Systematic review of hormonal strategies to improve fertility in rams.

Reviewing the current state of knowledge on reproductive performance and productive traits in rams has many advantages. First, the compilation of this information will serve as a literature resource for scientists conducting research around the world and will contribute to the understanding of the data collected and interpreted by researchers on the different hormonal strategies used to improve reproductive performance in rams. Second, it will allow scientists to identify current knowledge gaps and set future research priorities in ram reproduction. Rams play an important role in the global flock economy, but their reproductive analysis has been limited in the use of hormonal technologies to increase the productivity of sheep flocks. In this review, we cite the most important works on six hormones that, in one way or another, modify the hypothalamus-pituitary-gonadal axis, at different doses, in and out of the reproductive season, breeds, application methods, among other factors. The overall aim is to increase the reproductive efficiency of rams in different scenarios and, in some cases, of other species due to the lack of limited information on rams.

Rams' reproductive status during a controlled breeding period: effect of social dominance.

The aim of this study was to compare the variation in body weight, scrotal circumference (SC), and seminal quality during a breeding period in dominant and subordinate rams. Data were collected from 12 dyads of rams, each bred with 15 ewes for 7 weeks. Before joining, the dominance relationship between both rams from each dyad was determined. Body weight and SC were recorded weekly in the morning, and semen was collected by electroejaculation, determining the volume, sperm concentration, mass motility, and percentage of sperm with progressive motility. In addition, the total number of sperm and sperm with progressive motility ejaculated were calculated. Dominance had no direct effect or interaction with time on any of the variables analyzed. Body weight, seminal volume, sperm concentration, mass motility, percentage of sperm with progressive motility, and total ejaculated sperm varied with time (p < 0.05), and scrotal circumference and total ejaculated sperm with progressive motility tended to vary with time. In general, all indicators evaluated were affected in the first weeks, when most ewes were cycling, recovering as the breeding progressed. It was concluded that, at least under the conditions of this study, dominance position did not affect the profile of the reproductive variables evaluated, although all of them were affected during the breeding period.

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))

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)

Técnicas reversíveis de rufião em ovinos (Ovis aires) / Reversible techniques of ruffian in sheep (Ovis aires)

Background: The use of teaser rams is an essential practice for detecting estrus in ewes as well as for accelerating puberty and synchronizing ovulation in the animal. There are several methods for preparing teasers, and the method used should be based on an assessment of the producer’s requirements. The ideal technique should be low cost and safe, ensuring the non-fertilization of ewes. This study evaluated the feasibility and effectiveness of two reversible teaser preparation techniques using a reversible plastic clamp. The techniques were compared in terms of functionality, possible post-surgical complications, and hematological changes of the rams as well as durability and reversibility. Materials, Methods & Results: Twelve healthy rams, aged 14-20 months, were divided into two groups (G1 and G2). Blood samples were collected through the jugular vein to perform the following analyses: blood count, total plasma protein, and fibrinogen. Following local infiltrative anesthesia with 5.0 mL 2% lidocaine without vasoconstrictor, the procedure was performed as follows: in G1, the preputial ostium was partially closed, and in G2, sigmoid flexure was performed in the cranial region, approximately 5-8 cm immediately caudal to the scrotal sac. In the postoperative period, 20 mg/kg oxytetracycline and 2.2 mg/kg flunixinmeglumine were intramuscularly administered as a single dose. The wounds were dressed, sprayed with repellent, and allowed to heal for seven days. The procedures in both groups were simple to perform, low cost, and low risk; caused minimal tissue injury; enabled rapid recovery; promoted little or no stress to the animals; are reversible; and left no complications. The animals of both groups satisfactorily identified the females in estrus during the three-month experimental period, maintained libido, and failed to mate with any female. The blood count levels...

Caracterización morfológica y molecular de Oxalis tuberosa Mol. en el departamento de Boyacá / Morphological and molecular characterization of Oxalis tuberosa Mol. in the department of Boyacá

RESUMEN Dentro de los tubérculos andinos de mayor importancia, después de la papa, se encuentra la ibia (Oxalis tuberosa Mol.) ya que constituye un alimento básico para las comunidades campesinas. Boyacá es uno de los departamentos de Colombia, en donde todavía existe la tradición de cultivar y consumir tubérculos como los cubios, ullucus y rubas o ibias, sin embargo están amenazados por la erosión genética. No existen estudios sobre estos recursos fitogenéticos en Boyacá, por lo cual el objetivo de esta investigación fue colectar y caracterizar morfológica y molecularmente materiales de ibias en este departamento. El análisis morfológico mostró que las características más discriminantes fueron: color de los tallos aéreos, pigmentación de las axilas, color del follaje, color de la flor, color del pedúnculo y pedicelo, color predominante de la superficie del tubérculo, color secundario de la superficie del tubérculo, color predominante y secundario de la pulpa, distribución del color secundario de la pulpa y forma de los tubérculos. El análisis de similitud diferenció a los materiales en dos grandes grupos, de acuerdo principalmente a características morfológicas como el color y hábito de floración. El valor de heterocigosidad promedio para la población total fue de 0.39 el cual se considera alto al compararse con otros estudios de diversidad genética en ibias. El análisis de varianza molecular y el Fst (coeficiente de diferenciación genética) muestran que existe una alta variabilidad genética entre los materiales de ibias evaluados la cual debe ser conservada y aprovechada dentro de futuros programas de mejoramiento.

