Adult Rams Research Articles

Melatonin induces a stimulatory action on the scrotal skin components of Soay ram in the non-breeding season

Fifteen adult Soay rams were employed in this study to investigate the effect of melatonin on the scrotal skin using histological, histochemical, and morphometrical analysis. The results revealed that the melatonin treated group showed a significant increase in the thickness of the epidermis, the cross-sectional area of blood capillaries and nerve fibers compared with the control one. In addition, obvious hypertrophy and hyperplasia were detected in the sebaceous glands in association with a significant increase in the number and diameter of apocrine sweat glands with well-developed secretory activity. S100 protein and cytokeratin-19 strongly stained the basal cells of sebaceous glands in the melatonin treated group incomparable to the control group. Moreover, the nerve fibers were intensively immunoreacted for S100 and cytokeratin proteins in the melatonin treated group in contrast to the control one. A high number of telocytes (TCs) could be identified in the treated group around the nerve fibers and blood vessels in the dermis. The number of Langerhans cells showed a significant increase in the melatonin groups that were identified by MHC II and PGP 9.5 within the epidermal layer. Furthermore, a significant increase in the number of dendritic cells was identified in the melatonin group, which were distributed within the dermis, around hair follicles, sebaceous glands, and sweat glands and were strongly expressed PGP-9.5, MHC-II, VAMP, SNAP, keratin-5, and cytokeratin-19 immunoreactivity. Notably, Merkel cells showed a significant increase in the number in the melatonin group that could be stained against nestin, SNAP, and VAMP. On the other hand, the secretory granules in sweat glands were exhibited a strong positive reactivity for synaptophysin in melatonin group. The current study showed that the administration of melatonin induced a stimulatory effect on keratinocytes, non-keratinocytes, sebaceous and sweat glands, hair follicles, as well as the vascular, neuronal, and cellular constituents of the dermis.

The capacity of goat epidermal adult stem cells to reconstruct the damaged ocular surface of total LSCD and activate corneal genetic programs.

Epidermal adult stem cells (EpiASCs) have the potential for unlimited proliferation and differentiation, however, the ability of these stem cells to activate corneal genetic programs in response to corneal stroma stimulation needs to be further validated. Herein, a feasible strategy was developed to reconstruct the damaged corneal surface in a goat model with total limbal stem cell deficiency (LSCD) by transplanting EpiASCs, which had been explanted and cultured from the skin of an adult ram goat and were then purified by selecting single cell-derived clones and cultivating them on a denuded human amniotic membrane (HAM). These artificial tissues were then successfully transplanted into ewe goats with total LSCD. Binding of EpiASCs to the base membrane of an EpiASCs-HAM-Sheet (EHS) indicated their proliferating status. After transplantation, the EpiASCs could survive in the host tissue and they reconstructed the damaged ocular surface of total LSCD. The crystal reconstructed corneal epithelium expressed CK3 and Pax-6 similar to normal corneal epithelium and expressed the Sry gene after transplantation. These results demonstrated that EpiASCs could be induced to differentiate into corneal epithelial cell types in a corneal microenvironment and had the ability to activate corneal genetic programs. This work offer a foundation for promoting tissue-engineered cornea into clinical application.

Impact of sex and age on the bacterial composition in rumen of Tibetan sheep in Qinghai China

In this study, 18 Tibetan sheep were assigned into four groups by age and sex, namely young ewes (E1, n = 5), young rams (R1, n = 3), adult ewes (EA, n = 5) and adult rams (RA, n =5) to profile the bacterial composition of Tibetan sheep and to explore the impacts of sex and age on rumen microbiota using high-throughput sequencing method of amplicon of 16S rRNA V3–V4 region. The results indicated that all effective tags for 18 samples were clustered into 5429 OTUs (Operation taxonomic Units, OTUs) with an absolute abundance > 1 at a similarity level > 97%. The rumen bacterial composition of female adult sheep was more diverse than male adult sheep and yearling female sheep. SILVA database was used to annotate taxonomic information for each representative sequence using the RDP classifier, the results showed that Bacteroidetes, Firmicutes, Verrucomicrobia and Proteobacteria were the most dominant phyla, representing 56.68%, 28.81%, 6.71% and 1.73% of reads from all samples on average, respectively. At the genus level, Rikenellaceae_RC9_gut_group, and Prevotella_1 were most abundant with relative abundance of 12.98% and 11.19%, respectively. Relative abundance of Bacteroidetes in EA was greater than RA (p = 0.014), relative abundance of Firmicutes in RA was greater than EA (p < 0.001), and the relative abundance of Spirochaetae in E1 was greater than R1 (p = 0.038). Shifting genera between the sexes belonged to Order Bacteroidales and Clostridiales, both changed substantially across sex in adult sheep (p < 0.01). The abundance of Bacteroidetes in female sheep and Firmicutes and Planctomycetes in male sheep significantly increased with age (p < 0.05), while the abundance of Spirochaetae in female sheep and Cyanobacteria in male sheep decreased with age (p < 0.05). Shifting genera between age groups belonged to order Bacteroidales and Clostridial. The abundance of order Bacteroidales in female sheep increased with age (p = 0.046), and the abundance of order Clostridiales in male sheep increased with age (p = 0.006), while it decreased with age in female sheep (p = 0.030). In summary, this study profiled the bacterial composition characteristics of Tibetan sheep and concluded that physiological factors, e.g. sex and age, shaped the bacterial composition, which expands our knowledge of the rumen of ruminant animals residing on the Qinghai-Tibetan Plateau.

