Semen collection and ejaculation trigger changes in the blood flow of the reproductive system in rams.

The aim of this study was to determine if ejaculation modifies the testicular and accessory sex glands' blood flow after ejaculation, and if those changes differ according to the process that leads to ejaculation. Twelve adult Corriedale rams were used and assigned at random to the four procedures that lead to ejaculation: (G1) electroejaculation; (G2) artificial vagina; (G3) transrectal ultrasound-guided massage of the accessory sex glands; (G4) natural mating. Hemodynamic characteristics evaluation of the male reproductive system was conducted immediately before and at 30 and 90 min after ejaculation. The internal iliac artery peak systolic velocity (PSV) decreased (P=0.01) and supratesticular artery PSV increased (P=0.042) 90 min after ejaculation in all groups. In conclusion, ejaculation modifies the reproductive system's blood flow, with slight variations depending on the studied ejaculation methods. Additionally, ejaculation altered the internal iliac and supratesticular arteries PSV, and the supratesticular artery end-diastolic velocity (EDV) in rams. The supratesticular artery PSV was the only studied variable that differed according to the procedure that triggered the ejaculation.

Time of day modified the time required for semen collection with electroejaculation and slightly affected the quality of fresh semen in rams.

The aim was to compare some stress responses to electroejaculation (EE), and the quality of fresh semen, when ram semen is collected at dawn (06:00 h), noon (12:00 h), or evening (18:00 h). Twelve Corriedale rams were used, and semen was collected from four rams at each study time on three different days, with a Latin-square design. The time required for EE, the number of vocalizations emitted, heart rate, and rectal temperature were recorded, and fresh semen was evaluated. The time required for EE was shorter at evening than at dawn and noon (399.3 s, 480.6 s, and 460.2 s respectively; pooled SEM = 72.1; P = 0.03). The percentage of sperm with progressive motility was greater at noon than dawn (59.7% and 50.3%; pooled SEM = 5.8; P = 0.05). Curvilinear velocity was greater at dawn than evening (117.0 µm/s and 95.5 µm/s; pooled SEM = 7.1; P = 0.04), slow linear velocity was greater at evening than at dawn and noon (13.1 µm/s, 9.3 µm/s, and 8.5 µm/s respectively; pooled SEM = 1.7, P = 0.05), and the slow average path velocity was greater at evening than dawn and noon (16.2 µm/s, 11.7 µm/s, and 10.8 µm/s respectively; pooled SEM = 1.9, P = 0.05). In conclusion, the collection time modified the time required for electroejaculation and had only slight effects on the quality of fresh semen. Overall, the time of the day appears to have only slight effects on semen collection and quality.

Single or repeated immunization against GnRH fails to completely abolish spermatogenesis in dwarf bucks (Capra hircus).

In both captive wildlife and production animals is important to develop strategies for population control. Immunization against GnRH is an easy and inexpensive immunocastration method that reduces the concentration of testosterone and decreases sperm quality. However, its effectiveness depends on the species and repetition of the treatment. This study aimed to compare the effectiveness of a single treatment (initial immunization plus a booster with Improvac) vs repeated treatment (six doses of Improvac) to inhibit testicular function and maintain the contraceptive status during long periods in bucks. Three Dwarf bucks (Capra hircus) received two doses of Improvac, the first on Week 0, and the booster 4 weeks later (single immunization, group SI) while three Dwarf bucks received one dose of Improvac every 6 months during 3 consecutive years (repeated immunization, group RI). The other three Dwarf bucks remained untreated (control bucks, group CON). Bucks from RI had a greater decrease in scrotal circumference, testosterone concentration, male odor intensity, and sperm quality than SI bucks. However, there were no differences between SI and CON bucks in any of the variables studied. Overall, repeated treatment of Improvac decreased the testicular function of Dwarf bucks, although did not produce complete infertility. However, the repetition of the treatment produced more intensive negative effects, indicating that the strength of the effects of Improvac is rapidly lost in bucks.

Administration of carbetocin-a long-acting oxytocin analogue-before sperm collection by transrectal ultrasound-guided massage of the accessory sex glands in bucks (Capra hircus) and ibexes (Capra pyrenaica).

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.

Using transrectal ultrasound-guided massage of the accessory sex glands for buck semen collection yields semen with greater cryoresistance than electroejaculation alone during the breeding season.

