A single administration of carbetocin before electroejaculation increases the insemination doses produced from each ejaculate in rams.

The aim of the study was to determine the effect of carbetocin administration (a long-acting analog of oxytocin) 20 or 10 min before electroejaculation (EE) on the duration of semen collection procedure, quantitative and qualitative characteristics of the ejaculate, and stress biomarkers in rams. Semen was collected from 12 Corriedale rams (age, 2.5-5.5 years old) with EE, in a Latin-square design, administrating carbetocin (0.2 mg/100 kg of body weight i.v.) 20 or 10 min before EE, or without carbetocin administration (CB-20, CB-10, and CON treatments, respectively). Each treatment was applied to different rams every 3-4 days, allowing all the rams to receive all three treatments. Carbetocin administered 20 or 10 min before EE increased the number of sperm ejaculated (P = 0.01), the semen concentration (P = 0.02), the number of insemination doses collected in a single collection (P = 0.01), and the number of insemination doses collected/electrical pulses administered (P = 0.05) compared to control rams. Carbetocin administered 20 or 10 min before semen collection prolonged the time required for EE and the number of pulses administered during EE compared to CON rams (P < 0.03 for both). The CB-10 rams required the administration of more electrical pulses during ejaculation than CON rams (P = 0.001), and CB-20 treatment tended to require more electrical pulses than CON rams (P = 0.06). The volume of the ejaculate was greater in CB-10 than in CON rams (P = 0.01), and that of CB-20 treatment tended to be greater than CON rams (P = 0.08). The percentage of sperm with intact membrane was greater in CB-20 than in CON rams (P = 0.01). Total protein, albumin, and globulin concentrations were lower immediately after carbetocin administration 20 or 10 min before EE. The treatments did not affect cortisol concentration, glycemia, rectal and surface temperatures, heart rate, and facial expressions. Carbetocin administration before EE of rams improved the quantitative and qualitative characteristics of the ejaculate, duplicating the number of insemination doses collected. It can be a promising treatment to obtain a greater quantity of doses to inseminate with a lower frequency of semen collections, reducing the negative impacts of EE on animal welfare.

Frequent social regroupings affect corporal and reproductive development, metabolic status, and behavioral time budget in dairy heifers.

We aimed to evaluate the corporal and reproductive development, metabolic status, and behavioral patterns of resident dairy heifers subjected to social regroupings (SR). Holstein replacement heifers (153.3 ± 16.1 kg, 11 months old) were homogeneously allocated to: 1) Control (CON; n = 14 resident + 5 "permanent") and 2) Regrouped (RG; n = 14 resident + 5 "changing"). "Changing" heifers were exchanged for 5 novel heifers every 21 d over 205 days (total = 10 SR). At each SR, body weight, wither height, body condition score (BCS), eating, ruminating, lying and standing behaviors were recorded, and blood hormones and metabolites were determined. Behavior was registered through scan sampling every 10 min., and estrus was detected using patches. Wither height and BCS were greater in CON than RG at SR9 and 10, and at SR4 and 5, respectively (P < 0.01). Control heifers showed first heat earlier (93 ± 9 d vs 126 ± 14 d; P = 0.03) and presented greater insulin growth factor- 1 (171.3 vs 120.0 ng/ml ± 10.1; P < 0.01) and glucose (4.3 vs 4.2 mmol/L ± 0.07; P = 0.04) concentrations than RG, while non-esterified fatty acids concentration was greater in RG (0.35 vs 0.30 mmol/L ± 0.04 P = 0.04). Regrouped heifers were observed more times eating (0.45 vs 0.33 ± 0.01), but fewer times ruminating (0.21 vs 0.24 ± 0.01) than CON (P < 0.01). Frequent SRs negatively affected corporal and reproductive development and energy metabolism, and altered behavioral patterns in dairy heifers.

Administration of a single dose of long-acting injectable progesterone formulation reduces the stress response to weaning in anestrous ewes.

