freeze (77.1 ± 3.2%; P < 0.05), with longevity data consistent to prior results published for sperm collected via EEJ. These results suggest sperm quality and cryo-survival are not impacted by the collection method. Since EEJ was conducted after UC, sperm parameters could not be compared between the two methods. Results indicate that UC is a viable alternative to EEJ for sperm collection in rhinoceros.

Decreased genetic diversity and possible inbreeding depression have recently been documented in the last wild ocelot (Leopardus pardalis) population in the United States. One consequence of inbreeding depression in felids may be reduced semen quality which can adversely affect reproductive potential. Detailed assessments of reproductive parameters in wild individuals and populations can be conducted using assisted reproductive technologies, such as semen collection and analysis. For most felid species, semen has traditionally been collected via electroejaculation (EEJ22 Electroejaculation); however, an alternative method has been developed using alpha-2 agonist drugs to induce direct sperm release into the urethra, allowing collection by catheterization without requiring specialized equipment. The goal of this study was to characterize normative reproductive traits in free-ranging ocelots and co-occurring bobcats (Lynx rufus) in southern Texas and assess the effectiveness of urethral catheterization (UC33 Urethral Catheterization) for semen recovery in both species. For semen collection, free-ranging cats were live-captured and anesthetized using intramuscular ketamine and medetomidine/dexmedetomidine (alpha-2 agonist) with UC conducted 20–40 minutes post-induction. In ocelots only, EEJ was subsequently performed if UC failed to recover a viable sample. Semen collection was attempted in 31 felids (n=9 ocelots; n=22 bobcats), with sperm recovery by UC in seven of nine ocelots (78 %) and 14 of 22 bobcats (66 %), and by EEJ in four of five ocelots (80 %). For ocelots, the percentage of primary morphologic abnormalities was higher (p

Zoo managed cheetahs provide an insurance population for wild cheetahs that are under threat of extinction from habitat loss, lack of prey, competition, pet trade and poaching for skin and bones. Assisted reproductive techniques including artificial insemination, in vitro fertilization, and embryo transfer augment natural breeding programs but rely on good quality semen for best results. It is understood that anesthesia can affect semen characteristics such as ejaculate volume, total sperm count, sperm motility, and incidence of urine contamination. Thus, the aim of this study was to conduct a retrospective analysis of 23 years of data to investigate sperm parameters of semen collected under anesthesia using medetomidine in combination with butorphanol and midazolam or Telazol® alone. Electroejaculation records (Medetomidine, Butorphanol, and Midazolam anesthetized n = 59 ejaculates, from 30 cheetahs, Telazol® anesthetized, n= 169 ejaculates, from 72 cheetahs) were evaluated for incidence of urine contamination. Electroejaculation records (Medetomidine, Butorphanol, and Midazolam anesthetized n = 21 ejaculates, from 17 cheetahs, Telazol® anesthetized, n = 143 ejaculates, from 63 cheetahs) were evaluated for total sperm count, total motility, ejaculate volume, and testicle size. Telazol® treated cheetahs had a numerically higher total sperm count (Median ± SD: 42.58 ± 77.8 × 106 spermatozoa) compared to those treated with medetomidine (Median ± SD: 31.2 ±44.58 × 106 spermatozoa), and a significantly (p < 0.05) higher sperm motility (Median ± SD: 70.0 ± 9.71%) compared to medetomidine (Median ± SD: 53.0 ± 16.41%) treated cheetahs. The findings of this study indicate that medetomidine anesthesia results in significantly lower sperm motility and Telazol® anesthesia results in a higher total sperm count and motility, thus resulting in higher quality ejaculate. This information can aid in the veterinary management of the species when involved in genome resource banking and assisted reproductive technologies.

