Seasonal variations in sexual behavior, testosterone, testicular size and semen characteristics, as affected by social dominance, of tropical hair rams (Ovis aries)

The purpose of this study was to examine the effect of day length on seminal characteristics, testicular size, sexual behavior and testosterone concentration (T4) in dominant and subordinate Pelibuey rams. Six groups of three animals were evaluated every 2 weeks over a 12 month period. Dominant (D), medium (M) and subordinate (S) animals in each group were identified through a food competition test every 15 days. All rams ejaculated and produced semen throughout the year. A linear hierarchy was clearly established and maintained during the year. S rams had lower levels (P < 0.05) of semen volume, sperm concentration and testicular volume than D rams, regardless of season. Scrotal circumference was not influenced (P > 0.05) by their social position. D rams were more affected by the photoperiod, decreasing (P < 0.05) reaction times, increasing (P < 0.05) T₄ and producing more (P < 0.05) sperm per ejaculation than S rams during short days (14.65 ± 1.22 vs 26.92 ± 1.65 s; 8.68 ± 0.44 vs 7.37 ± 0.40 ng/mL and 3.37 ± 0.17 vs 2.04 ± 0.16 sperm/10⁹, respectively). Semen volume and sperm concentration were significantly (P < 0.05) greater during short days in all rams, regardless of their social status, with the exception of sperm concentration in D rams where no variation was found. M rams displayed variable values in the range between the D and S rams. It was concluded that the magnitude of the seasonal effects was not sufficient to prevent the rams being used for breeding throughout the year and that seasonal variation within the variables affected the social ranks differently.     

Zoonotic Cryptosporidium parvum in Romanian newborn lambs (Ovis aries)

This study was undertaken to investigate the occurrence and public health significance of Cryptosporidium species/genotypes and subtypes in a newborn lambs. A total of 175 diarrheic fecal samples from lambs (younger than 21 days) were collected in seven sheep flocks located in western Romania, and were microscopically examined for the presence of Cryptosporidium oocysts after staining with modified Ziehl–Neelsen technique. Twenty-four (13.7%) fecal samples were tested Cryptosporidium positive by microscopy and were subjected for molecular characterization. All positive samples were successfully amplified through a nested polymerase chain reaction (PCR) of the small subunit (SSU) rRNA gene (18S). Cryptosporidium species were determined by restriction fragment length polymorphism (RFLP) analysis of the secondary PCR products using the conventional SspI and VspI restriction enzymes. The identified species were: Cryptosporidium parvum (20/24), C. ubiquitum (2/24) and C. xiaoi (2/24), respectively. PCR-RFLP results for C. ubiquitum and C. xiaoi isolates were confirmed by DNA sequencing. Subsequently, subtyping of seven randomly selected C. parvum isolates, based on sequence analysis of the GP60 gene, revealed the presence of five different subtypes (IIaA17G1R1, IIaA16G1R1, IIdA20G1, IIdA24G1 and IIdA22G2R1) belonging in two zoonotic subtype families (IIa and IId). These findings may suggest the potential role of the newborn lambs as a source for human cryptosporidiosis. This is the first published report about the presence of C. ubiquitum and C. xiaoi in lambs from Romania.     

Use of naloxone challenge to predict sexual performance of young rams

The possibility of developing a hormone-based test to predict libido was evaluated using the response of LH and testosterone to naloxone. This test has been used to identify sexually active and inactive mature rams during the breeding season. The objective of this study was to determine whether the blood test could be used to detect differences in sexual activity of early postpubertal (29 +/- 0.1 wk) rams during the breeding season in November and again at 70 +/- 0.1 wk of age in August before the next breeding season. Rams were classed as sexually active or inactive using serving capacity tests (8 30-min observation periods to record sexual behaviors [mounts and ejaculations] of each ram individually exposed to three ewes in estrus) after the naloxone challenges. Naloxone (0.75 mg/kg of BW) was injected i.v. into 38 white-faced crossbred, 16 Polypay, and 49 Targhee rams. Blood samples were collected at 15-min intervals for 1 h before and 2 h after naloxone to measure LH and testosterone. Separate mixed-model analyses for repeated measures were used to analyze data for the same rams at 29 and 70 wk of age. Logistic regression procedures were used to model probabilities that rams were correctly predicted to be sexually active. A breed-type x sexual activity x time interaction for LH was observed (P < 0.05) after naloxone injection in 29-wk-old rams. At 70 wk of age, a breed-type x time interaction was detected (P < 0.001) for LH response to naloxone, but LH did not differ by sexual activity. At 29 wk of age, a breed-type x time interaction for testosterone response after naloxone was detected (P < 0.001), and at 70 wk of age, a sexual activity x time interaction was detected (P < 0.05) for testosterone after naloxone. Sexually active and inactive rams were not predicted accurately at 29 wk of age and were predicted 69 and 29% of the time for sexually active and inactive rams, respectively, at 70 wk of age. In conclusion, breed type at 29 and 70 wk of age can influence the naloxone challenge test, but the test cannot be used to discriminate between sexually active and inactive rams at 29 wk of age during the breeding season or at 70 wk of age immediately before the breeding season.     