High circulating progesterone concentrations decrease the responses to stressful situations in farm ruminants. We hypothesized that administering a single dose of long-acting progesterone formulation reduces the stress response of anestrous ewes to weaning. The aim of this study was to compare the behavioral, cortisol, and blood protein responses to weaning of anestrous ewes treated or not, with a single dose of a long-acting progesterone formulation. A complementary aim was to characterize the bioavailability of progesterone after this treatment. Thirty-six multiparous single-lambing Corriedale ewes and their lambs were used in this study. The lambs were weaned at 6:30 h when they were 65 d old (d0), and were taken to another paddock without chemical, visual or auditory contact with their mothers, while the ewes remained in the same paddock. Immediately before weaning, while 19 ewes received oil based long-acting progesterone formulation, other 17 ewes remained as controls, receiving sunflower oil. Twelve ewes from each group were used for behavioral recordings, the other 5 untreated ewes and 7 treated ewes were used to characterize the cortisol response and the progesterone profile. Behaviors were recorded every 10th min for 3 h in the morning and 3 h in the afternoon on day minus 3, d0 and d2, and expressed as percentage of recordings each animal displayed each behavior on each day. Treated ewes were recorded more times standing up than ewes on the day of weaning (P < 0.05), but the reverse result was found 2 d after (P < 0.01). Treated ewes were recorded more times lying down than control ewes on d2 after weaning (P < 0.0001), when treated ewes were observed fewer times walking than control ewes (P = 0.0004), as also happened the day of weaning (P < 0.0001). On the day of weaning, treated ewes grazed and paced less than control ewes (P = 0.001 and P = 0.009). Treated ewes ruminated more than control ewes on the day of weaning (P = 0.02) and 2 d later (P = 0.0002). Control ewes also secreted more cortisol than progesterone-treated ewes (P = 0.03), with no effect of the treatment on the concentration of total proteins, albumin or globulins. While control ewes never had luteal progesterone concentrations (>0.5 ng/mL), treated ewes achieved luteal progesterone concentrations 2 h after its administration, and remained well above luteal levels until 15 h after administration. The administration of a single dose of long-acting progesterone formulation appears to be a viable choice in reducing the stress to weaning in ewes. Ewes treated with progesterone coped better with weaning, returning to their normal basal behavior earlier than untreated ones, and secreting lower cortisol levels.

Daily variation of the cortisol and insulin concentration, glycemia, surface temperature, and behavior patterns due the administration of equine Chorionic Gonadotropin in rams.

Sex steroids can modulate the circadian rhythmicity of some physiological and behavioral functions of sheep. During the non-breeding season, Corriedale rams have a low testosterone concentration, but the administration of equine Chorionic Gonadotropin (eCG) stimulates its secretion. Therefore, the daily variation of cortisol and insulin concentrations, glycemia, behavioral pattern (eating, ruminating, standing, lying, and sleeping), and rostral and eye surface temperatures were compared in rams treated or not treated with eCG during the non-breeding season [between August (late winter) and September (early spring)]. Twenty Corriedale adult rams (BW 63.4 ± 7.3 kg; BCS: 3.5 ± 0.5; mean ± SD; age: 2-3 years old) were randomly assigned to 1 of 2 treatments: a) Treated: administration of 1000 IU of eCG intramuscularly and b) Control: control, without application of eCG. All the rams were managed under the same feeding and environmental conditions (individual pens outdoors). All variables were measured each hour for 26 h, except testosterone and insulin that were measured at 09:00, 10:00, 12:00, 15:00, 18:00, 21:00, 22:00, 24:00, 03:00, 06:00, 09:00 and 11:00 h. Testosterone and cortisol concentration were greater in Treated than Control rams (testosterone: 32.7 ± 0.7 nmol/L vs 5.8 ± 0.7 nmol/L; P <0.0001; cortisol: 11.3 ± 0.6 nmol/L vs 9.9 ± 0.6 nmol/L; P =0.02). The eye surface temperature was 0.9°C lower in the Treated than in the Control rams (P =0.004). The characteristics of the daily variation pattern of cortisol, glycemia, glycemia:insulin ratio, and surface temperature were not affected by the treatment, the acrophases being at ∼06:00 h, ∼18:00 h, ∼08:00 h, and ∼11:00 h, respectively. The acrophase of insulin concentration was observed earlier in Treated than in Control rams (13:02 ± 0:56 vs 15:47 ± 0:53; P =0.04). Additionally, the acrophase of the time spent eating was advanced in Treated rams (09:48 ± 00:16 vs 10:20 ± 00:16; P =0.04) and sleeping (21:39 ± 00:57 vs 02:02 ± 01:06; P <0.0001). Treated rams slept less time than Control rams (P =0.04). In conclusion, the administration of eCG modulated the daily variation pattern of insulin concentration and the behavioral pattern (period in which rams were observed eating and sleeping). Nevertheless, although there was a greater cortisol concentration and reduced eye surface temperature, the administration of eCG did not modify their daily variation pattern, suggesting that this is robust, limiting the effect of testosterone on their basal pattern.

Short- and long-term effects of temporary early cow-calf separation or restricted suckling on well-being and performance in zebu cattle.

