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Blood transcriptomics has emerged as a vital tool for tracking the immune system and supporting disease diagnosis, prognosis, treatment, and research. The present study was conducted to analyze the gene expression profile and potential biomarker candidates using the whole blood of mandarin fish (Siniperca chuatsi) infected with LPS or poly (I:C) at 0 h, 3 h, 6 h, and 12 h. Our data suggest that 310 shared differentially expressed genes (DEGs) were identified among each comparison group after LPS infection, and 137 shared DEGs were identified after poly (I:C) infection. A total of 62 shared DEGs were differentially expressed in all compared groups after LPS or poly (I:C) infection. Pathways analysis for DEGs in all different compared groups showed that cytokine-cytokine receptor interaction was the most enrichment pathway. The expression levels of genes C-X-C chemokine receptor type 2-like (cxcr2), chemokine (C-C motif) receptor 9a (ccr9a), chemokine (C-C motif) receptor 9b (ccr9b), chemokine (C-X-C motif) receptor 4b (cxcr4b), and interleukin 10 receptor alpha (il10ra) were significantly different in all compared groups and most enriched in cytokine-cytokine receptor interaction pathway. The protein-protein interactions analysis among all shared DEGs showed that cxcr4 was the hub gene with the highest degree. The biomarker candidates discovered in this study may, following validation, prove effective as diagnostic tools in monitoring mandarin fish diseases., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Background: Dietary kelp possesses a variety of useful biological qualities but does not have a toxic effect on the host. In this study, we examine how kelp dietary supplementation enhances the serum biochemistry, intestinal immunity, and metabolism of hybrid snakehead. A total of 810 juvenile hybrid snakeheads (Channa maculata ♀ × Channa argus ♂), with an initial average weight of 11.4 ± 0.15 g, were allocated randomly to three treatment groups (three replicates per group). The fish were fed for 60 days with isonitrogenous and isolipidic diets. The groups were the control group (C) (20% high-gluten flour), the medium replacement group (MR) (10% high-gluten flour and 10% kelp meal), and the full replacement group (FR) (0% high-gluten flour and 15% kelp meal)., Results: The results showed that dietary kelp increased the activity of serum antioxidant enzymes significantly and decreased the content of serum malondialdehyde (MDA) in hybrid snakeheads, with significant changes in the FR group (P < 0.05). The intestinal morphology results showed that dietary kelp helped to increase the specific surface area of intestinal villi, which was beneficial for intestinal digestion and absorption. According to transcriptome and quantitative real-time polymerase chain reaction (qRT-PCR) analysis, dietary kelp can improve the expression of intestinal immunity and metabolism-related pathways. Among them, immune-related genes MHC1 and HSPA1 were significantly up-regulated, and IGH, MHC2, and IL-8 were significantly down-regulated (P < 0.05). Lipid metabolism-related genes DGAT2, FABP2, RXRα, and PLPP1 were all significantly up-regulated (P < 0.05)., Conclusion: Dietary kelp can effectively improve the antioxidant function of hybrid snakeheads, improve intestinal morphology, reduce intestinal inflammation, and promote intestinal lipid synthesis and transportation, thereby improving intestinal immunity and metabolic functions. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Prolactin is a hormone conserved across all vertebrates and is renowned for its role in reproduction and parental care. Previous studies on prolactin in fish have primarily relied on administration of mammalian prolactin and have suggested that increases in prolactin lead to greater parental care. However, the influence of endogenous prolactin on fish parental care remains unknown. Here, we measure circulating concentrations of endogenous prolactin during parental care in a fish and link these concentrations to parental care behaviour. We provide evidence that male bluegill sunfish with higher circulating concentrations of prolactin provide more parental care to their offspring. Furthermore, we show that nesting males with experimentally reduced perceived paternity have lower circulating prolactin concentrations and perform fewer parental behaviours, facilitating an adaptive investment in offspring in response to paternity cues. Our findings not only confirm the role of endogenous prolactin in modulating parental care behaviour in a fish but also provide a mechanism underlying the adaptive changes in parental care made in response to perceived paternity., Competing Interests: Declaration of competing interest The authors declare no competing interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The content of membrane-bound methemoglobin (MtHb) in nucleated erythrocytes was studied in the black scorpionfish Scorpaena porcus (Linnaeus, 1758) in vitro. Spectral characteristics were determined for a whole hemolysate, a hemolysate obtained by stroma precipitation (a clarified hemolysate), and a resuspended stroma. The MtHb proportion in the erythrocyte stroma was found to exceed 80% (6.20 ± 0.59 µM). Clarified hemolysates were nearly free of MtHb (0.5 ± 0.2 µM). Membrane-bound ferric hemoglobin did not affect the erythrocyte resistance to osmotic shock. The osmotic fragility range was determined using a LaSca-TM laser microparticle analyzer (BioMedSystems, Russia) to be 102-136 mOsm/kg, much the same as in other bony fish species. A nitrite load (10 mg/L) significantly increased the MtHb content in the blood. However, the membrane-bound ferric hemoglobin content did not change significantly, amounting to 6.34 ± 1.09 µM (approximately 95%). The finding suggested a functional importance for MtHb present in the plasma membrane of nucleated erythrocytes. Membrane-bound MtHb was assumed to neutralize the external oxidative load and the toxic effect of hydrogen sulfide in bottom water layers, where the species lives., (© 2024. Pleiades Publishing, Ltd.)

