The Effects of Frequent Electroejaculation on the Semen Characteristics of a Captive Siberian Tiger (Panthera tigris altaica)

Artificial insemination (AI) can help to avoid inbreeding and genetic degeneration for sustaining genetically healthy populations of endangered species in captivity. Collection of a sufficient quantity of viable sperm is an essential first step in the AI process. In the present study, we examined the effects of frequent electroejaculation on semen characteristics in a Siberian tiger. We collected semen in all 17 trials during 6 breeding seasons (6 years). The mean number of sperm and the percentage of motile sperm were 294.3 ± 250.2×10⁶/ejaculate and 82.4 ± 11.4%, respectively. The number of motile sperm tended to increase during frequent electroejaculation in the same breeding season. Semen collection by electroejaculation can be performed effectively up to the fourth sequential ejaculate, which contained the most sperm in the study. In conclusion, frequent collection of sperm by electroejaculation from tigers may be effective for collection of a large number of motile sperm.     

Electroejaculation and artificial insemination in Vietnamese potbellied miniature pigs

A 15-month-old Vietnamese miniature boar was examined because of suspected infertility. A breeding soundness examination was conducted, using electroejaculation under anesthesia for semen collection. Semen values were normal despite a subpubic location of the testes. Artificial insemination of a gilt with extended semen resulted in the birth of a litter 111 days later. Vietnamese potbellied boars have small and sometimes nearly undetectable scrotal pouches, which may cause the producer to question the fertility of the boars.     

A comparison of quality parameters of fresh feline ejaculates collected by three different collection techniques

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.     

Evaluation of semen collected by electroejaculation from captive lesser Malay chevrotain (Tragulus javanicus).

Thirteen sexually mature captive male lesser Malay chevrotains (Tragulus javanicus) were each anesthetized twice with tiletamine-zolazepam for electroejaculation. Viable spermatozoa were collected from all animals. The semen was creamy, milky, pale yellowish, or watery. The mean values for ejaculate volume, sperm concentration, and percentages of sperm motility, normality and viability were 23.7 +/- 2.5 microl, 366.9 +/- 127.8 x 10(6) spermatozoa/ml, 40.0% +/- 3.1%, 71.4% +/- 1.6%, and 59.6% +/- 2.1%, respectively. Semen pH was 7-8. No adverse effects of electroejaculation were noted. These are the first reported values for semen of lesser Malay chevrotain. Electroejaculation should be usable for routine semen collection in this species.     

Ultrasonographic monitoring of artificially stimulated ejaculation in three rhinoceros species (Ceratotherium simum, Diceros bicornis, Rhinoceros unicornus).

Manual massage of the penis and rectal electroejaculation methods have been minimally effective for collecting semen from the rhinoceros. These two methods for stimulating ejaculation were evaluated by rectal ultrasonography. One individual each of three rhinoceros species (Ceratotherium simum simum, Rhinoceros unicornis and Diceros bicornis) was stimulated by manual massage of the penis and two black rhinoceroses (Diceros bicornis) were electroejaculated. On ultrasonography, neither manual massage nor rectal electroejaculation affected the accessory glands. The pelvic urethra remained empty during manual massage of unconditioned animals; however, this area was filled before commencement of massage of conditioned animals. The pelvic urethra also filled during electroejaculation. Semen accumulated in the pelvic urethra during pauses between electrical stimulations and was moved distally into the penile urethra by rectal and penile massage. The volumes of seminal fluid recovered from electroejaculation in this study exceeded previously reported attempts. This study demonstrated the potential of transrectal imaging for improving the recovery of semen by these methods.     

Semen collection techniques.

Semen collection techniques in the stallion have evolved considerably over the last 70 to 80 years and are used today primarily for artificial insemination. Semen can be collected from stallions that are otherwise unable to breed, allowing continued use of valuable animals. There are many options for collection of semen from stallions that present with ejaculatory dysfunction (see the article by McDonnell elsewhere in this issue.) Although there are many advantages to the use of artificial breeding, the collector must understand each step of the collection procedure as well as stallion preferences and proper use of an artificial vagina and mount source so that a representative semen sample is collected.     

