Semen Of Rams Research Articles

Effect of adding different levels of Artichoke (Cynara scolymus) aqueous extract to Tris extender on the Motility, Viability and some characteristics of rams semen stored at 5 °C

This study was conducted to show the effect of the aqueous extract of the leaves of Artichoke ( Cynara scolymus L.) on Tris extender and its effect on the semen traits of Awassi rams after preservation at a cooling of 5° C for different periods (0, 24, 48 and 72 hours). This study was conducted in the animal field and laboratories of the Department of Animal Production, College of Agricultural Engineering Sciences, University of Baghdad in Jadiriyah, from November 15, 2021 to April 25, 2022. Three local Awassi rams were used in this experiment. Their ages ranged from 2.5-3.0 years. The semen was collected from them by artificial vagina by one ejaculation/ram/week for 8 weeks. The semen of rams has been mixed in order to eliminate individual differences. The following tests were conducted to evaluate the semen characteristics for each motility, the percentage of viability sperm, total abnormalities, the integrity of the plasma membrane, and the acrosome's integrity. The current study showed that adding an aqueous extract from the leaves of the artichoke plant (Cynara scolymus L.) improved most of the studied traits when stored in preservation. The results indicated a highly significant (P<0.01) for the treatments T1, T2 and T4, which amounted to (87.00 ± 1.86, 85.50 ± 2.02 and 86.87 ± 0.78%) compared to the control group (C), which amounted to (77.25 ± 2.11%), While the previous treatments did not differ significantly with treatment T3, which amounted to (81.87 ± 2.94%) in the percentage of live sperm during the 0 hour preservation period. At the same time, the results indicated no significant differences between treatments T1, T2, T3 and T4 compared to the control treatment and for all periods (0, 24, 48 and 72 hours) in the individual motility of the sperm. As for measuring the biochemical traits of seminal plasma, The results of AST and ALT enzymes indicated that there was no significant increase between treatments during one period, while an arithmetic superiority occurred after 72 hours of preservation for treatments T1, T2, T3 and T4 compared with the 0 hour preservation period. It is concluded from the current study that adding an aqueous extract of (Cynara Scolymus) artichoke leaves to Tris extender improved the semen quality of Awassi rams after preservation. Keywords: Ram; Semen; Antioxidants; Cooling; Artichoke; Cynara Scolymus.

Sperm productivity of Texel breed rams depending on the period of seasonal activity and mode of use

The results of laboratory studies of freshly obtained and deep-frozen semen of Texel breeding rams in the non-mating and mating periods under different modes of using 4, 6, and 8 ejaculates per week are given. It has been proven that during the non-mating period, when receiving four ejaculates per week compared to 6 and 8 ejaculates, the ejaculate volume was 11.3 and 22.6 % higher, respectively. The sperm concentration in it was 5.1 and 12.2 %, sperm activity was higher by 1.7 and 3.2 %, and their resistance by 7.1 and 17.3 percent. In the mating period under the above regimen, the ejaculate volume was 9.9 and 19.7 % higher; sperm concentration was 4.1 and 5.9 % higher; sperm activity was higher by 5.7 and 8.3 %, resistance was higher by 7 .7 and 19.9 percent. Higher quality indicators were found in the mode of obtaining four ejaculates after sperm cryopreservation in both research periods, on average, according to the activity of deconserved sperm by 3.2 and 8.0 and 5.8 and 11.2 %, freezing resistance by 0.7 and 2,0 and 2.6 and 4.6 %, survival by 7.6 and 15.5 and 2.2 and 10.6%, absolute survival rate by 3.4 and 8.1 and 4.7 and 8.5 percent Based on the analysis of the investigated indicators of freshly obtained and de-preserved sperm, it was experimentally confirmed that in the mating period compared to the non-mating period, they were significantly higher on average in freshly obtained sperm from 7.7 to 40.1 %, in cryopreserved sperm – from 2.9 to 16.7 percent. The preservation of acrosomes of sperm in the non-mating and mating period, when receiving four ejaculates per week compared to receiving 6 and 8, was higher by periods by 3.5 and 6.0 %, and 2.5 and 5 %, respectively. The studies established significantly higher quantitative and qualitative indicators of freshly collected and deconserved sperm in both seasonal activity periods under the regime of obtaining four ejaculates per week from breeding rams. Under this regime, the sperm is biologically complete, and its quality indicators meet the requirements of the instructions for the artificial insemination of sheep.

