The fasted and post-prandial physiological responses of the Patagonian blennie Eleginops maclovinus.

Eleginops maclovinus is a native species with potential for Chilean aquaculture. Understanding the variations between the post-prandial and fasted metabolic responses can contribute to improving the aquaculture of this species. This study aimed to characterize variations in intermediate metabolism during the course of the day in the liver, serum, and gills of fed and unfed fish. For this, 72 fish were assigned to two experimental groups, "fed" and "fasted". The first group was fed "ad libitum" at 8.30, while the fasted group was not fed for 24 h. Samples were taken from both groups at 9:00, and every 2 h: 11:00, 13:00, 15:00, 17:00, and 19:00. In the fed group, food spent a long time in the gastrointestinal tract, with a large increase in stomach size and without evidence of complete emptying of the stomach at 19:00 (10.5 h post-feeding). In serum, the levels of amino acids, glucose, and triglycerides presented significant differences with peak levels at different times of day in the fed group. The cortisol in the fasted group presented a diurnal pattern with high levels during the morning and very low levels after 13:00, while in the fed group, the high cortisol variability did not allow a clear pattern to be established. In the liver, the effect of time on the enzymatic activity of the intermediary metabolism was greater compared to the effect of feeding. In the liver, enzyme activity decreased at later hours of the day, while glycogen levels increased at later hours of the day in both groups: but its levels were higher in the fed group. In gills, as well as in the liver, time had a greater effect than feeding on intermediate metabolism, since feeding only had a significant effect on the levels of hexokinase, lactate, and amino acids, suggesting an effect on carbohydrate metabolism. Meanwhile, time significantly affected the levels of Na+, K+-ATPase, glutamate dehydrogenase, aspartate aminotransferase, amino acids, and proteins, suggesting an effect on amino acid metabolism. In conclusion, the intermediate metabolism of E. maclovinus presents variations according to the time of day, with an increased metabolism during the morning and decreased metabolism as the day progresses, especially at the hepatic level. The gill tissue, despite not being a metabolic organ, presents feeding-dependent variations in its metabolism. Additional studies will be required to corroborate if coordinating a feeding strategy during the first hours of the day when metabolism is greater would improve the growth of E. maclovinus.

Intestinal variation of serotonin, melatonin, and digestive enzymes activities along food passage time through GIT in Salmo salar fed with supplemented diets with tryptophan and melatonin.

In teleosts, peripheral serotonin (5-HT) and melatonin (MEL) are synthesised in the gastrointestinal tract (GIT) and regulate secretion and motility processes. Their production is regulated by diet and the passage of food through the GIT. This study aimed to evaluate how intestinal 5-HT, melatonin, and the activity of digestive enzymes varied with food passage time through GIT in Atlantic salmon (Salmo salar). We fed fish diets supplemented with tryptophan and melatonin (L-Trp 2.5% and MEL 0.01%) and measured the activity of digestive enzymes (amylase, lipase, and total protease) in the pyloric caeca, midgut, and hindgut at different times after feeding. 5-HT levels increased in all GIT portions and diets at 120 min post-intake and were highest in the pyloric caeca. Intestinal enzymatic activity was varied with diet, post-intake time and in different intestinal portions. In conclusion, food passage time directly affects GIT 5-HT secretion and digestive enzyme activity in S. salar, and diet composition regulates S. salar GIT function.

Effect of Flavobacterium psychrophilum on the neuroendocrine response of rainbow trout (Oncorhynchus mykiss) in a time course experiment.

