Rodolfo Ungerfeld

Physiological and Biochemical Parameters in Response to Electroejaculation in Adult and Yearling Anesthetized Pampas Deer (Ozotoceros bezoarticus) Males

The aim of this study was to characterize changes in physiological and biochemical parameters during electroejaculation (EE) under general anaesthesia in adult and yearling pampas deer males (Ozotoceros bezoarticus). The relation between heart rate, pulse rate, respiratory rate and oximetry with EE voltages was studied. The changes in cortisol, alkaline phosphatase, creatine kinase, aspartate aminotransferase and rectal temperature were determined before and after electroejaculation (BEE and AEE). Heart rate and pulse rate values increased during EE, with a greater increase in heart rate in adults (p < 0.01). Respiratory rate and SpO(2) were not affected by EE or category. The rectal temperature decreased in adults and yearlings during EE (p = 0.0001). Alkaline phosphatase, creatine kinase and aspartate aminotransferase concentrations increased AEE (p < 0.001, p < 0.01 and p < 0.05, respectively). Alkaline phosphatase and creatine kinase concentrations were greater in young than in adult males (p < 0.05). In conclusion, the EE under general anaesthesia in the pampas deer provoked increases in heart and pulse rates, as well as in creatine kinase, aspartate aminotransferase and alkaline phosphatase concentrations, and a decrease in rectal temperature in both categories. Oximetry values were especially low during the procedure. Heart rate increase was greater in adult males, and alkaline phosphatase and creatine kinase concentrations were greater in yearlings.

Reproduction in a semi-captive herd of pampas deer Ozotoceros bezoarticus

Abstract Information on reproductive traits of semi-captive breeding of the pampas deer Ozotoceros bezoarticus arerunguaensis, an endangered species, was registered during 20 years. Birth events were monitored every 1-2 days from 1981 to 2000, and birth date, mother and father identities, sex and mass of fawns were all registered. Although births occurred throughout the year, a seasonal pattern with a peak in spring was observed. Dispersion of births throughout the year increased in multiparous females that previously gave birth four times or more. The interval between two consecutive parturitions was ∼10 months, independently of the mother parity and of the sex of the fawn, but differed according to the timing of the previous birth. Individual variability in interbirth intervals was significantly higher in primiparous than in multiparous females. Hinds born in semi-captivity had their first birth earlier in their life than hinds that were originally captured (at <1 month of age) from the wild. Single fawns were observed in all but one birth, in which twins were observed. Sex ratio was not influenced by the female parity, the season, or by the father identity. Only the mother identity influenced offspring birth mass.

Endocrine and Ovarian Changes in Response to the Ram Effect in Medroxyprogesterone Acetate-primed Corriedale Ewes During the Breeding and Nonbreeding Season