Gamete collection, gamete preservation, hormone analysis, and artificial insemination have become integral parts of in situ and ex situ conservation programs for threatened and endangered species. Although these methods have been used to assist conservation in many different vertebrate groups, limited work has been done in reptiles. For example, semen collection in lizards has only recently been described and determined to be safe. The purpose of this study was to determine effective doses of human chorionic gonadotropin (hCG) for increasing plasma testosterone concentrations in the veiled chameleon (Chamaeleo calyptratus), as well as to determine whether the rise in testosterone impacts semen collection. A crossover design was used. Initially, chameleons were given injections of 100, 200, and 300 IU of hCG per animal, and serial plasma testosterone measurements were collected over 24 h. The 100-IU/animal dose was determined to increase plasma testosterone concentrations at a similar level to that of the 200- and 300-IU/animal doses. Next, we determined weekly injections of hCG (100 IU/animal) would maintain elevated testosterone concentrations over 30 days. In addition, we determined that elevated testosterone secondary to repeated injections of hCG decreased testicular size as determined by ultrasound. Repeated hCG injections and long-term elevation of plasma testosterone concentrations did not increase the likelihood to collect a semen sample with electroejaculation or improve ejaculate quality. Further research is needed to exogenously stimulate spermatogenesis and increase ejaculate quality to perform timed semen collections in male reptile species. [ABSTRACT FROM AUTHOR]

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Simple Summary: The Bengal tiger (Panthera tigris tigris) is critically endangered, so artificial insemination (AI) is an important conservation tool. Electroejaculation (EE) protocols have been optimized to obtain sufficient amounts of viable sperm for artificial insemination in many felid species; however, less attention has been paid to animal wellbeing during the process. This study examined the effects of three EE protocols (Low, 2–5 volts; Medium, 3–6 volts; and High, 4–7 volts) on semen quality, testicular size, serum creatine kinase (CK, as a proxy for muscle damage), serum testosterone, and cortisol (as a proxy for stress) concentrations, and seminal plasma and sperm pellet malondialdehyde (MDA, as a measure of oxidative stress) measured after each EE series. 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 suggesting the potential for muscle damage (p < 0.05). However, no significant differences were observed for serum cortisol, testosterone, or MDA concentrations across voltage groups. Results suggest the Medium voltage protocol produced good quality samples at lower voltages and with no increase in CK compared to the High protocol, which might be better for animal welfare. 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. [ABSTRACT FROM AUTHOR]

BackgroundFilarial infections have been understudied in bats. Likewise, little is known about pathogens associated with the reproductive system in chiropterans. While semen quality is critical for reproductive success, semen-borne pathogens may contribute to reproductive failure.MethodsFor the first time we performed electroejaculation and used computer-assisted semen analysis to provide baseline data on semen quality in a parti-coloured bat (Vespertilio murinus).ResultsThe semen quality values measured in the V. murinus male appeared high (semen concentration = 305.4 × 106/mL; progressive and motile sperm = 46.58 and 60.27%, respectively). As an incidental finding, however, microfilariae were observed in the bat semen examined. At necropsy, eight adult filarial worms, later genetically identified as Litomosa sp., were found in the peritoneal cavity, close to the stomach, of the same particoloured bat male dying as a result of dysmicrobia and haemorrhagic gastroenteritis in a wildlife rescue centre. Histopathology revealed microfilariae in the testicular connective tissue and the epidydimal connective and fat tissues. A PCR assay targeting cytochrome c oxidase subunit 1 confirmed that adult worms from the peritoneal cavity and testicular microfilariae were of the same filarial species. Mildly engorged argasid mite larvae attached to the bat skin proved negative for filarial DNA and the adult filarial worms proved negative for endosymbiont Wolbachia.ConclusionWhile the standard filarial life cycle pattern involves a vertebrate definitive host and an invertebrate vector, represented by a blood-sucking ectoparasite, our finding suggests that microfilariae of this nematode species may also be semen-borne, with transmission intensity promoted by the polygynous mating system of vespertilionid bats in which an infected male mates with many females during the autumn swarming. Presence of microfilariae may be expected to decrease semen quality and transmission via this route may challenge the success of reproductive events in females after mating. Further investigation will be necessary to better understand the bat-parasite interaction and the life cycle of this filarial worm.