The aim of this study was to examine the effects of Flavobacterium psychrophilum, a pathogen that is economically important in the aquaculture sector, on the neuroendocrine response of Oncorhynchus mykiss during a time course experiment with sampling at 0.5, 1, 2, 6, 10, and 30 days post injection (dpi). In the brain, serotonin (5HT) content increased in the infected group at all the measured time points, a similar pattern was observed for 5-hydroxyindole-3-acetic acid (5HIAA). Infected fish presented an increase in brain dopamine levels on day 0.5 and 1 dpi. A non-significant variation in noradrenaline levels was observed on all treatment days. Foregut 5-HT and 5-HIAA content in the infected group presented the highest 5-HT concentrations with 248.6 and 983.5 ng/g tissue at 0.5 dpi respectively. Midgut 5-HT and 5-HIAA levels presented the highest 5-HT concentrations, 486.9 ng/g tissue and 1006.4 ng/g tissue respectively, at the beginning of the experiment (0.5 dpi). 5-HT levels in the hindgut presented the highest concentrations with 233.9 ng/g tissue at 0.5 dpi, while 5-HIAA presented the highest concentrations, 690.5 ng/g tissue, at the same time point. After injection with F. psychrophilum the neuroendocrine response in rainbow trout was tissue dependent. Brain levels of 5HT and 5HIIA indicate that the neuroendocrine response increased together with dopamine following intramuscular infection. These increases are in line with reports from other authors, indicating an early response of catecholamines as neurotransmitters to stressful stimulus. In addition the intestinal response was also increased, implying that there could be a possible relationship between the serotonergic system at the intestinal level and the immune system.

The effect of alterations in salinity and temperature on neuroendocrine responses of the Antarctic fish Harpagifer antarcticus.

Increased levels of tissue monoaminergic neurotransmitters, as well as circulating catecholamines, appear to play a role in the regulation of the physiological responses of teleost fish. Harpagifer antarcticus is a stenothermic, Antarctic notothenioid fish. The aim of this study was to determine the effect of increased seawater temperature and decreased salinity on the levels of 5-HT, 5-HIAA, DA, and Noradrenaline in the brain, stomach, and gut of H. antarcticus. Wild-gathered fish were acclimatized to habitat conditions (2 °C, 33 PSU) prior to placement in aquaria with 4 temperatures (2, 5, 8 and 11 °C) and 3 salinities (23, 28 and 33 PSU) for 10 days. Fish exposed to 11 °C had higher levels of the brain neurotransmitters than those at 2 °C. Concomitant exposure to low salinity exacerbated the effect of exposure to 11 °C. At lower temperatures, concomitant alterations in salinity induced differential effects on brain neurotransmitters. When fish were exposed to 28 PSU, 5-HIAA, DA, and Noradrenaline levels at 5 and 8 °C presented no significant differences with those at 2 °C. In contrast, only 5HT and 5-HIAA levels in fish at 33 PSU were elevated at 5 and 8 °C respectively. Fish at 28 and 33 PSU had lower Gut 5HT levels at the 3 elevated temperatures, meanwhile fish at 23 PSU showed a biphasic effect when exposed to elevated temperatures. 5-HIAA levels decreased at 5 and 8 °C at 33 PSU. Stomach 5HT levels also showed a differential response at the 3 salinity levels when exposed to increased temperatures. At 11 °C, 5HT levels were markedly higher than those at 2 °C for fish at 33 PSU, moderately elevated for fish at 28 PSU, and lower for fish at 23 PSU, meanwhile 5-HIAA levels only increased with temperature at 33 PSU. These findings indicate that rapid exposure to alterations in temperate with or without concomitant changes in salinity is associated with differential responses in tissue monoaminergic neurotransmitter levels. The relatively high changes in neurotransmitter levels in fish exposed to moderate salinity and high temperature changes may indicate the physiological plasticity of H. antarcticus to possible changes in ocean temperature and salinity.

The effects of intraperitoneal administration of Francisella noatunensis subsp. noatunensis on hepatic intermediary metabolism and indicators of stress in Patagonian blennie Eleginops maclovinus.