AbstractTwo experiments were performed to determine the endocrine and ovarian changes in medroxyprogesterone acetate (MAP)-primed ewes after ram introduction. Experiment 1 was performed during the mid-breeding season with 71 ewes primed with an intravaginal MAP sponge for 12 days. While the control (C) ewes (n = 35) were in permanent contact with rams, the ram effect (RE) ewes (n = 36) were isolated for 34 days prior to contact with rams. At sponge withdrawal, all ewes were joined with eight sexually experienced marking Corriedale rams and estrus was recorded over the next 4 days. The ovaries were observed by laparoscopy 4–6 days after estrus. Four weeks later, pregnancy was determined by transrectal ultrasonography. In eight ewes from each group, ovaries were ultrasonographically scanned; FSH, LH, and estradiol-17β were measured every 12 hours until ovulation or 96 hours after estrus. The response to the rams was not affected by the fact that ewes had been kept or not in close contact with males before teasing. No differences were found in FSH, LH, estradiol-17β concentrations, growth of the ovulatory follicle, onset of estrus, ovulation rate, or pregnancy rate. Experiment 2 was performed with 14 ewes during the nonbreeding season. Ewes were isolated from rams for 1 month, and received a 6-day MAP priming. Ovaries were ultrasonographically scanned every 12 hours, and FSH, LH, estradiol-17β, and progesterone were measured. Ewes that ovulated and came into estrus had higher FSH and estradiol-17β levels before introduction of the rams than did ewes that had a silent ovulation. The endocrine pattern of the induced follicular phase of ewes that came into estrus was more similar to a normal follicular phase, than in ewes that had a silent ovulation. The follicle that finally ovulated tended to emerge earlier and in a more synchronized fashion in those ewes that did come into estrus. All ewes that ovulated had an LH surge and reached higher maximum FSH levels than ewes that did not ovulate, none of which had an LH surge. We conclude that (a) the effect of ram introduction in cyclic ewes treated with MAP may vary depending on the time of the breeding season at which teasing is performed; (b) patterns of FSH, and estradiol-17β concentrations, as indicators of activity of the reproductive axis, may be used to classify depth of anestrus; and (c) the endocrine pattern of the induced follicular phase, which is related to the depth of anestrus, may be reflected in the behavioral responses to MAP priming and the ram effect.SammanfattningBaggeffektens inverkan på könshormonnivåer och follikeldynamik hos Corriedale tackor som behandlats med medroxyprogesteronacetat (MAP) under och utanför parningssäsongen. Experiment 1 genomfördes under parningssäsongen med 71 tackor som behandlats intravaginalt med MAP under 12 dagar. De tackor som fungerade som kontroller (n=35) var i ständig kontakt med baggar medan tackorna i försöksgruppen (n=36) hölls isolerade från baggar i 34 dagar. Efter avslutad behandling med MAP sammanfördes kontroll- och försöksgrupperna och 8 sexuellt erfarna baggar. Tecken på brunst övervakades under de 4 följande dagarna och hos de tackor som visade brunst undersöktes äggstockarna med laparaskopi 4-6 dagar efter visad brunst. Fyra veckor senare gjordes en dräktighetsundersökning med ultraljud. Hos 8 tackor från kontroll- och försöksgrupp som visat brunst undersöktes äggstockarna med ultraljud var 12 timme samtidigt som blodprov togs för bestämning av FSH, LH och östradiol 17ß. Detta pågick fram till ägglossning eller till 96 timmar efter visad brunst. Av resultaten kunde man inte utläsa någon skillnad i om tackorna varit isolerade från baggkontakt eller inte. Det framkom inte heller några skillnader mellan grupperna i hormonkoncentrationer, tillväxt av den ovulatoriska follikeln, när brunsten startade, ovulationsfrekvens eller dräktighetsresultat.Experiment 2 genomfördes med 14 tackor utanför parningssäsongen. Tackorna hölls isolerade från baggar under 1 månad. De sista 6 dagarna innan baggar återfördes till gruppen behandlades tackorna under 6 dagar intravaginalt med MAP. Innan baggarna återförts till gruppen, påbörjades undersökning av äggstockarna med ultraljud var 12 timme och blodprov togs för bestämning av FSH, LH, östradiol17ß och progesteron. Resultaten visade att de tackor som kom i brunst hade högre koncentrationer av FSH och östradiol17ß innan baggarna återfördes till gruppen. Det indikerar att dessa båda hormoner skulle kunna användas för att bedöma hur djupt anöstral en tacka är utanför parningssäsongen. Den follikelfas som inducerades av baggeffekten liknade hormonellt en normal follikelfas under parningssäsong. Hos alla tackor som ovulerade föregicks ägglossningen av höga halter av LH och FSH.

Sperm characterization and identification of sperm sub-populations in ejaculates from pampas deer (Ozotoceros bezoarticus)

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

Physiological responses and characteristics of sperm collected after electroejaculation or transrectal ultrasound-guided massage of the accessory sex glands in anesthetized mouflons (Ovis musimon) and Iberian ibexes (Capra pyrenaica)