The round goby (Neogobius melanostomus) is a fish native to the Ponto-Caspian region that is highly invasive through freshwater and brackish habitats in northern Europe and North America. Individual behavioural variation appears to be an important factor in their spread, for example a round goby's personality traits can influence their dispersal tendency, which may also produce variation in the behavioral composition of populations at different points along their invasion fronts. To further analyze the drivers of behavioral variation within invasive round goby populations, we focused on two populations along the Baltic Sea invasion front with closely comparable physical and community characteristics. Specifically, this study measured personality within a novel environment and predator response context (i.e., boldness), and directly analyzed links between individuals' personality traits and their physiological characteristics and stress responses (i.e., blood cortisol and lactate, brain neurotransmitters). In contrast to previous findings, the more recently established population had similar activity levels but were less bold in response to a predator cue than the older population, which suggests that behavioral compositions within our study populations may be more driven by local environmental conditions rather than being a result of personality-biased dispersal. Furthermore, we found that both populations showed similar physiological stress responses, and there also appeared to be no detectable relationship between physiological parameters and behavioral responses to predator cues. Instead, body size and body condition were important factors influencing individual behavioral responses. Overall, our results reinforce the importance of boldness traits as a form of phenotypic variation in round goby populations in the Baltic Sea. We also highlight the importance of these traits for future studies specifically testing for effects of invasion processes on phenotypic variation in the species. Nonetheless, our results also highlight that the physiological mechanisms underpinning behavioural variation in these populations remain unclear., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Teleost fishes are diverse and successful, comprising almost half of all extant vertebrate species. It has been suggested that their success as a group is related, in part, to their unique O 2 transport system, which includes pH-sensitive hemoglobin, a red blood cell β-adrenergic Na + /H + exchanger (RBC β-NHE) that protects red blood cell pH, and plasma accessible carbonic anhydrase which is absent at the gills but present in some tissues, that short-circuits the β-NHE to enhance O 2 unloading during periods of stress. However, direct support for this has only been examined in a few species of salmonids. Here, we expand the knowledge of this system to two warm-water, highly active marine percomorph fish, cobia (Rachycentron canadum) and mahi-mahi (Coryphaena hippurus). We show evidence for RBC β-NHE activity in both species, and characterize the Hb-O 2 transport system in one of those species, cobia. We found significant RBC swelling following β-adrenergic stimulation in both species, providing evidence for the presence of a rapid, active RBC β-NHE in both cobia and mahi-mahi, with a time-course similar to that of salmonids. We generated oxygen equilibrium curves (OECs) for cobia blood and determined the P 50 , Hill, and Bohr coefficients, and used these data to model the potential for enhanced O 2 unloading. We determined that there was potential for up to a 61% increase in O 2 unloading associated with RBC β-NHE short-circuiting, assuming a - 0.2 ∆pH a-v in the blood. Thus, despite phylogenetic and life history differences between cobia and the salmonids, we found few differences between their Hb-O 2 transport systems, suggesting conservation of this physiological trait across diverse teleost taxa., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The morphology, cell size and relative number of peripheral blood cells in two teleosts, Synechogobius hasta and Sebastes schlegelii, were compared using different staining methods. The results showed significant differences in cell size. The percentage of monocytes in S. hasta was greater than that in S. schlegelii (P < 0.01); however, the opposite results were obtained in the percentage of lymphocytes and thrombocytes. The two fishes shared common cytochemical-staining results, which showed that both erythrocytes were negative for all cytochemical staining; monocytes were strongly positive for PAS and positive for SBB and NAE; lymphocytes were negative for SBB, POX and NAE; neutrophils were positive for SBB and NAE; and thrombocytes were negative for SBB, ALP, POX and NAE. However, species specificity existed in the cytochemical properties. For S. hasta, monocytes were strongly positive for ALP and ACP; lymphocytes were strongly positive for ALP and weakly positive for ACP; neutrophils were strongly positive for ACP and POX; and thrombocytes were weakly positive for PAS and positive for ACP. Unlike S. hasta, monocytes were strongly positive for ACP and positive for ALP in S. schlegelii; lymphocytes were positive for ALP and partially positive for PAS; neutrophils were positive for ACP and POX; and thrombocytes were negative for PAS and ACP. The POX activity of monocytes in S. hasta was greater than that in S. schlegelii (P < 0.05), while the POX activity of neutrophils and the NAE activity of monocytes in S. hasta were significantly greater than those in S. schlegelii (P < 0.01). The results of this study can be used as a reference for the construction of haematological parameters in S. hasta and S. schlegelii for the assessment of fish health and can provide a research basis for fish diseases and immunity., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Our study showed differential expression of the arginine vasotocin ( avt )/ isotocin ( it ) in the brain and pituitary gland of the euryhaline black porgy ( Acanthopagrus schlegelii ) during osmotic stress. A decrease in serum osmolality and increased cortisol levels were observed after acute transfer from seawater (SW) to freshwater (FW). The increased expressions of avt , avt receptor ( avtr : v1a ), and isotocin receptor ( itr : itr1) transcripts on day 1 and it and itr transcripts on days 7 and 30 were found in the brains and pituitary glands of FW fish. Increased levels of avt mRNA in the diencephalon and avtr mRNA in the pituitary together with serum cortisol on day 1 of FW exposure indicated activation of the hypothalamic-pituitary-interrenal (HPI) axis. The expression levels of avtr and itr after FW transfer were increased in the pituitary on days 7 and 30. Furthermore, in situ hybridization demonstrated spatially differential expression of avt and itr transcripts in nucleus preopticus parvocellularis of pars gigantocellularis (PMgc), magnocellularis (PMmc), and parvocellularis (PMpc) of the preoptic area (POA). Positive signals for avt and it were highly abundant in PMpc after FW exposure. The data suggest involvement of neurohypophysial hormones in the brain (telencephalon and diencephalon) and pituitary for osmotic stress.