Comparison of Five Protocols of Estrous Synchronization on Reproductive Performance of Hu Sheep.

The purpose of this study is to compare five protocols of estrous synchronization for Hu ewes to obtain the most effective and economical protocol, to apply the advantageous scheme in large-scale sheep farming. Healthy multiparous Hu ewes (n = 150) were randomly divided into five groups, and all ewes were administered fluorogestone acetate (FGA, 45 mg) vaginal sponge. The sponges of the first three groups (Groups I, II, and III) were removed on the 11th day, and 0.1 mg of PGF2α was injected intramuscularly on the ninth day. Group I received 6 μg of gonadotropin-releasing hormone (GnRH) by intramuscular injection at 36th h after withdrawal of the sponge. Group II was injected 330 IU of pregnant mare serum gonadotropin (PMSG) on the ninth day. The combination of 6 μg of GnRH and 330 IU of PMSG was treated in Group III at the same time as Group I and Group II. The sponges of the latter two groups (Groups IV and V) were removed on the 13th day, and 330 IU of PMSG was injected intramuscularly simultaneously. PGF2α (0.1 mg) was administered on the 12th day in Group IV. All ewes were detected for estrus at 24, 36, 48, 60, and 72 h after the sponge removal. The loss of sponge and vaginitis was recorded when the sponge was withdrawn. Cervical artificial insemination (AI) was performed with fresh semen of Dorper rams diluted with skimmed milk. After 30 days of insemination, the conception was detected with a veterinary B-ultrasound scanner. The lambing status of all ewes and the cost of drugs for estrous synchronization in each group were recorded. The results showed the following: (1) on the whole, the average percentage of estrous ewes in the period of 24–36 h and 36–48 h after removal was significantly higher than other three periods and that of the period of 60–72 h was significantly lower than the first three periods after removal; (2) there was no significant difference in percentages of estrous ewes in any of the five time periods, sponge loss rate, vaginitis rate, total percentage of estrous ewes, conception rate, single lambing rate, twinning rate, and multiple lambing rate of ewes among five protocols; (3) total percentage of estrous ewes and conception rate were more than or equal to 80% in the Groups II and III, and the twinning lamb rate of the Group II protocol was 70%; (4) there was no difference in lambing rate of ewes among Groups II, III, IV, and V; (5) the Group III had the highest drug cost of 22.5 CNY. In conclusion, considering the lambing rate, twinning lamb rate, and drug cost for estrous synchronization, Group II was the most advisable for application and promotion in large-scale sheep farms among these five protocols of estrus synchronization.

Analysis of external features and live weight of woolly-meaty sheep (ovis aries)

This article addressed the development and the growth of offspring, the external characteristics and the productivity at different breeding periods of woolly-meaty sheep of Uzbekistan. It was observed that wool productivity was higher in sheep rich in wool-meat than ordinary sheep; sheared wool was 1.26 kg (49.6%), after washing it was 0.97 kg (67.8%), wool length was 2.13 (19.2%) higher. Live weights of lambs from ewes in the experimental groups were determined at birth, at 30 days of age, and 3 months of age. From September 1 to September 20 of the year, the live weight, body size and wool productivity of early inseminated ewes were higher than those of inseminated ewes from late October to November 10. The live weight of lambs obtained from artificially inseminated sheep with frozen semen of typical rams in world gene pool considered semi-fine wool in the meat-wool direction was higher than sheep naturally inseminated with pedigree rams. Accordingly, it was heavier by 00.4 kg (9.3%) at the first day of birth, followed by 0.5 kg (6.8%) at 10 days, and 0.4 kg (1.3%) at 3 months of age.