The cryoresistance of bucks' semen collected by transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) and electroejaculation (EE) was compared during the breeding season (Study 1) or the late non-breeding season (Study 2). Semen was collected from 10 Gabon bucks with both methods in each season and then frozen according to a standardized protocol. The time required for ejaculation, the number of electrical pulses applied, the sum of the electrical pulses applied∗voltage and the total number of vocalizations emitted during the collection by each animal were recorded. In Study 1, TUMASG required a longer time (P = 0.0006) but fewer electrical pulses and a lower sum of pulses∗voltage than EE (P < 0.0001 for both comparisons). Bucks vocalized fewer times during TUMASG than during EE (P < 0001). Semen collected with TUMASG had greater sperm concentration, sperm mass motility, total number of motile sperm and of sperm with progressive motility, and tended to have greater total number of sperm with functional membrane than semen collected with EE (P = 0.02; P = 0.003; P = 0.02; P = 0.02; P = 0.06, respectively). After the freezing-thawing process, sperm collected with TUMASG had a greater quality of the motility, a total number of motile sperm and sperm with progressive motility, a greater percentage of motile sperm and sperm with functional membrane, and tended to have a greater total number of sperm with normal morphology than semen collected with EE (P = 0.04; P = 0.04; P = 0.03; P = 0.02; P = 0.04; P = 0.06, respectively). In Study 1 sperm collected with TUMASG had greater cryoresistance for almost all the variables considered than when it was collected with EE. In Study 2, the number of electrical pulses and the sum of pulses∗voltage were greater with EE than with TUMASG (P < 0001; P = 0.0002, respectively). There were no differences in any sperm variable in fresh or thawed samples collected with TUMASG or EE. However, the other seminal characteristics evaluated did not differ according to the method, and there were no differences in the cryoresistance of any variable. Overall, TUMASG affected animal welfare less than EE and was suitable for collecting semen of good quality with high cryoresistance during the breeding season in buck.

Seasonality of the stress response in goat bucks when there is use of electroejaculation for semen collection.

Electroejaculation (EE) is frequently used to collect semen, but this procedure is both stressful and painful. In seasonal-breeding species the sensitivity to stressors might vary with season of the year when semen collection occurs, therefore, the present study was conducted to compare the responses when imposing the EE procedure, stress response to EE and fresh semen characteristics in goat bucks throughout the year. Semen was collected using EE from goat bucks once in autumn (mid-breeding season), winter (transition to the non-breeding season), spring (non-breeding season), and summer (beginning of the breeding season). The number of pulses applied during ejaculation was larger in the autumn and spring than winter and summer months. The number of vocalizations/pulse while bucks were ejaculating was greater in the summer than winter and spring months. The increase in heart rate was greater in spring than summer months. Cortisol concentration was least in the autumn, and concentration of creatine kinase (CK) was greatest in summer. Sperm mass motility was greater in autumn than spring months. Spermatozoa concentration, total number of spermatozoa ejaculated, motile spermatozoa ejaculated, and progressively motile spermatozoa were greater in the autumn and spring than in the winter and summer months. The bucks were less stressed by imposing EE procedures in the autumn, and buck responses were greater in the spring-summer period. Bucks had a greater incremental increase in serum CK and number of vocalizations in the summer months, indicating there was a greater muscular damage and quite likely pain associated with lesser semen quality.

Sperm collection by electroejaculation in small ruminants: A review on welfare problems and alternative techniques.

There are different perspectives on whether there should be use of electroejaculation (EE) for semen collection because it can be stressful and painful for the males when this technique is imposed. In the present review it is examined 1) the effects of EE on animal welfare and semen quality in domestic and wild small ruminants, 2) benefits and limitations of administering anaesthetics and sedatives prior to EE, 3) advantages/disadvantages of transrectal ultrasonic-guided massage of the accessory sex glands (TUMASG) as an alternative to EE, and 4) benefits of administering hormones, such as oxytocin or PGF2α analogues (which stimulate the contractility of the male accessory sex glands), prior to EE and TUMASG. In general, the administration of anaesthetics, sedatives or hormones reduces the pain and stress caused by EE, and can improve sperm quality, but results may vary depending on the species. The use of anaesthetics is, however, not devoid of risks and pre-EE administration of sedatives, or oxytocin or PGF2α analogues, can aid sperm collection mitigate risks. The TUMASG is less stressful than EE, but its effectiveness varies greatly among species, and it can only be performed by trained personnel. Prior administration of the hormones may also result in a reduction in the period needed to induce ejaculation with use of TUMASG procedures.

Daily administration of a GnRH analogue enhances sperm quality in bucks during the non-breeding season.