Attainment of Puberty in the European Mouflon (Ovis gmelini musimon) and the Domestic Manchega Ewe (Ovis aries)

The onset of puberty was assessed from plasma progesterone profiles for Mouflon (n = 8) and Manchega (n = 7) female lambs born in April. Four Mouflons exhibited their first ovulation at a mean date of 27 November ± 1.4 days, the mean age being 248.5 ± 3.9 days and the mean body weight 23.8 ± 0.6 kg. The remaining four Mouflons, which had a slower growth rate, attained puberty during the autumn of the following year. Five of seven Manchega lambs attained puberty in their first year at a mean date of 5 Oct ± 3.3, the mean age being 185.6 ± 2.6 days and the mean body weight 41.8 ± 2.0 kg. The mean live‐weight at the onset of puberty was 65.1 and 82% (p < 0.05) of adult body weights for Manchega and Mouflons, respectively. Overall, the results indicate that there is a threshold of body‐weight necessary for the attainment of puberty in the first breeding season for spring‐born lambs, which is very different between both Mouflon and Manchega. When body weight was below 23.8 ±0.6 and 41.8 ± 2.0 kg for Mouflon and Manchega lambs, respectively, first ovulation did not occur until the beginning of the next breeding season. The onset of puberty during the second year of life occurred within the specific reproductive seasons for Mouflon (October–April) and Manchega (July–March) ewes, despite minimal further growth.     

A note on the effect of number (single or twin) and sex of contemporary siblings on male-like play behavior of lambs (Ovis aries)

This study evaluates the effect of the number and sex of contemporary siblings on male-like play behavior of Suffolk lambs (Ovis aries). Fourteen males and 14 females, all single born, plus 26 ram lambs born co-twin to another male (n = 14) or female (n = 12) as well as 26 ewe lambs born co-twin to another male (n = 12) or female (n = 14) were observed daily for 7 weeks from 1 week of age. Two observers sitting outside a sand-bedded pen (10 × 5 m) recorded received and performed behavior (butts, nudging, attempts to mount, and mounts) by the lambs in the pen from 08:00 to 09:00 and 16:00 to 17:00 hours. Animals were in the pen only during the observation periods; the rest of the day they remained with their mothers in individual pens. Male-like play behavior patterns were displayed by both male and female lambs. Males were more active (P < 0.05) than females. Butting was the most frequent activity and performed mostly by ram lambs, recipients were generally males, and most resulted in reciprocal simultaneous encounters (sparring). Single males performed most of the nudging activity, which was the least frequent of the observed behaviors. Males with twin brothers mounted significantly (P < 0.05) more than males with twin sisters, and females with twin sisters performed the lowest amount of gutting and nudging. Lower (P < 0.05) levels of activity were found during the first 2 weeks. Overall, it was concluded that male lambs performed and received more male-like play behaviors than female lambs and the influence of sex of contemporary sibling was restricted to some behaviors: butting and nudging in females and mounting in males.     

Maternal behaviour in sheep (Ovis aries) following administration of opioid agonists

AIMS: As kappa(kappa)-opioids have marked effects on pain threshold in sheep during oestrus, late pregnancy and after birth, a study was undertaken to determine if kappa -opioids also had other roles during these states. METHODS: Opioid agonists (GR89696, DAMAGO) were administered into either the lateral hypothalamus (LH) or amygdala of non-pregnant sheep (n = 5) or pregnant sheep (n = 15) in late pregnancy (n = 5) within 12 h of birth (n = 5) or in the third week after birth (n = 5). Cortisol, behavioural and electrocardiographic responses to an auditory stimulus were assessed with and without drug administration, as was feed consumption and suckling events. RESULTS: In non-pregnant sheep, DAMAGO injected into either the LH or amygdala had little effect on the response to the auditory stimulus but when injected into the LH did decrease food consumption. GR 89696 had no obvious effects. In late pregnancy, cortisol and heart rate responses to the auditory stimulus were significantly reduced compared to the non-pregnant animals irrespective of treatment. This was also the case in the third week after birth. Immediately post-partum cortisol responses were closer to that seen in the non-pregnant animal. In late pregnancy and both post-partum periods, GR 89696 in the LH significantly increased feed consumption while DAMAGO had no significant effects. GR 89696 injected into the amygdala reduced the behavioural response to the auditory stimulus in late pregnancy and both post-partum periods. CONCLUSION: Changes in sensitivity to opioids occur in both the amygdala and LH with pregnancy and after birth. These may influence maternal behaviours and feed intake.     

