Blood serum triiodothyronine in Karagouniko and Chios rams as affected by season in a temperate environment

The current research investigated triiodothyronine concentrations in Karagouniko and Chios rams in a temperate climate zone, considering the periods January-June and July-November, characterized by a gradual increase and decrease of daylight length, respectively. The animals raised in the farm of the Artificial Insemination Center of Karditsa (39021΄18΄΄N, 21054΄19΄΄E), Greece, and the meteorological variables air temperature, relative humidity, and sunshine, as well as photoperiod, were considered for a more detailed analysis. T-tests were used to detect possible changes in the concentration of triiodothyronine in both sheep breeds and in the abovementioned meteorological variables. January-June coincided with significantly higher (P < 0.05) triiodothyronine concentrations in Karagouniko and Chios rams (0.82 ng/ml and 0.77 ng/ml, respectively) compared to July-November (0.72 ng/ml in Karagouniko and 0.67 ng/ml in Chios rams). Air temperature and sunshine followed the opposite pattern (P < 0.05). Karagouniko rams showed higher triiodothyronine concentrations than Chios rams, irrespective of the season. Our findings shed light on an important aspect of the thyroid gland function in Karagouniko and Chios sheep under temperate environmental conditions.

Group-training of rams at puberty for artificial vagina-mediated semen collection and its influence on semen quality and sexual behavior

There is a paucity of information with respect to group-training for artificial vagina and its influence on semen characteristics and sexual behavior of young untrained rams. A total of 18 healthy Najdi rams (with an initial body weight of 40-45 Kg and 7-8 month-old) were consequently used herein to test the usefulness of group-training for artificial vagina-mediated semen collection during the breeding season. Rams were randomly segregated into three groups (n = 6 rams per protocol), and the whole experiment was lasted for 10 weeks. The 1st group was subjected to a training protocol where one untrained ram was placed for 20 min with a teaser ewe, while the 2nd group were subjected to a protocol where one untrained ram was placed for 20 min with one trained ram and a teaser ewe, whereas the 3rd group were subjected to a protocol where three untrained rams were placed for 20 min with one trained ram and a teaser ewe. The obtained results clearly (P < 0.05) showed that training young rams in group has increased their sperm concentration and sexual stimulation, shortened the period of their training time, and descriptively had a complete training efficiency. The sexual stimulation of young untrained rams was intensified by the competition between rams in the co-presence of a trained ram. Collectively, these data may suggest that group-training of rams at puberty is a better protocol for AV-mediated semen collection compared to individual training. Some shortcomings were noted herein, but research dealing with this subject may very well improve the reproductive performance of young untrained rams.(AU)

Seasonal variation of thyroxine concentration in Karagouniko and Chios rams in temperate environmental conditions

It is well known that the concentration of the thyroid hormone thyroxine increases as day length increases, and the other way around, in Karagouniko and Chios ewes but based on openly international English literature, nothing is known about this hormone on the rams of the breeds mentioned above concerning a meticulous analysis of their aerial environment. So from this perspective, the current research was conducted to investigate the thyroxine concentrations of Karagouniko and Chios rams, taking into account two periods, January-June and July-November, when the day length was increasing and decreasing, respectively. Apart from photoperiod, the meteorological variables air temperature, relative humidity, and sunshine were taken into consideration in the farm of the Artificial Insemination Center of Karditsa (39021΄18΄΄N, 21054΄19΄΄E), Greece (temperate climate zone), where the experimental animals raised. The possible change of thyroxine concentration in both sheep breeds and the possible changes of the abovementioned meteorological variables between the examined periods were detected using t-tests. Higher thyroxine values (P < 0.05) were confirmed in JanuaryJune (55.11 ng/ml in Karagouniko and 47.72 ng/ml in Chios rams), in comparison to July-November (49.03 ng/ml in Karagouniko and 44.14 ng/ml in Chios rams). Air temperature and sunshine followed a reverse course (P < 0.05). In both periods, Karagouniko rams were characterized by higher concentrations of thyroxine than Chios rams (P < 0.01). Our results add more insight into the physiology of Karagouniko and Chios sheep related to thyroxine and temperate aerial environment.

Critérios e metodologias para produzir sêmen de caprinos e ovinos em nível de fazenda / The criteria and methodologies used to produce ram and goat semen on the farm

A criopreservação de sêmen é uma importante estratégia para o armazenamento de material genético de reprodutores ovinos e caprinos, considerados superiores do ponto de vista fenotípico, mas especialmente, pelos seus méritos genéticos. Hoje no Brasil apenas uma central regulamentada pelo MAPA está ema atividade, o que significa que existe pouca dose de sêmen comercial à disposição para programas de melhoramento genético. Programas de banco de sêmen podem ser realizado nas fazendas, entretanto, tem seu uso limitado a propriedade onde está o reprodutor. Neste caso, apesar de maiores limitações de estrutura e equipamentos para a execução do serviço, o domínio das variações da técnica de congelação de sêmen e o cuidado com as questões sanitárias dos reprodutores possibilitará ao Médico-Veterinário a execução do serviço com níveis adequados de segurança e qualidade das doses de sêmen produzidas. O objetivo deste estudo é apresentar e discutir critérios e metodologias para congelação de sêmen de pequenos ruminantes em diferentes condições, fruto de diversas experiências e trabalhos desenvolvidos por nosso grupo e outros, nos últimos vinte anos, e que tem possibilitado resultados satisfatórios.(AU