The aim of this study was to determine if daily administration of a GnRH analogue (buserelin acetate) to bucks during the non-breeding season increases testosterone concentration and improves sperm quality. Five bucks received a daily dose of buserelin for 10 days, starting on Day 0 (first administration), and another five bucks remained as controls. Testosterone concentrations were greater in treated than in control bucks during the first hours after buserelin administration (P = 0.05), but greater in controls 10 h later (P < 0.01). Sperm mass motility and percentage of motile sperm were greater in treated (3.9 ± 0.6 and 70.1 ± 7.9%, respectively) than in control bucks (1.0 ± 0.6, P < 0.01; 45.0 ± 7.9%, P < 0.05 respectively) on Day 4. Percentage of sperm with normal morphology tended to be greater in treated than in control bucks (81.8 ± 6.2% compared with 63.5 ± 6.4% respectively, P = 0.08). The treatment decreased the percentage of sperm with mid piece defect and with bent tail (7.0 ± 1.5% compared with 12.0 ± 1.5%; 8.0 ± 1.7% compared with 13.5 ± 1.7%, treated and control bucks, respectively, P = 0.05 for both). The square root percentage of sperm with loose but heads with normal structures tended to be less in treated than control bucks (1.3 ± 0.3% compared with 0.4 ± 0.3% respectively, P = 0.06). It was concluded that daily administration of buserelin during the non-breeding season led to a rapid increase in testosterone concentration and improved sperm quality.

Rearing bucks isolated from females affects negatively their sexual behavior when adults.

In some domestic ruminants, contact with females is necessary for normal development of sexual behavior. The aims of this experiment were to determine if rearing bucks isolated from does affects negatively their sexual behavior when adults, and if this negative effect is overcome after four short contacts with females. Sixteen Saanen male kids were maintained during one year in two groups: kids reared in permanent direct contact with four adult goats (FEM; n = 7), and kids that remained isolated from females (ISO; n = 9). When bucks were 12 mo-old, females were removed and both groups were joined in the FEM pen. Nine months later all bucks were individually exposed four times to estrual females for 20 min every 10 days, recording courtship and mounting behaviors. Bucks that were reared with females displayed more courtship behaviors, ejaculations and total mounts (mount attempts, mounts with and without ejaculation; P < 0.0001 for all) than ISO bucks. The combined effect of number of bucks that ejaculated and the time at which they first ejaculated in the first and second tests favored FEM bucks (P < 0.03). It was concluded that the lack of contact with females during the rearing period affects negatively adult bucks' sexual performance, an effect that could not be overcome after repeated exposures to estural does.

Administration of cloprostenol and oxytocin before electroejaculation in goat bucks reduces the needed amount of electrical stimulation without affecting seminal quality.

Electroejaculation (EE) is a widely used semen collection technique; but, it is stressful and painful for the animals. Considering these concerns, it may be important to develop practices to decrease the negative implications of EE on animals. Oxytocin and prostaglandin-F2alpha (PGF2α) stimulate the contractions of the muscles of the male genital tract. Therefore, the aim of this study was to analyze the effectiveness of the administration of oxytocin and/or a PGF2α analogue (cloprostenol) to bucks in relation to their stress response and sperm parameters before semen collection by EE was performed. Semen was collected with EE from 12 Gabon bucks in a 2 × 2 factorial arrangement (factors: with or without oxytocin, with or without cloprostenol). Each treatment was applied to different animals every 3 to 6 days, allowing all the animals to receive all the four treatments. The treatments applied to bucks before EE were as follows: 1) control (ConT), bucks received no hormonal treatment; 2) oxytocin (OxyT), bucks received 10 IU of oxytocin intramuscularly (IM) 30 s before beginning the EE; 3) PGF2α (PgT), bucks received 250 µg of cloprostenol IM 5 min before beginning the EE; and 4) oxytocin plus PGF2α (OxPgT), animals received treatment with both OxT and PgT. The number of electrical pulses, time length needed to achieve ejaculation, number of vocalizations, creatine kinase (CK) concentration and sperm parameters in goat bucks were recorded. The administration of cloprostenol and oxytocin before EE shortened the procedure and decreased the number of pulses and the pulse/voltage applied (P = 0.02 for all). This treatment also tended to decrease the number of vocalizations (P = 0.067). There were no treatment effects in the initial values; neither were there increases in heart rate and rectal temperature, or CK concentration. Seminal variables were not affected by the treatments. In conclusion, it would be important to consider the combined application of cloprostenol and oxytocin before EE, as it can shorten the process, reducing the electrical stimulus with positive effects on animal welfare and without affecting seminal quality in goat bucks.