Chameleons represent one of the most endangered groups of lizards, with most being characterized as threatened or near-threatened. Anthropogenic impacts on their native habitat are responsible for their declines. To protect these animals, we must develop functional assisted reproductive programs. However, a full understanding of their basic reproductive biology is required to develop such a program. This study represents the first attempt at characterizing the reproductive cycles of male chameleons. Sixteen veiled chameleons (Chamaeleo calyptratus) and 16 panther chameleons (Furcifer pardalis) being housed in captivity under static conditions were used for this study. Chameleons were anesthetized twice monthly for semen collection via electroejaculation, blood sampling to measure plasma testosterone concentrations, and ultrasonographic testicular measurements. Plasma testosterone concentrations were found to be significantly different between species (P= 0.02) and in the interaction of species and month (P= 0.04). In veiled chameleons, plasma testosterone concentrations were significantly elevated during October through December, while for panther chameleons plasma testosterone concentrations were elevated in July through September, and December. Electroejaculation success was found to be seasonal in both species. In veiled chameleons, semen collection was highest in April, May, and June, while in panther chameleons semen collection success was highest in March, April, May, and June. Median spermatozoa concentrations for veiled and panther chameleons were 6.66 and 7.52 × 107 sperm/ml, respectively. Motility was lower than expected for both veiled (median 23%) and panther chameleons (median 25%). Spermatozoa clumping was a common finding. The majority of the spermatozoa (mean 51%) were found to have normal morphology. The results of this study confirm that veiled chameleons have a prenuptial reproductive strategy, while panther chameleons have a postnuptial reproductive strategy. Additionally, this study demonstrated that chameleons can undergo repeated electroejaculations under anesthesia, twice monthly, for a year without significant mortality or clinical disease associated with the procedures. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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 0.05). 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.

Developing protocols that maximize the total sperm output during a semen collection from an endangered species is essential for strategic genome resource banking practices. Exogenous oxytocin has been administered to domestic bovid species prior to semen collection to increase the total sperm count from a single collection. The hypothesis tested in the present study was that the administration of exogenous oxytocin prior to semen collection by electroejaculation (EEJ) would significantly increase the total sperm count of the ejaculate obtained from banteng. Banteng were administered exogenous oxytocin (50 i.u.) intravenously ∼10 min prior to EEJ. Ejaculates from 13 banteng (n = 6 oxytocin treated, n = 8 untreated) were assessed for volume, pH, osmolarity, sperm concentration, total sperm count, percentage of motile sperm, status (scale of 0–5; 0 = no forward progression, 5 = rapid straight forward progression). In banteng ejaculates, significant (p

In 1995, eight female pumas from Texas (Puma concolor stanleyana) were introduced into five areas in South Florida to remediate morphological and biomedical correlates of inbreeding depression that threatened the long-term survival of Florida panthers (P. c. coryi). Initial analysis of the results of this genetic introgression initiative has highlighted its success, including growth of the population, increased genetic heterozygosity, and reduced frequencies of physical traits associated with inbreeding, including cryptorchidism. However, panthers remain isolated from other puma populations and natural gene flow is not currently plausible. Using data collected from 65 panthers in the post-genetic introgression era (1998–2020), we assessed the benefits accrued to male panther reproductive parameters via this management initiative, the effect of genetic variables, and compared results to historic data. Microsatellite data were used to determine individual heterozygosity levels and qualify panther genetic ancestry as admixed (i.e., panthers descended from crosses with female Texas puma) or canonical (i.e., panthers that comprised most of the population prior to genetic introgression). A total of 47 admixed and 10 canonical panthers were identified; mean individual heterozygosity of canonical panthers (0.39) was significantly different from admixed panthers (0.63; t54 = 4.7, P-value < 0.001). Admixed panthers exhibited higher mean values for total testicular volume, structurally normal spermatozoa, and total sperm in comparison to canonical panthers. The progressive improvement of sperm metrics demonstrates that the benefits of genetic introgression continue to have a positive impact on the population >25 years since the release of the Texas pumas. However, the Florida panther population remains small, isolated, and vulnerable to deterministic and stochastic events that warrants continued monitoring to assess when additional genetic introgression initiatives will need to be implemented in the future. [ABSTRACT FROM AUTHOR]

