Fluctuations of serum cortisol,insulin and non-esterified fatty acid concentrations in growing ewes over the year

Background

The physiological levels of endocrine and metabolic parameters in Slovene autochthonous breeds of sheep are not yet well known, nor are the mechanisms of their adaptability and responses to climate and environmental factors.Therefore, the aim of this study was to evaluate fluctuations of cortisol, insulin and non-esterified fatty acids (NEFA) in growing ewes over an one-year period. Blood samples were collected monthly from 10 yearling Jezersko-Solchava, 10 Bovec and 10 Istrian ewes. Serum cortisol, insulin and NEFA were measured with commercial kits.

Results

Mean monthly cortisol values fluctuated with low levels in summer and high levels in autumn. Significant peaked cortisol values of 25.69 ± 6.89, 14.67 ± 2.43 and 21.11 ± 7.25 μg/L in Jezersko-Solchava, Bovec and Istrian breed, respectively, were found in September (Bovec breed) and October (Jezersko-Solchava and Istrian breed). Mean monthly insulin values increased during the observation period. The highest levels of 14.60 ± 3.15, 16.03 ± 5.35 and 12.56 ± 2.52 μIU/mL in Jezersko-Solchava, Bovec and Istrian breed, respectively, were observed in the last sample collection in May. NEFA concentrations were found to be low except in some autumn and spring months. The peak values were observed in March for Jezersko-Solchava and Istrian breed (0.60 ± 0.05 and 0.66 ± 0.10 mmol/L), and in April for Bovec breed (0.71 ± 0.11 mmol/L).

Conclusions

Monthly fluctuations of cortisol, insulin and NEFA were measured in all observed sheep breeds, but between-breed differences in monthly values of examined parameters were insignificant. Significantly increased serum cortisol levels were found in autumn for all breeds and were probably associated with the onset of puberty and low environmental temperature. A gradual increase of insulin level in the examined ewes was in parallel with their growth. Significantly higher NEFA values in spring suggest qualitatively insufficient feed supply during that period.     

Development of a whole-body cortisol extraction procedure for determination of stress in golden shiners, <Emphasis Type="Italic">Notemigonus crysoleucas</Emphasis>

Baitfish such as golden shiners are subjected to stress during harvesting, grading, and transport. Their small size makes it difficult to measure the stress response with the biological indicator cortisol using conventional assay methods for plasma. This paper examines the development and validation of methods for whole-body cortisol extraction from individual baitfish. Three types of extracts were tested: (1) an ethyl ether unaltered extract (UA); (2) an extract reconstituted in phosphate buffered saline (PBS); (3) an extract that had been increased in volume by the addition of food-grade vegetable oil (VO). These extracts were evaluated using validation tests with radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA). The UA extract produced inadequate volumes of extract for multiple assays and could not be used for the determination of cortisol in a single fish. The PBS reconstitution method failed the precision recovery of serial dilutions (62.3%), linearity (R ²: 0.7864), and parallelism validation tests. The VO volume-boosting method passed all validation tests [intra-assay coefficent of variation (%CV): 16.3 for ELISA and 5.9 for RIA; inter-assay %CV: 10.3; spiked recovery: 102.0%; dilution recovery: 93.0%; linearity R ²: 0.9435; log of serial dilutions was parallel] and provided enough extract for multiple assays from an individual baitfish. Based on these results, we conclude that the VO volume-boosting method presents a means for determining cortisol from individual baitfish using either RIA or ELISA assays.     

Intermittent Stress in Pigs: Behavioural and Pituitary-Adrenocortical Reactivity

Abstract

Ninetysix pigs, half females and half castrated males from 12 litters, were housed in 24 groups of 4 litter mates (2 females, 2 castrates). The pigs were weaned at 4 weeks of age and from the age of 115 days half of the groups were subjected to a schedule of unpredictable, inescapable electroshocks for 33 days, and half served as control. The adrenocortical reactivity to ACTH stimulation and to an emotional stressor (the procedure associated with initiation of blood collection) was measured in females, whereas the behavioural and pituitary-adrenocortical reactivity to an open field test was measured in castrates, as was the behavioural reactivity in a social test. Intermittent stress increased adrenocortical reactivity to ACTH stimulation significantly. This effect was present within the first week of intermittent stress (p < 0.06), but no difference was found after 4–5 weeks of stress. In contrast, the adrenocortical reactivity to additional emotional stressors was unaffected by stress after one week of intermittent stress, but increased after 4–5 weeks. In the open field test, centre location increased after one week of intermittent stress, whereas exploration decreased and walking and ambulation increased after 4–5 weeks of stress. In addition, 4–5 weeks of intermittent stress decreased aggressiveness in a social test. In conclusion, the change in reactivity to an additional acute stressor during intermittent stress appeared in different variables and contexts at different times during the course of intermittent stress. Thus, assessment of stress based on changed reactivity has to include several distinct measures of behavioural and hormonal reactivity.     