Sperm cryopreservation is an important strategy for the storage of genetic material from ovine and goat breeders, considered superior from the phenotypic point of view, but especially for their genetic merits. Today in Brazil only one center regulated by the Ministry of Agriculture, Livestock and Supply is in operation, which means that there is little dose of commercial semen available for genetic improvement programs. Semen bank programs can be carried out on farms; however, their use is limited to the property where the breeder is located. In this case, despite greater limitations of structure and equipment for the execution of the service, mastering variations in the semen freezing technique and taking care of the health issues of the breeders will enable the Veterinarian to perform the service with adequate levels of safety and quality of semen doses produced. This study aims to present and discuss criteria and methodologies for freezing semen from small ruminants under different conditions, the result of several experiences and works carried out by our group and others, in the last twenty years, which have enabled satisfactory results.(AU))

Paternal effects on fetal programming

Paternal programming is the concept that the environmental signals from the sire's experiences leading up to mating can alter semen and ultimately affect the phenotype of resulting offspring. Potential mechanisms carrying the paternal effects to offspring can be associated with epigenetic signatures (DNA methylation, histone modification and non-coding RNAs), oxidative stress, cytokines, and the seminal microbiome. Several opportunities exist for sperm/semen to be influenced during development; these opportunities are within the testicle, the epididymis, or accessory sex glands. Epigenetic signatures of sperm can be impacted during the pre-natal and pre-pubertal periods, during sexual maturity and with advancing sire age. Sperm are susceptible to alterations as dictated by their developmental stage at the time of the perturbation, and sperm and seminal plasma likely have both dependent and independent effects on offspring. Research using rodent models has revealed that many factors including over/under nutrition, dietary fat, protein, and ingredient composition (e.g., macro- or micronutrients), stress, exercise, and exposure to drugs, alcohol, and endocrine disruptors all elicit paternal programming responses that are evident in offspring phenotype. Research using livestock species has also revealed that sire age, fertility level, plane of nutrition, and heat stress can induce alterations in the epigenetic, oxidative stress, cytokine, and microbiome profiles of sperm and/or seminal plasma. In addition, recent findings in pigs, sheep, and cattle have indicated programming effects in blastocysts post-fertilization with some continuing into post-natal life of the offspring. Our research group is focused on understanding the effects of common management scenarios of plane of nutrition and growth rates in bulls and rams on mechanisms resulting in paternal programming and subsequent offspring outcomes. Understanding the implication of paternal programming is imperative as short-term feeding and management decisions have the potential to impact productivity and profitability of our herds for generations to come.(AU)

Reproductive behavior of Pantaneiros sheep, Mato Grosso do Sul, Brazil

The objective was to evaluate fertility, cyclicality behavior, and sexual performance of Pantaneiro sheep throughout the year's photoperiods in Mato Grosso do Sul - Brazil. Two experiments were conducted. In experiment one, the ewes used aged between 2 and 4 years, distributed in three homogeneous groups of 54 animals and subjected to reproductive evaluation in three different predefined luminosity periods, consisting of autumn (11h 45 min.), spring (12h 30 min.), and summer (12h 55 min.). The characteristics studied were body weight (BW), body condition score (BCC), estrus distribution (EST), percentage of pregnant females per cycle (%P), plasma progesterone (PP), and fertility (F) in the three predefined seasons. The mating system used was controlled natural nocturnal and lasted for 45 days. Experiment two, evaluated the ram sexual activity. The males (10) were individually exposed to four ovariectomized ewes. The ewes cycled during evaluated seasons, and the highest fertility was observed in animals with BCC >2.75. The PP concentration differed among periods, and higher conception rates (F) were observed in autumn and spring. The rams (75%) had a medium of sexual performance when they performed more than two ejaculations in 20 min throughout the seasons. Pantaneiro sheep of both genders had a similar fertility index, with early and constant activity throughout the year, displaying no photoperiodism or seasonality.

Post-thaw quality of ram sperm frozen with different concentrations of low-density lipoproteins associated with non-enzymatic antioxidants

The cryopreservation reduces ram sperm quality, decreasing the pregnancy rate of ewes inseminated with thawed sperm. Hence, we aimed to improve the post-thaw quality of ram sperm replacing egg yolk on Tris-Glucose extender with different concentrations of LDL (2 or 8%), associated with the addition of 10 mM non-enzymatic antioxidants (ascorbic acid, hydroxytoluene butylate, ascorbyl palmitate, and trehalose). Semen samples were collected from six rams, split into different treatments, and frozen. After thawing, kinematic (CASA), structural (propidium iodide and carboxyfluorescein diacetate) and functional (hypoosmotic test) sperm membrane integrity was assessed. Total motility, VCL, and LIN were also assessed in thawed samples during 3 h of incubation (38 °C). The results showed that hydroxytoluene butylate at 10 mM in Tris-Glucose extender with 8% LDL improved velocity parameters immediately post-thaw compared with Tris-Glucose egg yolk extender, as well as prevented the reduction of total motility and VCL after incubation. There was no benefit of adding ascorbic acid and trehalose. Moreover, for the first time, it was shown the motility impairment promoted by ascorbyl palmitate to ram sperm.(AU)

Addition of orange, pineapple and beet juices as extenders for cryopreservation of ram semen / Adição de sucos de laranja, abacaxi e beterraba em diluidor para criopreservação de sêmen de carneiros