The objective was to characterize the stress response and the seminal parameters obtained with electroejaculation (EE) or transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) in two captive but nondomestic ruminants, the mouflons and the Iberian ibex under general anesthesia. In mouflons, the physiological responses (heart and respiratory rate, rectal temperature, cortisol, creatine kinase, potassium and glucose concentrations) changed similarly with both procedures. The TUMASG procedure was faster than EE in mouflons (21.7 ± 1.4 vs. 12.4 ± 1.2 minutes, P < 0.01). In ibexes, respiratory rate, cortisol and creatine kinase concentration changes were greater with EE than with TUMASG (final respiratory rate: 62.7 ± 5.5 vs. 38.1 ± 5.6 breaths/min [P < 0.05]; final cortisol: 51.4 ± 5.1 vs. 25.3 ± 5.6 ng/mL [P < 0.001]; and final creatine kinase: 300.9 ± 99.9 vs. 87.1 ± 16.9 U/L [P < 0.001]). Electroejaculation provided better results in some sperm parameters (mouflons: sperm score: 3.4 ± 0.3 vs. 2.6 ± 0.2 [P < 0.01]; total number of sperm ejaculated: 982.4 ± 299 vs. 710.0 ± 542.2 [P < 0.05]; ibexes: sperm with progressive motility: 47.7 ± 6.2 vs. 20.5 ± 8.3 [P < 0.05]). The transrectal ultrasound-guided massage of the accessory sex glands appears to be an alternative technique to collect sperm from wild ruminants, reducing the need for electrical stimuli and thus decreasing the undesired responses of EE in the more sensitive species. On the other hand, better fresh sperm may be collected with EE. However, TUMASG provides practical advantages in animal welfare, firstly in these wild species more sensible to stress management and capture myopathy.

Vocalizations During Electroejaculation in Anaesthetized Adult and Young Pampas Deer (Ozotoceros bezoarticus) Males

The aim of this study was to characterize the vocalizations produced during electroejaculation under general anaesthesia in pampas deer males and to determine whether the characteristics of those vocalizations differ in adult and young pampas deer males. Electroejaculation was applied to 13 adults (AM) and 13 young (YM) males under general anaesthesia. Vocalizations were digitally recorded, and the number and duration of vocalizations, the latency in relation to each voltage, the total time vocalizing, and the structure of the fundamental frequency (F0) [initial frequency (F(start)), maximal frequency (F(max)), minimal frequency (F(min)) and final frequency (F(end))] were analysed. No male vocalized with 0 V; the number of animals that vocalized increased at 2 and 3 V and increased again at 4, 5 and 6 V (p < 0.05). The latency time from the beginning of each series (each voltage) decreased until 4 V (p < 0.01). The number of vocalizations/voltage increased from 4 V (p < 0.05). The length of each vocalization and the total time during which animals vocalized were greater in YM than AM (p = 0.02 and p = 0.01, respectively). Similarly, the fundamental frequencies were higher in YM than AM (p ≤ 0.05). Overall, we concluded that the vocalizations emitted during electroejaculation in pampas deer under general anaesthesia are related to the voltage applied during the process. Young males vocalize more time, probably due to a greater sensibility to the electric stimulation. The differences in the characteristics of the vocalizations between adult and young males may be related to the anatomic differences in the neck of adult or young males.

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.

Early mother–young relationship and feeding behaviour of lambs are unaffected by low pasture allowance until the beginning of the last third of gestation in single-bearing ewes

Nutrition during gestation influences ewes’ bodyweight (BW), the bond with their lamb, milk yield and lamb development, and thus feeding behaviour of lambs during postnatal rearing. The aim of this study was to determine if the level of native pasture allowance from before conception until the beginning of the last third of gestation of single-bearing ewes influences the ewe–lamb relationship at birth and the feeding behaviour of lambs during postnatal rearing. A second aim was to determine whether the nutritional treatments affect ewes’ milk yield and their lambs’ bodyweight. Eleven multiparous Corriedale ewes grazed a high pasture allowance [10–12 kg of dry matter (DM)/100 kg of BW/day; HPA treatment], and another 11 grazed a low pasture allowance (5–8 kg of DM/100 kg of BW/day; LPA treatment) for 145 days beginning 40 days before conception. After the end of this treatment all ewes grazed on Festuca arundinacea and received rice bran and crude glycerine until 3 days before birth. Ewes’ BW was recorded monthly before and during gestation. Latency from parturition until the ewe licked her lamb, latency for lamb to stand up and suckle were recorded, and Maternal Behaviour Score was determined 12–24 h after birth. All the variables recorded during postpartum were determined approximately every 30 days. Grazing, ruminating and suckling events of the lambs were recorded from Day 23 until 142 days of age. Milk yield was determined from 32 until 140 days after lambing. Lambs’ BW was recorded from birth until 140 days of age. On Day 41 the BW change tended to be greater in HPA ewes than in LPA ewes, and on Days 79 and 105 it was greater in HPA ewes than LPA ewes (P = 0.006 and P = 0.005 respectively). Treatments did not affect the behaviours at birth of ewes and lambs, milk yield, ruminating and suckling events of lambs, ewes’ and lambs’ BW. The lambs from HPA ewes tended to present a greater frequency of grazing than the LPA lambs. Different levels of native pasture allowances from before conception until the last third of gestation in single-bearing ewes, followed by an increase in nutrient supply, did not affect ewe–lamb behaviours at birth, milk yield, lambs’ growth, and their feeding behaviours during rearing.