ABSTRACT Among the most important Andean tubers, after potato, is the ibia (Oxalis tuberosa Mol.) because it is a staple food for peasant communities. Boyacá in one of the departments of Colombia, where there is still tradition to grow and consume tubers like cubios, ullucus and rubas or ibias; however, they are threatened by genetic erosion. There are no studies on these plant genetic resources in Boyacá, therefore the goal of this research was to collect and morphological and molecularly characterize ibias in this department. Morphological analysis showed that the most discriminate characteristic were color aerial stems, pigmentation armpits, foliage color, flower color, color peduncle and pedicel, predominant color of the tuber surface, secondary color of the surface tuber, dominant and secondary color of the pulp, distribution of secondary pulp color and shape of tubers. The similarity analysis discriminate the materials in two groups according mainly to morphological characteristics such as color and flowering habit. The value of average heterozygosity for the total population was 0.39, which is considered high when compared with other studies of genetic diversity in ibias. The analysis of molecular variance and Fst (coefficient of genetic differentiation) show that existed a high genetic variability among the ibias evaluated which it should be maintained and exploited in future breeding programs of Andean tubers.

Effect of γ-oryzanol on testicular degeneration induced by scrotal insulation in rams.

The present study assessed the effects of daily supplementation with 33 mg/metabolic weight (MW) of γ-oryzanol on testicular degeneration induced by scrotal insulation in rams. Eight animals were divided into two groups: Control (subjected to scrotal insulation without treatment) and Gamma (subjected to scrotal insulation and γ-oryzanol treatment). The rams were subjected to scrotal insulation by covering the scrotum with a thermal bag for 72 h. Animals in the Gamma group received 33 mg/MW oral γ-oryzanol once-daily, beginning 7 days before insulation and continuing during insulation and for 20 days afterward, for a total treatment period of 30 days. Samples of semen and blood were collected during the experiment to perform biochemical evaluations of oxidative stress, seminal kinetics and morphology, and plasma testosterone concentrations. Ultrasound examinations of the testicular parenchyma and clinical evaluations of its consistency and the scrotal perimeter were also performed at weekly intervals. Testicular tissue was collected for biochemical analyses of oxidative stress parameters at the end of the experiment by orchiectomy. The results showed that testicular degeneration was induced by scrotal insulation, as was demonstrated by the reduced scrotal perimeter and increased in testicular flaccidity immediately after insulation. Moreover, a delayed increase in the number of hyperechoic points in the parenchyma and a delayed reduction in sperm motility were observed at 10 weeks after insulation by ultrasonography. Treatment with γ-oryzanol reduced levels of reactive oxygen species (ROS) levels in the testes, and increased the total antioxidant potential (assessed based on the ferric reducing ability (FRAP)) in week 10 and levels of lipid peroxidation (TBARS). It also increased the number of intact spermatozoa in week 3, but increased the total number of sperm defects from week 5 onwards. Although γ-oryzanol protected the semen and testes by reducing the levels of the parameters of oxidative stress evaluated herein, the other parameters studied were not improved by the treatment. In addition, supplementation with γ-oryzanol led to more morphological abnormalities in the sperm. This study presented new information on the oral administration of γ-oryzanol to rams with testicular degeneration, and described potential therapies for this pathology, which currently has no established treatment and has important impacts on reproductive health.

Impact of heat stress, nutritional stress and their combinations on the adaptive capability of Malpura sheep under hot semi-arid tropical environment

A study was conducted to assess the cumulative effects of combined stresses (heat and nutritional) on physiological adaptability, blood biochemical and endocrine responses in Malpura rams. The study was conducted for a period of 45 days. Twenty eight adult Malpura rams (average BW 66.0 Kg) were used in the present study. The rams were divided into four groups viz., CON (n=7; control), HS (n=7; heat stress), NS (n=7; nutritional stress) and CS (n=7; combined stress). The animals were stall fed with a diet consisting of 60% roughage and 40% concentrate. The CON and HS ewes were provided with ad libitum feeding while NS and CS rams were provided with restricted feed (30% intake of GI ewes) to induce nutritional stress. The HS and NS rams were kept in climatic chamber @ 42ºC and 55% RH for six hours a day between 10:00 hr to 16:00 hr to induce heat stress. The parameters studied were respiration rate (RR), pulse rate (PR), rectal temperature (RT), scrotal volume, sweating rate scrotum, sweating rate skin, haemoglobin (Hb) and packed cell volume (PCV), plasma cortisol, T3 (tri-iodo-thyronine), and T4 (thyroxin) level. Combined stresses significantly (P<0.01) influenced all adaptive parameters studied. The study shows that Malpura rams possess the adaptive capability to two stresses simultaneously. This is evident from the non-significant changes in RR, RT, sweating rate of scrotum and skin and Hb concentration between HS and CS groups. Further, the capability to adjust the cortisol level to minimum possible increase to elicit the heat stress relieving effects also proves the superior adaptive capability of Malpura rams to the effects of combined stresses.

