Fish faced with stressful stimuli launch an endocrine stress response through activation of the hypothalamic-pituitary-interrenal (HPI-) axis to release cortisol into the blood. Scientifically validated biomarkers to capture systemic cortisol exposure over longer periods of time are of utmost importance to assess chronic stress in governmental, wildlife, aquaculture and scientific settings. Here we demonstrate that cortisol in scales of common carp (Cyprinus carpio L.) is the long-sought biomarker for chronic stress. Undisturbed (CTR) and daily stressed (STRESS) carp were compared. Dexamethasone (DEX) or cortisol (CORT) fed fish served as negative and positive controls, respectively. Scale cortisol was quantified with a validated ultra-performance liquid chromatography tandem mass spectrometry method. An increase in scale cortisol content was found in STRESS and CORT but not in CTR and DEX fish. Scale cortisol content reflects its accumulation in a stressor and time dependent manner and validates the scale cortisol content as biomarker for chronic stress. Plasma analyses confirmed that (i) CTR, DEX and CORT treatments were effective, (ii) plasma cortisol of STRESS fish showed no signs of chronic HPI-axis activation, and (iii) plasma cortisol is a poor predictor for chronic stress. The expression of HPI key genes crf, pomc, and star were up-regulated in STRESS fish in the absence of a plasma cortisol response, as was the target gene of cortisol encoding subunit α1 of the Na+/K+-ATPase in gills. When lost, scales of fish regenerate fast. Regenerated scales corroborate our findings, offering (i) unsurpassed time resolution for cortisol incorporation and as such for stressful events, and (ii) the possibility to investigate stress in a well defined and controlled environment and time frame creating novel opportunities for bone physiological research. We conclude that the cortisol content in ontogenetic and regenerated scales is an innovative biomarker for chronic stress offering ample applications in science and industry.

Demand for meat from free-range broiler chickens is increasing in several countries. Consumers are motivated by better animal welfare and other product attributes such as quality and taste. However, scientific literature is not unanimous about whether free-range access influences quality, composition, and taste of the meat. Because chickens normally do not use free-range areas optimally, it is possible that provision of more suitable shelter will lead to more pronounced differences between chickens raised indoors and outdoors. In this study, an experiment with 2 production rounds of 600 slow-growing broilers each was performed. In each round, 200 chickens were raised indoors (IN), 200 had free-range access to grassland with artificial shelter (AS), and 200 had free-range access to short-rotation coppice with willow (SRC). Free-range use, feed intake, and growth were monitored, and after slaughter (d72) meat quality, composition, and taste were assessed. Free-range use was higher in SRC than in AS chickens (42.8 vs. 35.1%, P < 0.001). IN chickens were heavier at d70 than AS and SRC chickens (2.79 vs. 2.66 and 2.68 kg, P = 0.005). However, feed intake and conversion did not differ. Breast meat of chickens with free-range access was darker (P = 0.021) and yellower (P = 0.001) than that of IN chickens. Ultimate pH was lower (5.73 vs. 5.79; P = 0.006) and drip loss higher (1.29 vs. 1.09%; P = 0.05) in IN versus AS chickens. The percentage of polyunsaturated fatty acids was higher in AS than in IN meat (35.84 vs. 34.59%; P = 0.021). The taste panel judged breast meat of SRC chickens to be more tender (P = 0.003) and less fibrous (P = 0.013) compared to that of AS and IN chickens, and juicier compared to the IN chickens (P = 0.017). Overall, freerange access negatively affected slaughter weight, but positively affected meat quality, taste, and composition. Only a few differences between AS and SRC were found, possibly due to limited differences in free-range use.

When added to the feed of broiler breeder hens, dietary polyunsaturated fatty acids (FA) can be incorporated into the yolk and therefore become available to the progeny during their early development. The mechanism involved in lipid metabolism and deposition in the egg may be influenced by breeder age. Before the effect of an elevated concentration of certain polyunsaturated FA on the embryo can be investigated, the effect at breeder level and egg quality must be further assessed. The aim of the present experiment was to evaluate the effects of dietary n-6/n-3 ratios and dietary eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) ratios, provided to broiler breeder hens, in terms of their zoo technical performance, egg quality, and yolk FA composition. Starting at 6 wk of age, 640 Ross-308 broiler breeder hens were fed 1 of 4 different diets. The control diet was a basal diet, rich in n-6 FA. The 3 other diets were enriched in n-3 FA, formulated to obtain a different EPA/DHA ratio of 1/1 (EPA = DHA), 1/2 (DHA), or 2/1 (EPA). In fact, after analysis the EPA/DHA ratio was 0.8, 0.4, or 2.1, respectively. Dietary EPA and DHA addition did not affect the performance of the breeder hens, except for egg weight. Egg weight was lower (P < 0.001) for all n-3 treatments. Dietary EPA improved number of eggs laid in the first 2 wk of the production cycle (P = 0.029). The absolute and relative yolk weight of eggs laid by EPA = DHA fed hens was lowest (P = 0.004 and P = 0.025, respectively). The EPA and DHA concentrations in the yolk were highly dependent on dietary EPA and DHA concentrations with a regression coefficient equal to 0.89. It can be concluded that dietary EPA and DHA can be incorporated in the breeder egg yolk to become available for the developing embryo, without compromising the performance and egg quality of the flock.

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