A 6-wk trial was performed with 40 late-lactation Comisana ewes, which were either exposed to or protected from solar radiation and fed either in the morning (EXPM, PROM) or afternoon (EXPA, PROA) during summer in a Mediterranean climate. Behavioral traits of ewes were recorded once per week from 0800 to 2000 h. Rectal temperature (RT) and respiration rate (RR) were measured twice weekly at 1430 h. The phytohemagglutinin (PHA) skin test was performed to induce nonspecific delayed-type hypersensitivity at d 10, 20, and 32 of the experiment. Jugular blood samples were taken from ewes at the beginning and at d 21 and 42 of the experiment. Ewe milk yield was recorded daily. Individual milk samples were analyzed weekly for milk composition, coagulating properties, somatic cell count (SCC) and polymorphonuclear neutrophil leukocyte counts (PMNLC) and every 2 wk for bacteriological characteristics. Solar radiation and the interaction of solar radiation x time of feeding had significant effects on rectal temperatures. EXPM ewes had higher rectal temperatures than EXPA ewes, which in turn exhibited higher RT compared with PROM and PROA ewes. EXP groups also had significantly higher respiration rates than PRO groups. Immune response was lower in EXPM ewes at d 10 and in EXPM, EXPA, and PROM animals at d 20 compared with PROA ewes. Exposure to solar radiation resulted in decreased plasma concentrations of alanine amino-transferase, alkaline phosphatase, potassium, and magnesium, as well as in increased levels of nonesterified fatty acids and aspartate amino-transferase. Milk yield and composition were not changed by exposure to solar radiation and time of feeding, but the EXPM treatment resulted in lower yields of casein and fat and reduced clot firmness compared with the three other treatments. Milk SCC was similar across treatments, but PMNLC was higher in EXPM than in PROM and PROA milk. EXPM animals also had the greatest amounts of total and fecal coliforms and of Pseudomonadaceae as well as the highest number of mastitis related pathogens in their milk. Results suggest that provision of shaded areas can play a major role in helping lactating ewes to minimize the adverse effects of high ambient temperatures on thermal balance and energy and mineral metabolism. Changing the time of feeding to late afternoon may be beneficial to exposed ewes in lowering their heat loads during the warmest hours of the day, thereby reducing the detrimental impact of thermal stress on immune function and udder health.

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The existence of a relationship between cortisol levels, after an acute stress, and behavioral activities, immunological profile, and production performance in sheep was studied. An initial flock of 30 Comisana ewes was involved in the experiment, and each of the 30 ewes was individually subjected to an isolation test in a novel environment. Subsequently, from the initial flock, 2 groups of 8 Comisana ewes were each retrospectively selected, and the animals were divided, according to their cortisol concentration 10 min after the isolation test, into high cortisol (HC) ewes, having a peak of cortisol concentration >90 ng/mL (average: 119.3 ng/mL +/- 11.8), and low cortisol (LC) ewes having a peak of cortisol concentration <80 ng/mL (average: 52.4+/-11.8). During the isolation test, the behavior of each animal was video-recorded and behavioral activities were registered. Blood samples were collected before the isolation test, immediately after the test (10 min), and at 60, 120, 300 min, 24 h, and 48 h after the test to evaluate percentages of T-helper (CD4(+)) and T-cytotoxic (CD8(+)) cells, CD4(+)/CD8(+) ratio, and IL-1beta and IL-6 levels. The ewes were milked for 3 d after the isolation test to determine cortisol levels and IL-1beta and IL-6 concentrations in whey. Milk yield was recorded at each milking, and milk samples were analyzed for pH, nutritional parameters, renneting properties, and somatic cell count. During the isolation test, HC ewes exhibited a shorter duration of movement and fewer bleats than LC ewes. The average plasma IL-1beta concentration was higher in HC than in LC ewes. The average whey IL-1beta and IL-6 concentrations were higher in whey from HC ewes than in LC ewes. A positive correlation emerged between plasma and whey IL-1beta concentrations. The average CD4(+)/CD8(+) ratio in blood was lower in HC than in LC ewes. Time from isolation affected the CD4(+)/CD8(+) ratio: at 120 min, the CD4(+)/CD8(+) ratio increased compared with that at 10 min after isolation and then decreased until 300 min after isolation. On average, ewes with low cortisol concentrations showed higher milk production and lower SCC than ewes with high cortisol concentrations. Results suggest that plasma cortisol concentration is connected to the behavioral response and immune competence of dairy ewes and cytokine concentrations. Both whey IL-1beta and IL-6 can be considered reliable indicators of the magnitude of hypothalamic-pituitary-adrenal axis activation. The stress-induced changes in CD4(+)/CD8(+) ratio are critical for controlling disease incidence and planning appropriate vaccination programs. High reactivity of the hypothalamic-pituitary-adrenal axis is also associated with a reduction in milk production and an increased predisposition to develop intramammary inflammatory processes.

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