Searching for improvements in semen cryopreservation, natural substances are commonly studied focusing to improve the sperm quality. The aim of this study were evaluated the effect of adding orange, pineapple, and beet juices in different concentrations and combinations to the ram semen cryopreservation extender. Five ejaculates from five adult rams were used. The semen pool was diluted in egg yolk-based extender and mixed with the following 15 treatments (at a final concentration of 400.106 sptz/mL): orange 10% (O10) and 15% (O15); pineapple 10% (P10) and 15% (P15); beet 10% (B10) and 15% (B15); pineapple + orange 10% (PO10) and 15% (PO15); pineapple + beet 10% (PB10) and 15% (PB15); beet + orange 10% (BO10) and 15% (BO15); pineapple + beet + orange 10% (PBO10) and 15% (PBO15); and the control group (CON). Post-thaw in 0.25 mL straws semen quality analysis of cryopreserved semen was performed by CASA and flow cytometry. Analysis of variance (PROC GLM) was carried out and the averages were compared using the SNK test. Pearson's correlation test was also performed. No effect was noted in the addition of juices to the semen extender prior to cryopreservation. Post-thawed, although, statistically similar to the control group, the total motility of the B10 group reached acceptable standards of total motility. In addition, B10 group showed the highest values (p< 0.05) of progressive motility than control group or other treatments. The addition of 10% beet juice to the ram semen extender can improve the cryopreservation of sperm motility.

Em busca de melhorias na criopreservação do sêmen, substâncias naturais são comumente estudadas com o objetivo de melhorar a qualidade do sêmen. O objetivo deste estudo foi avaliar o efeito da adição de sucos de laranja, abacaxi e beterraba em diferentes concentrações e combinações ao diluidor de criopreservação de sêmen ovino. Foram utilizados cinco ejaculados de cinco carneiros adultos. O pool de sêmen foi diluído em diluente à base de gema de ovo e misturado com os seguintes 15 tratamentos (na concentração final de 400x106 sptz/ml): laranja 10% (O10) e 15% (O15); abacaxi 10% (P10) e 15% (P15); beterraba 10% (B10) e 15% (B15); abacaxi + laranja 10% (PO10) e 15% (PO15); abacaxi + beterraba 10% (PB10) e 15% (PB15); beterraba + laranja 10% (BO10) e 15% (BO15); abacaxi + beterraba + laranja 10% (PBO10) e 15% (PBO15); e o grupo controle (CON). Pós-descongelação em palhetas de 0,25 ml a análise da qualidade do sêmen criopreservado foi realizada pelo CASA e citometria de fluxo. A análise de variância foi realizada e as médias comparadas pelo teste SNK. O teste de correlação de Pearson também foi realizado. Nenhum efeito foi observado na adição de sucos ao diluidor de sêmen antes da criopreservação. Após o descongelamento, embora estatisticamente semelhante ao grupo controle, a motilidade total do grupo B10 atingiu padrões aceitáveis de motilidade total. Além disso, o grupo B10 apresentou os maiores valores (p<0,05) de motilidade progressiva que o grupo controle ou os outros tratamentos. A adição de 10% de suco de beterraba ao diluente de sêmen ovino pode melhorar a criopreservação da motilidade espermática.

Polymorphisms of the melatonin receptor 1A (MTNR1A) gene affect the sexual performance of Rasa Aragonesa rams without changing their social dominance

This study aimed to investigate whether a specific genotype of the MTNR1A gene influences social dominance and reproductive performance in rams. Thirty-one rams were genotyped for Rsal and Mnll polymorphism of the gene: CC (n=19), CT (n=5), and TT (n=7), and GG (n=20), GA (n=6), or AA (n=5), respectively. Maintenance (standing still, lying down, drinking, feeding, walking, or stereotyping) and social behavior (agonistic and antagonistic interactions among rams) were recorded by direct observations involving instantaneous scan sampling and continuous behavior sampling. For each animal, his Index of Success (IS) and Displacement (ID) were calculated. One week after the social-behavioral observations, rams were tested in individual serving-capacity tests involving three ewes that were synchronized into estrus in a 15-m2 pen for 20 min. The genotypes did not differ in the number of aggressions they performed (CC: 88.95±12.85; CT: 106.20±21.00; TT: 70.43±8.50; GG: 76.05±8.72; GA: 119.00±16.16; AA: 95.80±36.39) or received (CC:75.79±5.82; CT:92.40±8.68; TT: 86.14±15.05; GG: 79.25±6.58; GA: 86.83±11.51; AA: 79.80±12.74), and their active and passive behaviors were similar. The genotypes did not differ significantly in IS or ID, and the proportion of rams in each success category was similar among groups. For the Rsal and Mnll polymorphism, TT and GG rams were responsible for 54% and 56% of recorded sexual events (P < 0.05), respectively. Results of this study confirm that the best sexual performance of rams was among those that carried certain genotypes of the MTNR1A gene. Still, it was not correlated with differences in social dominance.

DNA methylation level of gene SIRT1 in ram spermatozoa and relationship with fertilizing ability according to breed and age