МЕРИНОСОВАЯ ПОРОДА ОВЕЦ ДЛЯ ГОРНО-ОТГОННОЙ СИСТЕМЫ РАЗВЕДЕНИ

Dagestan is the leading Republic of Russia in terms of the number of sheep and sheep breeding products. The zoned target sheep breed in Dagestan is the Dagestan mountain breed. However, this breed produces thin but non-merino wool. Developing the Merino sheep breed suitable for mountain breeding is an urgent problem. The new Merino sheep breed was developed for the foothill zone of Dagestan, where summer Alpine and winter low-lying pastures are located at a distance of no more than 150 km from each other, and summer Alpine pastures are located at an altitude of up to 3000 meters above sea level. The research and stock breeding work was carried out at the farm production enterprise “Krasniy Oktyabr’”, the Kazbek district, in the period between 2005 and 2019. The work resulted in the development of the Arthlukh Merino sheep breed. (Patent for selection achievement No. 10112 “State Commission of the Russian Federation for Selection Achievement Tests and Protection”). The breed was developed by crossing the Stavropol sheep rams with the Dagestan mountain sheep, with the subsequent improvement of crossbreeds by the Manych Merino breed. A distinctive feature of the new Merino breed from the basic herdmates of the Dagestan Mountain breed is the high live weight, which is 15.3–16.5% higher in different sex and age groups. Besides, the amount of washed wool shorn is 16.2–26.0% higher. The ewes’ wool fineness is 22.9 microns (19.4–23.0) – 64 qualities. That of adult rams amounts to 23.3 microns (19.9–23.3) and has 60 qualities. The difference of the wool fineness on the side and thigh does not exceed one quality. The coat length on the side of ewes is not less than 9.0 cm, rams – 9.5 cm, gimmers (young ewes) – 9.5 cm and replacement rams – 10.0 cm. The wool strength is 7.0 cN/Tex. The belly wool cover is good. The suint is stable, of a white and light cream color. The average yield of washed wool excluding the lowest grades is 64%. The fertility of ewes ranged from 125% to 135%.

A comparison between Platelet-rich plasma and low-level laser therapy for the treatment of second degree burn in sheep

Background and aims: This experiment was conducted in order to compare the effect of Platelet-rich plasma (PRP) and low-level laser (LLL) therapy on healing of second-degree burn wound in sheep. Materials and Methods: Fourteen adult rams were divided into four groups of equal numbers. A second-degree burn was generated on the back of all animals. The burned area in the first and second group was exposed and autologous PRP and LLL (6 J/cm2), respectively were applied on burned area continuously for five days. Fourteen days later, samples were collected from all animals for histopathological examinations. Results: Histopathological evaluations on the second week showed the burn healing to be better in the PRP than LLL and both of them were better with respect to the control group. Conclusion: The application of PRP for five days is effective than LLL application in healing burn-related skin wounds in sheep model.

111 Fertilizing ability of frozen and freeze-dried semen following intracytoplasmic sperm injection of invitro-matured sheep oocytes