Information on weaning techniques in the tropics is scarce, particularly regarding the long-term effect of temporary early cow-calf separation or restricted suckling. Therefore, we studied the effects of these two handling practices on well-being and performance at 150 days postpartum in fifteen zebu cow-calf pairs randomly assigned to three treatments. Continuous suckling (CS) where calves remained with their dams from birth to weaning; restricted suckling (RS) calves were allowed to suckle 30 min/day from Day 34 until weaning at Day 150 and kept separated the rest of the time; temporary separation (TS) calves were separated for 72 h from their dams from Day 33 to 36 but remained with their dams the rest of the time. Blood samples and behavioral data were collected on Days 32-36 (1st period) and 149-153 (2nd period). In the 1st period, a greater percentage of RS and TS calves were observed close to the fence line (<10 m) that separated them from their dams (P < 0.0001) and vocalized more than CS calves (P < 0.0001), while in the 2nd period, RS calves had the highest cortisol concentration and vocalization rate (P < 0.05). Similarly, during the 1st period, a greater percentage of RS and TS cows were observed close to the fence line than CS cows (P < 0.0001), with TS cows vocalizing the most (P = 0.001). In the 2nd period, RS cows had greater cortisol concentration than TS (P = 0.037) and CS cows (P = 0.003). More TS and CS cows than RS were observed close to the fence line (P = 0.03 and P = 0.05). On Day 150, TS calves and cows vocalized more than RS and CS animals (P < 0.0001). Before calf-cow separation, 27 out of 45 cows were cycling (CS = 10; RS = 6; TS = 11). After separation, 12 of the remaining 18 cows resumed ovarian activity (CS = 3; RS = 5; TS = 4), and all cows were cycling after estrous synchronization treatment. The pregnancy rate was similar between CS, RS, and TS (60, 53, and 60% respectively). In conclusion, temporary separation increased calf distress response to definitive weaning even four months later, while restricted suckling seemed to reduce it.

Evidence of cooperative calves' care and providers' characteristics in zebu cattle (Bos indicus) raised under extensive conditions.

Communal rearing has been reported in several mammals, including wild ungulates. However, until now, there was no evidence of any alloparental care in the Bos genus. To test the hypothesis that calves' groups are formed under the care of specific cows, a herd of 31 peri-partum zebu cows raised under pasture conditions were used. Groups of ≥ 3 individuals within a 10-m diameter were estimated using aerial pictures taken every other day at 7:00, 10:30, 13:30, 16:00, and 18:00 h, during 6 weeks. Temperament (exit speed, flight distance, intensity of reaction), age, and parity of each cow were registered. A total of 142 groups were observed, and in all of them, at least one calf was present. A total of 75% of the groups were more calves than cows, and in 65.4% of the cases, there were 1 to 3 cows with 2 to 32 calves. While there were no groups integrated only by cows, there were 3.5% integrated only with calves. The most frequent group was formed by 2 calves and 1 cow (14.8%). Parity was positively related with the number of times that a cow was observed in a group (R2 = 0.19, P = 0.01) and with the number of groups in which her calf was present (R2 = 0.16; P = 0.03). The results of the temperament tests were unrelated to the total number of groups in which a cow was observed nor the number of groups with or without its calf. It was concluded that zebu calves congregate in newborn groups in companion with one or more adults, preferably high parity cows, regardless of their temperament.

Chronic use of a GnRH agonist (deslorelin) or immunization against GnRH: effects on testicular function and sperm quality of bucks.

Chronic use of GnRH agonists and immunization against GnRH have been used as reversible contraceptive methods. The aim of this study was to compare the effectiveness of both treatments to inhibit reproductive function of adult bucks, in terms of strength and duration of the effects. We used 9 control untreated bucks (CON), 7 bucks treated chronically with a GnRH agonist (subcutaneous implants with 7.4 mg of deslorelin, Suprelorin, Virbac) (AGO), and another 7 bucks were immunized against GnRH (dose of 2 mL of Improvac-Zoetis with 300 µg of a synthetic incomplete analog of natural GnRH; 300 mg of diethylaminoethyl-dextran; and 2.0 mg of chlorocresol) (IMM). Testicular and sperm evaluations, testosterone concentrations, and male odor were determined from 4 wk before applying the treatments until 17 mo of their application. Scrotal circumference of CON (21.0 ± 0.1 cm) and IMM (21.2 ± 0.2 cm) was greater than that of AGO bucks (19.9 ± 0.2 cm) (P < 0.05 for each), without difference between CON and IMM bucks. Pixels' color intensity of testicular ultrasound images was not affected by treatment (general mean ± SEM: 116.0 ± 1.8). Testosterone concentration was greater in CON than AGO and IMM in months 3 and 4, greater in CON and IMM than AGO bucks in months 15 and 16, and greater in IMM than CON and AGO bucks in month 17 (P < 0.05 for all comparisons). Male odor was greater in CON (1.5 ± 0.0) than IMM bucks (1.3 ± 0.0) and greater in IMM than AGO (1.1 ± 0.0) bucks (P < 0.05 for each). Treatment negatively affected all the sperm variables: the total number of sperm in the ejaculate, sperm motility, sperm with normal morphology and sperm with integral membrane function. It was concluded that both treatments were effective in inhibiting the reproductive axis; however, neither of them produced azoospermia or decreased testosterone concentrations to undetectable levels. With both treatments, there were individual males exhibiting characteristics of fertility in all periods of the study. However, chronic use of a GnRH agonist seemed to be the most effective treatment in terms of duration and strength.

