The aims of this study were to compare male and female sea trout (Salmo trutta) with respect to their hypo-osmoregulatory ability over a simulated migration to seawater and their clinical and physiological response to salmon louse (Lepeophtheirus salmonis) infection in seawater and over a simulated pre-mature return to freshwater. For this purpose, 2-year-old hatchery-reared male and female brown trout (F1 offspring of wild caught anadromous fish) were infected with salmon lice and measured for changes in plasma ions, glucose, lactate and osmolality and relative heart, liver and gonad sizes during a simulated seawater migration and thereafter a premature return to freshwater after 4 weeks in seawater (pre-adult louse). Un-infected trout served as control. Male trout used longer time to develop full hypo-osmoregulatory ability in seawater and showed a stronger response in plasma glucose and lactate following simulated premature return to freshwater, compared to female trout. Response to salmon louse was stronger in female trout, shown by stronger osmotic stress by chalimus (plasma Cl-) and pre-adult louse (plasma osmolality) and elevated relative liver size (hepatosomatic index) by pre-adult louse in female compared to male trout. Moreover, high plasma cortisol in infected female and low plasma cortisol in infected male trout produced a significant treatment-sex interaction on plasma cortisol. Lice infection intensity was initially higher in male (0.18 lice g-1) compared to female trout (0.11 lice g-1) at the chalimus stage, but equal between sexes at the pre-adult stage (male 0.15 and female 0.17 lice g-1). This study showed that female trout were better adapted for changes in water salinity, while male trout were more robust against salmon louse infection. These results suggests that the elevated salmon louse infection pressure generated by salmon farming have strong and unexplored negative effects on wild sea trout populations. Further research on this topic is vital for the conservation of wild sea trout populations.

The current investigation was carried out with an aim to evaluate disparity in haematological and serum biochemical parameters of healthy male and female Schizothorax labiatus with respect to its four reproductive phases, i.e. preparatory (autumn), pre-spawning (winter), spawning (spring) and spent (summer). The study indicated that significant (p < 0.05) alterations were seen in gonadosomatic index (GSI) of S. labiatus with maximum value for both sexes noted in spring while minimum value in autumn. Contrary to this, the highest value of hepatosomatic index (HSI) for both sexes was noted in the autumn season and the lowest value in spring. The haemoglobin (Hb) content, red blood corpuscle (RBC) and white blood corpuscle (WBC) counts and haematocrit amount of S. labiatus also revealed significant (p < 0.05) variations with peak values noted in summer while least values were recorded during winter. It was also observed that male fishes comprised significantly (p < 0.05) maximum level of all these parameters amongst all seasons of the year, except WBC count. Erythrocyte indices specifically mean corpuscular haemoglobin (MCH), MCH concentration (MCHC) and mean corpuscular volume (MCV) exhibited insignificant (p > 0.05) alterations between the sexes. The lowest levels (p < 0.05) of serum glucose, cholesterol and urea were recorded in the pre-spawning period, i.e. winter, while as the highest values (p < 0.05) were seen in the summer because of the cessation of reproductive activities and greater availability of food. Similar trend was also noted in the amount of total protein, albumin and globulin of the fish, although insignificant (p > 0.05) variation was observed between sexes.

Immunity is an important component of invasion success since it enables invaders\' adaptation to conditions of the novel environment as they expand their range. Immune response of invaders may vary along the invasion range due to encountered parasites/microbial communities, conditions of the local environment, and ecological processes that arise during the range expansion. Here, we analyzed changes in the immune response along the invasion range of one of the most successful aquatic invaders, the signal crayfish, in the recently invaded Korana River, Croatia. We used several standard immune parameters (encapsulation response, hemocyte count, phenoloxidaze activity, and total prophenoloxidaze) to: i) compare immune response of the signal crayfish along its invasion range, and between species (comparison with co-occurring native narrow-clawed crayfish), and ii) analyze effects of specific predictors (water temperature, crayfish abundance, and body condition) on crayfish immune response changes. Immune response displayed species-specificity, differed significantly along the signal crayfish invasion range, and was mostly affected by water temperature and population abundance. Specific immune parameters showed density-dependent variation corresponding to increased investment in them during range expansion. Obtained results offer baseline insights for elucidating the role of immunocompetence in the invasion success of an invertebrate freshwater invader.