Fertility and flow cytometry evaluations of ram frozen semen in plant-based extender supplemented with Mito-TEMPO

Semen cryopreservation is an effective strategy for distributing spermatozoa for artificial insemination, but this process reduces the fertility potential of post-thawed spermatozoa. The current study was conducted to assess effects of the novel mitochondria-targeted antioxidant “Mito-TEMPO” on ram sperm quality and fertility potential during the freeze-thawing process in a plant-based extender. Semen samples were diluted in extenders supplemented with 0, 0.5, 5, 50 and 500 µM Mito-TEMPO and then frozen using a standard protocol. Motility, abnormal morphology, acrosome integrity, membrane integrity, mitochondria membrane potential, viability, apoptotic-like changes, lipid peroxidation, DNA fragmentation, H2O2 concentration and fertility potential were assessed after thawing. Results indicated that with the 5 and 50 μM Mito-TEMPO there was a greater (P ≤ 0.05) percentage of sperm total motility, progressive motility, acrosome integrity and viability as well as less (P ≤ 0.05) lipid peroxidation and late apoptotic-like changes. Membrane integrity and mitochondria membrane potential were greater (P ≤ 0.05) with the 50 μM Mito-TEMPO extender supplementation. Furthermore, with 0.5, 5 and 50 μM Mito-TEMPO supplementations there was a greater (P ≤ 0.05) average path velocity and lesser (P ≤ 0.05) percentages of spermatozoa with DNA fragmentation and relatively greater H2O2 concentration. Results from the fertility experiment indicated the 5 and 50 μM Mito-TEMPO treatments resulted in greater pregnancy, parturition and lambing rates. It, therefore, is concluded that Mito-TEMPO may enhance quality and fertility potential of post-thawed semen of rams.

Detection of Toxoplasma gondii infection in semen of rams used for natural mating in commercial sheep farms in Tunisia.

Toxoplasmosis, a parasitic infection, disrupts sexual functions resulting in decline in reproductive and economic performance in sheep. Lacking discernible symptoms, toxoplasmosis is difficult to detect and diagnose in infected animals. Here, to estimate the prevalence of natural infection by Toxoplasma gondii, we used PCR to amplify and detect T. gondii DNA in semen from 92 rams of three breeds from four regions in Tunisia and seroprevalence was determined using a commercial ELISA kit. From the PCR amplifications, 51.09 ± 10.21% of the rams were tested positive for T. gondii with an overall seroprevalence of 39.13 ± 9.97%. Risk factors including ram location and number of accomplished mating seasons significantly (p < .05) affected the sero- and molecular prevalence of T. gondii in semen but, there was a fair concordance between sero- and molecular prevalence (Kappa = 0.33). Sequences of T. gondii from five positive samples were 100% identical (same haplotype). Comparison of these sequences with those archived at the GenBank showed a sequence similarity range between 95 and 100%. The haplotype defining the five Tunisian sequences was similar to the one observed in chicken, cats, European pole cat and humans from Brazil, St Kitts and Nevis, Great Britain and Tunisia, respectively. This indicates its wide geographic distribution and non-species specificity. Our findings suggest a high prevalence of toxoplasmosis in Tunisian matting rams; further studies concerning its venereal transmission capacity are needed prior to recommending a systematic screening of T. gondii DNA in rams' semen used for both natural breeding and artificial insemination.

Neospora caninum Natural Infection in Tunisian Rams: Serological Study and Molecular Identification of Infection in Semen.

The aim of this study was to estimate the seroprevalence and molecular prevalence and perform a molecular identification of Neospora caninum in semen of Tunisian rams. A total of 92 blood samples were collected from four farms located in four Tunisian governorates (Jendouba, Kairouan, Zaghouan and Ben Arous) and samples were screened with a commercial ELISA kit for N. caninum antibodies. For the same rams, semen samples were collected and tested for the presence of N. caninum ITS1 gene using PCR. Five amplicons were randomly selected for sequencing. A phylogenetic tree was constructed to compare the partial sequences of the ITS1 gene with sequences deposited in GenBank. The seroprevalence of N. caninum infection was 25% (23/92) and PCR revealed that the molecular infection prevalence in semen was 11.95% (11/92). Kappa test showed an average agreement between seroprevalence and parasite prevalence in semen (κ = 0.44). The highest molecular prevalence was for rams that accomplished more than two mating seasons (21.0 ± 12.1%) compared to those performed less than two mating seasons and yearling individuals (4.0 ± 5.5%) (P = 0.01). There were no differences in N. caninum molecular prevalence according to either breed or locality. Comparison of the partial sequences of the ITS1 gene revealed 99-100% similarity with those deposited in GenBank. To the best of our knowledge, this is the first detection and molecular identification of N. caninum in semen from rams in North Africa. Our findings indicate that N. caninum infection rate was high in rams.

Growth performance and meat characteristics of the first filial Awassi-Rambouillet callipyge ram lambs.