Background: Effect of the epigenetic factors on the male fertility is well proofed. Sperm acts as a carrier of genetic material, and its DNA methylome can affect maternal pregnancy rate and offspring phenotype. However, the research on the DNA methylation in the spermatozoids of livestock males, in particular rams, is still limited. To best of our knowledge the data about as a global as well as gene specific DNA methylation in ram spermatozoa from different breeds and ages are missed in the scientific literature. The present study was designed to analyze the relationship between methylation levels of the important for spermatogenesis gene SIRT1 in spermatozoa and fertilizing ability of sperm in rams from different breeds and ages. Materials, Methods & Results: The ejaculates of 16 rams from Lacaune, East Friesian and Assaf breeds at age between 18 to 96 months were evaluated. The kinematic parameters of 2 semen samples from each animal were estimated by CASA. The separated spermatozoa were used for DNA extraction followed by bisulfite conversion. The DNA methylation of SIRT1 was detected through quantitative methylation-specific PCR using 2 sets of primers designed specifically for bisulfite-converted DNA sequences to attach methylated and unmethylated sites. The breed and age effect on the gene SIRT1 methylation by ANOVA was estimated. Experimental females included 393 clinically healthy milk ewes (Lacaune, n = 131; East Friesian sheep, n = 100 and Assaf, n = 162) in breeding season. Reproductive performances (conception rate at lambing, lambing percentage and fecundity) of ewes, inseminated by sperm of the investigated rams, were statistically processed. ANOVA showed that the animal breed influences significantly on the level of DNA methylation of gene SIRT1 in ram spermatozoa (P = 0.002) An average value of DNA methylation of SIRT1 in ram sperm from Lacaune breed was significantly higher than in Assaf and East Friesian (81.21 ± 15.1% vs 36.7 ± 14.2% and 38.3 ± 18.6 respectively, P < 0.01). The highest percent of SIRT1 methylation was observed in old animals compared to the young and middle-age. Moderate and strong correlations (r from 0.44 to 0.71, P < 0.05) between the methylation level of the SIRT1 gene in rams' sperm and reproductive parameters of inseminated ewes in all breeds were established. Discussion: Our data are the first message about the effect of breed on the specificity of DNA methylation of gene SIRT1 in ram spermatozoa. These results demonstrated an existence of the sheep breeds with high and low level of DNA methylation of gene SIRT1 in ram sperm. Although the effect of age on the methylation level in sperm is still discussable, our results showed a moderate correlation between age and methylation level of SIRT1 in spermatozoa of rams. Taking into account that DNA methylation in sperm is stabilized with puberty onset and is a heritable epigenetic modification, it can be a promising marker of sperm quality in animal breeding. In all investigated breeds the rams with relatively high level of DNA methylation of gene SIRT1 in spermatozoa (50-68%) demonstrated a high conception rate at lambing (> 70%). In conclusion, the DNA methylation level of the SIRT1 gene in ram spermatozoa is determined by both the breed and the age of the animals and correlates with fertilizing ability of sperm.

Three superovulation protocols for in vivo embryo production in Santa Inês sheep / Três protocolos para produção in vivo de embriões em ovelhas Santa Inês

There is still no consensus regarding the best protocol for in vivo embryo production in sheep despite increasing studies in this area. Moreover, there is variability in the response of ewes to superovulation (SOV). An approach to mitigate this inconsistency is to initiate gonadotropin administration under favorable ovarian conditions. The present study compared three treatments in a crossover design: a traditional SOV protocol (TRAD) and "Day 0" D0 SOV protocol with (D0+GnRH), or without Lecilerin (D0-GnRH). Fifteen Santa Inês ewes received 200 mg of FSH at six decreasing doses and PGF2α with the fifth dose of FSH. They were naturally mated with fertile rams and subjected to surgical embryo collection. The number of viable embryos was similar among the different treatments (TRAD = 6.0 ± 4.7; D0-GnRH = 3.8 ± 6.4; D0+GnRH = 7.5 ± 6.5). Regardless of the treatment method, ewes with follicles ≤ 4 mm, at the first FSH dose, produced more viable embryos (9.6 ± 6.0, P < 0.05) compared to ewes that had follicles > 4 mm at the beginning of the SOV (2.9 ± 3.1, viable embryos). Both the TRAD and D0+GnRH groups had fewer animals with large follicles (> 4 mm) at the first FSH dose than the D0-GnRH group (P < 0.05). In conclusion, both the TRAD and D0+GnRH treatments induced a more favorable ovarian condition (follicles ≤ 4 mm) for adequate SOV; although, all three treatments exhibited similar efficacies in Santa Inês sheep.

Ainda não há consenso sobre qual é o protocolo mais apropriado para a produção in vivo de embriões em ovinos, apesar do crescente conhecimento. Uma abordagem para mitigar a variabilidade de resposta de ovelhas à superovulação (SOV) é iniciar a aplicação de gonadotrofinas em uma condição ovariana favorável. O presente estudo comparou três tratamentos em delineamento do tipo crossover: protocolo de SOV tradicional (TRAD) e "Dia 0" D0 SOV sem (D0-GnRH) ou com GnRH (D0+GnRH). Quinze ovelhas Santa Inês foram superovuladas com 200 mg de FSH em seis doses decrescentes e receberam PGF2α na quinta dose de FSH. As ovelhas foram submetidas a monta natural com carneiros férteis e os embriões colhidos por via cirúrgica. O número de embriões viáveis não diferiu entre os tratamentos (TRAD = 6,0 ± 4,7; D0-GnRH = 3,8 ± 6,4; D0+GnRH = 7,5 ± 6,5). Independentemente do tratamento, ovelhas com folículos ≤ 4 mm na primeira dose de FSH produziram mais embriões viáveis (9,6 ± 6,0; P < 0.05) quando comparadas aos animais que apresentavam folículos > 4 mm no início da SOV (2,9 ± 3,1 embriões viáveis). Os grupos TRAD e D0+GnRH apresentaram menor número de animais com folículos grandes (> 4 mm), no momento da primeira dose de FSH, quando comparados ao grupo D0-GnRH (P < 0,05). Em conclusão, os protocolos TRAD e D0+GnRH induziram uma condição ovariana mais favorável (folículos ≤ 4 mm) para a SOV. No entanto, os três tratamentos apresentaram eficiência semelhante em ovelhas Santa Inês.