The waste recycling of lemon peel, as a functional feed additive in aquafeed was evaluated by estimating the effects of fermented lemon peel (FLP) supplementation in diet on growth performance, innate immune responses, and susceptibility to Photobacterium damselae of grouper, Epinephelus coioides. A basal diet was added FLP at 0%, 1%, 3%, and 5%. Four tested diets were each fed to juvenile grouper (initial weight: 15.89 ± 0.10 g, triplicate groups) in a recirculation rearing system for eight weeks. Fish fed diets with 0%-3% FLP exhibited higher (p < 0.05) final weight, weight gain, and feed efficiency than fish fed the 5% FLP-diet. After challenge test, fish fed the 3% FLP-diet appeared the lowest mortality, followed by fish fed the 1% FLP-diet, and lowest in fish fed 0% and 5% FLP-diets. Plasma lysozyme activities were higher in fish fed diets with FLP than in fish fed the FLP-free control diet before challenge test. After challenge, fish fed diets with 1% and 3% FLP showed highest lysozyme activities, followed by fish fed the diet with 5% FLP, and lowest in fish fed the control diet. Hepatic malondialdehyde content was higher in fish fed the control diet than in fish fed diets with 1%-3% FLP. Results found that diets supplemented with 1%-3% fermented lemon peel can enhance lysozyme activity and resistance to pathogen P. damselae of grouper., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Hypoxia has gradually become common in aquatic ecosystems and imposes a significant challenge for fish farming. The loss of equilibrium (LOE), 50% lethal time (LT50), plasma cortisol, glucose, red blood cells (RBC), hemoglobin (Hb), gill histological alteration, and related parameters (lamellar length [SLL] and width [SLW], interlamellar distance [ID], basal epithelial thickness [BET], lamellar surface area [LA], and gill surface area [GSA]); respiratory rate; the proportion of the secondary lamellae available for gas exchange (PAGE); and hypoxia-inducible factor (hif-1α, hif-2α) mRNA expression were determined during progressive hypoxia and reoxygenation (R-0, R-12, R-24 h) to illustrate the underlying physiological response mechanisms in black rockfish Sebastes schlegelii. Results showed that the DO concentration significantly decreased during progressive hypoxia, while DO at LOE and LT50 were 2.42 ± 0.10 mg L -1 and 1.67 ± 0.38 mg L -1 , respectively. Cortisol and glucose were significantly increased at LOE and LT50, with the highest levels observed at LT50, and then gradually recovered to normal within reoxygenation 24 h. RBC number and Hb results were like those of glucose. Hypoxia stress resulted in lamellar clubbing, hypertrophy, and hyperplasia. Respiratory frequency significantly increased at LOE and decreased at LT50. Lamellar perimeters, SLL, ID, LA, GSA, and PAGE, significantly increased at LOE and LT50, with the highest values observed at LT50. However, SLW and BET significantly decreased at LOE, LT50, and R-0. These parameters recovered to nearly normal levels at R-24 h. hif-1α mRNAs in gill and liver were significantly upregulated at LOE and LT50, and recovery to normal after reoxygenation 24 h. hif-2α mRNAs in gill was similar to that of hif-1α, whereas hepatic hif-2α mRNAs remained unchanged during hypoxia-reoxygenation. These results indicated that progressive hypoxia stress elevated RBC number, Hb, cortisol, and glucose levels, induced the alteration of gill morphology, increased LA and GSA, stimulated respiratory frequency and PAGE, and upregulated the transcription of hif-1α and hif-2α in gill and liver. Reoxygenation treatment for 24 h alleviated the stress mentioned above effects. These findings expand current knowledge on hypoxia tolerance in black rockfish Sebastes schlegelii., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