Sperm collection by transrectal ultrasound-guided massage of the accessory sex glands is less stressful than electroejaculation without altering sperm characteristics in conscious goat bucks.

In anesthetized non-domestic ruminants transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) is an alternative method to collect semen slightly less stressful than electroejaculation (EE). However, some sperm characteristics are better when semen is collected with EE than with TUMASG. As anesthesia reduces the response to stressors, the advantages of TUMASG may be reduced in anesthetized animals, and thus, TUMASG may be even more advantageous in conscious animals. Therefore, the aim of the present study was to compare the stress response and the characteristics of the sperm collected with TUMASG and EE in conscious goat bucks. Semen was collected in 10 bucks with both procedures. During each procedure, the time required for ejaculation, the number of electric pulses applied and the number of vocalizations were recorded. Rectal temperature, heart rate, serum cortisol concentration, biochemical and hematological parameters were measured before and after each procedure. Sperm characteristics [ejaculated volume, sperm concentration, sperm mass motility (scale 0-5), sperm vigor (scale 0-5), the percentages of motile and progressive motile sperm, of sperm vitality, of sperm with plasma membrane integrity, and with acrosome damage and morphological abnormalities] were also determined. Electroejaculation required more electric pulses than TUMASG (P < 0.0001), but TUMASG took more time than EE (P < 0.0001). The EE provoked more vocalizations (P = 0.02) and a greater increase of cortisol concentrations than TUMASG (P = 0.04). Heart rate also tended to be greater with EE than with TUMASG (P = 0.07). The sperm characteristics did not differ between TUMASG and EE. In conclusion, TUMASG was less stressful and probably less painful than EE without affecting the semen quality. Thus, although it required more time, TUMASG is an alternative procedure to decrease the welfare concerns raised by EE in conscious goat bucks.

Continuous contact with females in estrus throughout the year enhances testicular activity and improves seminal traits of male goats.

The "female effect" consists in the stimulation of males' reproductive activity by different signals emitted by females. This stimulation leads to endocrine and behavioral changes that may modify the seasonal pattern of male ruminants. The aims of this experiment were (1) to describe the local reproductive seasonal pattern of Gabon bucks and (2) to determine if continuous chemical, auditory, and visual contact with does in estrus enhances bucks' testicular activity and improves seminal traits throughout the year and modify their seasonal pattern. We used 16 adult Gabon bucks assigned to two experimental groups: nine bucks remained continuously isolated from females (isolated bucks, group IB) and seven bucks were in continuous chemical, auditory, and visual contact through a fence line with does in estrus (stimulated bucks, group SB). During 13 months, scrotal circumference and testosterone concentration were measured weekly and testicular echogenicity was measured every 2 weeks. Also, sperm motility mass and percentage of abnormal spermatozoa were determined, and sperm concentration and total number of motile spermatozoa were calculated every 2 weeks. Testicular echogenicity was greater in IB than that in SB bucks (P < 0.0001), but there were no differences in scrotal circumference. Overall, testosterone concentration was greater in IB than that in SB bucks (P = 0.04), but from late winter to mid-summer, when testosterone concentration presented basal concentrations, SB bucks had greater values than IB bucks (P = 0.004). Sperm concentration (P = 0.05) and sperm mass motility (P = 0.01) were greater in SB than that in IB bucks, and the total number of progressive motile spermatozoa tended to be greater in SB than in that IB bucks (P = 0.1). The percentage of abnormal spermatozoa was lower in SB than in IB bucks in several time points (P < 0.0001). Testicular and seminal traits were better from the end of the spring until mid-autumn. We concluded that does in estrus stimulated bucks' testicular activity, including better seminal quality and a greater increase of testicular fluid content than bucks isolated from females. However, the general seasonal pattern was not modified by stimulation with does in estrus.

Influence of seasonality and stimulus of oestrous does in bucks' aggressiveness.

Aggressiveness is directly related to testosterone concentration, which varies with seasons and in response to female stimulation. The aim was to determine if the frequency and pattern of agonistic interactions between bucks varies seasonally and in response to oestrous female stimulation. In the first study we characterized the pattern of agonistic interactions during feeding throughout a whole year in groups of Iberian ibex bucks; and in the second study, we determined the influence of oestrous does on the frequencies of agonistic behaviours between Gabon bucks in different seasons. In Iberian ibex bucks, the frequency of agonistic behaviours was maximum in summer and testosterone concentration in late autumn. In Gabon bucks, total agonistic interactions increased in bucks stimulated by females in late summer and autumn. In late autumn there were more interactions without physical contact in stimulated than in isolated bucks. Aggressiveness in bucks varied along the year, but the seasonal pattern was related to the context in which behaviours were recorded. Stimulation with oestrous females induced an increase of aggressiveness during the breeding season. At least in the conditions of these studies, testosterone concentrations seemed not to be the main influence on the frequency in which aggressive behaviour was displayed.