Impact of heat stress, nutritional stress and their combinations on the adaptive capability of Malpura sheep under hot semi-arid tropical environment

A study was conducted to assess the cumulative effects of combined stresses (heat and nutritional) on physiological adaptability, blood biochemical and endocrine responses in Malpura rams. The study was conducted for a period of 45 days. Twenty eight adult Malpura rams (average BW 66.0 Kg) were used in the present study. The rams were divided into four groups viz., CON (n=7; control), HS (n=7; heat stress), NS (n=7; nutritional stress) and CS (n=7; combined stress). The animals were stall fed with a diet consisting of 60% roughage and 40% concentrate. The CON and HS ewes were provided with ad libitum feeding while NS and CS rams were provided with restricted feed (30% intake of GI ewes) to induce nutritional stress. The HS and NS rams were kept in climatic chamber @ 42ºC and 55% RH for six hours a day between 10:00 hr to 16:00 hr to induce heat stress. The parameters studied were respiration rate (RR), pulse rate (PR), rectal temperature (RT), scrotal volume, sweating rate scrotum, sweating rate skin, haemoglobin (Hb) and packed cell volume (PCV), plasma cortisol, T3 (tri-iodo-thyronine), and T4 (thyroxin) level. Combined stresses significantly (P<0.01) influenced all adaptive parameters studied. The study shows that Malpura rams possess the adaptive capability to two stresses simultaneously. This is evident from the non-significant changes in RR, RT, sweating rate of scrotum and skin and Hb concentration between HS and CS groups. Further, the capability to adjust the cortisol level to minimum possible increase to elicit the heat stress relieving effects also proves the superior adaptive capability of Malpura rams to the effects of combined stresses.(AU)

Modeling of behavioral responses for successful selection of easy-to-train rams for semen collection with an artificial vagina.

The aim of this study was to analyze the reproductive behavioral responses in Australian Merino rams, to identify those related to a faster training for semen collection with an artificial vagina. Eight Australian Merino rams, aged 1.5 years and with no prior sexual experience, were randomly selected from an extensively grazed flock. One immobilized ewe with no hormone stimulation was used for rams to sexually interact and mount. The frequencies of approaching, sniffing, flehmen, pushing, pawing with chin resting, and tongue flicking were recorded during eight training and three post-training assessments periods. In addition, the duration of sniffing and flehmen responses, as well as the time from when the ram started to approach the ewe until the mount with ejaculation (completed mount) were recorded. Descriptive, correlation, and modeling analyses were performed. Amongst the rams, four mounted the ewe and ejaculated for the first time during the training phase, and three mounted and ejaculated for the first time after the training phase. The remaining ram mounted the ewe and ejaculated for the first time during the post-training evaluation in the following year. A great variability in the behavior repertoire was observed among rams. The correlation analysis indicated that the completed mount was associated with the behaviors during the approaching response. The expression of the sniffing response decreased between the training phase and post-training evaluation, while the responses of pushing the ewe and tongue flicking ceased to occur. Pawing the side of the ewe with the chin resting on the back of the ewe and flehmen responses, however, continued between the training and post-training phases. This led to a decrease in the time from when the ram started to approach the ewe until the completed mount. It is concluded that the responses of approaching the ewe, pawing the side of the ewe with chin resting on the ewe, and sniffing of the ewe (the latter occurring only during the training phase) are behavioral indicators that could be used for selection of easy-to-train rams for purposes of semen collection with an artificial vagina.

Paraoxonase 1 (PON1) activity in serum, follicular fluid and seminal plasma of sheep.

This work aimed to describe the activity of paraoxonase 1 (PON1) in serum, follicular fluid and seminal plasma of sheep. Average serum PON1 activity was 286.8 ± 96.2 U/ml in females and 237.6 ± 18.9 U/ml in males. There was a positive correlation between PON1 activity in serum and follicular fluid in females, being twice higher in serum than in follicular fluid (148.8 ± 15.7 U/ml). PON1 activity in males' serum was 10-fold higher than in seminal plasma (21.18 ± 14.2 U/ml), and there was no correlation between PON1 activity in both compartments. Finally, this work suggests that PON1 activity of in sheep is higher compared to other mammalian species, and there is an association between PON1 in serum and follicular fluid only.

Effect of extreme severe heat stress on respiratory rate in unshorn and shorn Australian Merino rams from Northern Patagonia / Efeito do estresse por calor extremamente intenso sobre a taxa respiratória de carneiros Merino Australiano com lã e tosquiados da Patagônia Norte, Argentina

The aim of this study was to determine the effects of severe extreme thermal stress on the respiratory rate (panting) of unsheared and sheared rams in standing and lying position, and to analyse two temperature-humidity indices (THIs). Six mature Australian Merino rams from Northern Patagonia, three unshorn and three shorn, were subjected to 40 hours (8 hours each day for five days) of gradual increase in temperature from 25°C to 40°C, guaranteeing 4h daily at 40°C in a heat chamber. Respiratory rate (RR) was registered continuously by counting flank movements, and the ambient air temperature and relative humidity were recorded every five minutes inside the heat chamber. From 1413 RR measurements, descriptive and modelling analyses were performed. Panting as a response variable was distributed into five categories, and fixed effects such as THI, wool (unshorn/shorn), and position (standing/lying) were considered. No significant differences were observed between the panting score in the unshorn (standing or lying) and standing (unshorn or shorn) sheep, but significant differences were observed in shorn and lying rams. The explanations could be that heat production from muscle activity was lower and the transfer and heat lost by conduction to the floor was easier in the rams with shorter fleece (at least 0.7 inches long). However, in the unshorn rams, their wool acted as an insulator, both with air as well as the floor. Australian Merino rams from Northern Patagonia are heat tolerant to an environment between 31.5°C and 42°C, and 32% to 48% humidity for 40 hours over five days. The rams were in the first phase of panting, and the normal rectal temperatures registered daily when leaving the heat chamber confirmed that regulation of body temperature was possible. The LPHSI's THI was adjusted to improve the analysis of the RR more than the National Research Council's THI.(AU)