The neuroendocrine system of fish responds to low temperature via regulating hormones. To explore the adaptability of Larimichthys crocea to low temperature, the levels of the plasma cortisol, thyroid stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), total cholesterol (TC), and glucose were determined after exposure to low temperature and during subsequent rewarming. Furthermore, the mRNA expression of the glucocorticoid receptor (GR) gene was analyzed under the stress. We found that the levels of the plasma cortisol, TSH, T3, glucose, and TC increased under the low temperature stress, suggesting that elevated hormones may be conducive to promoting the mobilization of the glucose and lipid in L. crocea exposed to low temperature. During the rewarming period, the plasma cortisol level decreased, whereas the T3 level was still significantly higher than that in the control group. Notably, the plasma T4 level was unaffected by the temperature changes. Furthermore, the sequence alignment and phylogenetic tree analysis revealed that the GR protein of L. crocea had high homology and a similar protein structure with those from other teleosts. Under the low temperature stress, the GR mRNA expression increased in the brain and head kidney, whereas it basically returned to the control level following rewarming. These findings revealed the changes of the hormones and the potential function of the GR gene in L. crocea following exposure to low temperature, providing some insights into breeding low temperature-resistant varieties of L. crocea., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This study aims to explore the mechanism on how aggressive interaction alters reproductive physiology by testing whether aggressive interaction can activate the reproductive neuroendocrine function via the hypothalamus-pituitary-gonadal (HPG) axis in black rockfish (Sebastes schlegelii). The expressions of the androgen receptor gene (ar) and gonadotropin-releasing hormone genes (gnrhs), the concentration of plasma androgens, and GSI (the ratio of testes mass to body mass) were compared between the interaction group (dominant males or subordinate males) and the isolation group in male black rockfish after 3 weeks. A full-length cDNA encoding an androgen receptor (AR) of 766 amino acids was isolated. Transcripts encoding this AR were detected at a high relative abundance in the liver, kidney, testis, ovary, muscle, and intestine tissue. Further evaluation of brain genes transcripts abundance revealed that the mRNA levels of gnrh I and ar genes were significantly different between the interaction group and the isolation group in the hypothalamus. However, no significant difference was detected in testosterone, 11-keto-testosterone, and GSI between these two groups. This study indicates that a long-term aggressive interaction affect the expression of hypothalamic gnrh I and ar but may not change the physiological function of the HPG axis in an all-male condition.

In fishes, the spleen can function as an important reservoir for red blood cells (RBCs), which, following splenic contraction, may be released into the circulation to increase haematocrit during energy-demanding activities. This trait is particularly pronounced in red-blooded Antarctic fishes in which the spleen can sequester a large proportion of RBCs during rest, thereby reducing blood viscosity, which may serve as an adaptation to life in cold environments. In one species, Pagothenia borchgrevinki, it has previously been shown that splenic contraction primarily depends on cholinergic stimulation. The aim of the present study was to investigate the regulation of splenic contraction in five other Antarctic fish species, three red-blooded notothenioids (Dissostichus mawsoni Norman, 1937, Gobionotothen gibberifrons Lönnberg, 1905, Notothenia coriiceps Richardson 1844) and two white-blooded "icefish" (Chaenocephalus aceratus Lönnberg, 1906 and Champsocephalus gunnari Lönnberg, 1905), which lack haemoglobin and RBCs, but nevertheless possess a large spleen. In all species, splenic strips constricted in response to both cholinergic (carbachol) and adrenergic (adrenaline) agonists. Surprisingly, in the two species of icefish, the spleen responded with similar sensitivity to red-blooded species, despite contraction being of little obvious benefit for releasing RBCs into the circulation. Although the icefish lineage lost functional haemoglobin before diversifying over the past 7.8-4.8 millions of years, they retain the capacity to contract the spleen, likely as a vestige inherited from their red-blooded ancestors., (© 2020 Fisheries Society of the British Isles.)

Plant proteins are suitable and alternative to fish meals (FMs), with less cost compared with that of all other types of fish feeds. In recent years, soy protein concentrate (SPC) has emerged as a cost-effective alternative to FM; however, little is known regarding the effects of dietary SPC on general fish physiology and well-being. This study aimed to perform comprehensive physiological and transcriptomic analysis for testing the applicability of SPC as fish feeds in hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂) [SPC replaced 0% (CK), 30% (SPC30), and 75% (SPC75) of FM protein]. Generally, SPC30 promoted fish survival and had less effects on the phenotype, while SPC75 reduced fish survival, promoted inflammation, and regulated multiple physiological responses. Thousands of differentially expressed genes (DEGs) by SPC were identified in the intestine, liver, and muscle, which were enriched in biological regulation, cellular process, metabolic process, single-organism process, cell, cell part, membrane, binding, and catalytic activity based on RNA-seq. Notably, some DEGs involved in amino acid and lipid metabolism in the digestive system highlighted the modulatory effect of SPC on these metabolic processes, consistent with the physiological responses including enzyme activities. The enriched aspects of these predominant DEGs might be directly related to the different effects of SPC30 and SPC75 on fish growth, digestibility, and underlying enzyme activities and histology. In conclusion, the comprehensive physiological and transcriptomic comparative analysis of CK, SPC30, and SPC75 was also effective in testing the applicability of SPC as fish feeds and in designing a proper diet with the best impact on the growth performance and health of fish in hybrid grouper.