Performance and endocrine responses of group housed weaner pigs exposed to the air quality of a commercial environment

The growth performance and endocrine responses of male weaner pigs (3 to 8 weeks of age) was evaluated in two different environments (clean and dirty) and housing (single or groups of 10 pigs/pen) conditions. The dirty environment contained significantly elevated ammonia, carbon dioxide and dust levels compared with the clean environment. Pigs grew faster and consumed more feed in the clean environment and this was associated with reduced plasma cortisol concentrations compared with pigs in the dirty environment. Pigs housed in groups in the dirty environment had increased β-endorphin and decreased IGF-I concentrations compared to group housed pigs in the clean environment. Feed conversion efficiency did not differ due to environment or group housing. Plasma concentration of cortisol, β-endorphin, IGF-I and IGF-II did not differ between single and group housed pigs. Activity of the hypothalamic-pituitary-adrenal (HPA) axis was greater in response to environmental conditions than group housing, and this was associated with reduced growth in weaner pigs.     

Molecular genetics of the adrenocortical axis and breeding for robustness

The concept of robustness refers to the combination of a high production potential and a low sensitivity to environmental perturbations. The importance of robustness-related traits in breeding objectives is progressively increasing toward the production of animals with a high production level in a wide range of climatic conditions and production systems, together with a high level of animal welfare. Current strategies to increase robustness include selection for "functional traits," such as skeletal and cardiovascular integrity, disease resistance, and mortality at various stages. It is also possible to use global evaluation of sensitivity to the environment (eg reaction norm analysis or canalization), but these techniques are difficult to implement in practice. The glucocorticoid hormones released by the adrenal cortex exert a wide range of effects on metabolism, the cardiovascular system, inflammatory processes, and brain function, for example. Protein catabolism toward energy production and storage (lipids and glycogen) supports their pivotal role in stress responses aiming at the adaptation and survival of individuals under strong environmental pressure. Large individual variations have been described in adrenocortical axis activity, with important physiopathological consequences. In terms of animal production, higher cortisol levels have negative effects on growth rate and feed efficiency and increase the fat:lean ratio of carcasses. On the contrary, cortisol has positive effects on functional traits and adaptation. Intense selection for lean tissue growth and more generally high protein output during the past decades has concomitantly reduced cortisol production, which may be responsible for the negative effects of selection on functional traits. In this paper, we review experimental evidence suggesting that the balance between production and functional traits was modified in favor of improved robustness by selecting animals with higher adrenocortical axis activity, as well as the molecular genetic tools that can be used to fine-tune this objective.     

The Effect of Different Physical Exercise on Plasma Leptin,Cortisol, and Some Energetic Parameters Concentrations in Purebred Arabian Horses

Leptin is a hormone that coordinates food intake, energy expenditure, and metabolic rate; it is, however, unknown how exercise influences plasma leptin concentration in horses. The aim of the study was to evaluate the exercise-induced changes in plasma leptin concentration in purebred Arabian horses competing in races and endurance rides. A total number of 26 horses (12 purebred Arabian racehorses aged 3–5 years and 14 endurance horses aged 7–14 years) were studied during performing routine physical exercise. From each horse, blood samples were collected at rest and immediately after the exercise session. Concentrations of plasma leptin, cortisol, lactic acid, uric acid, triacylglycerols, glycerol and free fatty acids were determined. Exercise-induced increase in cortisol and free fatty acid values were higher in endurance horses than in racehorses. Neither endurance nor race exercises influenced the plasma leptin concentration. Further research is needed to elucidate the effects of different types of exercise on leptin synthesis and secretion in horses.     

Validation,use, and disadvantages of enzyme-linked immunosorbent assay kits for detection of cortisol in channel catfish,largemouth bass,red pacu,and golden shiners