Freeze-drying is a novel technique that permits the storage of semen at room temperature for long time periods, retaining their fertilizing capacity. The main objective of this work was to compare the fertilization ability of frozen-thawed (FT) and freeze-dried (FD) ram semen following intracytoplasmic sperm injection (ICSI) of invitro-matured (IVM) sheep oocytes. Oocytes were recovered by slicing the ovaries of slaughtered sheep. Selected cumulus-oocyte complexes (COCs) were IVM for 24h in tissue culture medium 199 (TCM-199) supplemented with 10% heat-treated oestrous sheep serum (ESS), 0.36mM pyruvate, FSH (1IUmL−1), and luteinising hormone (LH; 1IUmL−1) under mineral oil in a humidified atmosphere of 5% CO2, at 38.5°C. Semen was collected from fertile adult rams using an artificial vagina and processed for (1) freezing and thawing (Khalifa and Lymberopoulos, 2013 Cell Tissue Bank 14, 687-698; https://doi.org/10.1007/s10561-012-9357-6) or (2) freeze-drying and rehydration according to Arav et al. (2018 J. Assist. Reprod. Genet. 35, 1149-115; https://doi.org/10.1007/s10815-018-1145-1) protocols. For FD protocol, sperm samples were diluted in a sugar solution of trehalose and sorbitol (LyoB) and dehydrated for 24h. Later, the samples were rehydrated in a warming solution and diluted in TCM-199 before ICSI. After maturation, metaphase II (MII) oocytes with a polar body were injected with FT or FD sperm. Briefly, oocytes were transferred into groups of six in an ICSI dish containing 6-µL drops of holding medium (TCM-199 + 5% fetal bovine serum) and 3-µL drops of PVP for the sperm samples. Injection was carried out with an inverted microscope (Olympus IX73) connected to a micromanipulation system (Narishige) using ICSI pipettes with 7-µm internal diameter. Within 1h, ICSI oocytes were activated with 5 µM ionomycin for 4min and invitro cultured in modified synthetic oviductal fluid medium (Bogliolo et al. 2011 Reprod. Fertil. Dev. 23, 809-817; https://doi.org/10.1071/RD11023). After 17-21h, injected oocytes were fixed and stained in a solution of ethanol Hoechst 33342 and classified as FPN (one female pronucleus and one condensed sperm head), MPN (one male pronucleus and one MII), 2PN (two pronuclei, male and female), 3PN (three or more pronuclei), and NPN (no pronuclei). Data were analysed using analysis of variance (two-way ANOVA) followed by Tukey post hoc test with SAS software, version 9.4. The ICSI-FD group had a higher number of NPN and a lower number of 2PN than did the ICSI-FT group (P&amp;lt;0.05). We think that more technical advances in the FD process as well as the rehydration procedure are necessary to improve the application of FD ovine semen for invitro fertilization by ICSI in sheep, but in any case these results have showed that FD could be a useful tool for the future of invitro embryo production. Table 1.Pronuclear formation at 17-21h post-injection1 Treatment n FPN MPN 2PN 3PN NPN FT 71 9.66±4.12 4.26±1.48 48.13±2.79a 5.97±4.16 31.98±6.75a FD 65 6.16±2.26 1.39±1.39 20.15±4.14b 10.57±6.59 61.73±6.89b a,bValues in the same column with different superscript letters differ significantly (P&amp;lt;0.05). 1Data are presented as mean±s.e.m. FPN=female pronucleus, MPN=one male pronucleus and one metaphase II oocyte, 2PN=two pronuclei, male and female, 3PN=three or more pronuclei, NPN=no pronuclei. Funding was provided by Spanish MINECO Grant AGL2017-85837-R, Spanish MECD pre-doctoral grant FPU15/00773, and Spanish MECD mobility grant EST18/00472 to Irene Menéndez Blanco.

Ultra-Sonographic Evaluation and Growth Factors Measurement of Using Tendon Derived Hydrogel and Hyaluronic Acid in Tendon Repair in Rams

The present study was planned to evaluate the efficacy of tendon derived hydrogel and hyaluronic acid in repair of experimentally induced superficial digital flexor tendon defect in rams, ultrasonographically with detection of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in blood. Thirty six adult rams were divided randomly into three equal groups: First group (control group) rams were subjected to partial removal of the superficial digital flexor tendon (SDFT), second group (tendon derived hydrogel group): rams were subjected to the same procedure as mentioned in first group except that the tendon defect was filled with tendon derived hydrogel directly, while third group (hyaluronic acid group): same procedure as in first group but the tendon defect was filled with hyaluronic acid directly. Clinically there was no pain, no infection, no signs of rejection and all wounds had healed completely in all rams, normal body temperature, except for the swelling which appeared in the first group in one ram at 30 days post operatively and two rams in third group at 60 days post operatively, all animals were able to stand and showed normal gait. Ultrasonographic results at 120th day post operation in first group showed, hypoechoic to hyperechoic regions at the site of the defect. At the same period for the second group the sonogram showed, well-aligned echoic tendon structure, while in the third group at the same period there were hypoechoic areas at the site of the defect. The results of bFGF and VEGF detection showed significant differences between the periods in first group at P≤0.05, while in second and third groups there were significant differences at P≤0.01for bFGF. The results of detection VEGF showed, significant differences in the mean values among the periods at P≤0.01in first group, also second group there was significant differences at P≤0.01. And in third group there was a significant difference at P≤0.05. Also there were significant differences among all groups P≤0.01in at 60th day postoperatively. In conclusion the tendon derived hydrogel matrix and the hyaluronic acid have a regenerative effect in repair of tendons injuries, but the superiority is for tendon derived hydrogel matrix.