O objetivo deste estudo foi determinar o efeito do estresse térmico extremamente severo sobre a frequência respiratória (ofego) em carneiros com lã e tosquiados, em pé ou deitados, e analisar dois índices de temperatura e umidade (ITU). Seis carneiros Merino Australiano da Patagônia Norte, três com lã e três tosquiados, foram expostos durante 40 horas a aumento gradual de temperatura de 25 a 40°C (oito horas durante cinco dias), garantindo quatro horas diárias a 40°C em uma câmara de calor. A frequência respiratória foi registrada continuamente mediante a contagem dos movimentos do flanco. A temperatura e a umidade relativa ambiente foram registradas a cada cinco minutos dentro da câmara. A partir de 1.413 frequências respiratórias registradas, uma análise descritiva foi realizada e um modelo calculado. A variável de resposta do ofego foi dividida em cinco categorias e os efeitos fixos considerados foram: ITU, lã (com lã ou tosquiado) e posição (em pé ou deitado). Não se observaram diferenças significativas nas frequências de ofego nos carneiros com lã (em pé ou deitados) nem entre os carneiros em pé (com lã e tosquiados), mas foram observadas diferenças significativas nos carneiros tosquiados deitados. Essas diferenças podem ser atribuídas à perda de calor com o solo, facilitada nos carneiros com mecha mais curta (pelo menos 1,8 cm), e à baixa produção de calor de atividade de músculo. Nos carneiros lanados, a lã atua como isolante tanto com o ar quanto com o solo. Os carneiros Merino Australianos do Norte da Patagônia foram adaptados a temperaturas ambiente entre 31,5 e 42°C e 32 e 48% de umidade durante 40 horas em cinco dias. Os carneiros permaneceram na primeira fase do ofego e as temperaturas retais diárias, que se registraram ao deixar a câmara de calor, permaneceram dentro da normalidade, o que demonstra que eles puderam regular a temperatura corporal.[...] (AU)

Effect of extreme severe heat stress on respiratory rate in unshorn and shorn Australian Merino rams from Northern Patagonia / Efeito do estresse por calor extremamente intenso sobre a taxa respiratória de carneiros Merino Australiano com lã e tosquiados da Patagônia Norte, Argentina

The aim of this study was to determine the effects of severe extreme thermal stress on the respiratory rate (panting) of unsheared and sheared rams in standing and lying position, and to analyse two temperature-humidity indices (THIs). Six mature Australian Merino rams from Northern Patagonia, three unshorn and three shorn, were subjected to 40 hours (8 hours each day for five days) of gradual increase in temperature from 25°C to 40°C, guaranteeing 4h daily at 40°C in a heat chamber. Respiratory rate (RR) was registered continuously by counting flank movements, and the ambient air temperature and relative humidity were recorded every five minutes inside the heat chamber. From 1413 RR measurements, descriptive and modelling analyses were performed. Panting as a response variable was distributed into five categories, and fixed effects such as THI, wool (unshorn/shorn), and position (standing/lying) were considered. No significant differences were observed between the panting score in the unshorn (standing or lying) and standing (unshorn or shorn) sheep, but significant differences were observed in shorn and lying rams. The explanations could be that heat production from muscle activity was lower and the transfer and heat lost by conduction to the floor was easier in the rams with shorter fleece (at least 0.7 inches long). However, in the unshorn rams, their wool acted as an insulator, both with air as well as the floor. Australian Merino rams from Northern Patagonia are heat tolerant to an environment between 31.5°C and 42°C, and 32% to 48% humidity for 40 hours over five days. The rams were in the first phase of panting, and the normal rectal temperatures registered daily when leaving the heat chamber confirmed that regulation of body temperature was possible. The LPHSI's THI was adjusted to improve the analysis of the RR more than the National Research Council's THI.(AU)

O objetivo deste estudo foi determinar o efeito do estresse térmico extremamente severo sobre a frequência respiratória (ofego) em carneiros com lã e tosquiados, em pé ou deitados, e analisar dois índices de temperatura e umidade (ITU). Seis carneiros Merino Australiano da Patagônia Norte, três com lã e três tosquiados, foram expostos durante 40 horas a aumento gradual de temperatura de 25 a 40°C (oito horas durante cinco dias), garantindo quatro horas diárias a 40°C em uma câmara de calor. A frequência respiratória foi registrada continuamente mediante a contagem dos movimentos do flanco. A temperatura e a umidade relativa ambiente foram registradas a cada cinco minutos dentro da câmara. A partir de 1.413 frequências respiratórias registradas, uma análise descritiva foi realizada e um modelo calculado. A variável de resposta do ofego foi dividida em cinco categorias e os efeitos fixos considerados foram: ITU, lã (com lã ou tosquiado) e posição (em pé ou deitado). Não se observaram diferenças significativas nas frequências de ofego nos carneiros com lã (em pé ou deitados) nem entre os carneiros em pé (com lã e tosquiados), mas foram observadas diferenças significativas nos carneiros tosquiados deitados. Essas diferenças podem ser atribuídas à perda de calor com o solo, facilitada nos carneiros com mecha mais curta (pelo menos 1,8 cm), e à baixa produção de calor de atividade de músculo. Nos carneiros lanados, a lã atua como isolante tanto com o ar quanto com o solo. Os carneiros Merino Australianos do Norte da Patagônia foram adaptados a temperaturas ambiente entre 31,5 e 42°C e 32 e 48% de umidade durante 40 horas em cinco dias. Os carneiros permaneceram na primeira fase do ofego e as temperaturas retais diárias, que se registraram ao deixar a câmara de calor, permaneceram dentro da normalidade, o que demonstra que eles puderam regular a temperatura corporal. Além disso, foram analisados comparativamente os ajustes à frequência respiratória dos ITUs: LPHSI e National Research Council.(AU)

