Skin Mucus as a Relevant Low-Invasive Biological Matrix for the Measurement of an Acute Stress Response in Rainbow Trout (Oncorhynchus mykiss)

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Abstract

Skin mucus is a non-lethal and low-invasive matrix appropriate to assess fish welfare as it contributes to their defence against external aggressions and reflects changes in fish health status. However, more information on the response of this matrix to specific stressors is needed. In this study, rainbow trout (Oncorhynchus mykiss) specimens were subjected to an acute stress by air exposure and sampled after 1, 6, and 24 h post-stress. Blood and skin mucus were collected, and a battery of biochemical biomarkers were measured in both matrices. Cortisol and glucose values showed the expected classical stress response in plasma, increasing after the acute stress. The same pattern was observed in skin mucus, corroborating previous data in fish, and allowing us to confirm that skin mucus can be a useful complementary matrix for stress assessment in fish. The results showed sensitivity to hypoxic stress in skin mucus for cortisol, glucose, alkaline phosphatase (ALP), aspartate transaminase (AST), alanine aminotransferase (ALT), creatinine kinase (CK), and calcium. From the 15 parameters evaluated, 12 did not show statistically significant changes between plasma and mucus; therefore, using skin mucus cannot replace the use of plasma. Finally, the principal component analysis in skin mucus revealed a complete separation between the two experimental groups, being ALP, AST, glucose, cortisol, and CK, the biomarkers that contributed the most to this separation.

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Yoav Benjamini, Yosef Hochberg (1995)

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Exploration of the hypothalamic-pituitary-adrenal function as a tool to evaluate animal welfare.

Bonne Beerda, Stéphane Andanson, Pierre Mormède … (2007)

Measuring HPA axis activity is the standard approach to the study of stress and welfare in farm animals. Although the reference technique is the use of blood plasma to measure glucocorticoid hormones (cortisol or corticosterone), several alternative methods such as the measurement of corticosteroids in saliva, urine or faeces have been developed to overcome the stress induced by blood sampling itself. In chronic stress situations, as is frequently the case in studies about farm animal welfare, hormonal secretions are usually unchanged but dynamic testing allows the demonstration of functional changes at several levels of the system, including the sensitization of the adrenal cortex to ACTH and the resistance of the axis to feedback inhibition by corticosteroids (dexamethasone suppression test). Beyond these procedural aspects, the main pitfall in the use of HPA axis activity is in the interpretation of experimental data. The large variability of the system has to be taken into consideration, since corticosteroid hormone secretion is usually pulsatile, follows diurnal and seasonal rhythms, is influenced by feed intake and environmental factors such as temperature and humidity, age and physiological state, just to cite the main sources of variation. The corresponding changes reflect the important role of glucocorticoid hormones in a number of basic physiological processes such as energy metabolism and central nervous system functioning. Furthermore, large differences have been found across species, breeds and individuals, which reflect the contribution of genetic factors and environmental influences, especially during development, in HPA axis functioning. Usually, these results will be integrated with data from behavioral observation, production and pathology records in a comprehensive approach of farm animal welfare.

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Mucosal Immunity and B Cells in Teleosts: Effect of Vaccination and Stress

David Parra, Felipe E. Reyes-López, Lluis Tort (2015)

Fish are subjected to several insults from the environment, which may endanger animal survival. Mucosal surfaces are the first line of defense against these threats, acting as a physical barrier to protect the animal but also functioning as an active immune tissue. Thus, four mucosal-associated lymphoid tissues (MALTs), which lead the immune responses in gut, skin, gills, and nose, have been described in fish. Humoral and cellular immunity, as well as their regulation and the factors that influence the response in these mucosal lymphoid tissues, are still not well known in most fish species. Mucosal B-lymphocytes and immunoglobulins (Igs) are key players in the immune response that takes place in those MALTs. The existence of IgT as a mucosal specialized Ig gives us the opportunity of measuring specific responses after infection or vaccination, a fact that was not possible until recently in most fish species. The vaccination process is influenced by several factors, being stress one of the main stimuli determining the success of the vaccine. Thus, one of the major goals in a vaccination process is to avoid possible situations of stress, which might interfere with fish immune performance. However, interaction between immune and neuroendocrine systems at mucosal tissues is still unknown. In this review, we will summarize the latest findings about B-lymphocytes and Igs in mucosal immunity and the effect of stress and vaccination on B-cell response at mucosal sites. It is important to point out that a limited number of studies have been published regarding stress in mucosa and very few about the influence of stress over mucosal B-lymphocytes.