Curcumin exhibited numerous key activities including antioxidant, anti-inflammatory, and immunostimulatory effects in fish. This study evaluated the protective effects of curcumin (CUR) against lead (Pb)-induced toxicities in fish. Healthy Cyprinus carpio L. individuals were segregated into control, Pb only, Pb+CUR, and CUR only groups. Pb groups were exposed to 1 mg L -1 of Pb, and CUR groups were fed a basal diet supplemented with 15 g kg-1 of CUR. After eight weeks, growth performance, Pb accumulation in tissues, various haemato-biochemical parameters, immune responses, and cytokine gene expression were measured. Dietary CUR effectively decreased Pb accumulation in tissues and increased the survival of Pb-exposed fish. Co-treatment with Pb and CUR reversed alterations in haemato-biochemical parameters, ameliorated Pb-induced oxidative stress, enhanced immune responses, and restored intestinal enzymatic activities. Dietary CUR reversed changes in intestinal microbiota in Pb-exposed fish. Pb-induced upregulation of NF-κBp65 and HSP70 was inhibited by dietary CUR. CUR supplementation upregulated the mRNA levels of SOD, Nrf2, IL-10, and CYP450 1A and attenuated Pb-induced degradation of I κB-α mRNA levels. Overall, CUR antagonizes Pb-induced negative impacts in fish. Thus, dietary CUR had several beneficial effects on immune responses, decreased Pb accumulation in tissues, and reversed Pb-induced oxidative stress in fish. Therefore, CUR plays a protective role in Pb-induced immune toxicity in fish, and, as such, may be suitable as an aqua feed additive for use in carp aquaculture.

Parasite distribution among hosts is a fundamental aspect of host-parasite interactions. Aggregated parasite distributions within and across host species are commonly reported and potentially influenced by many factors, whether host or parasite specific, or related to host-parasite encounter and compatibility. Yet, the respective role of each in observed parasite distributions are often unclear. Here, we documented the distribution of the acanthocephalan parasite Pomphorhynchus laevis sensu lato (s.l.) in two replicate fish host populations. Aggregated distributions were observed in both populations, within and across fish host species. We found a positive abundance-prevalence relationship across fish species, suggesting that resource availability (fish host biomass density) was the main driver of P. laevis s.l. distribution. This was supported by further positive associations between mean parasite load and fish biomass density. We found little evidence for intensity-dependent regulation within host (i.e. intra-host competition among co-infecting parasites). Furthermore, P. laevis s.l. infection had no detectable effect on fish condition indices, except on the body condition of female barbel (Barbus barbus). Therefore, P. laevis s.l. tended to accumulate with size/age within fish species, and with fish biomass density among fish species, with apparently negligible limitations due to intra-host intensity-dependent regulation of parasite, or to parasite-induced morbidity in fish. The relative availability of final hosts for trophic transmission thus appears to be the main driver of P. laevis s.l. distribution among fish.

Muscle and liver tissues of Lophius vomerinus off the coast of Namibia were analysed to investigated the influence of MeHg on the biological parameters of L.vomerinus by (i) determining if the variability in total MeHg concentrations is influenced by length, maturity status and sex, and (ii) assessing if there is a relationship between biological indices (Condition factor (K), Gonadosomatic Index (GSI) Hepatosomatic Index (HSI)) and MeHg concentrations. Correlations between total MeHg concentrations and fish length, K and HSI were observed. A weak positive correlation was observed between total MeHg and GSI for combined sex. Total MeHg concentration in tissues of L. vomerinus is significantly dependent on the maturity stages (p < 0.05). K was significantly inversely correlated with total MeHg in tissues of L. vomerinus. The evidence presented in this study suggests that MeHg in L. vomerinus tissues could be detrimental to both its physiology and population dynamics.

Wildfires are a common phenomenon in Mediterranean regions that is becoming increasingly frequent and severe, causing several environmental concerns, of which ash runoff represents an important source of disturbance for aquatic organisms, in particular for fishes. Studies on the behavioural response of fishes to wildfire ash runoff are scarce and seldom include cyprinid species. The goal of this study was to investigate in a 3-artificial flume channel mesocosm, the behavioural and hepatosomatic condition responses of a native widespread potamodromous fish, the Iberian barbel (Luciobarbus bocagei), previously exposed for 24 h to different concentrations of wildfire ashes: 0.0 g/L (the control, no ash), 1.0 g/L (low concentration) and 2.0 g/L (high concentration). Behavioural parameters included i) routine activity, ii) boldness and iii) shoaling cohesion. The hepatosomatic index (HSI) was further determined to assess the health condition of fish. Significant differences on fish behaviour parameters were detected between the control and the high concentration of ash. Accordingly, i) an increasing proportion of fish were found on resting activity (56.2% vs 30.6% in the control), whereas the proportion of fish on searching behaviour (58.4% in the control) decreased (41.5%); ii) the proportion of bolder individuals was found to decrease (42.5% in the control vs. 29.4%) and iii) the same trend was detected for shoaling cohesion (61.3% in the control to 33.8%, of all fish within a body length of each other). Such differences were paralleled by an increase in the HSI from 1.62% (control) to 2.40% (high concentration). The present study shows that even short duration exposure to ash-loaded runoff can alter fish behaviour and hepatosomatic condition and highlights the need to maintain an unfragmented river network, or, when this is not possible, to prioritize the removal or retrofitting of barriers to increase movement dispersal and provide conditions for species recovery from fire-disturbances.