Implementing alternatives methods to dorsal massage (e.g., electroejaculation) for recovering semen from fighting rooster, known to be very stressful due to its aggressiveness, has become a priority for breeders of this cock breed in Ecuador. Therefore, the objective of the present study was to evaluate two semen collection techniques in fighting roosters, one by electroejaculation (EE) and another by dorsal massage (DM) on seminal quality parameters. For this purpose, thirty attempts of semen recovery from six adult Spanish fighting roosters were carried out using DM (n = 12) and EE (n = 18). Electroejaculation was performed previous sedation, applying five stimulation cycles (of 2 s) generated from a handmade electroejaculation probe (9 to 12 V). The results showed that the EE produced lower response (P < 0.01) to semen ejaculation than the DM (44.4 % vs. 100.0 %, respectively). However, semen samples obtained by EE had better (P < 0.05) spermatic kinetic with greater values of straight-line velocity (VSL, µm/s), average path velocity (VAP, µm/s), and beat-cross frequency (BCF, Hz) as well as higher percentages (P < 0.01) of wobble and linearity compared to DM, irrespective of sperm viability. In addition, the number of urates present in the ejaculates obtained by EE was lower (P < 0.05) than those obtained by DM. In conclusion, electrical stimulation with prior sedation produced a low semen ejaculation response in fighting cocks. However, EE yielded semen ejaculates with better spermatic kinetic compared with the conventional dorsal massage technique.

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.

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. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the morphological and morphometric characteristics of guinea pig sperm (Cavia porcellus), using an image analysis system. Semen from collected from five type 1 guinea pigs, four months old (1.19 ± 0.15 kg) by electroejaculation method, between 3 and 5 times for each animal, making a total of 22 adequate collections. The volume, pH, sperm concentration, total motility and percentage of live sperm were determined. The samples for the analysis of sperm morphology and morphometry were stained with Spermac® and analyzed using the Motic Image Plus® software. The morphology was analyzed according to the dynamics of its acrosomal reaction, distinguishing 4 classes. The length, width, area, perimeter, ellipticity, elongation, regularity and rugosity of the head of the guinea pig sperm were determined, as well as the midpiece length and the tail of the spermatozoa. Guinea pig sperm morphology was analyzed with a complete random design, and the morphometric parameters by random blocks, using Duncan's test to compare means in both cases. In relation to morphology, significant differences (P

ABSTRACT: The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%. In ejaculate samples, higher values of total morphological defects were observed after 24 and 48 hours of refrigeration at 4°C (P

abstract: The Bornean sun bear (Helarctos malayanus euryspilus), endemic to Borneo island, is the subspecies of the Malayan sun bear. The species is at risk, not just because of anthropogenic threats, but also slow reproduction in the wild. In captivity, due to poor reproductive performance, assisted reproductive technology is deemed a fundamental tool to propagate the depleting numbers of the Bornean sun bear. This is a pioneer study that presents the semen characteristics of the Bornean sun bear via conventional semen evaluation methods. Forty two semen samples from ten sun bears were collected via electroejaculation and evaluated. The electroejaculator probe (2.5 cm in diameter and 7.0 cm in length) was inserted rectally and positioned dorsal to the prostate gland. The optimum voltage used to obtain semen differed with each individual, but all showed hindlegs contraction and penile erection before ejaculation. The average combined testes volume in the Bornean sun bear was 23.37 ± 5.09 cm³ . The mean semen volume was 617.30 µL, with sperm concentration of 1034.40 × 10⁶ sperm/mL and pH 7.79. Sperm viability was 80.19% with a general motility of 79.13% and progressive movement of 70.20%. There were high sperm abnormalities at 70.67%. Sun bear sperm length was 61.28 ± 2.46 µm and consisted of an oval head, midpiece, and tail. From this study, good semen donors were identified from the captive Bornean sun bear population in Sabah, Malaysia. The fresh semen baseline data established in this study will provide crucial reference for assisted reproduction programs in the Bornean sun bear.