Aim:This study was designed to introduce the callipyge (CLPG) and 50% of Rambouillet sheep genes to improve meat quality and quantity of Awassi (AW) sheep.Materials and Methods:The CLPG mutation was introduced into the AW sheep through frozen semen of homozygous Rambouillet rams for the CLPG mutation. Four ram lambs from the first-generation Rambouillet callipyge Awassi (F1-RCA) and five from pure AW were recruited for a fattening trial conducted in individual pens using standard ration, following which ram lambs were slaughtered for carcass and meat evaluation.Results:Final body weight, dry matter intake, average daily gain, and feed conversion ratio were significantly higher in F1-RCA than AW. Hot and cold carcass weights and the other carcass cuts’ weights, except for the fat tail, were heavier in F1-RCA than AW. There was no difference in dressing percentage between the two genetic groups (p>0.05). All non-carcass components’ weights, except spleen, kidney, and testis, were higher in F1-RCA. Total lean, total bone, and intermuscular fat weight were greater in F1-RCA, but bone-to-lean ratio was lower in F1-RCA when compared with AW (p<0.01). No differences (p>0.05) were observed in all meat quality parameters for muscle longissimus with the exception of pH, redness color, and tenderness that were lower (p<0.05) in F1-RCA than AW. F1-RCA lambs had larger longissimus muscle area (30.9 vs. 16.9 cm2) and less leg fat depth (11.1 vs. 17.4 mm).Conclusion:The implications of this research show the potential of 50% of Rambouillet genes and the CLPG mutation to improve growth and meat characteristics in AW-Rambouillet crosses and can be used further to develop a meat-type AW with improved productivity and muscle mass.

The influence of dietary sunflower oil, rich in n-6 polyunsaturated fatty acids, in combination with vitamin C on ram semen parameters, sperm lipids and fertility.

Dietary fats, fed to ruminant animals, have been observed to improve reproductive function. Sunflower oil is one of the richest sources of polyunsaturated fatty acids (PUFAs), over 60% of which is linoleic acid (C18:2). With regard to recommendations for increased consumption of PUFAs, special emphasis has been given to the importance of consuming more antioxidants to prevent lipid peroxide formation. This study evaluated the effects of dietary n-6 PUFAs and vitamin C (VC) in rams-in particular, on semen quality, blood metabolites, sperm lipids and overall fertility. Diets supplemented with SFO and VC increased the proportion of motile sperms and their progressive motility improved (P < 0.01). Rams fed a diet containing SFO had an increased proportion of sperms with normal acrosomes in their semen samples (P < 0.01). The highest levels of lactate dehydrogenase activity (LDH) enzyme were found in control rams (2.60 U mL-1 ). Feeding SFO significantly affected blood triglyceride and cholesterol concentrations (P < 0.05). Diets containing SFO increased the proportion of C18:2c, C20:4, C20:3 and C22 in the sperm lipid composition (P < 0.05). Semen of SFO and VC-fed rams was more fertile than that of control rams when it was artificially inseminated to ewes. Feeding rams with supplemental SFO and VC increased semen quality and improved fertility rates. © 2019 Society of Chemical Industry.

Detection of Mycobacterium avium subsp. paratuberculosis in reproductive tissue and semen of naturally infected rams

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis (PTB), disease that causes a syndrome of bad nutrient absorption, weight loss and eventually death. The intestine is the main target organ where the infection develops; however, there is evidence of infection by MAP in extra-intestine sites of sheep, including mesenteric nodes and semen. The aim of the study was to identify the presence of MAP in reproductive tissue and semen of infected Pelibuey rams in clinical state of PTB. Seven rams were used in clinical PTB state and a non-infected ram by MAP of the Pelibuey breed, confirmed by serology, nPCR and bacteriological culture, with average weight and age of 57.23 ± 1.73 kg and 2.91 ± 0.17 years, respectively. The presence of MAP was identified in different tissue samples: spleen (1/7, 14.3% and 2/7, 28.6%), small intestine (3/7, 42.9% and 4/7, 57.1%) and mesenteric lymph nodes (3/7, 42.9% and 3/7, 42.9%), with nPCR and culture, respectively. It was also identified in epididymis tissue (1/7, 14.3%), Cowper gland (2/7, 28.6%) and prostate (1/7, 14.3%), using nPCR, although without detection in culture. It was identified in testicular tissue in 42.8% (3/7; culture or nPCR technique), but in 28.6% (2/7) with both techniques. Finally, the presence of MAP was identified in 42.9% (3/7) of semen samples with nPCR; however, it was not detected through culture. In conclusion, the presence of MAP was identified in lymphatic, digestive tissue, and semen; the presence of MAP was reported for the first time in epididymis, Cowper gland, prostate and testicles of infected Pelibuey rams.