Electroejaculation and semen cryopreservation of free-ranging Japanese black bears (Ursus thibetanus japonicus)

The Japanese black bear (Ursus thibetanus japonicus) is endangered for extinction in some areas of Japan, and semen collection and cryopreservation are an important means to preserve genetic resources. The aim of this study was to characterize and cryopreserve semen of free-ranging Japanese black bears. Semen was collected by electroejaculation procedure from 4 free-ranging Japanese black bears at the capture point in the field. Ejaculates containing motile sperm were recovered from all of the animals and ejaculate volume, total sperm count, % motility (percentage of motile spermatozoa), % viability (percentage of spermatozoa that excluded eosin) and % abnormal morphology (range (mean)) were 0.65-2.20 (1.51) ml, 99-1082 (490) x 10(6), 5-100 (31), 42-97 (66) and 20-87 (53), respectively. Three of the 4 ejaculates were diluted with an egg yolk-TRIS-citrate-glucose extender and cryopreserved in liquid nitrogen. Motile spermatozoa were observed after freezing and thawing in all cases. This study showed that electroejaculation would be a useful method for collecting semen from free-ranging Japanese black bears and that at least motile spermatozoa would be obtained by freezing the thus collected electroejaculates.     

Comparison between the Sperm Morphology in Semen Samples Obtained from Yearling Beef Bulls by Transrectal Massage of the Ampullae and Cauda Epididymal Dissection

As electroejaculation (EEJ) is prohibited for use on unanaesthetized animals in Sweden, there is a need for an alternative method of semen collection from bulls in the field. The aim of the present study was to evaluate the use of transrectal massage (TM) of the ampullae to collect semen from yearling beef bulls under field conditions in Sweden. Transrectal massage was performed on 52 yearling beef bulls. Volume of semen collected, duration of procedure, percentage progressively motile sperm, and sperm concentration were measured. Smears were prepared for sperm morphology examination. Semen samples were obtained from 47 of 52 bulls. Mean volume was 3.2 ml (SD +/- 3.7), mean duration of collection was 7.4 min (SD +/- 2.8), mean percentage progressively motile sperm was 43.5% (SD +/- 29.2) and mean concentration was 201.9 x 10(6) spermatozoa/ml (SD +/- 278 x 10(6)). Twenty-three of the 52 bulls were slaughtered 3-4 days after semen collection and aliquots of the cauda epididymal contents were collected for sperm morphology examination. The percentages of proximal droplets, abnormal tails and abnormal midpieces were significantly higher (p < 0.05) and the percentage of detached heads was significantly lower (p < 0.05) in the post-mortem samples compared with those in the TM samples. However, importantly there was no significant difference between the two sample types in the percentages of abnormal heads. This study demonstrates that semen can be collected from yearling beef bulls by TM. We think that TM constitutes a useful tool, when semen collection with EEJ or artificial vagina (AV) is not possible under field conditions, when included in the bull breeding soundness evaluation (BBSE) protocol. However, further studies are needed, and presently being carried out, to evaluate if semen samples collected by TM are comparable with semen samples collected by AV.     

Semen collection by urethral catheterization and electro-ejaculation with different voltages,and the effect of holding temperature and cooling rate before cryopreservation on semen quality in the Japanese macaque (Macaca fuscata)

In the Japanese macaque, semen has been collected by electro-ejaculation (EE), using the higher voltage stimuli compared to other species including genus Macaca. Semen coagulates immediately after ejaculation, which makes difficult to produce high-quality semen for artificial insemination. Recently, semen collection using urethral catheterization (UC) has been reported in carnivore and this technique may allow semen collection without coagulation in a less invasive manner. Further, the temporal preservation temperature and cooling rate of semen during cryopreservation affect post thawing sperm quality. In this study, to improve semen quality and quantity, as well as the animal welfare, semen collection was performed by EE with high (5–15 V) or low (3–6 V) voltage, UC and a combination of the two (EE-UC). It has been suggested that a high voltage is necessary for semen collection, but 10 V stimulation was effective enough and 15 V is for additional sperm collection. Also, liquid semen was collected by EE-UC and this could increase the total number of sperm. Further, to improve the post thawing sperm motility, semen was kept at four temperatures (4, 15, 25 and 37°C) for 60 min, and processed with two cooling procedures (slow cooling before second dilution and fast cooling after second dilution). Holding semen at 25°C and fast cooling after the second dilution maintained progressive motile sperm rate. The present results will contribute to the improvement of semen collection and animal welfare of Japanese macaques.     

COLLECTION OF SEMEN FROM STALLIONS AT STUD

Semen was collected from 222 stallions of 13 breed or colour types in southern Queensland and northern New South Wales. A total of 648 collection attempts were made, using an artificial vagina, during 4 consecutive breeding seasons (1974/5 to 1977/8). Modifications were made to the techniques used by previous workers because collections were made at commercial studs using minimal animal restraint.
Of all collection attempts, 621 (96%) were successful, while at least one semen sample was collected from each of 216 stallions (97%). There were no significant relationships between stallion collection failures and breeding season, time of year or age and breed of stallion. Time of year (seasons and months) was the only factor having a significant relationship with collection failures; highest failure rates occurring in autumn and winter.
The techniques described are applicable for breeding soundness examinations of untrained stallions and for collection of semen for artificial insemination.     