Francisellosis is a disease produced by Francisella spp. which affects farmed fish. Eleginops maclovinus specimens can be caught close to salmon farming centers, feeding on un-consumed pellet, making the transmission of pathogens such as Francisella noatunensis possible. The aim of this study was to evaluate the effect of F. noatunensis on liver intermediary metabolism in E. maclovinus. 144 fish were injected intraperitoneally with F. noatunensis at a low dose LD (1 × 101 cells/µL), medium dose MD (1 × 105 cells/µL), high dose HD (1 × 1010 cells/µL), or with culture medium C (control), and sampled at 1, 3, 7, 14, 21 and 28 days post injection (dpi). No mortality was recorded during the experimental period, but there was a marked metabolic response in fish injected with high doses. Metabolites in plasma were lowest in the high bacterial dose. Cortisol levels were highest at day 7 in the high dose and then decreased from day 14 until the end of the study. Liver enzymes showed a similar pattern to plasma metabolites, with decreased enzymatic activity, mostly with the high bacteria dose. PK was the exception, with increased enzymatic activity in a dose-dependent manner over time. Liver metabolites were highly variable, except in the high bacterial dose where variability and total levels decreased significantly. Our results show that fish infection with F. noatunensis induces a clear stress response, especially with at the highest dose, shifting intermediary metabolism towards mobilization of energy and suggesting that E. maclovinus detects experimental infection of F. noatunensis as a stressor, which it is dependent on the bacterial dose.

Effect of ration level on growth performance, body composition, intermediary metabolism and serum parameters in juvenile Patagonian blennie Eleginops maclovinus.

Eleginops maclovinus is an endemic species of the southern cone with beneficial physiological characteristics for aquaculture. However, this species has a low growth rate under captive conditions, and the optimal feed ration together with the metabolic process is unknown. This study aimed to determine the optimum feed ration during 90 days based on growth performance, body composition, intermediary metabolism, and serum parameters. For this, fish were randomly assigned to rations of 0.5, 1, 2 and 4% of body weight. No fish mortality was registered, but all fish, developed fatty liver. The results of weight, length, growth performance (WGR, SGR), and body parameters (HSI, VSI and K) followed a similar pattern, with the lowest values observed in the 0.5% and no significant differences between rations of 1, 2 and 4%. The feed intake and feces increased with the feed ration. However, the percentage of food consumed by the fish decreased with the ration size and the feed conversion ratio was lowest in the 1% ration. Total serum proteins and calcium were lowest in the 0.5% ration and presented no differences in the rations 1, 2 and 4%, while triglyceride content was significantly different only between the rations of 0.5 and 4%. Blood cortisol levels were significantly higher in the rations of 0.5 and 1%, and decreased in rations of 2 and 4%. The lipids, fiber, and energy of the total body mass increased with the feed ration, while dry matter, proteins, and ash of the body decreased to higher feed ration. In liver, triglyceride and protein levels decreased with a larger feed ration, amino acids increased in the rations of 0.5 and 4%, while glucose levels increased in rations of 2 and 4%. Liver enzymes Glucose-6-phosphate dehydrogenase and Glutamate dehydrogenase increased their activity at a higher feed ration, while Glycogen Phosphorylase, Aspartate aminotransferase and 3-Hydroxyacyl-CoA dehydrogenase presented their highest enzymatic activity only in the 4% ration. Fructose-1,6-bisphosphate showed low enzymatic activity in rations of 2 and 4% and Glycerol-3-phosphate dehydrogenase was significantly different only between the ration of 0.5 and 1%. Finally, our results suggests that optimal feed rations for E. maclovinus corresponds to 1% since this ration size produces the highest growth and metabolism with a minimum loss of food and feces present in the environment. Additionally, we recommend to reduce the percentage of fat in the diet to avoid the development of fatty liver.

Immunological response of the Sub-Antarctic Notothenioid fish Eleginops maclovinus injected with two strains of Piscirickettsia salmonis.