Seminal plasma proteins modify the distribution of sperm subpopulations in cryopreserved semen of rams with lesser fertility

Any physiological mechanism involved in sperm selection and semen improvement has effects on heterogeneous sperm populations. This is mainly due to the fact that sperm populations within a single ejaculate have considerable heterogeneity for many variables, such as motility which is meaningful in terms of understanding how some sperm cells possess fertility advantages as compared with other cells. In the present research, initially there was a multivariate and clustering analysis used to assess sperm motility data from cryopreserved ram semen to identify subpopulations and compare the distribution of these clusters between rams with lesser and greater fertility. There were four classifications made of sperm subpopulations (clusters): CL1 fast/linear/progressive sperm; CL2 fast/non-linear sperm; CL3 very fast/linear sperm with vigorous beating and CL4 slow/non-linear sperm. Rams with greater fertility had a lesser proportion of sperm considered as “hyperactivated” (CL2) and a greater proportion of slow and non-linear sperm (CL4) than sperm of rams with lesser fertility. In addition, the effects were assessed for the capacity of seminal plasma (SP) and interacting SP proteins (iSPP) that were present during different seasons of the year to improve the distribution of sperm within subpopulations of semen from rams with lesser fertility. The iSPP and SP were obtained by artificial vagina (AV) and electroejaculation (EE) during breeding and non-breeding seasons and added to thawed semen. All the aggregates had a significant effect on the distribution of sperm subpopulations and effects differed among seasons of the year and depending on collection method used. Even though, future studies are needed to assess the contribution of each subpopulation on ram sperm fertility, it is important that a multivariate analysis be used to evaluate the effect of a treatment on sperm quality variables.