The yellowtail clownfish (Amphiprion clarkii) is a hermaphrodite fish, whose sex differentiation and gonad development are closely related to its social status. The kisspeptin/KissR system is regarded as a key factor mediating social stress on reproductive regulation. In order to understand the effects of social rank stress on the yellowtail clownfish gonadal differentiation, full-length cDNAs of two paralogous genes encoding kisspeptin (kiss1 and kiss2) and KissR (kissr2 and kissr3) were cloned and characterized. The results of real-time PCR showed that kiss1 was primarily expressed in the hypothalamus, and kiss2/kissr2 were abundantly expressed in the liver, while kissr3 was almost exclusively concentrated in the cerebellum and pituitary. Moreover, both Kiss1-10 and Kiss2-10 peptides could initiate downstream signaling pathways by interacting with cognate receptors expressed in eukaryotic cells. Among the three social status groups, the mRNA levels of kiss2 in the hypothalamus and pituitary as well as kissr2 in the pituitary were significantly higher in subordinate individuals (nonbreeders) than dominate individuals (females and males); while the mRNA levels of kissr3 in the hypothalamus and gonad were low in subordinate individuals. Furthermore, the plasma estradiol (E 2 ) and testosterone (T) levels were higher in subordinate than dominate individuals. This study shows that kiss2 is involved in the regulation of social stress on the gonad development in the yellowtail clownfish, but not kiss1., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Meagre (Argyrosomus regius) undergo spermatogenesis and spermiation when reared in captivity, but often produce low milt volumes, sometimes with reduced quality and for a limited time period. In the present study we a) compared the efficiency of gonadotropin releasing hormone agonist (GnRHa) implants versus injections on testicular stimulation and spermiation enhancement, b) investigated the effect of GnRHa on the endocrine spermiation regulation (sex steroid hormones), and c) evaluated a commercial induced spawning simulation scenario. Firstly, males (n = 5) were injected with 15.0 ± 0.2 μg GnRHa kg -1 (Injections) or implanted with 51.0 ± 5.1 μg GnRHa kg -1 (Implant) and compared their sperm production response. Secondly, the best hormonal treatment (Implant) was tested treating males (n = 8) with 57.5 ± 7.5 μg GnRHa kg -1 every 3 weeks for a period of 70 days. Milt production was improved by the GnRHa implants with only minor sperm quality alterations (improved sperm motility percentage). Elevated plasma testosterone (T) and 11-ketotestosterone (11-KT) levels were recorded in response to GnRHa implants, while no significant difference for 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) was observed. In the commercial induced spawning simulation, it was shown that meagre females are capable of on-demand induction of spawning at random intervals (5-21 days) using GnRHa injections, over a period of at least 2.5 months. During this period, spermiation enhancement was achieved with GnRHa implants every 3 weeks, producing sperm with stable, in general, quality and motility parameters. Percentage of motile cells, motility duration and density fluctuated significantly, but remained within levels that are considered appropriate for high fertilization success in this species., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

An 8-week feeding trial was conducted to evaluate the effects of dietary xylanase supplementation on growth performance, digestive enzyme activity, intestinal morphology parameter, intestinal microbiome diversity, and carbohydrate metabolism for juvenile large yellow croaker (Larimichthys crocea). Four levels of xylanase were added to basal diets (0, 600, 1200, and 1800 U kg -1 ). The results indicated that fish fed the 1200 U kg -1 xylanase diet had higher weight gain than those fed the 0 and 600 U kg -1 xylanase diet. The highest intestinal folds and microvillous height were observed at fish fed the 1200 U kg -1 xylanase diet. High-throughput sequencing revealed that the majority of reads derived from the large yellow croaker digesta belonged to members of Proteobacteria followed by Chloroflex, Bacteroidetes, Spirochaetae, and Firmicute. Supplementation of xylanase in diets increased the relative abundance of Bacteroides and Gemmatimonadete. The higher hepatic glucokinase (GK) and glucose-6-phosphate dehydrogenase (G6PD) activities were observed in fish fed the xylanase supplementation diet. Accordingly, dietary xylanase supplementation upgraded the relative expressions of gk and g6pd genes in liver. In conclusion, optimum dietary xylanase supplementation (600-1200 U kg -1 ) could improve the growth performance, optimize the intestinal morphology structure and microbiota constitution, and enhance the ability of carbohydrate utilization of juvenile large yellow croaker.

