Effect of cortisol and triiodothyronine bath treatments on the digestive enzyme profile and growth of Catla catla larvae during ontogenic development

Digestive enzyme profile is a good indicator of the nutritional and health status of the fish. The present investigation aims to evaluate the effect of exogenous bath treatment of hormones, cortisol and triiodothyronine, on the digestive enzyme activities and growth of carp Catla catla (Ham.) during ontogenic development. Catla larvae (4 days old) were given bath treatment with cortisol (hydrocortisone, 0.2 mg L^?1), 3,5,3′‐triiodothyronine (T3, 2.5 mg L^?1) and a combination of cortisol and T3 for 30 min. Digestive enzyme profile was recorded on every third day and was continued for 30 days. Larvae were fed with live food for initial 14 days and then weaned to mix feeding of live food and prepared diet. Significantly (P < 0.05) higher amylase, total protease, trypsin, chymotrypsin, lipase, chitinase and chitinobiase activities were found in the hormone‐treated groups compared to the control one during ontogenic development. Among the treated groups, amylase activity was highest in cortisol‐treated larvae. Total protease, trypsin, chymotrypsin, lipase, chitinase and chitinobiase activities were significantly (P < 0.05) higher in larvae exposed at combined treatment of cortisol and T3 compared to the other two groups in most sampling days. Average length, weight and specific growth rate of treated larvae were higher compared to the control one. The combined bath treatment of cortisol and triiodothyronine influenced the digestive enzyme activities of catla larvae and thereby enhanced the growth at early developmental stage. This helps the larvae to overcome the problems associated with early developmental stage.     

Docosahexaenoic acid (C22:6n‐3) alters cortisol response after air exposure in Prochilodus lineatus (Valenciennes) larvae fed on enriched Artemia

This study investigated the effect of different proportions of dietary docosahexaenoic acid (DHA) and arachidonic acid (ARA) on the growth, survival and stress response of curimba (Prochilodus lineatus Valenciennes) larvae, an omnivorous freshwater species, fed on enriched Artemia. Six different experimental emulsions were formulated to have two levels of ARA (A‐0.5 and A‐1.0), two levels of DHA (D‐0.5 and D‐1.0) two combinations of ARA and DHA (AD‐3:1 and AD‐1:3). There were no significant effects of treatments on curimba standard length and survival at the end of the experiment. The results showed a negative correlation between dietary DHA and whole‐body cortisol after 60 s of air exposure. The resting concentration of cortisol was not affected by the diets (p = .424). After air exposure, all treatments except for D‐1.0 (p = .137) and D‐0.5 (p = .545) exhibited a marked cortisol rise. These results showed that DHA supplementation can have an attenuating effect on cortisol response after air exposure.     

Salivary cortisol and behavior in therapy dogs during animal-assisted interventions: A pilot study

Animal-assisted interventions (AAIs) have been associated with positive effects on human psychological and physiological health. Although the perception of quality standards in AAIs is high, only few investigations have focused on potential welfare implications for therapy dogs linked to their performance in AAIs. The standardized program “multiprofessional animal-assisted intervention (MTI)” has been carried out in adult mental health care, significantly improving patients' prosocial behaviors. In the present study, we monitored salivary cortisol and behavioral measures in therapy dogs that participated in MTI group therapy sessions in an in-patient substance abuse treatment facility. Work-related activity (lay, sit, stand, walk, and run), behavior (lip licking, yawning, paw lifting, body shake, tail wagging, and panting), response to human action (taking food treats and obeying commands), and salivary cortisol levels were analyzed over the course of 5 subsequent MTI working sessions in experienced therapy dogs (N = 5), aged 5.4 ± 2.8 years (mean ± standard deviation). Salivary cortisol levels decreased from presession to postsession in sessions 1, 2, and 3. However, only in session 4 and 5, postsession cortisol levels were significantly lower than presession levels (P = 0.043). There was no difference between salivary cortisol levels sampled on a nonworking day at home and work-related levels sampled at the therapy site. None of the behavioral parameters varied significantly over the course of the 5 MTI sessions. Both lip licking (P = 0.038) and body shake (P = 0.021) were positively correlated with the decline in cortisol during session 5. The study results suggest that trained dogs are not being stressed by repeated participation in in-patient substance abuse therapy sessions. Further investigation into the effects of animal-assisted therapy on dogs' physiological markers and behavior is warranted.     