Oxytocin administration before sperm collection by transrectal ultrasonic-guided massage of the accessory sex glands in mouflons and bucks.

Transrectal ultrasonic-guided massage of the accessory sex glands (TUMASG) is an alternative technique for sperm collection which decreases negative animal welfare concerns of electroejaculation. However, quality of collected sperm with TUMASG is less than that collected by electroejaculation. Utilisation of TUMASG may not, however, induce strong muscular contractions, so administration of oxytocin before the procedure may improve sperm quality. Therefore, two studies were designed to determine if administration of oxytocin before TUMASG improves the procedure and quality of the collected semen. The first study was performed with anaesthetised mouflons, and the second with non-anaesthetised Gabon bucks. Time length, number of electric pulses applied and numbers of vocalizations were recorded, and sperm characteristics were analysed. The number of electrical pulses and sperm characteristics did not differ with oxytocin administration in mouflons. However, oxytocin administration decreased time length (P=0.02) and tended to decrease the number of electric pulses needed for induction of ejaculation (P=0.08) in bucks. The number of vocalizations and sperm variables did not differ when the two procedures were used. In conclusion, oxytocin administration before TUMASG shortened the procedure and tended to decrease the number of electrical pulses needed to induce ejaculation in conscious bucks, thus, decreasing welfare concerns for the animals in which TUMASG is used. Therefore, its routine inclusion in this species should be considered.

Grouping previously unknown bucks is a stressor with negative effects on reproduction.

Grouping previously unknown animals produces social stress, which might have negative effects on reproduction. The aims of the experiment were to determine if grouping unknown bucks (1) triggers a stress response and produces changes in body weight; (2) affects scrotal circumference, testosterone concentration, and semen quality; and (3) has differential effects between resident and introduced bucks. One group of nine Saanen bucks was transported and introduced (introduced bucks, group IG) to a group of eight Saanen bucks (resident bucks, group RG). On day 0, cortisol concentration and rectal temperature were determined, and from day -7 to day 29, body weight, scrotal circumference, and testosterone concentration were recorded and semen quality was determined. The stress response was different between groups: on several moments on day 0, resident bucks had greater cortisol concentration (P < 0.0001), while introduced bucks had higher rectal temperature (P = 0.02). Body weight decreased similarly in both groups from day -7 to day 2 (P < 0.0001), but on day 29, IG bucks were lighter than RG bucks (P = 0.05). Also, the reproductive response differed between groups: introduced bucks had lower scrotal circumference (P < 0.01), lower testosterone concentration (P = 0.02), and lower percentage of motile spermatozoa in the ejaculate (P = 0.05). It was concluded that grouping unknown bucks was stressful and negatively affected the reproduction, being more serious for the introduced than the resident bucks.

Homosexual behavior in male goats is more frequent during breeding season and in bucks isolated from females.

Male homosexual behavior is observed in many ruminant species; seasonality and isolation from females may affect the display of male-male sexual behavior. Our objectives were to determine whether: 1) homosexual behavior has a seasonal pattern in male goats (Capra hircus); 2) isolation from female goats influences the frequency of the display of homosexual behavior; and 3) the frequency of homosexual behaviors is related to testosterone concentration. Bucks were housed in a pen adjacent to another pen with 3 estrous goats (FC group) or in complete isolation from females (IF group). Homosexual behaviors and testosterone concentrations were recorded in October, December, February and May. Frequencies of penile display and flehmen increased in May and were observed more frequently in the IF than in the FC group. Ano-genital sniffing was more frequent in February and May and was more frequent in IF than in FC bucks. Lateral approaches were more frequent in May, in which period this was more frequently observed in IF bucks. Mount attempts and mounts were only observed in IF bucks. Testosterone concentrations increased from December to February and decreased in May, although they were still greater than in October and December. In May IF bucks had greater testosterone concentrations than FC bucks. Testosterone concentrations were only related to the number of lateral approaches in IF bucks in December. In conclusion, homosexual behavior was more frequent during the late breeding season than during the early and the non-breeding seasons, and in males isolated from females than in males housed near estrous females. Testosterone concentrations appear to be unrelated to homosexual behavior.