Measurement of cortisol response is an important tool to asses stress in fisheries research. Radioimmunoassay (RIA) is a common method for the measure of cortisol in fish. Use of enzyme-linked immunosorbent assays (ELISA) to detect cortisol would eliminate health hazards, costs of handling radioisotopes, and the short stability time associated with RIA. Enzyme-linked immunosorbent assays have been used for the determination of cortisol in several fish species. However, the ELISA method of cortisol determination in fish lacks proper validation testing. We conducted validation procedures for multiple commercial cortisol ELISA kits and compared the results to RIA. The assays were tested for four species: (1) channel catfish Ictalurus punctatus, (2) largemouth bass Micropterus salmoides, (3) red pacu Piaractus brachypomus, and (4) golden shiners Notemigonus crysoleucas. We evaluated the ELISA methods against RIA, and determined that at least one kit is suitable (accuracy: mean recovery of spiked samples, 102.8%; reproducibility: interassay coefficient of variation < 10.5% for all species; precision: intra-assay coefficient of variation < 16.8% for all species; linearity: R ² > 0.96 for all species) for the measurement of cortisol response in fish and comparative determination of stress. All of the ELISA assay results varied by more than 10% from the cortisol concentrations detected by the RIA. The high variability of the kit results indicates that commercial ELISA kits could be utilized for qualitative determination of cortisol in fish, but should be fully validated in each laboratory for each species before being used for research.     

Live microbial feed supplement in aquaculture for improvement of stress tolerance

Two bacterial strains, Lactobacillus fructivorans, isolated from sea bream (Sparus aurata) gut, and Lactobacillus plantarum, isolated from human faeces, were administered simultaneously, during sea bream development, using Brachionus plicatilis and/or Artemia salina as vectors. The probiotic treatment significantly affected gut colonization. To test the probiotic influence on stress responsiveness, sea bream fry, 47 days post-hatching (p.h.), were subjected to pH stress (from 8.6 to 6.3) and cumulative mortality, cortisol levels and HSP70 gene expression were analysed. Cortisol was selected, since under stress conditions its level increases. HSP70 was selected with consideration of its wide involvement in response to a great number of injuries, and because it protects cells probably by binding and refolding damaged proteins. The results obtained indicated that the administration of probiotic to sea bream fry induced higher HSP70 levels, indicating a greater potentiality to respond to the harmful conditions possibly present in fish farms. This hypothesis is supported by the fact that the levels of cortisol found were significantly lower (P < 0.05) in both groups under probiotic treatment. When pH was used as a stressor, it induced a higher cumulative mortality in the control; the mortality was found to be significantly lower in both treated groups. Interestingly, a significant increase (P < 0.05) in HSP70 gene expression was observed in all stressed groups. These results suggest an improvement in tolerance to acute stress of fry fed with probiotics.     

Effect of cortisol infusion patterns and castration on metabolic and immunological indices of stress response in cattle

This study tested the hypotheses that: (1) either acute stress induced by Burdizzo castration, or cortisol infusion would modulate plasma glucose, insulin and growth hormone (GH) concentrations; and (2) immune modulation induced by cortisol would be dependent on the pattern, intensity and duration of circulating cortisol concentrations. Fifty 9.2-month-old Holstein×Friesian bulls (232±2.0 kg) were blocked by weight and randomly assigned to one of five treatments (n=10 per treatment): (1) sham handled control; (2) Burdizzo castration; (3) hydrocortisone infusion to mimic the castration-induced secretion pattern of cortisol; (4) hourly pulse infusion of hydrocortisone; and (5) sustained infusion of hydrocortisone for 8 h. Blood samples were collected intensively on day 0, and weekly from days 1 to 35. Castration acutely increased plasma cortisol, GH and haptoglobin concentrations, suppressed lymphocyte in vitro interferon-γ (IFN-γ) production, but had no effect on plasma glucose and insulin concentrations. Cortisol infusion to simulate the castration-induced secretion pattern of cortisol, and pulse infusion of cortisol did not suppress the IFN-γ production. A sustained infusion of cortisol resulted in the transient suppression of IFN-γ production. Moreover, the sustained cortisol infusion resulted in increased plasma glucose, insulin and GH concentrations. The overall 14-day feed intakes and 35-day growth rates were not affected by treatments. In conclusion, cortisol infusion to induce immune suppression in vivo occurred only at pharmacological doses. Within physiological ranges, cortisol was not associated with the suppression of immune function, indicating that during castration cortisol per se is not responsible for the suppression of in vitro IFN-γ production.     

子宫内膜炎患牛唾液生化变化的初步研究

[目的]探究子宫内膜炎患牛唾液生物标志物特点,[方法]研究收集试验组子宫内膜炎患牛和对照组健康奶牛唾液,针对所选10种生物标志物使用相关试剂盒在自动生化分析仪上进行检测,并将所得数据进行对比分析。[结果]相比健康奶牛,子宫内膜炎患牛唾液生物标志物的升高项为α唾液淀粉酶、皮质醇、乳酸和尿酸,而腺苷脱氨酶、乙酰胆碱酯酶水平、pH值、钠钾比则下降明显。[结论]说明应激、炎症和氧化应激在患子宫内膜炎奶牛体内的存在,钠营养状况略受影响。