Effect of seasonally changing aerial environment on testosterone in Karagouniko and Chios sheep

A well-established activity in Greece and other Mediterranean countries is sheep farming. Karagouniko and Chios sheep are promising breeds and targets for the reproductive physiologist's attention. Under this context, part of the reproductive physiology of Karagouniko and Chios was investigated, taking into account the environment. Specifically, four time periods were considered; April-June (Long days, increasing photoperiod length), July-September (Long days, decreasing photoperiod length), October-December (Short days, decreasing photoperiod length), and January-March (Short days, increasing photoperiod length). The effect of the period on the concentration of the testosterone (main male hormone) in rams of the breeds above was studied, by using repeated measures analysis of variance, also taking into account the environmental variables air temperature, relative humidity, sunshine, rainfall, and photoperiod, in the Artificial Insemination Center of Karditsa (39°21'18''N, 21°54'19''E), Greece. The lowering of photoperiod from April-June to July-September (long days) and afterward, even more to October-December (short days) coincided with a progressive increase of testosterone in both breeds. Photoperiod seems to have a major role in the concentration of Karagouniko and Chios breeds' testosterone compared to the other environmental variables. When considering the whole experimental period, Chios sheep had significantly higher (P < 0.05) testosterone (1.95 ng/ml) than the Karagouniko one (1.55 ng/ml). Our findings could contribute towards the more rational exploitation of the examined sheep breeds.(AU)

Novel missense variant L46Q of fatty acid synthase gene and fatty acids content in Awassi sheep

This study was conducted to investigate the association between the polymorphism of the FASN gene with fatty acid content in Awassi sheep. A total of 100 male Awassi sheep between the ages of one and two and a half years old were used in this study. Phenotypic measurement was recorded at slaughter, and from each animal, the longissimus dorsi (LD) muscle samples were taken to analyze the fatty acid profile. Genotyping, sequencing reactions, and in silico tools were performed to confirm the variants in amplified fragments. The result of genotyping revealed two genotypes (AA and AB) of the ovine FASN gene (exon 3). Novel SNP (L46Q) was discovered only within the FASN gene (AB genotype). All utilized in silico tools revealed remarkably deleterious effects for the L46Q on the mutant protein structure, function, and stability. Association analysis revealed that the AB genotype has significantly (p < 0.05) higher levels of animal length and monounsaturated fatty acids (MUFA) with lower amounts of saturated fatty acids (SFA) content than the AA genotype. In conclusion, novel SNP (L46Q) was discovered within the FASN gene (AB genotype), made the animals that has the AB genotype associated with good meat quality traits and this polymorphism may serve as markers for meat quality.(AU)

Meat quality of the Berrichon du Cher lambs and its cross-breeds with a native breed / Qualidade da carne dos cordeiros Berrichon du Cher e seus cruzamentos com raça nativa

This study evaluated the meat of lambs from crossbreeding of Polish lowland sheep, heather and rams of the Berrichone du Cher breed (B). The lambs were kept in a medium-intensive feeding system. During fattening, they received haylage and grass hay with the addition of clover, and a mixture of cereal grains with the addition of minerals and vitamins. During the fattening period, all feeds were given at will. The slaughter and cutting of the carcasses were performed in accordance with the technology applicable in the meat industry. In the longissimus lumborum muscle, the content of dry matter, total protein, total fat and ash was determined. In addition, the pH value of 45 minutes and 24 hours after slaughter, as well as the parameters of colour and tenderness of the meat were measured. The content of collagen protein, cholesterol and selected fatty acids was also determined. As a result of the conducted analyzes, the influence of the Berrichone du Cher breed on the improvement of quantitative parameters related to the slaughter value and some parameters related to the physicochemical composition of the hybrid meat was reported.

O objetivo deste estudo foi avaliar como o cruzamento da raça Berrichon du Cher com raças nativas de ovelhas polonesas de baixa altitude e ovelhas de charneca polonesa afeta os parâmetros qualitativos da carne .O maior teor de proteína (19,86 ± 0,36%, P <0,05) em cordeiros foi encontrado no grupo Berrichon du Cher e no grupo de cordeiros ovinos (19,23 ± 0,67%, P <0,05). Por outro lado, o maior teor de colágeno total (1,10 ± 0,21%) e a maior sensibilidade do tecido muscular (10,69 ± 0,74 N cm-2), que também apresentaram o menor diâmetro das fibras musculares (24,01 ± 4,62 µm, P < 0,01) foi encontrado nos cordeiros mestiços Berrichon du Cher e Polish Heath Sheep. Tanto a raça quanto o gênero afetaram significativamente o teor de colesterol na carne de cordeiro e o maior teor de colesterol foi encontrado na carne dos cordeiros Berrichon du Cher. Seu menor teor foi demonstrado na carne de Ovinos da Baixada Polonesa com a diferença de 9,1 mg 100g-1 de tecido muscular. Um dos níveis mais baixos do isômero CLA (C 18: 2 cis-9 trans-11) também foi encontrado no grupo dos cordeiros Berrichon du Cher. Este isômero evita a formação de colesterol. Dependência semelhante em relação ao gênero também foi observada. Os resultados obtidos indicam que as raças nativas são utilizadas em cruzamentos com a raça Berrichon du Cher destinada à engorda, principalmente para melhorar parâmetros quantitativos relacionados ao valor de abate e parâmetros selecionados de propriedades físicas e químicas da carne ovina.