BACKGROUND: Global trend of the introduction of high levels of relatively cheap carbohydrates to reduce the amount of costly protein in the aquatic animal feed production has affected the aquaculture of an economically important cyprinid fish, blunt snout bream (Megalobrama amblycephala). This dietary shift has resulted in increased prevalence of metabolic disorders, often causing economic losses. High dietary intake of carbohydrates, associated with obesity, is one of the major causes of non-alcoholic fatty liver disease (NAFLD) in humans.
RESULTS: We have conducted an eight-week feeding trial to better understand how a high-carbohydrate diet (HCBD) affects the liver health in this fish. Hepatosomatic index and lipid content were significantly (P < 0.05) higher in the HCBD group. Histology results also suggested pathological changes in the livers of HCBD group, with excessive lipid accumulation and indication of liver damage. Metabolomics and serum biochemistry analyses showed that a number of metabolites indicative of liver damage were increased in the HCBD group. This group also exhibited low levels of betaine, which is a metabolite crucial for maintaining the healthy liver functions. Transcriptomic and qPCR analyses indicated that HCBD had a strong impact on the expression of a large number of genes associated with the NAFLD and insulin signalling pathways, which may lead to the development of insulin resistance in hepatocytes, pathological liver changes, and eventually the NAFLD.
CONCLUSIONS: Transcriptomics, metabolomics and histology results all indicate early symptoms of liver damage. However whether these would actually lead to the development of NAFLD after a longer period of time, remains inconclusive. Additionally, a very high number of upregulated genes in the HCBD group associated with several neurodegenerative diseases is a strong indication of neurodegenerative changes caused by the high-carbohydrate diet in blunt snout bream. This suggests that fish might present a good model to study neurodegenerative changes associated with high-carbohydrate diet in humans.

Strictly carnivorous fish with high requirements for dietary protein, such as rainbow trout (Oncorhynchus mykiss) are interesting models for studying the role of amino acids as key regulators of intermediary metabolism. Methionine is an essential amino acid for rainbow trout, and works as a signalling factor in different metabolic pathways. The study investigated the effect of increasing dietary methionine intake on the intermediary metabolism in the liver of juvenile rainbow trout. For this purpose, five diets were formulated with increasing methionine levels from 0.60 to 1.29% dry matter. The diets were fed in excess for six weeks before three sampling campaigns carried out successively to elucidate (i) the hepatic expression of selected genes involved in lipid, glucose and amino acid metabolism; (ii) the postprandial ammonia excretion; and (iii) the postprandial plasma methionine concentrations. The transcript levels of enzymes involved in lipid metabolism (fatty acid synthase, glucose 6 phosphate dehydrogenase and carnitine palmitoyl transferase 1 a), gluconeogenesis (fructose-1,6-biphosphatase) and amino acid catabolism (alanine amino transferase and glutamate dehydrogenase) were significantly affected by the increase in dietary methionine. Changes in gene expression reflected to some extent the decrease in ammonia excretion (P=0.022) and in the hepatosomatic index (HSI; P<0.001) when dietary methionine increased. Postprandial plasma methionine concentrations correlated positively with the dietary level (P<0.001) at the different sampling points. The study shows that the expression of several genes related to the hepatic intermediary metabolism in rainbow trout responded in a dose-dependent manner to increasing levels of dietary methionine.

Over-winter mortality is an important selective force for warm-water fish (e.g., centrarchids) that live in temperate habitats. Inherent challenges faced by fish during winter may be compounded by additional stressors that activate the hypothalamic-pituitary-interrenal axis, either before or during winter, leading to negative sub-lethal impacts on fish health and condition, and possibly reducing chance of survival. We used experimental cortisol manipulation to test the hypothesis that juvenile largemouth bass (Micropterus salmoides) exposed to semi-chronic elevation in cortisol prior to winter would experience higher levels of over-winter mortality, physiological alterations and impaired immune status relative to control and sham-treated bass. Over-winter survival in experimental ponds was high, averaging 83%, and did not differ among treatment groups. Over the study period, bass exhibited an average increase in mass of 19.4%, as well as a slight increase in Fulton\'s condition factor, but neither measure differed among groups. Hepatosomatic index in cortisol-treated bass was 23% lower than in control fish, suggesting lower energy status, but white muscle lipid content was similar across all groups. Lastly, there was no difference in spleen somatic index or parasite load among treatment groups, indicating no long-term immune impairment related to our cortisol manipulation. The current study adds to a growing body of literature on glucocorticoid manipulations where field-based findings are not consistent with laboratory-based conceptual understanding of multiple stressors. This suggests that field conditions may provide fish with opportunities to mitigate negative effects of some stressors.