Social dominance affects intake rate and behavioral time budget in pre-pubertal dairy heifers allocated in continuous competitive situations.

In intensive feeding systems, competition may be high and social dominance may affect animal performance by changing dry matter intake (DMI) and behavioral time budgets. If competition level is maintain over time, the strategies developed by heifers of different social status are expected to differ. Thus, the aim of this study was to compare individual DMI, intake rate and eating, ruminating, lying and standing behaviors in dominant (DOM) and subordinate (SUB) pre-pubertal dairy heifers in a model study implying continuous competitive situations. A total of 16 Holstein and Jersey×Holstein pre-pubertal heifers (251±10 days old, weighing 208±14 kg; mean±SEM) were allocated into eight homogeneous dyads. Each dyad was maintained during 120 days (day 0=beginning of measurements) in pens, and received a total mixed ration from one feeder/dyad. The DOM and SUB heifers was determined (day 0, twice during the first month of the experiment and every month afterwards) by observation of the winner in agonistic interactions in each dyad after the feed was supplied. The general activity pattern (eating, ruminating, lying and standing) of each heifer was recorded by direct instantaneous scan-sampling, every 10 min for 12 h, in 7 days (days 1, 21, 35, 60, 75, 100 and 120). Individual DMI was estimated with the double marker technique, in three intervals (I=days 17-26; II=days 78-87 and III=days 112-120), while estimated intake rate (kg/min) was calculated for each interval as the DMI per total eating time. After the experiment was concluded, data of the first 5 and the last 6 h of the 12 h scan-sampling (related to the moment the feed was supplied) was grouped according to the moments of greater and lesser competition for feed on each day. During the first 5 h, where competition was expected to be highest, no differences in eating behavior were found between heifers of different social status, but DOM heifers spent more time ruminating and lying than SUB heifers, while SUB spent more time standing than DOM heifers. No differences were found on DMI between DOM and SUB, but SUB ate at a faster rate on interval II compared with DOM heifers. In conclusion, in this model study of heifer dyads, SUB heifers had greater intake rate with no differences in feed intake, spent less time ruminating and lying, and more time standing than DOM heifers during the first hours after feed delivery.

Vaginal biota number is smaller in tailed than docked hair ewes (Ovis aries), but is not affected by copulation.

A greater accumulation of feces could facilitate the entrance of bacteria or inert particles into the vagina, particularly while mating tailed ewes. Two experiments were performed to determine if the tail has an active role in the growth of bacterial flora and the entrance of inert material in the vagina after mating, during induced or natural estrus. Twenty ewes were used, 10 tailed and 10 docked. All ewes displayed estrus after CIDR treatment, and half of them; 5 tailed and 5 docked were exposed to rams during the first experiment. For the second experiment, the rest of the animals were mated 15-17 d later at spontaneous estrus. In the induced estrus, no differences in the number of CFU/mL in docked or tailed ewes were observed before mating, but after mating, CFU values tended to be greater in docked than tailed ewes, and the number of inert particles was greater in docked than tailed ewes (5.8 ± 1.3 vs 1.6 ± 0.6, P = 0.024). During natural estrus, docked ewes had less CFU/mL than tailed ewes (19.6 × 103 ± 0.4 vs 80.0 × 103 ± 0.0, P = 0.006) before mating, and a similar tendency was maintained after mating, when the number of inert particles was similar in docked and tailed ewes. Staphylococcus and Escherichia were observed in the induced estrus, while in the natural estrus, even though these were the more frequent, five other genera of bacteria were observed. It was concluded that the presence of the tail protects the vaginal environment against increases of the biota load.

Sedation or anaesthesia decrease the stress response to electroejaculation and improve the quality of the collected semen in goat bucks.