Frontiers in sheep reproduction - making use of natural responses to environmental challenges to manage productivity

This review addresses advances, directions and opportunities for research on sheep reproduction in the context of the global challenges of food security and climate change, and demand for 'clean, green and ethical' (CGE) animal management. The foundation of CGE management is an understanding of the physiological processes through which the reproductive system responds to changes in the animal's environment. These days, to the main environmental factors (photoperiod, nutrition, pheromones), we need to add stress from extreme weather events. With respect to nutrition in rams, we now have a deeper understanding of the responses of the brain centres that control gonadotrophin secretion (the kisspeptin system). At testis level, we have found that nutrition affects non-coding RNAs in Sertoli cells and germ cells, thus affecting the balance between cell proliferation and apoptosis. This proliferation-apoptosis balance is also affected during prenatal development, when undernutrition or stress in pregnant ewes seems to elicit epigenetic changes in developing gonads that could affect offspring fertility in adult life. With respect to nutrition in ewes, metabolic signals act directly on ovarian follicles, and thus change ovulation rate, but the variety of signals now includes the adipokines. An early concern was that nutritional supplements that increase ovulation rate would also increase embryo mortality but we now know that embryo survival is improved under field conditions. Finally, we had always thought that the efficiency gains from early puberty in lambs could only be achieved by accelerating fat accumulation, but we now know that faster muscle growth will achieve the same goal, offering two advantages in meat production systems. With respect to pheromones ('ram effect'), we have a deeper understanding of the brain responses (kisspeptin system) but, most importantly, a realization that the response of ewes to the ram signal involves cell division in memory centres. Many opportunities remain.(AU)

Effect of seasonally changing aerial environment on testosterone in Karagouniko and Chios sheep

A well-established activity in Greece and other Mediterranean countries is sheep farming. Karagouniko and Chios sheep are promising breeds and targets for the reproductive physiologist's attention. Under this context, part of the reproductive physiology of Karagouniko and Chios was investigated, taking into account the environment. Specifically, four time periods were considered; April-June (Long days, increasing photoperiod length), July-September (Long days, decreasing photoperiod length), October-December (Short days, decreasing photoperiod length), and January-March (Short days, increasing photoperiod length). The effect of the period on the concentration of the testosterone (main male hormone) in rams of the breeds above was studied, by using repeated measures analysis of variance, also taking into account the environmental variables air temperature, relative humidity, sunshine, rainfall, and photoperiod, in the Artificial Insemination Center of Karditsa (39°21’18’’N, 21054’19’’E), Greece. The lowering of photoperiod from April-June to July-September (long days) and afterward, even more to October-December (short days) coincided with a progressive increase of testosterone in both breeds. Photoperiod seems to have a major role in the concentration of Karagouniko and Chios breeds' testosterone compared to the other environmental variables. When considering the whole experimental period, Chios sheep had significantly higher (P < 0.05) testosterone (1.95 ng/ml) than the Karagouniko one (1.55 ng/ml). Our findings could contribute towards the more rational exploitation of the examined sheep breeds.

Effect of quercetin or butylated hydroxytoluene on cooled or frozen-thawed ram sperm quality / Efeito da inclusão de quercetina ou butil-hidroxitolueno na qualidade do sêmen de carneiro resfriado ou congelado

Cooling and freezing processes cause physical and chemical damage to sperm by cold shock and oxidative stress. This study aimed to evaluate the effect of two antioxidants on sperm parameters of cooled and frozen-thawed ram semen diluted in an egg yolk-based extender. Semen was collected from 30 rams and processed in two consecutive experiments to test the inclusion of different concentrations of quercetin and butylated hydroxytoluene (BHT) in an egg yolk-based semen extender. Dimethyl sulfoxide (DMSO) was added as a solvent to the semen extender in a ratio of 1 mL DMSO for 90 mg of quercetin and 1 mL DMSO for 880 mg of BHT. After collection, semen was diluted at 200 × 106 motile sperm/mL (control) and split into different groups in each experiment. In experiment 1, semen was diluted with the extender containing quercetin (Q5, 5 µg/mL; Q10, 10 µg/mL; Q15, 15 µg/mL) or DMSO alone (DMSO1, 0.055 µL DMSO per mL; DMSO2, 0.165 µL DMSO per mL). In experiment 2, semen was diluted with the extender with BHT (BHT1, 0.5 µg/mL; BHT2, 1 µg/mL; BHT3, 1.5 µg/mL) or DMSO alone (DMSO3, 0.375 µL DMSO per mL; DMSO4, 1.125 µL DMSO per mL). After dilution, the semen was divided into two aliquots. Treated ram sperm samples were also subjected to different storage methods. The first set of samples was cooled at 5 °C for 24 h, whereas the second set of samples was frozen-thawed. Sperm motility parameters and plasma membrane integrity (PMI) were evaluated immediately after dilution (0h) and 24 h after cooling and in the frozen-thawed samples via computer-assisted sperm analysis and epifluorescence microscopy, respectively. The inclusion of quercetin or BHT did not affect sperm motility parameters or PMI of fresh, cooled, or frozen-thawed sperm in this study (P < 0.05). However, further studies are needed to test the effects of these antioxidants on the fertility of cryopreserved ram semen.(AU)