ECONOMIC BASES OF GENE POOL OF LOCAL AND ENDANGERED BREEDS OF FARM ANIMALS SPECIES PRESERVATION IN UKRAINE

The aim was to determine the amount of state financial support for the full gene pool preservation of local and endangered breeds of farm animals in Ukraine for 2016-2020.&#x0D; Given the great strategic and economic importance of the problem of gene pool preservation and rational use to enhance food security, the level of state financial support for preservation of the gene pool of local and endangered breeds of farm animals was determined for using two methods of conservation: in situ (live animals) and ex situ (cryoconservation of their genetic materials).&#x0D; The level of budget support to preserve gene pool objects involves calculating the minimum (normative) amount of genetic resources (livestock females, bull semen, embryos) and rational choice of methodical approach to determining the level of budget support for the preservation of each type of genetic resources.&#x0D; The level of budget support for in situ maintenance of gene pool objects was determined based on level of budget subsidy per head and livestock number of each species, recommended to preserve (normative). The basis of the level of budget subsidies was determined as compensation amount of normative costs for feed produced at cost. Normative costs of feed were determined on the basis of scientifically grounded rations for species and breeds of animals taking into account their productivity.&#x0D; The full in situ preservation of dairy and dual-purpose breeds of cattle requires the holding herds of each breed which include animals of all main bloodlines, where at least 3 live bulls and 60 cows should be kept per bloodline. In gene pool subjects of beef breeds at least 3 bulls and 20 cows should be kept per bloodline. Breeding cattle of all breeds should be performed by linear purebred breeding and randomized fixing of bulls in lines. Under these conditions, it is possible to preserve purebred livestock number for 5-6 generations, or 20 years. To preserve in situ the gene pool of local and endangered breeds of pigs, sheep and poultry it is necessary to provide herd size not less than 25 boars and 100 sows, 20 rams and 200 ewes, 10 stallions and 50 mares, 50 geese and 200 fowl, 50 drakes and 250 ducks, 50 cocks and 250 hens.&#x0D; The constancy of gene pool of local and endangered breeds is fully possible to provide only by ex situ method, providing measures to preserve genetic fund of breeds, types, lines in an artificial environment (cryoconservation of gametes, somatic cells, zygotes, tissues).&#x0D; Ex situ preservation of the gene pool of local and endangered breeds is expedient as a "virtual" cryo-herds. For their expansion annually 1-3 thousand doses of bull semen of dairy, dual-purpose and beef breeds per each farm or genealogical line (depending on the number of bulls in a line) must be collected and stored in cryo-depositories in over 3 thousand doses of bull semen for each line. Annual accumulation and storage of semen of rams should be accordingly 1.2 and 2.4 thousand doses, boars – 2 thousand doses and stallions – 300 doses in both cases. The annual need for purchasing sires’ semen of farm animals is 35.7 thousand doses. For implementation of programs for preserving the gene pool of local breeds it is necessary to pass 10% of annual purchased semen to the Bank of Animal Genetic Resources of Institute of Animal Breeding and Genetics nd. a. M.V.Zubets of NAAS.&#x0D; The normative costs for an annual storage of sires’ semen in 2015 have been determined, which amount is UAH 4.54 and UAH 2.98 according to normative capacity of cryo-depositories (5 thousand doses and 20 thousand doses).&#x0D; The project of preserving populations of local and endangered breeds of cattle as "cryo-herds" (cryopreserved bovine embryos) during 2016-2020 requires the state support for obtaining 250 bovine embryos, prepared for long-term storage, of each breed and storing them in Bank of Animal Genetic Resources of Institute of Animal Breeding and Genetics nd. a. M.V.Zubets of NAAS.&#x0D; To provide the full preservation of the gene pool of local and endangered breeds of farm animals in Ukraine using two methods of conservation: in situ (live animals) and ex situ (“virtual cryo-herds”) during the next five years it is necessary to finance UAH 75,137.5 thousand from the State Budget. The implementation of the developed economic mechanism ensures the gene pool preservation of domestic autochthonous and local breeds of farm animals and their further use for breeding animals of specialized and dual-purpose breeds, the execution of commitments laid down by international agreements of Ukraine for the biodiversity preservation. These research results have been used in the development of the Programme of preservation of local and endangered breeds of farm animals in Ukraine for 2016-2020.

Presence of Mycoplasma agalactiae in semen of naturally infected asymptomatic rams

The purpose of the present study was to assess the presence of Mycoplasma agalactiae (Ma), the main causative agent of ovine contagious agalactia (CA), in semen of naturally infected rams. Therefore, semen samples from 167 rams residing in three different artificial insemination (AI) centers of a CA–endemic area were studied by microbiological and molecular techniques. In addition, serial ejaculates from the same rams were evaluated to determine the excretion dynamics of Ma. Of the 384 samples studied, Ma was detected in 56 (14.58%) which belonged to 44 different rams (26.35%). These findings confirm the ability of Ma to be excreted in semen of asymptomatic rams. Furthermore, these results also evidence the presence of these asymptomatic carriers of Ma in ovine AI centers, representing a serious health risk regarding the spread and maintenance of CA, especially in endemic areas. Moreover, the excretion of Ma in semen also points to the risk of venereal transmission of this disease. The current results highlight the need to implement control measures to prevent the admission of infected rams in AI centers and the necessity to continuously monitor semen samples to effectively detect infected individuals.

Effect of preservation time on the quality of frozen semen in indigenous rams

The study was carried out to observe the effects of preservation time on the quality of frozen semen of indigenous rams. Semen was collected using AV once a week from 4 rams. Tris based with 10% egg yolk and 7% glycerol extender was used to extend and freezing the semen. Fresh semen was evaluated for volume, density, mass motility and concentration, and mean values were observed as 0.8±0.2ml, 3.0±0.3, 3.2±0.7, 3.9±0.7×109/ml, respectively. Significant difference (p&lt;0.05) was found for all the parameters among the rams. Mean values of motility, viability and normal morphology percentages were 83.3±4.3%, 88.2±4.4%, 84.2±3.5% in fresh semen while those of chilled semen at 40C were 74.7±2.3, 78.8±4.9 and 79.2±2.9%, respectively. For all the parameters, significant (p&lt;0.05) difference was found among the rams. Frozen sperm motility was observed after thawing at 39-400C for 14-15 seconds. The mean motility, viability and normal morphology percentages after freezing for 24hrs, 7, 15 and 30 days of duration were 39.8±3.1, 41.1±4.3, 40.1±4.1 and 39.4±2.9%; 44.5±2.5, 45.3±2.8, 44.6±2.8 and 43.9±2.8%; 71.0±2.0, 71.7±1.5, 70.7±1.7 and 70.3±1.8%, respectively and values did not decrease significantly (p&gt;0.05) with the increasing time of preservation. Non significantly decrease of the semen quality with advance of preservation time indicates the suitability of the protocol used for freezing of indigenous ram semen in Bangladesh.DOI: http://dx.doi.org/10.3329/bjas.v44i1.23113 Bang. J. Anim. Sci. 2014. 44 (1): 10-15