Eleginops maclovinus is an endemic fish to Chile that lives in proximity to salmonid culture centers, feeding off of uneaten pellet and salmonid feces. Occurring in the natural environment, this interaction between native and farmed fish could result in the horizontal transmission of pathogens affecting the aquaculture industry. The aim of this study was to evaluate the innate and adaptive immune responses of E. maclovinus challenged with P. salmonis. Treatment injections (in duplicate) were as follows: control (100 µL of culture medium), wild type LF-89 strain (100 µL, 1 × 108 live bacteria), and antibiotic resistant strain Austral-005 (100 µL, 1 × 108 live bacteria). The fish were sampled at various time-points during the 35-day experimental period. The gene expression of TLRs (1, 5, and 8), NLRCs (3 and 5), C3, IL-1ß, MHCII, and IgMs were significantly modulated during the experimental period in both the spleen and gut (excepting TLR1 and TLR8 spleen expressions), with tissue-specific expression profiles and punctual differences between the injected strains. Anti-P. salmonis antibodies increased in E. maclovinus serum from day 14-28 for the LF-89 strain and from day 14-35 for the Austral-005 strain. These results suggest temporal activation of the innate and adaptive immune responses in E. maclovinus tissues when injected by distinct P. salmonis strains. The Austral-005 strain did not always cause the greatest increases/decreases in the number of transcripts, so the magnitude of the observed immune response (mRNA) may not be related to antibiotic resistance. This is the first immunological study to relate a pathogen widely studied in salmonids with a native fish.

Ectoparasite Caligus rogercresseyi modifies the lactate response in Atlantic salmon (Salmo salar) and Coho salmon (Oncorhynchus kisutch).

Although Caligus rogercresseyi negatively impacts Chilean salmon farming, the metabolic effects of infection by this sea louse have never been completely characterized. Therefore, this study analyzed lactate responses in the plasma, as well as the liver/muscle lactate dehydrogenase (LDH) activity and gene expression, in Salmo salar and Oncorhynchus kisutch infested by C. rogercresseyi. The lactate responses of Atlantic and Coho salmon were modified by the ectoparasite. Both salmon species showed increasing in plasma levels, whereas enzymatic activity increased in the muscle but decreased in the liver. Gene expression was overexpressed in both Coho salmon tissues but only in the liver for Atlantic salmon. These results suggest that salmonids need more energy to adapt to infection, resulting in increased gene expression, plasma levels, and enzyme activity in the muscles. The responses differed between both salmon species and over the course of infection, suggesting potential species-specific responses to sea-lice infection.

Oral administration of melatonin counteracts several of the effects of chronic stress in rainbow trout.

To assess a possible antistress role of melatonin in fish, we orally administered melatonin to rainbow trout for 10 d and then kept the fish under normal or high stocking density conditions during the last 4 d. Food intake; biochemical parameters in plasma (cortisol, glucose, and lactate concentrations); liver (glucose and glycogen concentrations, and glycogen synthase activity); enzyme activities of amylase, lipase, and protease in foregut and midgut; and content of the hypothalamic neurotransmitters dopamine and serotonin, as well as their oxidized metabolites, 3,4-dihydroxyphenylacetic acid and 5-hydroxy-3-indoleacetic acid, were evaluated under those conditions. High stocking density conditions alone induced changes indicative of stress conditions in plasma cortisol concentrations, liver glycogenolytic potential, the activities of some digestive enzymes, and the 3,4-dihydroxyphenylacetic acid-to-dopamine and 5-hydroxy-3-indoleacetic acid-to-serotonin ratios in the hypothalamus. Melatonin treatment in nonstressed fish induced an increase in liver glycogenolytic potential, increased the activity of some digestive enzymes, and enhanced serotoninergic and dopaminergic metabolism in hypothalamus. The presence of melatonin in stressed fish resulted in a significant interaction with cortisol concentrations in plasma, glycogen content, and glycogen synthase activity in liver and dopaminergic and serotoninergic metabolism in the hypothalamus. In general, the presence of melatonin mitigated several of the effects induced by stress, supporting an antistress role for melatonin in rainbow trout.