Although, the Red sea is highly rich with fish fauna but still the information known is so limited for the researchers especially about the fish physiology baselines. So, in the present study we investigated the heamto-biochemical parameters and cell characterization of two fish having the same feeding habitats. Fish specimens of Red Sea goatfish (Parupeneus forsskali) and Klunzinger's wrasse (Thalassoma klunzingeri) were captured from Hurghada, Egypt. Haematological and biochemical analysis as well as blood cells characterization were performed. The morphological and cytochemical aspects of peripheral blood cells of the two species were studied by light microscopy. Thalassoma klunzingeri showed lower Hct and Hb values and RBCs count. Fusiform to spindle shape thrombocytes were found only in the blood of Parupeneus forsskali while spiked thrombocytes were found only in the blood of Thalassoma klunzingeri. This investigation may be helpful as a tool to monitor the health status of the two species and will be used as biomarkers for clinical pathology., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Variation in food intake affects somatic growth by altering the expression of hormones in the somatotropic endocrine axis including insulin-like growth factor-1 (IGF-1). Here, we examined IGF-1 pathway responses to long- and short-term variation in food availability in copper rockfish (Sebastes caurinus), a nearshore Pacific rockfish important for commercial and recreational fisheries. Juvenile copper rockfish were raised under differing ration amounts (3% or 9% mass feed·g -1 fish wet mass·day -1 ) for 140 d to simulate 'long-term' feeding variation, after which some fish from both rations were fasted for 12 d to generate 'short-term' conditions of food deprivation. Rockfish on the 9% ration treatment grew more quickly than those on the 3% ration and were larger in mass, length, and body condition (k) after 152 d. Fish on the 9% ration had higher blood glucose than those on the 3% ration, with fasting decreasing blood glucose in both ration treatments, indicating that both long-term and short-term feed treatments altered energy status. Plasma IGF-1 was higher in rockfish from the 9% ration than those in the 3% ration and was also higher in fed fish than fasted fish. Additionally, plasma IGF-1 related positively to individual variation in specific growth rate (SGR). The positive association between IGF-1 and SGR showed discordance in fish that had experienced different levels of food and growth over the long-term but not short-term, suggesting that long-term nutritional experience can influence the relationship between IGF-1 and growth in this species. Rockfish on the 3% ration showed a lower relative abundance of gene transcripts encoding igf1 in the liver, but higher hepatic mRNAs for IGF binding proteins igfbp1a and igfbp1b. Fasting similarly decreased the abundance of igf1 mRNAs in the liver of fish reared under both the 9% and 3% rations, while concurrently increasing mRNAs encoding the IGF binding proteins igfbp1a, -1b, and -3a. Hepatic mRNAs for igfbp2b, -5a, and -5b were lower with long-term ration variation (3% ration) and fasting. Fish that experienced long-term reduced rations also had higher mRNA levels for igfbp3a, -3b, and IGF receptors isoforms A (igf1rA) and B (igf1rB) in skeletal muscle, but lower mRNA levels for igf1. Fasting increased muscle mRNA abundance for igfbp3a, igf1rA, and igf1rB, and decreased levels for igfbp2a and igf1. These data show that a positive relationship between circulating IGF-1 and individual growth rate is maintained in copper rockfish even when that growth variation relates to differences in food consumption across varying time scales, but that long- and short-term variation in food quantity can shift basal concentrations of circulating IGF-1 in this species., (Copyright © 2019 Elsevier Inc. All rights reserved.)

We compared baseline and maximal cortisol concentrations between predator exposure and prey blood samples in pumpkinseed Lepomis gibbosus, captured using a standardised fishing event underneath osprey Pandion haliaetus nests and away from osprey nests. We did not detect differences in cortisol or glucose between sites. These findings suggest that predictable sources of predation risk may not confer stress-related costs in teleosts., (© 2019 The Fisheries Society of the British Isles.)

We examined the influence of blood deposition on flesh quality of ordinary muscle in yellowtail. This study compared the flesh quality changes in upper and under-sides of yellowtails killed by two different methods: spinal-cord destruction (SCD) with blood removal and suffocation in air without blood removal (SA). The under-sides of the SA group showed the highest values for a*, cathepsin B and B + L activities, the lowest value in breaking strength and the greatest degradation of myosin heavy chain (MHC) among the four groups. However, the values of the SCD-upper group indicated the best flesh quality. In addition, the white blood cells presented the highest cathepsin B and B + L activities among the blood components. These results indicate that blood has the tendency to deposit downward in accordance with the direction of placement. This phenomenon influences the distribution of white blood cells which contain enzymes that accelerate the deterioration of flesh quality. The texture of fish muscle is an important part of the flesh quality. In captured fishery (purse-seine fishery and dragnet fishery), it is impossible to immediately and completely remove blood. Therefore, suffocation in air is the common method after the fish is caught. The commercial value of fish is decreased and the price varies greatly when these fish enter market circulation. In our study, we examined the influence of blood deposition on the flesh quality of yellowtail during storage. The degradation of structural proteins accelerated in the deposited blood which contain proteases. The movement and deposition of blood caused the difference of quality on both sides, which seriously affected the quality of fish during preservation. Our study has some theoretical guidance for muscle softening and give a better understanding of the adverse effect of blood during preservation., (© 2019 Wiley Periodicals, Inc.)