Effects of a topical anaesthetic formulation and systemic carprofen, given singly or in combination, on the cortisol and behavioural responses of Merino lambs to castration

Objective   To determine the effectiveness of a topical anaesthetic formulation (Tri-Solfen) with or without the administration of a non-steroidal anti-inflammatory drug (carprofen) on the pain and distress response associated with ring or surgical castration of ram lambs.
Procedures   Merino ram lambs (n = 78) were allocated to 10 treatment groups: 4 groups of knife-castrated lambs and 4 groups of ring-castrated lambs received carprofen (4 mg/kg SC) and Tri-Solfen; 2 control groups (sham) received carprofen at 0 or 4 mg/kg SC. Measurements included plasma cortisol and haptoglobin concentrations, haematology, and behaviour, including posture.
Results   Knife-castrated lambs had higher peak cortisol and integrated cortisol responses for the first 6 h after treatment and greater concentration s of circulating acute phase proteins than ring-castrated lambs, both of which were significantly different from the sham controls. Tri-Solfen applied to the knife castration wound significantly reduced both the peak plasma cortisol concentration and the integrated cortisol response for the first 6 h and improved lying behaviour in the first 12 h. Carprofen reduced the cortisol response to knife castration at 30 min, but elevated the cortisol responses at 24 and 48 h. Carprofen nearly halved the number of acute pain behaviours associated with ring castration. There were no significant additive or synergistic effects from combining the analgesic treatments. Tri-Solfen applied to the tail wound provided no detectible benefits during ring castration + tail docking.
Conclusions   The physiological and behavioural responses suggest that ring castration has less impact on the lamb than knife castration. The specific analgesic treatments can provide modest amelioration of the pain and discomfort associated with castration. Alternative doses or application methods may enhance their efficacy.     

Alternative matrices for cortisol measurement in fish

Plasma cortisol is the most commonly used indicator of stress in fish but, as the blood sampling procedure itself can be a source of stress, it would be helpful to measure cortisol using less invasive matrices. It is also necessary to find alternative matrices as stress indicators in dead fish in which blood sampling is impossible. In the present study, we investigated transport stress in three aquaculture species, European sea bass (Dicentrarchus labrax L.), common carp (Cyprinus carpio L.) and rainbow trout (Oncorhynchus mykiss Walbaum), by cortisol determination (radioimmunoassay) in plasma and other matrices (skin mucus, gut content, lateral muscle and caudal fin). Cortisol significantly increased after transport in all species and matrices, except in the sea bass gut content, where it remained unchanged. The three species responded to transport stress by producing different cortisol levels. In conclusion, the significant correlation found between plasma cortisol and most of the other matrices opens up the possibility of using them to evaluate stress in fish: mucus sampling is a less invasive method than blood sampling, and in addition to muscle and fin sampling, it can be used in postmortem fish.     

Changes in selected stress and immune‐related genes in Atlantic cod,Gadus morhua,following overcrowding

Using Atlantic cod, Gadus morhua, as a model for the stress response in gadoid fish, the changes in the expression of some stress and immune genes as well as the profiles of plasma cortisol were examined. Adult fish were kept at a density of ca. 100 kg m^?3 by lowering the water level in the rearing tank for 1 h and this short‐term crowding stress was repeated thrice over a 12‐h interval period. Blood samples were collected before exposure and at 2, 24 and 72 h post crowding. Plasma cortisol level significantly increased at 2 h post crowding but returned to pre‐crowding levels 24 h after exposure. The relative expression of the stress response genes, glucose transporter‐3 and a putative heat shock protein 70 significantly increased at 2 and 24 h post crowding respectively. Significant up‐regulation of the pro‐inflammatory cytokines, interleukin (IL)‐1β and IL‐8, as well as anti‐bacterial genes, g‐type lysozyme and bactericidal permeability‐increasing protein/lipopolysaccharide‐binding protein (BPI/LBP) was also observed at 2 h and the levels were maintained until 72 h post exposure, except for BPI/LBP which had maximum up‐regulation at 24 h. The present observations have implications with respect to fish welfare and assessment of the health status of the farmed fish.     