Distribution of lymphocytes, immunoglobulin-containing cells, macrophages, and dendritic cells in the accessory sex glands of rams experimentally infected with actinobacillus seminis / Distribuição de linfócitos, células carreadoras de imunoglobulinas, macrófagos e células dendríticas em glândulas sexuais acessórias de carneiros infectados experimentalmente com actinobacillus seminis

The distribution of cells involved in the immune response in accessory sex glands of rams experimentally infected with Actinobacillus seminis was studied. Twelve one-year old rams were experimentally infected by intraurethral (IU) (n=4) and intraepididymal (IE) (n=4) route, and four control (CON) animals were used. The animals were slaughtered 35 days post-inoculation, samples were taken from accessory sex glands, and bacteriology and histopathology tests were performed. The presence of CD4, CD8 and TCRγδ (WC1) lymphocytes, CD45RO cells, macrophages (CD14), dendritic cells (CD1b), IgA-, IgG- and IgM-containing cells (IgCC) was determined. Animals of the IE group developed clinical epididymitis. No lesions were seen in rams of the IU group; two of the intraepididymal inoculated CON developed small lesions in the epididymis. A. seminis isolates were achieved from 6:16 (37.5%) accessory sex glands in the IE group, but not in the IU and CON groups. In the CON group, IgA- and IgM- containing cells predominated in the bulbourethral glands and the disseminated prostate, and they were scarce or null in the vesicles and ampullae. A significant increase of IgA-, IgG- and IgM- containing cells was confirmed in the seminal vesicles, the ampullae and the bulbourethral glands in the IE group. In the IE and IU groups, an increase in CD4, CD8, WC1, CD45RO and CD14 was evidenced in the vesicles and ampullae. CD1b dendritic cells were present in the ampullae and vesicles with inflammatory processes. A. seminis triggered a local immune response in the IE and IU groups. These results indicate a different pattern of infiltrating immune cells in the accessory sex glands of infected A. seminis rams.

A distribuição das células envolvidas na resposta imune em glândulas sexuais acessórias de carneiros experimentalmente infectados com Actinobacillus seminis foi estudada. Doze carneiros de um ano de idade foram experimentalmente infectados via intrauretral (IU) (n=4) e via intraepididimal (IE) (n=4) e quatro animais controles (CON) foram utilizados. Os animais foram abatidos 35 dias após a inoculação, amostras foram retiradas das glândulas sexuais acessórias e testes bacteriológicos e histopatológicos foram realizados. A presença de linfócitos CD4, CD8 e TCRγδ (WC1), células CD45RO, macrófagos (CD14), células dendríticas (CD1b) e células contendo IgA, IgG and IgM (IgCC) foi determinada. Os animais do grupo IE desenvolveram epididimite clínica. Não foram visualizadas lesões nos carneiros do grupo IU, dois dos CON inoculados intraepididimalmente desenvolveram pequenas lesões no epidídimo. Isolados de A. seminis foram obtidos de 6:16 (37,5%) nas glândulas sexuais acessórias no grupo IE mas não nos grupos IU e CON. No grupo CON células contendo IgA and IgM predominaram nas glândulas bulbouretrais e na próstata e foram escassas ou ausentes nas vesículas e na ampola. Um incremento significativo de células contendo IgA, IgG and IgM foi confirmado nas vesículas seminais, na ampola e nas glândulas bulbouretrais no grupo IE. Nos grupos IE e IU foi evidenciado um aumento em CD4, CD8, WC1, CD45RO e CD14 nas vesículas e ampola. As células dendríticas CD1b estavam presentes na ampola e nas vesículas com processo inflamatório. A. seminis induziu uma resposta imune local nos grupos IE e IU. Estes resultados indicam um padrão diferente de células imunes infiltrantes nas glândulas sexuais acessórias de carneiros infectados por A. seminis.

Serologic and molecular evidence of Brucella ovis infection in ovine and caprine flocks in the State of Minas Gerais, Brazil.

BACKGROUND: Brucella ovis infection is one of the leading causes of sub fertility and infertility in ovine, been characterized mainly by epididymitis, orchitis and testicular atrophy in rams. This study aimed to determine the frequency of B. ovis positivity in rams and goats flocks in the State of Minas Gerais, Brazil, by agarose gel immunodiffusion (AGID), ELISA, Rose Bengal, PCR and bacteriological isolation as diagnostic tools. FINDINGS: Serum and urine samples were collected from properties with sheep or goat flocks, or from properties with mixed flock. Out of 50 sheep flocks, 6% (3/50) were seropositive by AGID while 4% (2/50) were positive by urine PCR for B. ovis. Out of five goat farms, 20% (1/5) were seropositive for B. ovis by AGID. Mixed flock farms had 11.1% (2/18) of positivity by AGID. By ELISA, 19.5% (8/41) of sheep properties and 61.1% (11/18) of the properties with mixed flocks were positive for B. ovis. No samples were positive in the test of Rose Bengal, ruling out exposure to smooth LPS Brucella species (particularly Brucella melitensis) and indicating that the positive in the ELISA was associated with Brucella spp. LPS rough (presumably B. ovis). No urine sample from sheep or goat was positive by bacteriological isolation. CONCLUSIONS: Our results demonstrate serologic or molecular evidence of B. ovis infection in several rams and billy goats from meso-regions of the State of Minas Gerais, Brazil. Also, this study report the indirect ELISA as an important tool for the diagnosis of B. ovis infection, as indirect ELISA in this study demonstrated to be the most sensitive diagnostic method adopted.