O resfriamento e o congelamento causam danos físicos e químicos aos espermatozoides por choque térmico e estresse oxidativo. Portanto, este estudo teve como objetivo avaliar o efeito da inclusão de dois antioxidantes em um diluente à base de gema de ovo sobre os parâmetros espermáticos do sêmen ovino resfriado e congelado. Trinta carneiros tiveram o sêmen coletado e processado em dois experimentos consecutivos para testar a inclusão de diferentes concentrações de quercetina e hidroxitolueno butilado (BHT) em diluente de sêmen à base de gema de ovo. O DMSO foi adicionado como solvente ao diluente de sêmen em uma proporção de 1 mL de DMSO parra 90 mg de quercetina e 1 Ml de DMSO para 880 mg de BHT. Após a coleta, o sêmen foi diluído a 200 × 106 espermatozoides móveis/mL (Controle) e dividido em diferentes grupos em cada experimento. Experimento 1, Quercetina (Q5, 5 µg / mL; Q10, 10 µg / mL; Q15, 15 µg / mL) ou DMSO (DMSO1, 0,055 µL de DMSO por ml; DMSO2, 0,165 µL de DMSO / mL) foram adicionados ao extensor. Experimento 2, BHT (BHT1, 0,5 µg / mL; BHT2, 1 µg / mL; BHT3, 1,5 µg / mL) ou DMSO (DMSO3, 0,375 µL de DMSO por ml; DMSO4, 1,125 µL de DMSO / mL) foram adicionados à o extensor. Após a diluição, o sêmen foi dividido em duas alíquotas. O primeiro foi resfriado a 5 ° C por 24h, enquanto o segundo foi congelado. Os parâmetros de motilidade espermática e integridade da membrana plasmática (PMI) foram avaliados, imediatamente após a diluição (0h) e 24h após o resfriamento e nas amostras congeladas, pelo CASA e microscopia de epifluorescência, respectivamente. A inclusão de quercetina ou BHT não afetou os parâmetros de motilidade espermática e PMI de espermatozoides frescos, resfriados ou congelados (P < 0,05). Portanto, a inclusão de quercetina e BHT não beneficiou os parâmetros espermáticos do sêmen ovino submetido a armazenamento líquido a 5 ° C por 24h ou protocolo de congelamento no presente estudo. No entanto, mais estudos são necessários para testar o efeito desses antioxidantes na fertilidade do sêmen ovino criopreservado.(AU)

Effect of 20 mg fluorogestone acetate (FGA) sponges on reproductive parameters of dairy ovine breeds in Southern Italy

Background: Intravaginal devices containing progestins are widely used for oestrus synchronization in sheep. Progestins give economic benefits to farmers but may have some limitations and the efficacy strictly depends on farm management. There are different devices, with different molecules (progesterone, medroxyprogesterone acetate, fluorogestone acetate), different dosages and long (12-14 days) and short-term (5-7 days) protocols. Experimental studies often include a limited number of animals and are held at different latitudes and with different system of management. To our knowledge, there are few reports in the literature on field application of the recently licensed 20 mg fluorogestone acetate (FGA) sponges in large ewe flocks, excluding the registration trials. Materials, Methods & Results: This study was designed to evaluate oestrous synchronization in 2 different breeds. A total of 1100 Lacaune (L) and 618 Sarda (S) were assigned to 8 groups, comparing multiparous (M) and nulliparous (N) and those synchronized in November (n) and May (m). The groups were: LMm (n. 556), LNm (n. 180), SMm (n. 70), SNm (n. 32), LMn (n. 242), LNn (n. 222), SMn (n. 440) e SNn (n. 76). The intravaginal sponge was inserted for 14 days. At sponge withdrawal, eCG (400 IU, IM) was injected, and rams were joined into flocks 30 h later with a male/female ratio of 1:8. Transrectal ultrasonography was performed for pregnancy diagnosis after 30 days. Fertility parameters such as oestrus (OR), pregnancy (PR), lambing rates (LR) and prolificacy were calculated. Fisher exact test was used to compare parameters of each group with the corresponding and significance was set at P < 0.01. The use of intravaginal sponges in this study was easily performed in all animals. In nulliparous groups, digital insertion of the sponge, instead of the applicator, was preferred to avoid discomfort. Sponge loss was observed in 2% of treated ewes. At the removal of the sponge, mild vaginal contamination was observed in 90% of the ewes. Ultrasound finding of embryo resorption, pseudopregnancy and pyometra were found in rates under 2%. The loss of pregnancy ranged from 0 to 3.6% without significant differences among groups. The SMn group showed the best performances (0.91 OR, 0.88 PR, 0.85 LR), significantly higher than the other groups. Prolificacy was maximum in the LMn (1.68) and minimum in the SNm group (1.06), without significant differences among groups. Discussion: The 14-day regimen based on 20 mg FGA-releasing intravaginal devices is an easy and satisfactory synchronization regimen to improve the productivity and the fertility of sheep farm, and this can be managed and optimized in different breeding conditions. In this study, Sarda breed shows a good adaption to the environment with adult ewes giving the best results in reproductive season. However young Sarda ewes were less productive especially in non-breading season. Lacaune showed good reproductive potential, young and adult ewes responded to progestins in reproductive and non-reproductive season. It was confirmed that the reproductive performance is affected by season, age, and breed. This study encourages the breeding of indigenous breeds by implementing the management with modern technologies. The imported highly productive breeds, if well managed, can give a production less influenced by seasonal variables and age.