Detection of Toxoplasma gondii DNA in fresh and frozen semen from rams in Brazil.

The aim of this study was to estimate the prevalence of genomic DNA of Toxoplasma gondii in semen samples from commercial rams in artificial insemination centres in Brazil, as well as in fresh semen from rams in the northeast of Brazil. In total, 108 semen samples were obtained from artificial insemination centres, and genomic DNA of T.gondii was detected in 24 of 108 (22.2%). The prevalence of antibodies anti-Toxoplasma gondii among sheep on rural properties was 9.2% (10/109), and 100% of the semen samples of these animals were positive in the PCR for T.gondii DNA. The molecular identity was confirmed through sequencing, which indicated 99.9% similarity with the T.gondii DNA sequences stored in the GenBank. This study reports the first occurrence of T.gondii DNA in the semen of rams, which came from artificial insemination centres in Brazil, as well as the occurrence of T.gondii DNA in the fresh semen of naturally infected rams in the northeast of Brazil.

Koçlarda Taze Sperma Parametreleri ve DNA Bütünlüğü Arasındaki İlişkilerin Araştırılması

Summary The aim of the present study was to evaluate the correlation between the routine semen analyses and sperm DNA integrity, as assesed by TUNEL in fresh semen of rams in Kivircik and Awassi breeds in breeding season. Semen was collected by electro-ejaculation five times every other day. For that purpose, 50 fresh ejaculates from 6 Kivircik and 4 Awassi rams were evaluated. The mean percentages of semen volume, concentration, mass activity, motility, acrosomal defects, HOST and DNA fragmentation of Kivircik and Awassi rams were 1.3 ml, 1.9 x109, 3.3 (0-5 scale), 72.6%, 13.2%, 73.6%, 16.2% and 1.0 ml, 1.9 x109, 3.3 (0-5 scale), 72.5%, 11.1%, 77.9%, 15.9%; respectively (P>0.05). Sperm DNA fragmentation was correlated adversely with semen volume (r = -0.329, P 0.05), mass activity (r = -0.349, P 0.05) and HOS test (r = -0.513, P<0.01). This study provided clear evidence that most of the parameters evaluated herein related to DNA fragmentation at one point or the other. Hence, DNA structure evaluation of sperm might be a useful tool for accurate prediction of the male fertility in individual rams.

PENGARUH MALTOSA SEBAGAI KRIOPROTEKTAN EKSTRASELULER DALAM MENINGKATKAN KUALITAS SEMEN BEKU GUNA MENDUKUNG KEBERHASILAN TEKNOLOGI INSEMINASI BUATAN

The research carried out to observe the effect of maltose addition on the quality of frozen semen of garut rams. Semen was collected once a week using artificial vagina from six mature garut rams. Semen was equilibrated at 5oC for four hours, frozen and stored in liquid nitrogen. The thawing was carried out at the temperature 30oC for 30 seconds. The result showed that percentages of viable sperm for addition of maltose 1,2 g / 100 ml extender (68,50 ± 0,84%) was significantly difference (P&lt;0,05) than control (54,83 ± 1,94%) and addition of maltose 0,6 g / 100 ml extender (65,67 ± 1,03 %). The percentages of progressive motile sperm and percentages of plasma membrane for addition of maltose 1,2 g / 100 ml extender (53% and 64,67%) were significantly difference (P&lt;0,05) than control (43% and 53,83%) but were not significantly different (P&gt;0,05) from addition of maltose 0,6 g / 100 ml extender (50,83% and 64,67%)respectively. In conclusion, the addition of maltose 1,2 g / 100 ml extender is optimal dose to improve the quality of frozen semen of garut rams.