Effect of Season on Hematologic, Biochemical, and Hormonal Analytes in Rams of Two Breeds

Abstract Clinicopathological investigations are essential for the evaluation of the health status of ruminants. Apart from species-specific reference intervals, the effect of common biological factors should be considered for an accurate interpretation of laboratory data. The aim of this study was to evaluate the effect of season on hematologic and biochemical analytes, and serum total thyroxine and cortisol in adult rams of two breeds. Four blood samples (one every season) were collected from each ram. Complete blood count was performed on the Advia 120 (Siemens Healthcare Diagnostics, USA), while the differential leukocyte count was manually conducted. Biochemical and hormonal analyses were performed on Flexor E (Vital Scientific, The Netherlands), AVL 9180 (Roche Diagnostics, Belgium), and Immulite 1000 (Siemens Healthcare Diagnostics, USA), respectively. Linear mixed effects models (R language) were employed for statistical analyses. Forty-three (26 Chios, 17 Florina), adult, clinically healthy rams were included. Statistically significant (p&lt;0.05), mostly breed-independent seasonal differences were observed in almost all of the analytes. However, when assessing these differences in view of the respective reference intervals, only a few of them were considered biologically important. Specifically, mild hyperglycemia and mild decrease in the concentration of total calcium and inorganic phosphorus were detected in winter, while a mild increase in thyroxine concentration (autumn) and creatine kinase activity (spring and summer) was also noted. In conclusion, seasonal effects should be considered when evaluating laboratory results in rams; however, season does not appear to have an essential effect on the clinicopathological profile of rams reared in the Mediterranean region.

Enhancing liquid-chilled storage and cryopreservation capacities of ram spermatozoa by supplementing the diluent with different additives.

ObjectiveIn the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 polyunsaturated fatty acid in the diluent on mitigating consequences of (a) liquid chilled- and (b) cryo-storage of ram spermatozoa.MethodsIn the first experiment, ejaculates (n = 30) were collected from 5 adult rams and were pooled, diluted (1:10) with Tris-citric acid (base diluent) and were split into 4 aliquots assigned for: control (untreated), caffeine (0.1 mM), melatonin (0.3 mM) or omega-3 fatty acids (0.3 mM) (T0). The diluted specimens were stored at 4°C for 48 h, during which sperm physical and cytological properties were evaluated along with oxidative stress indices (T24, T48). In the second experiment, 15 ejaculates (3 per male) were pooled, diluted with glycerolized base diluent (4% glycerol, v/v) and were split corresponding to the same previous treatment groups before being processed for cryopreservation. Post-thaw physical and kinematic sperm properties were assessed by a computer-assisted sperm analysis system.ResultsThe results clarified superiority of both melatonin and omega-3 supplementation on maintaining (p<0.05) sperm properties, while reducing (p<0.05) lipid peroxidase reaction and enzymatic activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of spermatozoa.ConclusionMelatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for application of artificial insemination or in vitro fertilization.

Histological and morphometrical study of the effect of season on layers and glands of skin in local Awassi sheep (Ovis ovis)

The study aimed to investigate the effect of season on the skin and cutaneous glands in local Awassi breed of sheep (Ovis ovis). The study was done on five healthy adult rams of Awassi breed in winter and summer at five anatomical regions (armpit, back, scrotum, ventral surface of fatty tail and dorsal surface of the fatty tail). The study explained that season had no significant effect on the histological structure of the sheep's skin, while there was a significant variation in the histological measurements of most structures of skin such as skin layers, sweat and sebaceous glands where the thickness of the corneal layer of the skin varied between the winter and summer seasons ,it was thicker in the winter than in the summer for most of the studied areas, the variation was higher and more significantly at scrotum , where the epidermis was thicker in winter than in summer for most of the few wool areas that included in the study, while in the dense wool areas the epidermis was thicker in summer than in winter. The study also showed that the season has a significant effect on the size of the sweat glands. The size of the sweat glands in the summer appeared to be greater than in the winter and for all areas of study, but the area of the scrotum and the skin of the ventral surface of fatty tail showed the opposite. the season had an effect on the depth of the sweat glands in the dermis. With the exception of the scrotum and the ventral surface of the fatty tail, the sweat glands were deeper in the summer than in the winter for the rest of the study areas. The lining of the sweat glands units was affected by the change of season. In the summer, the height of the cells of the sweat units was increased and for all regions of the study to reached 16.43 micrometers in the scrotum area for the summer while decreased in winter to reached 10.70 micrometers. So, we can say that the sweat glands of the study areas faced the warm climate not only by the increase in the size of the glands and their proximity to the surface in the summer but also by increased functional activity. The sebaceous glands in the area of the ventral surface of the fatty tail had a large difference between the winter and summer followed by the rest of the areas, either the armpit areas and the dorsal surface of the fatty tail did not show any significant difference between the two seasons. Finally, we found the season has been shown to play a significant role in influencing the histological measurements of skin and skin glands.