This study determined high temperature effects on ovarian development in a marine groundfish species, sablefish (Anoplopoma fimbria), with potential application in sex reversal or sterilization for aquaculture. Monosex female (XX-genotype) sablefish larvae (∼30 mm) were randomly divided into three groups and exposed to control (15.6 °C ± 0.8 °C), moderate (20.4 °C ± 0.5 °C), or high (21.7 °C ± 0.5 °C) temperatures for 19 weeks. Treated fish were then tagged and transferred to ambient seawater (11.2 °C ± 2.3 °C) for one year to determine whether temperature effects on reproductive development were maintained post-treatment. Fish were periodically sampled for gonadal histology, gene expression and plasma 17β-estradiol (E2) analyses to assess gonadal development. Short-term (4-week) exposure to elevated temperatures had only minor effects, whereas longer exposure (12-19 weeks) markedly inhibited ovarian development. Fish from the moderate and high treatment groups had significantly less developed ovaries relative to controls, and mRNA levels for germ cell (vasa, zpc) and apoptosis-associated genes (p53, casp8) generally indicated gonadal degeneration. The high treatment group also had significantly reduced plasma E2 levels and elevated gonadal amh gene expression. After one year at ambient temperatures, however, ovaries of moderate and high treatment fish exhibited compensatory recovery and were indistinguishable from controls. Two genotypic females possessing immature testes (neomales) were observed in the high treatment group, indicating sex reversal had occurred (6% rate). These results demonstrate that extreme elevated temperatures may inhibit ovarian development or trigger sex reversal. High temperature treatment is likely not an effective sterilization method but may be preferable for sablefish neomale broodstock production., (Published by Elsevier Inc.)

The thermal stability of the Antarctic Ocean raises questions concerning the metabolic plasticity of Antarctic notothenioids to changes in the environmental temperature. In this study, Notothenia rossii survived 90 days at 8 °C, and their condition factor level was maintained. However, their hepatosomatic (0.29×) index decreased, indicating a decrease in nutrient storage as a result of changes in the energy demands to support survival. At 8 °C, the plasma calcium, magnesium, cholesterol, and triglyceride concentrations decreased, whereas the glucose (1.91×) and albumin (1.26×) concentrations increased. The main energy substrate of the fish changed from lipids to glucose due to a marked increase in lactate dehydrogenase activity, as demonstrated by an increase in anaerobic metabolism. Moreover, malate dehydrogenase activity increased in all tissues, suggesting that fish acclimated at 8 °C exhibit enhanced gluconeogenesis. The aerobic demand increased only in the liver due to an increase (2.23×) in citrate synthase activity. Decreases in the activities of superoxide dismutase, catalase, and glutathione-S-transferase to levels that are most likely sufficient at 8 °C were observed, establishing a new physiological activity range for antioxidant defense. Our findings indicate that N. rossii has some compensatory mechanisms that enabled its long-term survival at 8 °C.

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., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The Siamese fighting fish, Betta splendens, and the blue gourami, Trichopodus trichopterus, are two closely related air-breathing anabantoid fishes. B. splendens is a sedentary facultative air breather frequenting often hypoxic waters, while T. trichopterus is a more active obligatory air-breather inhabiting better oxygenated waters. Despite their close taxonomic relationship, previous studies have shown inter-specific differences in both physiological and morphological plasticity. Consequently, we hypothesized that B. splendens would have the higher blood oxygen affinity characteristics typical of more hypoxia-tolerant fishes. Whole blood oxygen equilibrium curves were determined at 27 °C and pHs of 7.62, 7.44 and 7.25. At a pH of 7.62, the blood O 2 affinity (P 50 ) of B. splendens was just 2.9 mmHg, while that of T. trichopterus was ~5 times higher at 14.7 mmHg. There were no significant differences in P 50 between males and females in either species. The Bohr coefficient in B. splendens and T. trichopterus was -1.79 and - 0.83, respectively. B. splendens, unlike T. trichopterus, showed a large Root effect. Hills cooperatively coefficient, n, was ~2 in both species, indicating a significant binding cooperative between oxygen and hemoglobin. Collectively, these differences in blood O 2 transport characteristics in these two closely related species are likely correlated with the differing habitats in which they breed and inhabit as adults, as well as different activity levels. Finally, the very high blood O 2 affinity of B. splendens is not extraordinary among air-breathing fish, as revealed by a review of the literature of blood oxygen affinity in air-breathing fishes., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Low temperature influences multiple physiological processes in fish. To explore the adaptability of the large yellow croaker (Larimichthys crocea) to low temperature, the concentrations of glycerol, blood urea nitrogen (BUN), and triglycerides (TG) in plasma, as well as the expression levels of metabolism-related genes aqp7 and aqp10, were measured after exposure to low temperature stress and during subsequent rewarming. In addition, tissue samples from the intestine and liver were histologically analyzed. We found that the concentrations of plasma glycerol, BUN, and TG, decreased under low temperature stress, suggesting the metabolism of fat and protein slowed at low temperature. The expression levels of aqp7 and aqp10 mRNA were also downregulated under exposure to low temperature. Interestingly, above plasma indices and gene expression returned to basic levels within 24 h after rewarming. Furthermore, the liver and the intestine were damaged under continuous low temperature stress, whereas they were repaired upon rewarming. From the above results, we concluded that aqp7 and aqp10 genes were sensitive to low temperature, and that the decrease in their expression levels at low temperature might reduce energy consumption by L. crocea. However, the adaptation to low temperature was limited because the key metabolic tissues were damaged under continuous exposure to low temperature. Interestingly, the metabolism of L. crocea was basically back to normal within 24 h of rewarming, showing that it has high capacity of self-recovery.