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. •Fasting for 24 h prior semen collection by electroejaculation reduced the bladder size. •Increased the proportion of bulls with penis protrusion, erection and ejaculation. •No differences were detected in behavioral responses, volume of rectal fecal content, and ejaculate parameters compared with non-fasting beef bulls. [ABSTRACT FROM AUTHOR]

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 × 106 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. Highlights: Electroejaculation was used for the semen collection of giant anteaters.Teratospermia was found in few captive giant anteaters. [ABSTRACT FROM AUTHOR]

Assisted reproductive technologies can aid conservation efforts via support of ex situ population management and preservation of genetic material. Data from 38 sperm collection attempts from 17 polar bears (1–5 procedures/bear) were evaluated. Sample collections were attempted via electroejaculation (EEJ; n = 6), urethral catheterization (UC; n = 25), or sperm rescue (SR; n = 7) during the breeding season (Jan. 1-May 21; n = 27) and nonbreeding season (May 22-Dec. 31; n = 11). Sperm retrieval was successful in 1 EEJ (16.7%), 18 UC (72.0%) and 4 SR (57.1%) collections. Initial sperm motility and viability were 50.0% and 77.0% for EEJ, 64.3 ± 7.4% and 80.9 ± 3.8% for UC, and 56.7 ± 8.8% and 80.5 ± 0.5% for SR. UC and SR were more likely to be successful during the breeding season (84.2–100%) than the nonbreeding season (25.0–33.3%). Testicular tumors were observed in four males (57%) during SR. In total, 13 samples were cryopreserved (n = 1 EEJ, 9 UC, and 3 SR) with egg-yolk-based equine extender (EQ) or OptiXcell (OP). For both extenders, post-thaw motility and viability were reduced by 20–60% and 30–65%, respectively. Further efforts to optimize procedures are warranted, but this summary provides data useful for enhancing the success of polar bear sperm collection and cryopreservation.

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. [ABSTRACT FROM AUTHOR]

The giant armadillo (Priodontes maximus) is the largest living armadillo and is classified as Vulnerable by the International Union for Conservation of Nature. Little is known about the species biology and especially its reproduction. However, data such as the age at sexual maturity are employed for estimation of a species generation length, which is an important parameter for extinction risk assessment. Thus, the aim of this study is to estimate the age at sexual maturity for male P. maximus in the wild and to predict generation length for this species. During a long-term in situ study, eight male P. maximus were captured for reproductive evaluation considering both morphometric characteristics (e.g. body, penis and testes dimensions), and data on sperm production (assessed by electroejaculation). Based on observations of this study, it is estimated that male P. maximus reach sexual maturity at an age of 6.5–8 years. This is at least 5.5–7 years later than previously published. Consequently, generation length for P. maximus results in 11 years when applying standard formula including age at sexual maturity in males and an estimate of age at last reproduction of 20 years. However, the generation length is probably underestimated here as sexual maturity in females and age at last reproduction could be later than estimates used. Even these conservative estimates result in noticeable change for the timeframe used in extinction risk assessments for this species as it is 33 instead of 21 years. Thus, results from this study might affect the classification status of P. maximus particularly for regional assessments. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The current study aimed to evaluate motility and viability of chilled ram semen collected by artificial vagina and electroejaculation during the non-breeding season. A total of 18 ejaculates from clinically healthy rams in a non-breeding season were collected by artificial vagina (AV; n = 9) and electroejaculation (EE; n = 9) and submitted to preliminary evaluation. After that all ejaculates were diluted through a Tris-based extender containing low concentration (5%) of glycerol and egg yolk and stored at 5°C for 48 hours. Motility and viability of semen samples were evaluated at 0, 6, 24 and 48 h of storage. Estimation of motility was carried out by a microscopic digital system, and viability was assessed by the one-step eosin-nigrosin staining technique. Until 6 h of storage, the differences between motility and viability of semen collected by AV or EE were non-significant, while at 24 and 48 h the parameters were higher (P < 0.05) in semen collected by AV. The increasing time of storage correlated negatively with the evaluated parameters (P < 0.05). In conclusion, the chilled semen from ram in the non-breeding season collected by AV demonstrated better motility and viability until 48 h, compared with the semen collected by EE, and could be recommended for artificial insemination up to 24 h after storage at 5°C. The time of storage had a negative effect on sperm motility and viability (P < 0.05). [ABSTRACT FROM AUTHOR]