Distribution of lymphocytes, immunoglobulin-containing cells, macrophages, and dendritic cells in the accessory sex glands of rams experimentally infected with Actinobacillus seminis / Distribuição de linfócitos, células carreadoras de imunoglobulinas, macrófagos e células dendríticas em glândulas sexuais acessórias de carneiros infectados experimentalmente com Actinobacillus seminis

The distribution of cells involved in the immune response in accessory sex glands of rams experimentally infected with Actinobacillus seminis was studied. Twelve one-year old rams were experimentally infected by intraurethral (IU) (n=4) and intraepididymal (IE) (n=4) route, and four control (CON) animals were used. The animals were slaughtered 35 days post-inoculation, samples were taken from accessory sex glands, and bacteriology and histopathology tests were performed. The presence of CD4, CD8 and TCRγδ (WC1) lymphocytes, CD45RO cells, macrophages (CD14), dendritic cells (CD1b), IgA-, IgG- and IgM-containing cells (IgCC) was determined. Animals of the IE group developed clinical epididymitis. No lesions were seen in rams of the IU group; two of the intraepididymal inoculated CON developed small lesions in the epididymis. A. seminis isolates were achieved from 6:16 (37.5%) accessory sex glands in the IE group, but not in the IU and CON groups. In the CON group, IgA- and IgM- containing cells predominated in the bulbourethral glands and the disseminated prostate, and they were scarce or null in the vesicles and ampullae. A significant increase of IgA-, IgG- and IgM- containing cells was confirmed in the seminal vesicles, the ampullae and the bulbourethral glands in the IE group. In the IE and IU groups, an increase in CD4, CD8, WC1, CD45RO and CD14 was evidenced in the vesicles and ampullae. CD1b dendritic cells were present in the ampullae and vesicles with inflammatory processes. A. seminis triggered a local immune response in the IE and IU groups. These results indicate a different pattern of infiltrating immune cells in the accessory sex glands of infected A. seminis rams.(AU)

A distribuição das células envolvidas na resposta imune em glândulas sexuais acessórias de carneiros experimentalmente infectados com Actinobacillus seminis foi estudada. Doze carneiros de um ano de idade foram experimentalmente infectados via intrauretral (IU) (n=4) e via intraepididimal (IE) (n=4) e quatro animais controles (CON) foram utilizados. Os animais foram abatidos 35 dias após a inoculação, amostras foram retiradas das glândulas sexuais acessórias e testes bacteriológicos e histopatológicos foram realizados. A presença de linfócitos CD4, CD8 e TCRγδ (WC1), células CD45RO, macrófagos (CD14), células dendríticas (CD1b) e células contendo IgA, IgG and IgM (IgCC) foi determinada. Os animais do grupo IE desenvolveram epididimite clínica. Não foram visualizadas lesões nos carneiros do grupo IU, dois dos CON inoculados intraepididimalmente desenvolveram pequenas lesões no epidídimo. Isolados de A. seminis foram obtidos de 6:16 (37,5%) nas glândulas sexuais acessórias no grupo IE mas não nos grupos IU e CON. No grupo CON células contendo IgA and IgM predominaram nas glândulas bulbouretrais e na próstata e foram escassas ou ausentes nas vesículas e na ampola. Um incremento significativo de células contendo IgA, IgG and IgM foi confirmado nas vesículas seminais, na ampola e nas glândulas bulbouretrais no grupo IE. Nos grupos IE e IU foi evidenciado um aumento em CD4, CD8, WC1, CD45RO e CD14 nas vesículas e ampola. As células dendríticas CD1b estavam presentes na ampola e nas vesículas com processo inflamatório. A. seminis induziu uma resposta imune local nos grupos IE e IU. Estes resultados indicam um padrão diferente de células imunes infiltrantes nas glândulas sexuais acessórias de carneiros infectados por A. seminis.(AU)

Variabilidad interspecífica de duraznos (Prunus pérsica L. Batsch.) y ciruelos (Prunus domestica) usando RAMs / Intraespecific variability of peach (Prunus persica L. Batsch.) and plums (Prunus domestica) using RAMs

Se seleccionó una muestra de 41 materiales de Prunus de la colección de caducifolios de la Universidad Pedagógica y Tecnológica de Colombia, para evaluar su variabilidad genética usando ocho marcadores Microsatélites Amplificados al Azar (RAMs). Se generaron un total de 121 alelos con pesos moleculares entre 260 y 1000 Kb. Se formaron tres grupos, a un coeficiente de similitud de 0.75, de acuerdo a las características del fruto y a la especie, encontrándose en el grupo II a los materiales de ciruelo. El porcentaje de loci polimórficos varío entre 71 y 99% para los cebadores CGA y CCA, respectivamente. El valor promedio de heterocigosidad fue de 0.22, mucho más bajo que los encontrados en otros estudios de diversidad genética en el género Prunus. La técnica RAMs mostró ser una herramienta útil para evaluar la diversidad genética en frutales caducifolios, al discriminar a los materiales en tres grupos e identificar el alto grado de consanguinidad que existe entre las diferentes especies de Prunus lo cual debe ser aprovechado dentro de las estrategias de hibridación que busquen la obtención de nuevos y mejores materiales.