Sanitary Procedures for the Production of Extended Semen

Semen is collected and processed from a variety of animal species for use in artificial insemination breeding programmes. Because of the inherent nature of the semen collection process, bacterial contamination of the ejaculate is a common occurrence. Additionally, manipulation of the ejaculate during processing in the laboratory can expose the sample to possible introduction of bacterial contamination. If preventative measures at the stud fail to adequately control these risks, decreases in semen quality, dose longevity and fertility may occur. Multiple mammalian and non-mammalian sources have been identified as origins of contamination in the stud. Knowledge of these sources has aided the industries in developing strategies that help in controlling the introduction of contaminant bacteria in extended semen. A primary step in minimizing contamination is in the practice of good hygiene by stud personnel. Prudent general sanitation protocols should also be followed in the laboratory, animal housing and semen collection areas. Cleanliness and attention to the actual semen collection process can also aid in reducing bacterial load originating from the stud semen donor. Attentiveness to all of these steps significantly contributes to an overall reduction in the type and amount of bacterial contamination. However, their complete elimination stills remains unavoidable. To address residual bacteria load in the sample, antimicrobials are commonly used in semen extenders intended to promote in vitro sperm longevity beyond that of a few hours. Current research by the animal industries continues in the selection and prudent use of antimicrobials that will lead to the success and sustainability of this modality in controlling bacterial contamination.     

The value of sexed bovine semen

Semen sexing may be used in a variety of practical situations, where part (or all) of the herd may be inseminated with X‐ and Y‐chromosome‐enriched semen. Expressions are presented to calculate the net present value of progeny derived from a semen dose, dependent on the values of females and males, and on the efficiency of sexing. Sexed semen allows selling a higher proportion of the more valuable sex and also increases the value of animals retained for breeding through more intense selection. The expressions to economically evaluate sexed semen are quite general but numerical examples are presented for several practical situations of interest in Brazil.     

Methods of collection,extender type,and freezability of semen collected from creole bulls raised in the tropical highlands of Ecuador

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.

     

Post-mortem recovery,in vitro maturation and fertilization of fallow deer (Dama dama,Linnaeus 1758) oocytes collected during reproductive and no reproductive season

Habitat degradation leads to small and fragmented populations, lower genetic variability and fertility overtime. Assisted reproductive techniques represent important tools to cope with the dramatic loss of biodiversity. Fallow deer (Dama dama), beyond its high commercial value and wide distribution, may represent the most suitable model to study endangered cervids. In this study, oocytes were recovered post-mortem from fallow deer during the breeding and no breeding seasons and were in vitro matured (IVM). The ability of cryopreserved thawed sperm samples recovered by electroejaculation from four adult males was tested by in vitro fertilization of IVM oocytes. The number of oocytes collected per ovary did significantly vary across seasons from 6.2 ± 0.92 during breeding season to 10.4 ± 1.26 during no breeding season (p = .006). Oocytes collected during the breeding season showed higher in vitro fertilization rate compared to the no breeding season (p = .045). However, no embryos reached the blastocyst stage. Semen samples obtained by electroejaculation were successfully cryopreserved, although the cryopreservation process negatively affected most kinetic parameters, mainly at 2 hr post-thawing. Moreover, the percentage of rapid spermatozoa significantly decreased between fresh samples and at 2 hr post-thawing, whereas the percentage of slow spermatozoa increased across the same period (p < .05). Our study provides the logistic steps for the application of assisted reproductive techniques in fallow deer and might be of great interest for genetic resource bank planning.     

The Effect of Season on Spermatozoa Motility,Plasma Membrane and Acrosome Integrity in Fresh and Frozen–Thawed Semen from Xinong Saanen Bucks