Testosterone and luteinizing hormone responses to naloxone help predict sexual performance in rams

The first objective of this study was to determine whether LH and testosterone respond differently to a naloxone injection in relation to varying sexual performance in rams. If differences occurred, the second objective was to determine whether differences would predict variation in sexual performance. From a group of 1.5- to 3-yr-old rams, 20 sexually active and 39 sexually inactive rams were selected based on previously observed sexual behavior with estrual ewes. Each ram was exposed to three estrual ewes for 18 30-min sexual performance tests, and those found to be inactive were given two 30-min sexual partner preference tests. The final distribution was 28 sexually active, 22 inactive, and nine male-oriented rams. Rams were treated with 1.5 mg of naloxone/kg BW in December of Year 1 and again with either 0.75 or 1.5 mg of naloxone/kg BW in November of Year 2. Plasma concentrations of LH and testosterone were evaluated with mixed model analyses for repeated measures separately for each year to coincide with logistic procedures for modeling the probability that rams were sexually active. For Year 1, a sexual activity x age x time interaction for LH after naloxone was observed (P < 0.03). For testosterone, there was a sexual activity x time interaction (P < 0.03), with a similar, early increase for sexually active female-and male-oriented rams compared with a delayed, minimal increase for inactive rams. For Year 2, when all rams were over 2.5 yr of age, a sexual activity x time interaction for both LH and testosterone (P < 0.02) seemed more related to an earlier increase of both hormones for sexually active rams than the increase observed for inactive rams. In addition, sexually active rams had a greater increase in testosterone than inactive rams. No significant difference was observed between 0.75 and 1.50 mg of naloxone/kg BW. Testosterone and LH were used as explanatory variables and sexual activity was used as the response variable in logistic procedures. In Year 1, greatest prediction accuracy was 73.5% using testosterone at 60 min after naloxone injection. In Year 2, the greatest prediction accuracy was 85% using LH at 15 min multiplied by testosterone at 60 min after naloxone. Test repeatability for both years on the same rams was 76%. In conclusion, pattern and magnitude of naloxone-induced changes in endocrine function may facilitate identification of sexually active and inactive rams during the breeding season. Prediction accuracy of the naloxone-based test was 69 to 85%.     

Differences in leukocyte differentiation molecule abundances on domestic sheep (Ovis aries) and bighorn sheep (Ovis canadensis) neutrophils identified by flow cytometry

Although both domestic sheep (DS) and bighorn sheep (BHS) are affected by similar respiratory bacterial pathogens, experimental and field data indicate BHS are more susceptible to pneumonia. Cross-reactive monoclonal antibodies (mAbs) for use in flow cytometry (FC) are valuable reagents for interspecies comparative immune system analyses. This study describes cross-reactive mAbs that recognize leukocyte differentiation molecules (LDMs) and major histocompatibility complex antigens on DS and BHS leukocytes. Characterization of multichannel eosinophil autofluorescence in this study permitted cell-type specific gating of granulocytes for evaluating LDMs, specifically on neutrophils, by single-label FC. Evaluation of relative abundances of LDMs by flow cytometry revealed greater CD11a, CD11b, CD18 (β₂ integrins) and CD 172a (SIRPα) on DS neutrophils and greater CD14 (lipopolysaccharide receptor) on BHS neutrophils. Greater CD25 (IL-2) was identified on BHS lymphocytes following Concavalin A stimulation. While DS and BHS have similar total peripheral blood leukocyte counts, BHS have proportionately more neutrophils.     

Involución de Germenes Dentarios en la Oveja (Ovis aries)

In this study we examined some histologic and histochemical characteristics of the embryonic sheep dental epithelium in early odontogenesis. During the first trimester of development, a short-lived dental lamina was observed. Apparently in the sheep, the interactions between epithelial and ectomesenchymal cells required for tooth normal morphogenesis are altered.     