Electroejaculation (EE) is stressful and probably painful; thus the administration of anaesthesia is recommended to decrease those negative effects. However, anaesthesia has a direct risk of provoking death, but sedation is less risky than anaesthesia. At the same time, α2-adrenergic agonists may improve semen quality. Therefore, the aim of the present study was to compare the physiological and behavioural responses indicative of stress and possibly pain, and the semen quality in electroejaculated untreated, anaesthetised or sedated goat bucks. Semen was collected from eight bucks using three different procedures in all them (EE in untreated bucks, EE under sedation or EE under general anaesthesia). The number of vocalizations during EE and the behavioural pattern before and after procedures were recorded. Pain visual analogue scale (VAS) score was also determined during EE. Rectal temperature, heart rate, serum cortisol concentration, biochemical and haematological parameters were measured before and after each procedure, and sperm characteristics were determined. Bucks vocalised more often when untreated than sedated or anaesthetised (P<0.02). The pain VAS score was greater when bucks were untreated than sedated or anaesthetised (P<0.002). The rectal temperature, heart rate, total protein, albumin and haemoglobin concentrations were greater when bucks were untreated than anaesthetised or sedated (P<0.02). Serum cortisol increased after EE (P=0.0006), without differences between procedures. The frequency and duration of lying down after EE were greater when bucks were anaesthetised than sedated or untreated (P<0.05), and were also greater when bucks were sedated than untreated (P<0.05). The number of times that the animal tried to stand up after EE was greater when bucks were anaesthetised than sedated or untreated (P<0.02). The sperm mass motility was greater when bucks were anaesthetised or sedated than when they were untreated (P=0.048). When animals were sedated, the ejaculate contained more spermatozoa with functional plasma membrane (P=0.03) and morphologically normal (P=0.05) than when they were untreated. In conclusion, general anaesthesia and sedation decreased the stress and probably the pain response provoked by EE and especially sedation improved the quality of the semen collected.

Slow and ultra-rapid freezing protocols for cryopreserving mouflon (Ovis musimon) and fallow deer (Dama dama) epididymal sperm.

This study examines the effectiveness of two methods for cryopreserving post-mortem epididymal sperm - conventional slow freezing employing a short equilibration time with glycerol, and ultra-rapid freezing - from the wild ruminant species Ovis musimon (mouflon) and Dama dama (fallow deer). A Tris-citric acid-glucose (TCG) + 12% egg yolk-based medium was used for the conventional slow freezing of the fallow deer sperm, whereas a Tes-Tris-glucose (TEST) + 6% egg yolk-based medium was used for the mouflon sperm. Glycerol was added to a final concentration of 5% to both media. The same diluents were used for ultra-rapid freezing but replacing the glycerol with 100 mM of sucrose. Sperm variables (motility, viability, acrosome integrity, membrane integrity, and morphological abnormalities) were analyzed before and after cryopreservation. Although values were generally better after the thawing of the conventionally cryopreserved sperm, total sperm motility (38.40 ±â€¯4.44% in mouflon and 31.25 ±â€¯3.37% in fallow deer) and total live sperm (47.19 ±â€¯5.18% in mouflon and 43.13 ±â€¯2.43% in fallow deer) were acceptable for the ultra-rapidly cooled sperm. Independent of the cryopreservation method, membrane integrity, acrosome integrity and the percentages of dead sperm and sperms with a damaged acrosome were better for the cryopreserved mouflon sperm than the fallow deer sperm (P < 0.05). Despite exerting a more harmful effect on sperm variables than conventional freezing, ultra-rapid freezing may be a useful alternative for the cryopreservation of these species' epididymal sperm in the field, as this simple technique does not require sophisticated equipment and expertise.

Growing without a mother results in poorer sexual behaviour in adult rams.

The aim of this study was to determine if the absence of the mother during rearing has long-term effects on sexual behaviour and physiological reproductive parameters of adult rams. Two groups of rams were: (1) artificially reared, separated from their dams 24 to 36 h after birth (Week 0) and fed using sheep milk until 10 weeks of age (group AR, n=14); and (2) reared by their dams until 10 weeks of age (group DR, n=13). Sexual behaviour (tests of 20 min) and physiological reproductive parameters were analysed separately for the non-breeding (Weeks 42 to 64) and the breeding (Weeks 66 to 90) seasons. Body weight, scrotal circumference, gonado-somatic index, testosterone concentrations or sperm parameters were similar in both rearing conditions (AR v. DR) in both seasons. During the non-breeding season AR rams displayed fewer ano-genital sniffings (AR: 4.2±0.4 v. DR: 5.3±0.4, P=0.04) and matings (AR: 1.2±0.2 v. DR: 1.8±0.2, P=0.002) than DR rams. During the breeding season AR rams displayed fewer ano-genital sniffings (AR: 4.3±0.5 v. DR: 5.7±0.5, P=0.005), flehmen (AR: 0.7±0.2 v. DR: 1.1±0.2, P=0.03), mount attempts (AR: 1.4±0.2 v. DR: 2.1±0.2, P=0.04), and tended to mount less frequently (AR: 6.6±0.9 v. DR: 8.8±0.9, P=0.08) than DR rams. In conclusion, the absence of the mother during the rearing period negatively affected display of sexual behaviour towards oestrous ewes during a rams adult life in both breeding and non-breeding seasons. However, it did not affect testis size, testosterone secretion or sperm variables.