Assessment of eco-sustainability vis-à-vis zoo-technical attributes of soybean meal (SBM) replacement with varying levels of coated urea in Nellore sheep (Ovis aries).

The contemporary environmental-stewardship programmes primarily aimed at curbing the global warming potential by adopting a multidisciplinary approach. Manipulating the feeding strategies has great potential in reducing the environmental footprints of livestock production. This study intends to assess the effect of soybean meal (SBM) replacement with varying levels of coated urea (SRU) on both zoo-technical (nutrient digestibility, heat increment, and physio-biochemical parameters) and environmental attributes. The coated urea was used to replace the SBM at 0, 25, 50, and 75 percent levels. Eight adult rams (43.02 ± 0.76) maintained in a conventional shed were used in a replicated 4 x 4 Latin square design. Not all the physiological parameters viz. rectal temperature, pulse rate, and respiratory rate were affected (P>0.05)f by varying levels of SRU incorporation. The SRU fed animals had higher (P<0.05) crude protein digestibility compared to SBM fed animals; however, the replacements did not affect the nutrient digestibility coefficients of DM, OM, NFC, NDFap, ADF, and hemicellulose components. The SRU did not affect various biochemical parameters such as serum glucose, total protein, albumin, globulin, urea, creatinine, ALT, AST, Ca, P and T3, and T4 levels; however, post-prandial serum urea N (SUN) values showed a diurnal quadratic pattern (P<0.05) with a dose-dependent relationship. Further, the SBM replacements had no effect on the calcium excretion, while the SRU incorporation decreased the faecal phosphorous content, thereby abating the eutrophication phenomenon. Although the SBM replacements did not affect in vivo water variables and faecal solid fractions, they managed to decrease the land and virtual water requirement along with global warming potential (GWP) of the entire trial. The GWP-perceptual map unveils the fact that replacement of conventional feed ingredients with NPN compounds aids in eco-friendly livestock production. Further, the conjectural analysis of the carbon footprint methodology revealed that agricultural by-products consideration could cause a huge increase in the GWP share of feed consumed, thus compelling the importance of research pertaining to feed production perspective as equal as ruminal methane amelioration.

Long days in winter or the presence of adult sexually active rams did not influence the timing of puberty of autumn-born Rasa Aragonesa ram-lambs

ABSTRACT Forty autumn-born ram-lambs were used to study the effect of several rearing strategies on their sexual activity and fertility at 8 months of age. The animals were assigned to one of the following four groups (n = 10): FA, which were exposed to 2 months of long days (15 January–15 March); SAR, which were housed with three Sexually Activated adult Rams from 15 March to 20 June; NAR, housed with three Nonactivated adult Rams, and Control (C), kept isolated from adult rams throughout the experiment. In mid-June, individual and group ram serving-capacity tests were performed. Mean plasma testosterone levels (ng/ml) throughout the experiment were higher (P < 0.001) in the C (4.4 ± 0.2) and SAR (4.3 ± 0.2) groups than in the FA (3.4 ± 0.2) and NSA (3.4 ± 0.2) groups. In the serving tests, control ram-lambs executed a higher total mean number of sexual events than the other groups (P < 0.05). Fertility rates of SAR- (90%) or C-mated ewes (100%) were higher (P < 0.01) than ewes mated by the FA (50%) and NAR (40%) ram-lambs. In conclusion, autumn-born ram-lambs reared in isolation from adult congeners had the highest sexual activity and fertility at 8 months of age, which suggests that no special rearing strategy is necessary to induce puberty in these rams.

Hematologic, biochemical, hormonal, and redox status alterations during reproductive activity in adult rams of two breeds.