A sample of 41 Prunus materials from the deciduous collection of the Pedagogical and Technological University of Colombia was selected to evaluate its genetic diversity using eight primers for Random Amplified Microsatellite (RAMs). A total of 121 alleles were generated with molecular weights ranging between 260 and 1000 Kb. Three groups were formed, a similarity coefficient of 0.75, according to fruit characteristics and specie, found plum materials in group II. The percentage of polymorphic loci ranged from 71 to 99% for the primers CGA and CCA, respectively. The average value of heterozygosity was 0.33, much lower than values found in other genetic diversity studies in the genus Prunus. The RAMs technique showed to be a useful tool for assessing genetic diversity in deciduous fruit, discriminate the materials into three groups and identify the high degree of consanguinity between different Prunus species which should be exploited in hybridization strategies looking for obtaining new and improved materials.

Parâmetros anátomo-estruturais de orgãos reprodutivos de ovinos sem raça definida (SRD) nativos do Estado da Paraíba, com e sem bipartição escrotal: estudo do escroto e funículo espermático / Anatomical and structural parameters of reproductive organs of crossbred rams native from the state of Paraíba, Brazil, with and without scrotal bipartition: study of the scrotal skin and spermatic cord

Foram utilizados 42 ovinos sem raça definida, divididos segundo a configuração escrotal. Destes animais, 12 foram utilizados na investigação da biometria testicular e histologia da pele escrotal. Os demais foram destinados ao estudo do funículo espermático. Os animais foram agrupados em um grupo de 21 animais sem bipartição escrotal (GEI) e 21 com bipartição escrotal, (GEII), esta não atingindo 50% do comprimento do eixo longitudinal do escroto. Em cada grupo, em 6 animais foram coletados fragmentos da pele do escroto e em 5 do funículo espermáticos, e processados em rotina histológica e analisados em microscopia de luz; e em 10 foram injetados látex na artéria testicular para obtenção de moldes vasculares e obtenção do comprimento da artéria. Quando comparados os grupos GEI e GEII, não foram encontradas diferenças estatísticas significativas (p<0,05) entre a espessura do escroto (epiderme e derme), constituição histológica da pele escrotal, número de glândulas sudoríparas por área, comprimento do funículo espermático ou parâmetros biométricos testiculares. Entretanto, o comprimento total das artérias testiculares do GEI foi maior do que o GEII (p<0,05). Concluiu-se, com base nos parâmetros morfológicos analisados, que a bipartição escrotal em ovinos não influenciou na estrutura da pele, funículo ou biometria testicular quando comparado aos animais que não apresentavam esta característica. Outros estudos merecem atenção para desmistificar o porquê do aparecimento dessa característica em ovinos e se esta característica é ou não desejável para melhoria na produção desses animais em regiões de clima quente.(AU)

Forty-two crossbred rams were used, divided according to their scrotal configuration. Twelve of these animals were used to investigate the testicular biometry and scrotal skin histology. The other animals were used to study the spermatic cord. The animals were placed in a group of 21 animals without scrotal bipartition (GEI) and another with scrotal bipartition (GEII) which did not reach 50% of the length of the scrotum's longitudinal axis. In each group, scrotal skin fragments from six rams and spermatic cord fragments from 5 others were collected, processed by routine histological tests and analyzed with a microscope. In 10 rams latex was injected into the testicular artery to obtain vascular molds and the artery length. Comparison of groups GEI and GEII showed no significant statistical differences (p<0.05) between scrotal thickness (epidermis and dermis), histological constituency of the scrotal skin, number of sudoriferous glands per area, scrotal bipartition length or testicular biometric parameters. However, the total length of the testicular arteries of GEI was greater than of GEII (p<0.05). It was concluded from the morphological parameters analyzed that scrotal bipartition in rams did not influence the skin structure, scrotal bipartition or testicular biometry when compared with animals that did not present this characteristic. Further studies should be carried out to elucidate the appearance of this characteristic in rams and whether it is desirable or not for improved production of these animals in warm climate regions.(AU)

Parâmetros anátomo-estruturais de orgãos reprodutivos de ovinos sem raça definida (SRD) nativos do Estado da Paraíba, com e sem bipartição escrotal: estudo do escroto e funículo espermático / Anatomical and structural parameters of reproductive organs of crossbred rams native from the state of Paraíba, Brazil, with and without scrotal bipartition: study of the scrotal skin and spermatic cord

Foram utilizados 42 ovinos sem raça definida, divididos segundo a configuração escrotal. Destes animais, 12 foram utilizados na investigação da biometria testicular e histologia da pele escrotal. Os demais foram destinados ao estudo do funículo espermático. Os animais foram agrupados em um grupo de 21 animais sem bipartição escrotal (GEI) e 21 com bipartição escrotal, (GEII), esta não atingindo 50% do comprimento do eixo longitudinal do escroto. Em cada grupo, em 6 animais foram coletados fragmentos da pele do escroto e em 5 do funículo espermáticos, e processados em rotina histológica e analisados em microscopia de luz; e em 10 foram injetados látex na artéria testicular para obtenção de moldes vasculares e obtenção do comprimento da artéria. Quando comparados os grupos GEI e GEII, não foram encontradas diferenças estatísticas significativas (p<0,05) entre a espessura do escroto (epiderme e derme), constituição histológica da pele escrotal, número de glândulas sudoríparas por área, comprimento do funículo espermático ou parâmetros biométricos testiculares. Entretanto, o comprimento total das artérias testiculares do GEI foi maior do que o GEII (p<0,05). Concluiu-se, com base nos parâmetros morfológicos analisados, que a bipartição escrotal em ovinos não influenciou na estrutura da pele, funículo ou biometria testicular quando comparado aos animais que não apresentavam esta característica. Outros estudos merecem atenção para desmistificar o porquê do aparecimento dessa característica em ovinos e se esta característica é ou não desejável para melhoria na produção desses animais em regiões de clima quente.