The aim of this study was to evaluate whether the season of ejaculate collection influences seminal quality parameters of pre‐ and post‐freeze–thawing in Xinong Saanen bucks. Ejaculates were collected from eight bucks throughout the four seasons (spring, summer, autumn and winter) in a 12 months’ time period, identified in the Northern Hemisphere. Semen samples were evaluated by the combinations of conventional and Computer‐Assisted Sperm Analysis (CASA) when fresh and after frozen–thawed, respectively. The results clearly demonstrated that season of ejaculate collection influenced (p < 0.05) fresh semen quality. Highest semen quality was observed during autumn. On the contrary, undesirable indices (significantly lower, p < 0.05) were observed in winter as compared with the other remaining seasons. CASA has clearly shown the influences of seasonal variations on semen motility parameters. Furthermore, season of ejaculate collection was also found to influence sperm freezability. Semen characteristics after frozen–thawed followed a similar pattern with that of fresh ejaculate except in spring. The results revealed that sperm quality was higher (p < 0.01) in summer and autumn than in spring and winter. In conclusion, seasonal variation influences semen quality in Xinong Saanen bucks. In addition to summer and autumn, fresh ejaculates in spring can also be successfully used for AI. Sperm from ejaculates collected during summer and autumn are more suitable for cryopreservation. Hence, it is possible to increase the efficiency of goat breeding by manipulating the seasonal variations of semen quality for immediate AI and/or cryopreservation.     

胚胎移植蒙贝利亚种公牛在北方地区适应性研究

本研究于2008年7月～2011年3月,对进口胚胎移植出生并经选育的9头蒙贝利亚种公牛的环境适应性、生长发育、抗病能力、繁殖性能等进行了观察、测定和分析研究。结果显示,30月龄时平均体高151.89cm,睾丸周径42.11cm,体重922.67kg,平均日增重0.959kg。10月龄开始初次采精,24月龄正式采精;平均每次射精量6.20mL,鲜精活力0.732,精子密度13.083亿个/mL,平均利用率82.8%,均产冻精40 606剂;冻精活力≥0.4,精子畸形率<15%。精液质量达到牛冷冻精液国家标准(GB4143-2008)。适应性研究表明,胚胎移植蒙贝利亚种公牛对北方地区的环境条件较为适应,生长发育状况良好,抗病力强,具有良好的繁殖生产性能。     

Pregnancies from imipramine and xylazine-induced ex copula ejaculation in a disabled stallion.

Breeding or semen collection was attempted using: natural cover, manual stimulation, artificial vagina, pharmacologic induction of ejaculation, and electroejaculation. Sperm cells were recovered from the ductus deferens and epididymides post mortem. Only semen collected ex copula by imipramine and xylazine treatment resulted in conceptions (4/5). This is the first report of pregnancies in horses from ex copula semen collection.     

Evaluation of possible methods to reduce pain associated with electroejaculation in bulls.

Heart rate changes were recorded in attempt to objectively measure pain associated with 6 methods of semen collection, including transrectal massage, conventional electroejaculation, and electroejaculation after either intrarectal lidocaine, epidural lidocaine, epidural xylazine, or intravenous xylazine. Epidural anesthesia with lidocaine or xylazine tended to result in smaller heart rate changes.     

Strategies for the management of donkey jacks in intensive breeding systems

Donkeys are bred throughout the world and often play an important role in agriculture. They are also bred to produce mules. Traditionally, jacks are considered challenging to breed in domestic conditions, whether for natural breeding or semen collection using either jennies or mares. The donkey's natural sexual behaviour significantly differs from that of other domestic animals. This presents challenges for in‐hand donkey breeding, particularly on mule studs where normally only jacks and mares are kept for breeding. This article describes some of the authors' observations on sexual behaviour in donkeys and practical experience of some of the strategies employed to apply this knowledge to breeding management, in order to improve the success of using donkeys for both natural service and semen collection.     

Electroejaculation in the Chinchilla (Chinchilla lanigera): effects of anesthesia on seminal characteristics

Repetitive electroejaculation is probably extremely stressful in conscious animals and could adversely affect fertility. The present study was designed to (a) evaluate the effects of anesthesia (40 mg ketamine/kg body weight, i.m.) on a method of electroejaculation used previously in conscious chinchillas (Chinchilla lanigera), and (b) determine the quality of the electroejaculated semen obtained under anesthesia. In Experiment 1 (8 animals), a 4 x 4 Latin square design was used to study the effects of anesthesia and ejaculatory voltage on semen collection, ejaculate volume, sperm concentration, motility, viability, response to the hypoosmotic swelling test (HOST), and acrosomal status. In Experiment 2 (12 animals), the effects of differing voltages and the number of stimuli on ejaculation by conscious or anesthetized males was determined. In both experiments, all the conscious animals ejaculated, but only 60% ejaculated under anesthesia and they required more stimuli and higher voltages to achieve ejaculation. Ejaculate volume was significantly lower in anesthetized (<5 microl) than in conscious animals (>40 microl), but sperm concentration was unaffected. None of the indices of sperm quality were affected by anesthesia. The techniques we have developed in anesthetized domestic chinchillas should be applicable to endangered chinchillas in the wild, and though the number of sperm available is reduced, there are still sufficient for assisted reproduction.