Detection of Neospora caninum DNA in semen of experimental infected rams with no evidence of horizontal transmission in ewes

Recent reports from New Zealand indicate Neospora caninum has a possible role in causing abortions in sheep. Transmission of N. caninum via semen has been documented in cattle. This study aimed to investigate if horizontal transmission through semen was also possible in sheep. Initially, 6-month old crossbred ram lambs (n=32), seronegative to N. caninum, were divided into 4 equal groups. Group 1 remained uninoculated whilst the remainder were inoculated with N. caninum tachyzoites intravenously as follows: Group 2 – 50 tachyzoites; Group 3 – 103 tachyzoites; Group 4 – 107 tachyzoites. Semen samples were collected weekly for 8 weeks for the detection of N. caninum DNA and quantified using quantitative PCR (qPCR). Plasma collected 1 month post-inoculation was subjected to ELISA (IDEXX Chekit) and Western blot. At 2 weeks post-infection, three rams from Group 1 (uninoculated) and three rams from Group 4 (107tachyzoites/ml) were mated with two groups of 16 ewes over two oestrus cycles. Ewe sera collected 1 and 2 months post-mating were tested for seroconversion by ELISA and Western blot. All experimentally infected rams seroconverted by 1 month with ELISA S/P% values ranging from 11% to 36.5% in Group 2, 12–39.5% in Group 3 and 40–81% in Group 4. However, none of the ewes mated with the experimentally infected rams seroconverted. For the Western blot, responses towards immunodominant antigens (IDAs) were observed in ram sera directed against proteins at 10, 17, 21, 25–29, 30, 31, 33 and 37kDa. Rams in Group 2, 3 and 4 were noted to have at least 3 IDAs present. None of the ewes showed any of the 8 prominent IDAs except for the one at 21kDa which was seen in 30 out of 32 ewes in both groups. N. caninum DNA was detected intermittently in the ram's semen up to 5 weeks post-inoculation with the concentrations ranging from that equivalent to 1–889 tachyzoites per ml of semen. Low concentrations of N. caninum DNA were also detected in the brain tissue of two rams (Groups 1 and 4). These results suggest that although N. caninum DNA can be found in the semen of experimentally infected rams, the transmission of N. caninum via natural mating is an unlikely event.

Effects of vitamin B12 supplementation on the quality of Ovine spermatozoa

The present study was undertaken to investigate the effect of various levels of vitamin B12 in Tris extender on semen quality of Dallagh rams following the cooling and freeze/thawing process. Semen was collected from six healthy and mature rams with an average body weight of 60.0 ± 5.0 Kg using an electro ejaculator. High quality samples were mixed and diluted in Tris extender supplemented with different concentrations of vitamin B12 (0, 1, 2 and 3 mg/ml). The semen aliquots were cooled and preserved at 5°C and their qualities were evaluated during pre-freezing and then the cooled semen samples were packaged into 0.25 ml straws. Straws were frozen in the vapor of liquid nitrogen, and were then stored at -196ºC. Straws were thawed seven days later and the characteristics of spermatozoa were examined. Results of this study showed that the effect of vitamin B12 on characteristics such as viability, motility, progressive motility and normality of spermatozoa were significant in pre and post freezing conditions (P<0.05). In conclusion, for long term storage of semen of Dallagh rams, we recommend using 2 mg/ml of vitamin B12 in semen extender.

Laparoscopic insemination of sheep in Republic of Serbia

This paper describes a procedure of laparoscopic insemination (LAI) performed in 42 Ile de France ewes with deep-frozen rams? semen. Frozen semen doses was imported from France, originating from two rams (A and B), and used intermittently in three LAI cycles on different females. The ewes were examined ultrasonographically for pregnancy, two months following insemination and data of lambing were recorded at the end. Total achieved lambing rate was 61.90% (26/42) and total number of lambs was 49 (prolificacy 1.88; 1.16 lambs per sheep included in LAI protocol). Three labs from 2 ewes were stillborn and one dyed at 10th day after lambing. Lambing rates for rams A and B were 53.85% (14/26) and 75.00% (12/16), respectively. Introduction of laparoscopic insemination of sheep with imported deep-frozen semen of top quality rams, gaining technical experience and achieving conception rate over 65% could make this technique acceptable in improving genetic potential of top quality sheep flocks.