Asymmetrical size and functionality of the pampas deer (Ozotoceros bezoarticus) testes: Right testis is bigger but left testis is more efficient in spermatogenesis.

Information about gonadal asymmetries in ruminants is very scarce. In this work, we performed three complementary studies to compare characteristics of both testes: (i) weight and size of offspring and adult dead males; (ii) the tissue:fluid relationship determined by ultrasound scanning; and (iii) the spermatogenic status using fine needle aspiration cytology. The right testis was heavier than the left one in both offspring and adult animals and had greater width and volume in adult males than the left one. The ultrasound pixel intensity was similar in both testes. The right testis tended to have more spermatogonia (p = .06) and had a greater percentage of early spermatids (p = .004) than the left testis. On the other hand, the left testis had a greater percentage of spermatozoa (p = .05). The left testis had a greater spermatozoa/spermatogonia ratio (p = .02) and tended to have more spermatozoa/Sertoli cells ratio (p = .07). The spermatogenic index tended to be greater in the left than in the right testis (p = .06). Overall, we concluded that the right testis of pampas deer males is bigger but according to the cytology, it seems to be less spermatogenically effective than the left one, but these differences are not explained by different tissue:fluid ratio in each testis. Although differences were greater in adults than in offspring, asymmetry was observed even in just born offspring.

Competition for oestrous ewes between rams reared by their mothers or artificially reared: Effects on sexual behaviour and testosterone and cortisol serum concentrations.

The objective of this study was to determine how the social competition for an oestrous ewe affects the sexual behaviour and the endocrine response in two groups of rams, one reared by their mothers and another artificially reared. Thus, we compared the sexual behaviour and testosterone and cortisol changes in each group of rams in competitive and non-competitive tests, both during the first and second breeding seasons. Two groups of rams were: 1) artificially reared lambs, separated from their dams 24-36 h after birth (Week 0) and artificially fed with sheep milk until 10 weeks of age (group AR, n = 14); and 2) lambs reared by their dams until 10 weeks of age (group DR, n = 13). Rams were subjected to non-competitive and competitive tests for an oestrous ewe during their first and second breeding seasons, when they were 8 and 20 months old, respectively. Sexual behaviours toward an oestrous ewe were recorded during 20 min and the testosterone and cortisol concentrations were determined in serum samples collected immediately before the test, and 20, 40 and 60 min after it. During the first breeding season, the number of flehmen decreased in DR rams, and the number of flehmen and ano-genital sniffings also decreased in DR rams, but the frequency of some copulatory behaviours increased (matings and ejaculation/total mounts in DR rams, and total mounts in AR rams) in competitive tests. During the second breeding season, competition caused a decrease in the number of all the recorded behaviours (courtship and copulation) with the exception of flehmen in AR rams; however, in DR rams only the number of the copulatory behaviours decreased under competition. Competition did not affect the endocrine response during the first breeding season. During the second breeding season, while testosterone concentrations were greater in non-competitive than in competitive tests at 60 min (P = 0.0008) in AR rams, in DR rams it tended to be greater (P = 0.09). Competition did not affect cortisol concentrations in any group or season, but in all tests the concentrations increased at the end of the test (P < 0.05). In conclusion, the lack of the mother during rearing negatively affected the sexual motivation and the testosterone response of rams to oestrous ewes in competitive tests, effects that were more evident when adults. Neither the absence of the mother during rearing nor competition for oestrous ewes affected the stress response (evidenced by increase in cortisol concentration) in rams during both seasons.

Ejaculation does not contribute to the stress response to electroejaculation in sheep.

Electroejaculation procedures (EEPs) provoke stress; nevertheless, ejaculation produces physiological changes similar as those usually used to measure stress responses. The application of EEP to animals that cannot ejaculate-as ewes-may be useful to discriminate the responses induced by ejaculation from those provoked by EEP. The aim was to determine the stress response to EEP in rams and ewes. The EEPs were applied to 10 rams and 10 ewes during the non-breeding season, and the number of vocalizations, the heart rate, rectal temperature, serum cortisol concentration, biochemical and haematological parameters were measured. Overall, EEP provoked increases in cortisol concentration, glycaemia, rectal temperature and concentration of creatine kinase (all them: p < .0001) as well as relative concentration of granulocytes (p = .003) and absolute granulocyte concentration (p = .0002) in both, rams and ewes. Heart rate, relative concentration of lymphocytes (p = .001), haematocrit (p = .02) and haemoglobin (p = .045) decreased in animals from both genders after EEP. Besides, cortisol (p < .0001), rectal temperature (p = .002) and glycaemia (p = .001) were greater in ewes than rams, and creatine kinase also tended to be greater in ewes than rams (p = .054). On the other hand, the number of animals that vocalized (p = .006), white blood cells (p = .02) and absolute lymphocytes (p = .02) were greater in rams than ewes. The general trends show a similar pattern of stress responses in animals from both genders. Therefore, we concluded that ejaculation does not contribute to the stress response provoked by the EEP. This procedure also provokes muscular damage and probably pain.