Forty-two crossbred rams were used, divided according to their scrotal configuration. Twelve of these animals were used to investigate the testicular biometry and scrotal skin histology. The other animals were used to study the spermatic cord. The animals were placed in a group of 21 animals without scrotal bipartition (GEI) and another with scrotal bipartition (GEII) which did not reach 50% of the length of the scrotum's longitudinal axis. In each group, scrotal skin fragments from six rams and spermatic cord fragments from 5 others were collected, processed by routine histological tests and analyzed with a microscope. In 10 rams latex was injected into the testicular artery to obtain vascular molds and the artery length. Comparison of groups GEI and GEII showed no significant statistical differences (p<0.05) between scrotal thickness (epidermis and dermis), histological constituency of the scrotal skin, number of sudoriferous glands per area, scrotal bipartition length or testicular biometric parameters. However, the total length of the testicular arteries of GEI was greater than of GEII (p<0.05). It was concluded from the morphological parameters analyzed that scrotal bipartition in rams did not influence the skin structure, scrotal bipartition or testicular biometry when compared with animals that did not present this characteristic. Further studies should be carried out to elucidate the appearance of this characteristic in rams and whether it is desirable or not for improved production of these animals in warm climate regions.

Distribution of immune response cells in the pelvic urethra and the prepuce of rams / Distribuição de células da resposta imune na uretra pélvica e em prepúcio de carneiros

The pathogens of the reproductive system in the male can penetrate and establish by ascending route, from to the prepuce to the urethra, accessory glands, epididymis and testicles. The aim of this paper is determine the distribution and number of cells involved in the immune response in prepuce and pelvic urethra of rams, without apparent clinical alterations in testicle, epididymis and prepuce. [...] Significant differences were found in the total number of CD4, CD45RO, and WC1 lymphocytes, in CD14 macrophages, and CD1b dendritic cells, with mean values being greater in the fornix than in the urethra (p<0.05) in all cases. Only dendritic cells were found in the prepuce. No differences were found in the number of CD8 lymphocytes between both organs. The ratio between each cell type in the connective and the intraepithelial tissues and between organs was 10/1 for CD4 in the fornix (p<0.05), against 7/1 in the urethra (p<0.05), while CD8 had a 1/1 distribution in both mucosae. The WC1 ratio was 5/1 in both mucosae (p<0.05). CD45RO labeling was 19/1 in the prepuce (p<0.05) and 1/1 in the urethra. IgA-containing cells did not show differences in the total number of cells in both tissues. In the urethra, no IgG-containing cells were observed and IgM-containing cells were scarce; in contrast, both cell types were present in the prepuce, in amounts greater than in the urethra (p<0.05). IgA-, IgG-, and IgM-containing cells were located in both organs in the mucosal connective tissue. The presence of antigen-presenting cells, macrophages, and dendritic cells, as well as of lymphocytes CD4, CD8 TCR γδ (WC1), IgA-, IgG and IgM positive cells, and CD45RO cells suggests that both mucosae may behave as inductive and effector sites for the mucosal immune response.(AU)

Os patógenos do aparelho reprodutor do macho podem penetrar e se estabelecer por via ascendente, a partir do prepúcio à uretra, glándulas anexas, epidídimo e testículos. Neste trabalho foi quantificada a distribuição de algumas das células envolvidas na resposta imune, em nível de prepúcio e uretra pélvica, em quatro carneiros de um ano de idade, sem lesões aparentes no testículo, no epidídimo e no prepúcio.[...] Não foram encontradas diferenças significativas no número de linfocitos CD8 entre ambos os orgãos. A relação entre cada tipo celular no tecido conectivo e intra-epitelial e entre os diferentes órgãos, resultou para CD4 10/1 no prepúcio (p<0.05), contra 7/1 na uretra (p<0.05), entretanto os CD8 se distribuíram 1/1 em ambas as mucosas, não sendo significativa as diferenças. Os WC1 foram observados na relação 5/1 em ambas as mucosas (p<0.05). A célula CD45RO, no prepucio, foi observada de 19/1(p<0.05) e na uretra de 1/1, não sendo um resultado significativo. As CC-IgA não mostraram diferença significativa no total de células em ambos os tecidos. Na uretra não foram observadas as CC-IgG, e as CC-IgM foram escassas; em contrapartida, ambos os tipos celulares foram observadas no prepucio, em quantidades menores que na uretra (p<0.05). As CC-IgA, IgG e IgM foram observadas em ambos os tecidos conectivos da mucosa. A presença de células apresentadoras de antígenos, macrófagoss e células dendríticas, assim como de linfócitos CD4, CD8. TCR γδ (WC1), CC-IgA, IgG e IgM e células CD45RO, determinam que ambas as mucosas podem se comportar como locais de indução e promoção da resposta imune das mucosas.(AU)