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.

Microscopic sperm head damage and abnormalities as heat stress indicators in Australian Merino rams (Ovis aries) in Northern Patagonia, Argentina / Danos e anormalidades microscópicas das cabeças espermáticas como indicadores de estresse por calor em carneiros Merino Australiano (Ovis aries) da Patagônia Norte, Argentina

In Northern Patagonia, the mating season starts on March 15th, when rams are submitted to summer temperatures. Exposure of rams to heat stress increases the prevalence of microscopic damage to spermatozoa, morphological abnormalities, and reductions in fertility. This study assesses the adaptive capabilities of six unshorn and six shorn Australian Merino rams, half of which were treated in a heat chamber for eight hours for five days, gradually reaching a temperature of up to 40 °C. Microscopic damage, abnormalities and ultramicroscopic alterations of the plasma membrane and the acrosome of sperm head were analysed. There were significant differences in the percentage of tailless spermatozoa and proximal cytoplasmic droplets between post-treatment periods. Temperature primarily affected the shorn rams and the sperm heads during spermiogenesis. Submicroscopic alterations were observed when the plasma membrane was present in the anterior segment. These alterations can be intact, waved, or dilated. When the plasma membrane was absent, the acrosome might be intact, dilated, and waved. In addition, the outer acrosomal membrane may completely lose its contents or have a nude nucleus. The plasma membrane assumes a waved shape as a result of the effect of temperature on the epididymis. According to this study, the tailless head, proximal cytoplasmic droplets, and the ultramicroscopic categories studied were robust indicators of semen heat stress. After ten weeks, the sperm head recovered its normal shape. Unshorn rams are better adapted to summer heat stress than shorn ones. Microscopy and transmission electron microscopy alterations have been shown to be excellent indicators of thermal stress in Australian Merino rams and may be useful tools to help sheep farmers choose when to begin the mating season, which will vary depending on the environmental conditions of the summer.(AU)

Na Patagônia Norte, os ovinos têm sua estação de acasalamento iniciada em 15 de março, portanto, ficam sujeitos às temperaturas do verão. A exposição de carneiros a estresse térmico aumenta a prevalência de danos microscópicos e anomalias morfológicas nos espermatozoides, que implica uma redução na fertilidade. Este trabalho avaliou a capacidade adaptativa de carneiros Merino Australiano com lã (N = 6) e tosquiados (N = 6): metade ficou ao ar livre e outra metade foi mantida em uma câmara climática por oito horas, durante cinco dias, chegando gradualmente a uma temperatura máxima de 40 °C. Foram analisados danos microscópicos, anormalidades e alterações ultramicroscópicas da membrana plasmática e do acrossoma da cabeça dos espermatozoides. Os resultados microscópicos confirmaram a existência de diferença significativa na porcentagem de espermatozoides sem cauda e com gota citoplasmática proximal, entre os ejaculados pós-tratamento. A temperatura afetou os carneiros tosquiados, principalmente a cabeça de seus espermatozoides, durante a espermatogênese. Alterações submicroscópicas foram observados na membrana plasmática quando ela estava presente no segmento anterior: quando não intacta, ficava ondulada ou dilatada. Quando a membrana plasmática estava ausente, o acrossoma podia se apresentar ondulado ou dilatado. Além disso, sob efeito do calor, a membrana acrossomal externa pode perder completamente seu conteúdo ou apresentar núcleo desnudo. A membrana plasmática assume uma forma ondulada pelo efeito da temperatura no epidídimo. Depois de dez semanas, a cabeça dos espermatozoides recuperou sua forma normal. Como demonstrado neste estudo, a cabeça sem cauda, as gotas citoplasmáticas proximais e as categorias ultramicroscópicas estudadas são indicadores do efeito do estresse térmico no sêmen, e os carneiros com maior cobertura de lã se adaptam melhor ao estresse por calor. Alterações de microscopia e de microscopia eletrônica de transmissão têm se mostrado excelentes indicadores de estresse por calor em carneiros Merino Australiano e podem ser ferramentas úteis para ajudar criadores de ovelhas a escolher quando começar a época de acasalamento, o que irá variar de acordo com as condições ambientais do verão.(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)

Comparative study of toxicity and biochemical responses induced by sublethal levels of the pesticide azinphosmethyl in two fish species from North-Patagonia, Argentina.

Biochemical effects of azinphosmethyl (AZM), an organophosphate pesticide, were determined in gill, brain and muscle tissues of Odontesthes hatcheri and Jenynsia multidentata. The 96-h toxicity was first assessed, estimating lethal concentrations fifty (LC50) of 7 and 30µgL(-1) AZM for O. hatcheri and J. multidentata, respectively. Considering the LC50, sublethal 96-h static exposures were designed for O. hatcheri (0.1-0.5µgL(-1) AZM) and J. multidentata (5-10µgL(-1)AZM) to determine biochemical endpoints. Brain acetylcholinesterase (AchE) was inhibited by AZM in both species, while the buffer enzyme carboxylesterase (CarbE) was not affected in this tissue. Conversely, muscular AchE was not affected but CarbE was augmented by AZM. The enzymes glutathione reductase, glutathione-S-transferase and CarbE were significantly inhibited in O. hatcheri gills but none of them was affected by AZM in J. multidentata gills compared to control. GSH levels were augmented in gills of both species in exposed fish compared to controls and in addition, lipid peroxidation was significantly increased in O. hatcheri gills. Ex vivo histochemical analysis of ROS by fluorescence microscopy was also performed in J. multidentata gills, indicating a significant increase upon exposure to 10µgL(-1) AZM. Principal component analyses (PCA) were applied, both to the species together or separately. The general analysis demonstrated a clear separation of responses in the two species. For O. hatcheri, the variable that explains the major variation in PC1 is gill catalase and brain AchE in PC2. In J. multidentata in turn, the variable that explains the major variation in PC1 is brain AchE and total oxyradical scavenging capacity in PC2. The toxicity data and biomarker responses obtained for both species were compared to environmental concentrations of AZM detected in superficial water from different points in the Alto Valle region and risk quotients (RQ) were calculated. This approach indicated probable acute effects for O. hatcheri in river and irrigation channels (RQ>0.1), while the risk was unacceptable in drainage superficial water (RQ>1). In contrast, J. multidentata showed minimal risk in river or channel water (RQ<0.1) and probable risk in drainage water (RQ=0.75). We conclude that not only the differential susceptibility of both species to AZM is environmentally relevant, but also that the different biomarkers responding in each case underlie particular pathways stressed by this agrochemical.