The tail in tropical hair ewes (Ovis aries) that are in estrus is used as a proceptive signal and favors ram’ copulation

Two experiments were performed to determine if the tail has a role: 1) in proceptive behavior, and 2) in facilitating copulation in hair sheep. The movements and position of the tail of estrous and non-estrous ewes in response to male courtship were compared in the first study. In the second study, the courtship of rams and mating behavior directed towards tailed or tail-docked, in estrus or diestrus ewes, were compared. Both experiments were conducted with restrained ewes during 3 min assessment periods. In Experiment 1, tail movements occurred in short episodes in response to physical contact of the male, but the rest of the time the tail of non-estrous ewes was drawn inward to the body, while in estrous ewes it simply hung naturally straight downward. Ewes in estrus had many tail moving episodes (P <0.001) with more movements per episode (P <0.001), that were of greater amplitude (P <0.001) and greater elevation in angle from the body (P <0.001) than in non-estrus ewes. In Experiment 2, ejaculation latencies with non-estrous ewes were longer than with estrous ewes (P <0.0001), and rams had more attempts to mount (P = 0.008), and more completed mounts (P = 0.0003) in non-estrous than estrous ewes. In addition, rams ejaculated more times (P <0.0001), and had a greater ejaculations/(mounts + ejaculations) ratio in estrous than non-estrous ewes (P <0.0001). More rams also had anal intromissions in docked than in intact tail-ewes (P = 0.0002). In conclusion, movements and position of the tail of estrous ewes is a primary component of the proceptive behavior that facilitates mating and serves as a physical barrier to avoid anal intromissions in hair sheep.

Vaginal mucus from ewes treated with progestogen sponges affects quality of ram spermatozoa

The use of intravaginal sponges (IS) to synchronize estrous onset in ewes provokes vaginitis, an increase in the vaginal bacterial load, and growth of bacterial species that are not present during spontaneous estrous behavior. The objective of the study was to compare the functional sperm parameters after incubating it with mucus collected from the vagina of ewes during spontaneous estrus or estrous synchronized with IS. Pooled spermatozoa were co-incubated with: (1) vaginal mucus collected from ewes in spontaneous estrus; (2) vaginal mucus collected from ewes in estrus pretreated with progestogen-impregnated IS; (3) synthetic mucus; and (4) medium without mucus as a control group. Sperm samples were evaluated after incubating it for 30 and 90 minutes. The number of colony-forming units (CFUs/mL), pH, and osmolality were greater in the mucus collected from ewes treated with IS than from those untreated (P = 0.046; P < 0.0001, and P < 0.0001, respectively). The percentage of sperm with progressive motility was lower after incubation with vaginal mucus collected from estrous ewes treated with IS than in the other three treatments both, 30 and 90 minutes after incubation (P = 0.0009 and P < 0.0001, respectively). The sample incubated for 30 minutes with mucus from ewes treated with IS had a lower percentage of sperm with intact plasma membrane than all the other treatments (P < 0.0001). The percentage of sperm with functional membrane was significantly lower in the sample incubated for 30 minutes with vaginal mucus from ewes treated with IS than in the other three treatments (P < 0.0001). After 90 minutes, the percentage was still lower than that in the sample collected from ewes during their spontaneous estrus (P = 0.0005). The lowest percentages of sperm with acrosome damage were observed in sperm incubated with mucus collected from sheep in spontaneous estrus for 30 and 90 minutes (P < 0.0001 and P = 0.008, respectively). The percentage of apoptotic spermatozoa was greater in samples incubated during 30 minutes with vaginal mucus collected from ewes treated with IS than in the other three groups (P = 0.0005). The functionality and the viability of ram sperm is negatively affected by the cervical mucus of ewes pretreated with progestagen-impregnated IS used in estrous synchronization treatments. This may partially explain the decrease in conception rate obtained with treatments with IS.