Sperm limitation is a concern for a number of heavily fished decapods; however, work to assess this concern is sometimes hampered by a lack of simple techniques to quantify sperm transferred during reproduction. Our primary goal was to determine if DNA measurements could be used to quantify the sperm content of spermatophores and thus facilitate investigations of sperm limitation in American lobsters (Homarus americanus H. Milne Edwards, 1837). This was achieved by measuring the amount of DNA in a sample and then calibrating those values by using flow cytometry to count the number of individual sperm present in the sample. Our results show that the DNA quantification technique provides a fast and accurate way to quantify sperm. We then demonstrated the utility of the method by using it to examine the rate at which males can produce sperm under simulated conditions of repeated mating events, a situation that might lead to a reduction in the number of sperm per spermatophore. While spermatophores obtained from male lobsters at three-day intervals varied substantially in the number of sperm they contained (range 427,090–5,028,996; mean 2,306,473), there was no clear decline in sperm count over time. These results suggest that male lobsters replenish their sperm supplies rapidly, and that sperm recharge rate is unlikely to be a factor that could lead to sperm limitation in American lobster populations. [ABSTRACT FROM AUTHOR]

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This study was carried out to evaluate the effect of electroejaculation (EEJ) on the serum concentrations of the acute phase proteins (APPs) serum amyloid A (SAA) and haptoglobin (Hp), and on the bone metabolism biomarkers osteocalcin (OC), bone-specific alkaline phosphatase (b-ALP) and pyridinoline cross-links (PYD). Twenty sexually mature apparently healthy male camels were assigned to EEJ. Parallel, 8 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 h after (T2) EEJ or natural mating camels. The serum concentrations of the APPs and bone biomarkers were determined. The serum concentrations of SAA increased significantly at T1 compared to T0. In none of the 2 groups was the serum concentration of Hp differed significantly. The serum concentrations of OC increased significantly at T1 compared to T0. In none of the 2 groups was the serum concentration of b-ALP differed significantly among T0, T1 and T2. It is concluded that acute phase reaction, manifested by significant increases of SAA, had occurred in male camels as a result of EEJ. In addition, EEJ increased serum concentration of the bone formation biomarker OC.

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 (106/ml), general motility (%), progressive motility (%), and % morphology of normal spermatozoa are 0.86±0.18 ml, thin milky to milky, 1194.2±346.1 106/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®.

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

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 × 106 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. [ABSTRACT FROM AUTHOR]