Reproduction is a crucial, energy-consuming physiologic activity, which can temporarily disturb the homeostatic balance of the organism. Although rams are commonly placed in intense breeding programs on contemporary sheep farms, the clinicopathologic and redox status alterations have not been adequately studied during reproductive activity. The objective was to investigate the clinicopathologic and redox status changes in rams during reproductive activity. Thirty-seven (19 Chios and 18 Florina breed) adult, healthy rams were individually mated with ten estrus-synchronized ewes of the same breed. Blood samples were collected prior to mating, during mating, and postmating. CBC (Advia 120), cytologic blood smear evaluation, biochemical analyses (Flexor E, AVL 9180 Electrolyte Analyzer), and thyroxine measurements (Immulite 1000) were performed. The reactive oxygen metabolites (d-ROMs) test, the biological antioxidant potential (BAP) test, and oxidative stress index were used to assess redox status. The linear mixed effects model (statistical language R) was used for statistical analyses. Many, mostly breed-independent, clinicopathologic changes were detected during reproductive activity. Conversely, redox status alterations were found to be breed-dependent. Intense reproductive activity was related to mild, acute muscle damage in both breeds and mild oxidative stress in Florina rams, whereas Chios rams exhibited oxidative stress resistance. The observed clinicopathologic and redox status alterations were generally consistent with those reported during various forms of physical activity and exercise in animals and humans. Intense reproductive activity does not seem to be physiologically innocuous; however, it appears to be a relatively mild muscle-damaging physical activity.

Organic breeding in Sarda ewes: Utilization of the ram effect in an artificial insemination program

The current study assesses the ram effect as an alternative to conventional hormonal treatment for estrus synchronization prior to artificial insemination. Two groups of 50 anoestrus Sarda ewes were induced to ovulate with adult rams (RE group) or synchronized using intravaginal sponges impregnated with progestogen and equine chorionic gonadotropin (eCG) (PRO group). Ewes in the RE group were isolated from rams for 6 weeks. After the isolation period, 5 vasectomized rams were introduced into the flock at a ratio of 1 ram/10 females (day 0) to induce estrus and ovulation. Ewes in both experimental groups were inseminated via the cervical route with fresh semen (400 million spermatozoa/0.25 ml) 24 h after the onset of estrus (day 15–24). The pregnancy rate, assessed by ultrasound 35 days after insemination, was 48.9% (24/49) and 43.47% (22/50) for sheep in RE and PRO groups respectively. Lambing rates were identical to pregnancy rates in both groups, while the prolificacy rate was 120% (59/49) and 130% (65/50) in RE and PRO groups respectively. No significant differences were found between the 2 groups. The data supports the conclusion that the ram effect can be used successfully to synchronize estrus in organic farming operations.

Reproductive behavior in hair sheep rams under tropical conditions.

The reproductive efficiency of sheep herds depends to a great extent on the ram. Male reproductive evaluation allows to select for the best and eliminate those with reproductive problems. The objective was to evaluate the effect of breed and age group on the reproductive behavior of hair sheep rams in the tropics of Mexico. Pelibuey (n = 42), Blackbelly (n = 30), Dorper (n = 44), and Katahdin (n = 30) rams of two age groups: young (n = 74, 1-1.5years old) and adult (n = 72, 2-4years old) were evaluated. Serving capacity (10-min duration) and breeding soundness evaluation (BSE) tests were carried out in each ram. In the first test, rams were classified as suitable and unsuitable, and in the BSE test, they were classified as satisfactory, questionable, or unsatisfactory. The response variables were analyzed using chi-square test or analyses of variance (ANOVA). ANOVA included the fixed effects of breed, age group, and their interaction. In the serving capacity test, 79.5% of the rams were considered suitable, with no breed differences (P > 0.05). Adult rams (90.3%) had the highest proportion of suitable rams (P < 0.05). In the BSE test, 80.2% of the rams were satisfactory; only breed being significant (P < 0.05). Pelibuey breed had the highest proportion of satisfactory rams (91.4%). Breed × age interaction was no significant for any trait. After serving capacity and BSE tests, a high proportion of rams was found not suitable for reproduction (36.3%), which is expected to cause low fertility in the flock.

Influence of omega‐3 incorporation in sperm preservation medium on physical and kinematic properties of chilled and cryopreserved ram spermatozoa