Validation of enzyme-linked immunosorbent assay for measurement of faecal cortisol in fish

Quantification of glucocorticoid (GC) levels in faeces has become an established method for the non-invasive assessment of adrenocortical activity. These hormones are frequently determined in plasma samples as parameters of adrenal activity and response to stress. Because GCs are metabolized and excreted with both intact hormone and their metabolites present in faeces, the concentration of GCs can be measured in excreta. Faecal samples present the advantages of easy collection, no stress to the animal and elimination of the issue of potentially misleading acute GC spikes. The aim of this study was to determine if an enzyme-linked immunosorbent assay (ELISA) for cortisol was appropriate for monitoring adrenocortical activity in faecal casts of fishes. Performance of the cortisol ELISA was validated by comparison to high-performance liquid chromatography, which is an established method for measuring free GCs and GC metabolites in faeces. Parallelism and sample extraction efficiency were compared for the two methods. Pearson's correlation across samples for these methods was 0.996. Results demonstrated that the ELISA was an efficient, sensitive and reliable method for cortisol measurement in faecal extracts, which should permit integration of non-invasive stress monitoring into studies of fish behaviour and physiology.     

Benefits of using the probiotic Efinol®L during transportation of cardinal tetra, Paracheirodon axelrodi (Schultz), in the Amazon

The objective of this experiment was to test the probiotic Efinol^®L during transportation of cardinal tetra, Paracheirodon axelrodi (Schultz). For the transportation, fish were distributed in 18 plastic tanks, of which nine contained the Efinol^®L (10 mg L⁻¹; probiotic treatment) and the remaining had no probiotic (control treatment). Transport lasted 24 h and three different boxes of each treatment were sampled at 3, 12 and 24 h. Up to the 12-h sampling period, no significant difference in the survival was observed; on the other hand, survival was higher at the end of the transport (24 h) in the probiotic treatment. No significant difference was found in dissolved oxygen and temperature between treatments. Conductivity, pH and alkalinity increased along the transport, but without a difference between treatments. Ammonia increased in all treatments, although it was significantly lower in the probiotic group at 24 h. Cortisol levels were significantly higher in all transported fish when compared with the basal values. At 24 h, cortisol levels in control fish were significantly higher than those in the probiotic treatment. With the observed results, we are able to conclude that the probiotic Efinol^®L is efficient during cardinal transport, lowering the mortality and helping maintain water quality possibly by lowering metabolic wastes.     

Physiological responses of juvenile wedge sole Dicologoglossa cuneata (Moreau) to high stocking density

Physiological responses to a high stocking density were tested in juvenile wedge sole Dicologoglossa cuneata (Moreau). Fish were kept at low (1 kg m⁻²), medium (3 kg m⁻²) and high (9 kg m⁻²) stocking densities for 22 days. No differences in the weight, length, survival and hepatosomatic index were observed among treatments. Basal plasma cortisol and osmolality were found to be directly and positively related to stocking density. A mild increase in plasma glucose was seen at medium density, and plasma protein was elevated at medium and high densities. The liver glucose and glycogen content was inversely related to stocking density. The liver triglyceride level was significantly elevated at the highest density, and the α-amino acid content decreased at the highest density. In muscle, glucose levels were significantly higher in fish kept at the lowest density; the α-amino acid content was elevated in fish kept at high density. In conclusion, plasma cortisol levels indicated an increasing stress level depending on the culture density, but significant changes in energy reserves did not occur in tissue (mainly liver and muscle glycogen and glucose reserves were significantly affected).