Social dominance in prepubertal dairy heifers allocated in continuous competitive dyads: Effects on body growth, metabolic status, and reproductive development.

The objective of this study was to compare the body weight (BW) and size, metabolic status, and reproductive development of dominant and subordinate prepubertal dairy heifers allocated in competitive dyads. Sixteen Holstein and Jersey × Holstein prepubertal heifers (means ± SEM; 250.8 ± 9.8 d; 208.5 ± 13.9 kg of BW) were assigned to 8 homogeneous dyads according to breed, age, and BW. Dyads were housed in pens separated 1 m from each other during 120 d, receiving a total mixed ration on a 5% restriction of their potential dry matter intake, and had access to the same feeder (60 cm) throughout the experiment. Dominant and subordinate heifers were defined based on the winning agonistic interactions in each dyad. Body development was recorded every 20 d in all heifers, and blood samples were collected on the same days to determine endocrine and metabolic status. The maximum follicle diameter, number of follicles >6 mm, and the presence of corpus luteum were observed weekly by ultrasound. Heifer BW (269.3 vs. 265.3 ± 1.5 kg) and average daily gains (0.858 vs. 0.770 ± 0.02 kg/d) were greater in dominant than subordinate heifers. On d 30, 37, and 53, dominant heifers had more follicles than subordinate heifers, and maximum follicle diameter was greater in dominant than in subordinate heifers (10.0 vs. 9.0 ± 0.3 mm). Dominant heifers achieved puberty earlier than subordinate heifers (313.9 ± 4.9 vs. 329.6 ± 5.7 d) with similar BW (279.4 ± 2.6 vs. 277.4 ± 5.8 kg). Glucose concentrations were greater in dominant than subordinate heifers (89.2 vs. 86.8 ± 1.2 mg/dL), but cholesterol concentrations were greater in subordinate than dominant heifers (86.1 vs. 90.2 ± 2.6 mg/dL). We concluded that, under continuous competitive situations, dominant heifers were more precocious than subordinate ones, achieving an earlier puberty. Dominant heifers had greater body growth and glucose concentrations than subordinate heifers, which may be responsible, at least in part, for the differences on reproductive development between heifers of different social status.

Low pasture allowance until late gestation in ewes: behavioural and physiological changes in ewes and lambs from lambing to weaning.

Low pasture allowance during gestation affects ewes' BW at parturition, the bond with their lamb, lamb development, and thus also may affect their responses to weaning. The objectives were to determine if native pasture allowance from before conception until late pregnancy affects ewe-lamb behaviours at lambing, ewes' milk yield, lambs' BW, and the behavioural and physiological changes of ewes and lambs at weaning. From 23 days before conception until 122 days of pregnancy, 24 ewes grazed on two different native pasture allowances: high (10 to 12 kg of dry matter (DM)/100 kg of BW per day; HPA treatment; n=12) or low (5 to 8 kg of DM/100 kg of BW per day; LPA treatment; n=12). Thereafter, all ewes grazed on Festuca arundinacea and received rice bran and crude glycerine. Ewes' body condition score (BCS) and BW were recorded during pregnancy and postpartum periods. Milk yield was determined on days 32, 41 and 54 after lambing. Lambs' BW was recorded from birth until 72 days after lambing. Latency from parturition until the ewe licked her lamb, maternal behaviour score (a test that evaluates maternal attachment to the lamb) and latency for lamb to stand up and suckle were determined. The behaviour of the lambs and ewes was recorded before and after weaning (at 65 days). The ewes' serum total protein, albumin and globulin concentrations were measured before and after weaning. The HPA ewes presented greater BW (P<0.005) and BCS (P<0.005) than the LPA ewes during pregnancy and postpartum (P<0.04), and had a greater milk yield than the LPA ewes (P<0.03). Treatments did not influence any behaviour at lambing, lambs' BW, neither the ewes' behavioural and physiological changes at weaning. HPA lambs paced and vocalized more than LPA lambs (P<0.0001). The variation of albumin concentration before and after weaning was greater in the HPA lambs than in the LPA lambs (P<0.0001). In conclusion, although ewes' BW, BCS and milk production were affected by pasture allowance until late pregnancy, this did not affect the behaviours that lead to the establishment of the mother-young bond, nor the ewes' behavioural responses at weaning. Lambs reared by ewes that grazed on low pasture allowance during pregnancy presented fewer behavioural changes and a lower decrease of albumin concentration after weaning. Lambs' BW was not affected by the feeding received by their mothers.