Two experiments were carried out to investigate the efficiency of supplementing sperm preservation medium with omega-3 polyunsaturated fatty acids on improving liquid-chilled storage and cryopreservation capacity of ram spermatozoa. Ejaculates (n=100) were collected from five adult rams, Ovis aries, by an artificial vagina twice weekly throughout the period February-April, 2017. After initial evaluation, ejaculates of each collection session from the same males were pooled, diluted (1:10) with Tris-citric acid egg yolk extender, and were further split into five aliquots using a split-sample technique. The first aliquot served as control (omega-free), whereas the other four portions were supplemented with 0.1, 0.2, 0.3 or 0.4mM omega-3, respectively (T0 ). Thereafter, the diluted specimens were stored at 4°C for 48hr, during which sperm physical and morphometric properties were evaluated along with oxidative stress indices (T24 , T48 ). Omega-3 levels that efficiently mitigated the detrimental effects of chilled preservation, and maintained preservation aptitude of spermatozoa were further investigated for sperm cryosurvival against control (untreated). Post-thaw physical and kinematic properties of spermatozoa, in all groups, were objectively evaluated by a computer-assisted sperm analysis (CASA) system. The results showed that, at 48hr of chilled storage, supplementing preservation medium with 0.4mM omega-3 was positively correlated (p<0.01) with each of progressive motility, live sperm, intact acrosome and intact cell membrane (r=0.83, 0.85, 0.85, 0.89, respectively). Furthermore, a positive correlation (p<0.01) was observed between inclusion of omega-3 in cryopreservation medium and each of post-thaw total sperm motility, progressive motility, live sperm, normal sperm, intact acrosome, intact cell membrane, VCL, VSL, VAP, ALH and STR (r=0.76, 0.84, 0.79, 0.90, 0.89, 0.91, 0.61, 0.73, 0.65, 0.78 and 0.60, respectively). These results accentuate efficiency of supplementing the diluent with omega-3 fatty acids on improving chilled and cryopreservation aptitude of ram spermatozoa.

Beneficial and harmful effects of cyclodextrin-vitamin E complex on cryopreserved ram sperm

Vitamin E is a potent molecule, especially when loaded in cyclodextrin, in modulating oxidative stress during the freeze-thawing process. The present study aimed to investigate the effect of different concentrations of cyclodextrin-vitamin E complex (CD-Vit E) on cryopreserved ram sperm. Ejaculates collected from five adult rams were pooled and divided into four aliquots. All aliquots were treated in Tris-extender (Tris-glucose-citric acid) containing 2 mg cholesterol-loaded methyl-β- cyclodextrin/120 × 106 spermatozoa and either 0 (Control), 2, 4 or 6 mg CD-Vit E/120 × 106 spermatozoa, corresponding to 0, 0.5, 1 or 1.5 of pure vitamin E, respectively. After incubation at 22 °C for 15 min and the addition of Tris-extender containing glycerol and egg yolk (v/v), all aliquots were frozen in liquid nitrogen. After thawing, motility (computer aided sperm analysis), viability (eosin staining), membrane integrity (HOST), acrosome integrity (Coomassie G-250 staining) and lipid peroxidation (Thiobarbituric acid assay) were evaluated. Compared to control, 2 mg CD-Vit E had a significant positive effect on total motility, progressive motility, movement linearity (LIN%), viability and lipid peroxidation. At 4 mg, however, CD-Vit E had a significant negative effect on total motility, progressive motility, membrane functionality and acrosome integrity. At a greater concentration (6 mg), the negative effects were greater as compared with inclusion of 4 mg in the cryoprotectant and the percentage of rapidly and moderately motile gametes and viability were also altered. In conclusion, the effect of CD-Vit E on cryopreserved ram sperm was concentration-dependent with the 2 mg amount having a beneficial effect while greater concentrations (4 and 6 mg) had a harmful effect on sperm motility and gamete integrity but without affecting oxidative stress status.