Pharmacokinetics of flubendazole and its metabolites in lambs and adult sheep (Ovis aries)

Flubendazole (FLU) is indicated for control of helminthoses in pig and avian species (monogastric animals) and its corresponding pharmacokinetics are well known. The information on FLU's pharmacokinetic behavior in animal species with forestomach (ruminants) has been limited although the use of FLU in these species could be beneficial. The aim of this study was to investigate the pharmacokinetics of FLU and its main metabolites in sheep. The effects of animal age (sexually immature and mature ones) and gender were also studied. FLU was orally administered in a single experimental dose (30 mg/kg of body weight) in the form of oral suspension. Treated immature animals (aged 3 months) and 5 months later the same mature individuals (aged 8 months) were kept under the same conditions (food, water and management) and treated with FLU. Within 72 h after FLU administration, plasmatic samples were collected and FLU and its Phase I metabolites were quantified using high-performance liquid chromatography. FLU was detected in very low concentrations only, reduced FLU (FLU-R) was identified as the main metabolite, and hydrolyzed FLU (FLU-H) as the minor one. Formation of FLU-R was stereospecific with (+)-FLU-R domination. The plasmatic concentrations of (+)-FLU-R reached 10–15 times higher values than those of FLU, (−)-FLU-R and FLU-H. A significant gender effect on pharmacokinetics of FLU or (+)-FLU-R metabolite in the mature animals was found and a wide significant difference between lambs and adult sheep in FLU including both metabolites has been proved.     

Anatomic and radiographic studies of the lacrimal drainage system in sheep (Ovis aries)

The lacrimal drainage system of the sheep (Ovis aries) was studied, using radiographic and anatomic techniques. The straight course of the nasolacrimal duct and the adequate diameter of different parts of the system facilitates radiography and catheterization of the lacrimal drainage system.     

Procedure for successful interspecific embryo transfer from mouflon (Ovis gmelini musimon) to Spanish Merino sheep (Ovis aries).

Embryos from five anesthetized mouflons (Ovis gmelini musimon), superovulated with FSH-o (Ovagen) were transferred into preselected Spanish Merino sheep (Ovis aries). Myorelaxation was complete in four of five donor mouflons. The status of the uterus of potential recipients was evaluated by transrectal ultrasonography, and those ewes with fluid in the uterine horn were rejected. The corpus luteum in each ewe was assessed ultrasonographically the day before surgery. Plasma progesterone levels and the quality of the corpora lutea were the criteria for selection of recipients. Ten embryos were transferred to the five selected Spanish Merino recipients, resulting in four pregnancies and seven live-born lambs, including three sets of twins. This study shows that determination of plasma progesterone levels combined with ultrasonographic assessment of the corpus luteum provides information useful for screening of potential recipients.     

Differential pulmonary immunopathology of domestic sheep (Ovis aries) and bighorn sheep (Ovis canadensis) with Mycoplasma ovipneumoniae infection: A retrospective study

Mycoplasma ovipneumoniae is a respiratory pathogen that impacts domestic sheep (Ovis aries; DS) and bighorn sheep (Ovis canadensis; BHS). BHS are reported to be more susceptible than DS to developing polymicrobial pneumonia associated with M. ovipneumoniae infection. Using formalin-fixed paraffin-embedded tissues, we performed a retrospective study investigating the pulmonary immune response of DS and BHS to M. ovipneumoniae infection. M. ovipneumoniae infected DS exhibited a more robust and well-organized BALT formation as compared to BHS. Digital analysis of immunohistochemical chromogen deposition in lung tissue was used to quantitate T cell marker CD3, B cell markers CD20 and CD79a, macrophage markers CD163 and Iba1, and cytokine IL-17. A significant interaction of species and infection status was identified for CD3, CD163, and IL-17. BHS had a greater increase in bronchiolar CD3 and bronchiolar and alveolar CD163 with infection, as compared to DS. BHS had an increase in bronchiolar associated lymph tissue (BALT) and alveolar IL-17 with infection, while these remained similar in DS regardless of infection status. IL-17 in respiratory epithelium of bronchi and bronchioles comparatively decreased in DS and increased in BHS with infection. These data begin to define the interspecies differential immune response to pulmonary M. ovipneumoniae infection in DS and BHS and provide the first investigations of respiratory epithelium-associated IL-17 in ovine.     