Gross Anatomy of Pampas Deer (Ozotoceros bezoarticus, Linnaeus 1758) Mouth and Pharynx.

The aims of this study were to describe the anatomy of the mouth and pharynx of the pampas deer, and to consider its evolutionary feeding niche according to those characteristics. Gross dissections of the mouth and pharynx were performed in 15 animals, 10 adult females and five young animals under 1 year (three males and two females), all dead by causes unrelated to this anatomical region. The upper lip entered in the constitution of a pigmented nasolabial plane. The masseter muscles weighed 43.8 ± 3.5 g and represented 0.23% of body weight, which corresponds to ruminants of feeders intermediate to grazers and browsers. Parotid glands represented 0.08% of the body weight, characteristic that also categorize the pampas deer as belonging to the intermediate feeding group. The dental formula was the same of the domestic ruminants. The upper incisors and canines were absent, and instead of them, there was a dental pad (Pulvinus dentalis). The upper canine teeth were present only in the deciduous dentition. The existence of a brachydont dentition turns Ozotoceros very vulnerable to continuous use as there is no compensatory teeth growth. The particular anatomy of the mouth and lips of this animal was adapted to a very selective feeding, taking highly nutritious sprouts beyond plant category. In conclusion and in addition to previous studies of anatomy of the digestive organs in this species, pampas deer may be categorized as belonging to the intermediate type of feeding.

Antler velvet is thicker in adult than in yearling pampas deer (Ozotoceros bezoarticus): a histological study.

BACKGROUND: Antlers are lined by soft velvet tissue during antler growth. Later, the velvet is shed before rut onset. There are no detailed histological descriptions of the growing velvet, nor whether the velvet changes according to stag age. Our aims were to: 1) describe the basic histology of pampas deer antler velvet from adult and yearling males; and 2) determine the influence of age and time of antler growth on velvet's tissues morphometry. MATERIALS AND METHODS: Samples were collected from 10 stags allocated in two groups, either adult (3-5 years old, n = 5) or yearling males (2 years old, n = 5). The day of antler cast was recorded for each animal. In spring, the stags were anaesthetised and velvet samples were collected from the third tine's distal end. Samples were described qualitatively and a restricted morphometrical analysis of the antler velvet was performed. RESULTS: The number of keratinocyte layers and the thicknesses of: total epidermis, corneum, intermediate and basale epidermal strata, total dermis, superficial and deep dermis were determined. Age and days after antler casting positively influenced in conjunction epidermal thickness (p = 0.037), and tended to influence both stratum intermedium (p = 0.076) and stratum corneum (p = 0.1) thicknesses. Age influenced stratum corneum thickness (p = 0.04). The pampas deer antler velvet lacked both sweat glands and arrector pili muscles. CONCLUSIONS: The deep dermis was densely irrigated but displayed abundant and well developed collagen bundles. Both total epidermal and stratum corneum thicknesses related positively to the age of the animals but were not to the time since antler cast.

Heart rate patterns during courtship and mating in rams and in estrous and nonestrous ewes ().

The aim of this study was to compare the heart rate (HR) patterns in rams mated with estrous or nonestrous ewes and in mated estrous and nonestrous ewes () during courtship and mating. For this purpose, HR and behavior were recorded using a radio telemetry recording system and a closed-circuit television system with video recording, respectively. Rams were joined with either an estrous ( = 10) or a nonestrous ( = 10) ewe that was restrained in a stanchion by the neck. Data were continuously recorded until each ram performed 3 ejaculations. Eight days later, the HR of the 10 estrous and 10 nonestrous ewes was recorded during mating. Although the time between entrance into the yard and the first ejaculation was similar across rams, rams that mounted estrous ewes were faster at attaining their second (3min5s ± 17 s vs. 5min28s ± 18 s) and third (7min58s ± 45 s vs. 12 min ± 1min14s) ejaculations (all < 0.05). By contrast, no differences in HR were observed between rams that interacted with estrous versus nonestrous ewes. In all cases, HR reached maximum values immediately after each ejaculation and the HR pattern was similar across ejaculations (first, second, and third). Although HR was similar between estrous and nonestrous ewes before mating, nonestrous ewes had higher HR ( < 0.05) during mating. In summary, 1) rams that mated estrous ewes displayed shorter interejaculation periods but HR did not differ between groups of rams during any ejaculation (first, second, or third), 2) HR for both groups of rams peaked shortly after each ejaculation, and 3) HR increased more in nonestrous than in estrous ewes while mating.