Safety of Antitheilerial Drug Buparvaquone in Rams

Background: Theileriosis is a tick-borne disease caused by Theileria strains of the protozoan species. Buparvaquone is the mostly preferred drug in the treatment theileriosis, while it is safety in sheep, has not been detailed investigated. It has been hypothesized that buparvaquone may show side effects and these effects may be defined some parameters measured from blood in sheep when it is used at the recommended dose and duration. The aim of this research was to determine the effect of buparvaquone on the blood oxidative status, cardiac, hepatic and renal damage and bone marrow function markers.Materials, Methods &amp; Results: In this study, ten adult (&gt; 2 years) Akkaraman rams were used. Healthy rams were placed in paddocks, provided water ad libitum, and fed with appropriate rations during the experiment. Buparvaquone was ad­ministered at the dose of 2.5 mg/kg (IM) intramuscularly twice at 3-day intervals. Blood samples were obtained before (0. h, Control) and after drug administration at 0.25, 0.5, 1, 2, 3, 4 and 5 days. The blood samples were transferred to gel tubes, and the sera were removed (2000 g, 15 min). During the study, the heart rate, respiratory rate, and body temperature were measured at each sampling time. In addition, the animals were clinically observed. Plasma oxidative status mark­ers (Malondialdehyde, total antioxidant status, catalase, glutathione peroxidase, superoxide dismutase), serum cardiac (Troponin I, creatine kinase-MBmass, lactate dehydrogenase), hepatic (Alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, gamma glutamyltransferase, total protein, albumin, globulin) and renal (Creatinine, blood urea nitrogen) damage markers and hemogram values (white blood cell, red blood cell, platelet, hemogram, hematocrit) were measured. Buparvaquone caused statistically significantly (P &lt; 0.05) increases in the troponin I and blood urea nitrogen levels and fluctuations in alkaline phosphatase activity, but there was no any statistically significance difference determined in the other parameters.Discussion: In this study, buparvaquone was administered two times at a dose of 2.5 mg/kg (IM) at 3-day intervals. Al­though the result was not statistically significant (P &gt; 0.05), it was determined that buparvaquone gradually increased the levels of the main oxidative stress marker, MDA, by approximately 2.8 fold. CAT and GPX levels were also found to have decreased by 2.2 fold. Buparvaquone may cause lipid peroxidation by producing free radicals. Some other antiprotozoal drugs may affect the oxidative status and may increase MDA level and decrease SOD level. In this study, MDA, which is an indicator of lipid peroxidation in vivo, was used to partially detect developing lipid peroxidation. Changes in the levels of reduced GPX and CAT enzymes could be attributed to their use in mediating the hydrogen peroxide detoxification mechanisms. The absence of significant changes in the TAS levels in this study suggests that buparvaquone may partially induce oxidative stress by producing hydrogen peroxide, but no significant changes occurred in the oxidative stress level because of the high antioxidant capacity of sheep. In this study, buparvaquone caused a statistically significant increase (P &lt; 0.05) in the level of Tn-I, which is a marker of specific cardiac damage (P &lt; 0.05), whereas there was no statistically (P &gt; 0.05) significant increase in CK-MBmass. Tn-I and CK-MB levels, which are used to define heart damage in humans, have been successfully used to determine heart damage in sheep. In this research study, the statistically significant increases in Tn-I but not CK-MBmass levels could be considered indicative of mild cardiac damage.Keywords: ram, buparvaquone, safety.

Computer assisted semen analysis of deccani ram semen presevability at 5ºC

To investigate the preservability of Deccani ram semen in three different extenders - SC (Sodium Citrate), T (Tris), C (Coconut milk) egg yolk extenders after dilution and chilling (5ºC) at 1, 24 and 48 hr. of storage. Eight Deccani adult rams (aged 2-4 years) were selected and ejaculate from each ram was divided into 3 aliquots and diluted with SC, T and C extenders (final concentration - 400 million spermatozoa/0.2 ml semen) and stored at 5ºC. Semen evaluation was done by computer assisted semen analyser (CASA). Analysis of Variance (ANOVA) was used to test the significance difference between extenders. S: Sodium citrate (SC) and Tris (T) - Egg yolk extender was significantly (P less than 0.05) better as compared to Coconut milk extender (C). No significant difference (P&gt;0.05) was observed between SC and T. Sperm kinematics assessment by CASA revealed Sodium citrate-based egg-yolk extender (SC) as better extender for preservation of Deccani Ram semen upto 48 hr. of storage as compared to T and C.

Isolation and enrichment of putative spermatogonial stem cells from ram (Ovis aries) testis

The present study aimed to isolate and enrich putative SSCs from ram testes, which are positive for promyelocytic leukaemia zinc-finger protein (PLZF). The putative SSCs were isolated using a combination of enzymes with different concentrations, collagenase (1 and 2 mg/ml), hyaluronidase (1 mg/ml) and trypsin (0.25 and 0.5 mg/ml). The isolated SSCs were purified using an extracellular matrix such as laminin (20 μg/ml), DSA-lectin (5 μg/ml) and gelatin (0.2%) in combination with BSA (0.5 mg/ml). The number of putative SSCs/ tubule was significantly (p < 0.05) higher in prepubertal (3.1 ± 0.51) and adult (3.45 ± 0.58) than the number of gonocytes/tubule in neonatal (0.59 ± 0.03) testis. Optimum enzyme combinations required for isolation of putative SSCs from prepubertal testis (collagenase; 2 mg/ml and trypsin; 0.5 mg/ml) were different from adult testis (collagenase; 1 mg/ml, trypsin; 0.25 mg/ml and hyaluronidase; 1 mg/ml). Though the number of putative SSCs/tubule was comparable in prepubertal and adult animals, a significantly (p < 0.05) higher percentage of putative SSCs (7.33 Vs 0.47%) were isolated from prepubertal testis than the adult. Differential plating using laminin along with BSA resulted in a significantly (p < 0.05) higher number of putative SSCs. The enzyme combinations suitable for isolation of putative SSCs from prepubertal testis are different from adult ram testis and the laminin has been found to be effective for purification of putative SSCs from testicular cells isolates.