CD11b of Ovis canadensis and Ovis aries: molecular cloning and characterization

Leukotoxin (Lkt) is the primary virulence factor secreted by Mannheimia haemolytica which causes pneumonia in ruminants. Previously, we have shown that CD18, the beta subunit of beta(2) integrins, mediates Lkt-induced cytolysis of ruminant leukocytes. CD18 associates with four distinct alpha subunits giving rise to four beta(2) integrins, CD11a/CD18 (LFA-1), CD11b/CD18 (Mac-1), CD11c/CD18 (CR4), and CD11d/CD18. It is not known whether all the beta(2) integrins serve as a receptor for Lkt. Since PMNs are the leukocyte subset that is most susceptible to Lkt, and Mac-1 expression on PMNs exceeds that of other beta(2) integrins, it is of interest to determine whether Mac-1 serves as a receptor for Lkt which necessitates the cloning of CD11b and CD18. In this study, we cloned and sequenced the cDNA encoding CD11b of Ovis canadensis (bighorn sheep) and Ovis aries (domestic sheep). CD11b cDNA is 3455 nucleotides long encoding a polypeptide of 1152 amino acids. CD11b polypeptides from these two species exhibit 99% identity with each other, and 92% with that of cattle, and 70-80% with that of the non-ruminants analyzed.     

Dietary restriction reduces the rate of estradiol clearance in sheep (Ovis aries)

Three experiments were designed to test the effect of dietary restriction on clearance of 17beta-estradiol (E(2)) in sheep. A preliminary experiment examined the effect of a 4-d fast on the rate of E(2) clearance in wethers. The second experiment tested the hypothesis that either long-term restriction (7 wk) or a 5-d fast would increase steroid-binding capacity of serum by increasing the concentration of sex hormone-binding globulin (SHBG) in the blood of ovariectomized ewes. In Exp. 3, we hypothesized that nutrition-dependent regulation of E(2) clearance by the liver would result in divergence in biliary extraction of E(2) in fed and fasted wethers receiving comparable levels of exogenous E(2). A marked difference in E(2) clearance between fed and fasted wethers was noted in the preliminary study. Relative to ad libitumfed wethers, a 4-d fast decreased E(2) clearance by 52%. Serum concentrations of SHBG were increased in long-term energy-restricted and fasted ewes, relative to the concentration in maintenancefed ewes (P = 0.015). Furthermore, a 5-d fast nearly doubled serum steroid-binding capacity in wethers. The E(2) concentration in bile was 2 times greater in fasted than in fed wethers. This fasting-dependent increase in biliary E(2) may be reflective of the increased serum E(2) in fasted animals, because each 1 pg/mL increase in serum E(2) increased bile E(2) by 0.86 +/- 0.12 pg/mL, independent of nutrition (P = 0.002). Our results demonstrate that the rate of clearance of E(2) is decreased during nutritional restriction. Additionally, these data indicate that altered SHBG expression, enterohepatic recirculation, or both are involved in the decreased E(2) clearance during dietary restriction.     

Isolation and genetic characterization of Toxoplasma gondii from alpaca (Vicugna pacos) and sheep (Ovis aries)

Alpacas are important to the economy of several countries. Little is known of Toxoplasma gondii infection in alpacas worldwide. In the present study, T. gondii was isolated and genetically characterized from alpacas for the first time. Alpacas (n?=?16) and rams (n?=?12) pastured on a farm in Virginia, USA, were examined at necropsy. Antibodies to T. gondii were determined by the modified agglutination test (MAT, 1:25) and found in 6 of 16 alpacas with titers of 1:100 (2 alpaca), 1:400 (2 alpacas), 1:800 (1 alpaca), and 1:1,600 (1 alpaca), and 5 of 12 rams in titers of 1:50 in one, 1:400 in one, 1:800 in one, 1:1,600 in one, and 1:3,200 in one. Tissues of all 16 alpacas were bioassayed in mice or in cats. Muscles (heart, skeletal muscle) of nine alpacas with MAT titers of 1:25 were fed to T. gondii-free cats; the cats did not shed oocysts. Viable T. gondii was isolated from tissues of two of six seropositive alpacas by bioassay in mice. Viable T. gondii was isolated from three of three seropositive sheep by bioassay in mice. Genotyping using cell-cultured tachyzoites revealed four genotypes, including one for ToxoDB PCR-RFLP genotype #2 (type III), one for genotype #3 (type II variant), one for genotype #170, and two for a new genotype designated as ToxoDB PCR-RFLP genotype #230. Thus, four of the five T. gondii isolates in the present study belonged to different genotypes. These results indicate a higher genetic diversity among T. gondii isolates circulating in the USA than previously realized.