Highlights • Normal ECG values with reference ranges of the Egyptian Baladi goat bucks were determined. • Electroejaculation (EE) induced sinus tachycardia with changes in waves, amplitudes and durations denoting cardiac stress. • Cortisol, P 4 , testosterone, cardiac troponin I, glucose, sodium, potassium and triglycerides levels were affected by EE. • The marked increase in HR indices and cortisol between the first and last weeks might indicate lack of habituation to EE. Abstract This study aimed to evaluate the stress response to electroejaculation (EE) through electrocardiogram (ECG) indices alongside hormonal and biochemical indices. Lead II base-apex ECG was recorded at rest (control, T0) and immediately after EE (T1), repeated in four consecutive weeks. The collected ejaculates were evaluated for standard semen characteristics (ejaculate volume, mass motility, individual motility, sperm cell concentration and total sperm count per ejaculate). Blood samples were collected at rest (T0), 10 (T2) and 120 min (T3) after EE for determination of biochemical and hormonal stress indices. EE induced sinus tachycardia with simultaneous marked increase in P waves amplitude and decrease in all waves and intervals durations (except for QRS complex and PR interval). Similarly, significant variation between the first and the last EE sessions were observed in heart rate (HR) indices (mean RR, mean, maximum and minimum HR), some wave amplitudes and segment durations. While the stress index significantly increased in the 3rd week only. This was associated with significant short term increases in serum level of cortisol and progesterone (P 4) with significant decrease of glucose and globulins at T2. Furthermore, at T3 there was significant increase in cardiac troponin I (CTnI) concentration, while free triiodothyronine (FT 3), potassium (K+), calcium (Ca2+) and glucose levels decreased significantly. There were significant increases between the first and the last EE sessions in cortisol, FT 3 and significant decrease in Lactate dehydrogenase (LDH) concentrations. On contrary, there were no significant changes in time related to EE and weekly repetitions in the corrected QT interval (QTc), root mean square of sum of squares of normal to normal difference (RMSSD), standard deviation of the normal to normal (SDNN) vasovagal tonus index (VVTI) and semen characteristics. In conclusion, although repeated EE causes acute stress response which increases with repetition, the response appeared transient and not deleterious to the cardiac activity and semen characteristics evidenced by non-persistent significant increases in CTnI and absence of significant changes in the standard semen characteristics of the bucks. Additionally, ECG might be beneficial in assessing the welfare of electroejaculated bucks. Further assessment of heart rate variability (HRV) indices of bucks after EE using frequency and non-linear indices in a larger animal population is recommended. [ABSTRACT FROM AUTHOR]

Contents: 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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.

Couples with a spinal cord injured male partner require assisted ejaculation techniques to collect semen that can then be further used in various assisted reproductive technology methods to achieve a pregnancy. The majority of men sustaining a spinal cord injury regardless of the cause or the level of injury cannot ejaculate during sexual intercourse. Only a small minority can ejaculate by masturbation. Penile vibratory stimulation and electroejaculation are the two most common methods used to retrieve sperm. Other techniques such as prostatic massage and the adjunct application of other medications can be used, but the results are inconsistent. Surgical sperm retrieval should be considered as a last resort if all other methods fail. Special attention must be paid to patients with T6 and rostral levels of injury due to the risk of autonomic dysreflexia resulting from stimulation below the level of injury. Bladder preparation should be performed before stimulation if retrograde ejaculation is anticipated. Erectile dysfunction is ubiquitous in the spinal cord injured population but is usually easily managed and does not pose a barrier to semen retrieval in these men. Semen analysis parameters of men with spinal cord injury are unique for this population regardless of the method of retrieval, generally presenting as normal sperm concentration but abnormally low sperm motility and viability. When sperm retrieval is desired in this population, emphasis should be placed on initially trying the simple methods of penile vibratory stimulation or electroejaculation before resorting to more advanced and invasive surgical procedures.

This study was carried out to evaluate the effect of electroejaculation (EEJ) on the serum concentrations of the acute phase proteins (APPs) serum amyloid A (SAA) and haptoglobin (Hp), and on the bone metabolism biomarkers osteocalcin (OC), bone-specific alkaline phosphatase (b-ALP) and pyridinoline cross-links (PYD). Twenty sexually mature apparently healthy male camels were assigned to EEJ. Parallel, 8 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 h after (T2) EEJ or natural mating camels. The serum concentrations of the APPs and bone biomarkers were determined. The serum concentrations of SAA increased significantly at T1 compared to T0. In none of the 2 groups was the serum concentration of Hp differed significantly. The serum concentrations of OC increased significantly at T1 compared to T0. In none of the 2 groups was the serum concentration of b-ALP differed significantly among T0, T1 and T2. It is concluded that acute phase reaction, manifested by significant increases of SAA, had occurred in male camels as a result of EEJ. In addition, EEJ increased serum concentration of the bone formation biomarker OC. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract: 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]