Congenital cataract in a Blackbelly lamb

Background: A cataract is an opacity of the crystalline structure that results in impaired vision. The congenital form manifests itself at birth or shortly thereafter and might also be inherited and therefore capable of passing on to descendants. Cataracts can be caused by systemic diseases, medications, toxic substances, radiation, metabolic alterations, dietary deficiencies, inflammation, traumatic injuries, age, or genetic factors. The few Blackbelly sheep herds are located in the northeast and north regions of Brazil and are considered rare, which could result in high levels of consanguinity. In this context, we report a case of congenital cataract in a Blackbelly lamb and its possible etiology. Case: A 3-month-old lamb presented with ophthalmic alterations since birth, with white and cloudy spots in both eyes and impaired vision. In the same herd, 3 elderly sheep showed similar ophthalmic alterations. The lamb was able to follow its dam, but when walking, bumped into small objects or very close to his vision field. The lamb managed to follow the herd and dodge large objects, suggesting partial vision loss. During a physical examination, both lens showed opacity and reduced corneal reflex, pupillary reflex to direct light, pupillary reflex to consensual light, and threat reflex. Ultrasonographic examination revealed that both lens presented hyperechogenicity. Hematological values were within the reference limits. In the same herd, three elderly sheep presented bilateral cataracts (2 rams and 1 ewe) in previous years, which at that time was attributed to natural aging. One ram was the lamb's grandfather. The other ram was the father of the female, both with cataracts. Based on history, physical examination, and complementary examinations, the lamb was diagnosed with bilateral congenital cataracts with a probable hereditary condition. Discussion: Multiple factors can be related to the etiology of cataracts, and it can be difficult to establish the correct etiology. Regarding the age of onset, cataracts can be classified mainly as congenital and senile. Senile cataract is a bilateral opacification process that involves the entire lens, with slow progression and gradual loss of vision with increasing age. In adult sheep, the high proportion of eyes affected by spontaneously arising cataracts could be related to age, increased exposure to sunlight, increased genetic susceptibility, or a combination of these factors. In this case, the herd had three adult elderly sheep with cataracts previously characterized as senile. However, after reviewing the genealogy, it was found that all animals had some degree of parentage, suggesting a hereditary factor. Congenital cataracts are expressed soon after birth, resulting from the malformation of fibers in the lens, and are generally nonprogressive. The congenital form may or may not be associated with hereditary factors. Inheritance cataracts have been reported in several breeds of dogs and usually occur as an autosomal recessive trait. Blackbelly sheep are rare in Brazil, favoring consanguinity, so we believe that cataracts are inherited in this herd. To control this ophthalmic alteration, all animals with crystalline opacities were excluded from reproduction, and the herd should be monitored in future cases.

Sperm cell capacitation status of ram semen after cooling

Background: The use of conventional artificial insemination (AI) in sheep production is usually associated with lower fertility rates when frozen semen is used. Cooled ram semen has been an alternative over frozen semen due to the higher viability, seminal quality and fertility rates following AI. The semen preservation process promotes sperm cell modifications similar to capacitation (capacitation-like) that causes cell damage affecting viability and seminal quality, but such effects are unclear for cooled semen. The aim of this study was to determine the status of sperm cell capacitation (CA) and acrosome reaction (AR) during ram semen processing and cooling under different extenders, dilution factors, and aerobiosis conditions as a function of storage time at 5o C. Materials, Methods & Results: Two consecutive ejaculates per day per male were collected from 2 adult rams by artificial vagina at 48-72 h intervals, in three replications. After macro- and microscopic evaluations, semen was segregated into groups under 3 extenders (Tris-egg yolk or TY, citrate-egg yolk or CY, skimmed milk or SM), 2 dilution factors (1 x 109 or Bi, 100 x 106 or Mi cells/mL), and 2 aerobiosis conditions (aerobic or A, semi-anaerobic or SA). Diluted semen was cooled to 5ºC and stored for up to 72 h, with evaluations every 24 h. Aliquots of fresh ejaculates and of each cooled diluted subgroup, according to extender, dilution, and aerobiosis, were collected at times T0 and T72 for determination of acrosome status and membrane integrity by the chlortetracycline (CTC) and trypan blue-Giemsa stainings, respectively. No differences were detected in sperm cell motility (M) and motility vigor (V) between fresh and diluted semen. After cooling, a significant decrease in M was observed after 48 h in CY and SM compared with fresh semen and 0 h of cooling, while V started to decrease after 24 h in CY compared with TY. Likewise, M/V from different dilutions and aerobic conditions decreased more significantly after 48 and 24 h of cooling, respectively. The sperm capacitation status did not show differences in the proportion of non-capacitated (NCA), CA and AR sperm cells between TY, CY, and SM extenders (NCA: 75.0%, 71.3%, 74.0%; CA: 15.7%, 17.2%, 15.9%; AR: 9.3%, 11.5%, 10.2%) or between Bi and Mi dilutions (NCA: 74.0%, 72.9%; CA: 15.9%, 16.6%; AR: 10.1%, 10.5%), respectively. However, differences (P < 0.05) were observed between A and SA aerobic conditions, with CA (17.0% vs. 15.5%) and AR (11.9% vs. 8.7%) rates being higher in A than SA, respectively, with no differences in NCA (71.1% vs. 75.8%), irrespective of the storage time. Sperm cell viability decreased after 48 h, especially in CY (P < 0.05). Discussion: Ram sperm cells can suffer irreversible damage due to thermal shock during cooling. Egg yolk-based extenders provide phospholipids and cholesterol to protect the sperm cell membrane during the thermal shock caused by the change in temperature. In this study, sperm cells had irreversible decreases in M/V, with increase in acrosome and plasma membrane damage after cooling to 5ºC. The largest and smallest decreases in M and V over time were observed in the CY and TY extenders, respectively. In addition to the extender type, the semen preservation method and storage time promoted changes in the capacitation status, AR and in sperm cell viability, which per se were associated with a decrease in semen fertility. In fact, the proportions of CA and/or AR sperm cells gradually increased over time after dilution and storage at 5ºC, with a negative correlation between sperm cell viability and M/V over time. In summary, extender and cooling time affected mostly M/V, while aerobiosis condition and dilution factor were more associated with acrosome status and sperm survival, with the extender having less impact on the acrosome status as a function of time.