Nonylphenol (NP) and Cadmium (Cd) are two common contaminants that can be detected in aquatic environments. Nevertheless, the combined toxicity of NP and Cd at environmentally relevant concentrations in aquatic organisms has not been thoroughly characterized to date. In the present study, the interactions between NP and Cd on male Sebastiscus marmoratus were studied. After 21 days of exposure, the brain aromatase activity was observed to be significantly induced by 100 ng/L NP and 40 μg/L Cd, whereas all of the concentrations of co-treatment resulted in an increase in brain aromatase activity. Additionally, NP could also reduce plasma testosterone concentration, while NP, Cd and their mixture could induce plasma 17β-estradiol (E2) concentration and VTG concentration. The interactions between NP and Cd on the reproductive physiology were antagonism. Our results also support the notion of using these indicators as biomarkers for exposure to EDCs and further extend the boundary of biomonitoring to environmental levels., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The newly emerging tissue microbiota hypothesis suggests that bacteria found in blood and tissues play a role in host health, as these bacterial communities have been associated with various noncommunicable diseases such as obesity, liver disease, and cardiovascular disease. Numerous reports have identified bacteria in the blood of healthy finfish, indicating bacteremia may not always indicate disease. Current research priorities in aquaculture include the development of technologies and practices that will allow for an effective reduction in antibiotic use for the prevention and treatment of disease. Overall, a better understanding of fish health is needed, particularly among species selected for commercial-scale production. This study investigated blood characteristics of cultured Red Drum Sciaenops ocellatus with the tissue microbiota hypothesis in mind. Bacterial assemblages within the blood were characterized using next-generation sequencing and compared with other various blood characteristics, including innate immune function enzymes, between two fish cohorts reared in aquaculture. A total of 137 prokaryotic operational taxonomic units (OTUs) were identified from the blood of Red Drum. Microbiota diversity and structure varied greatly among individuals, for which the number of OTUs ranged from 4 to 58; however, predicted metagenomic function was highly similar between individuals and was dominated by the metabolism of carbohydrates and amino acids and membrane transport. Communities were dominated by Proteobacteria, followed by Bacteroidetes, Firmicutes, and Actinobacteria. The most commonly identified genera included Acinetobacter, Bacillus, Corynebacterium, and Pseudomonas. Three genera previously identified as containing marine fish pathogens were detected: Corynebacterium, Pantoea, and Chryseobacterium. A subset of bacterial OTUs were positively correlated with superoxide dismutase activity and negatively correlated with lysozyme activity, indicating a relationship between blood microbiota and the innate immune system. The results of this study provide further evidence for the tissue microbiota hypothesis and demonstrate the potential for these bacterial communities to be linked to immunological characteristics often used as biomarkers for fish health., (© 2018 American Fisheries Society.)

In Seriola species, exposure to a long photoperiod regime is known to induce ovarian development. This study examined photoperiodic effects on pituitary gene expression and plasma levels of follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh) in previtellogenic greater amberjack (Seriola dumerili). The fish were exposed to short (8L:16D) or long (18L:6D) photoperiod. The water temperature was maintained at 22 °C. Compared with the short-photoperiod group, plasma Fsh levels were higher on days 10 and 30 in the long-photoperiod group, but plasma Lh levels did not significantly differ. On day 30, pituitary Fsh- and Lh-β subunit gene expressions were also higher in the long-photoperiod group than the short-photoperiod group, whereas α-subunit gene expressions were higher on days 20 and 30. Throughout the experiment, average gonadosomatic index and plasma E2 levels did not significantly differ between the two groups. This study clearly demonstrated that a long photoperiod induced Fsh release in the previtellogenic fish followed by upregulation of pituitary Fsh and Lh subunit gene expressions. An increase in plasma Fsh levels may be a key factor that mediates the photoperiodic effect on the initiation of ovarian development., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Cottonseed protein concentrate (CPC) has similar amino acid composition compared with fish meal, and has the characteristics of low gossypol and low toxicity. The present study was conducted to investigate the growth performance, antioxidant capacity and different intestinal segments immune responses of hybrid grouper to replacement dietary fish meal ofCPC. Six iso-nitrogenous (50% crude protein) and iso-lipidic (10% crude lipid) diets were formulated: a reference diet (FM) containing 60% fishmeal and five experimental diets (12%, 24%, 36%, 48 and 60%) in which fishmeal protein was substituted at different levels by CPC to feed fish (initial body weight: 11 ± 0.23 g) for 8 weeks. Thena challenge test with injection of Vibrio parahaemolyticus was conducted for 7 days until the fish stabilized. The results showed that specific growth rate (SGR) was the highest with 24% replacement level and feed conversion ratio (FCR)was significantly increased when the replacement level reached 48% (P < 0.05). The content of malonaldehyde (MDA) in the serum was significantly increased when the replacement level reached 36% (P < 0.05). The plica height in the proximal, mid and distal intestine were significantly decreased with the replacement level up to 48% (P < 0.05). Hepatic fat deposition wasaggravatedwhen the replacement level reached 36% (P < 0.05). The expression of IL-6, TNF-α, and IL-1β mRNAs were significantly up-regulated (P < 0.05). The hepcidin mRNA expression was significantly down-regulated (P < 0.05). In proximal intestine (PI) and mid intestine (MI), IFN-γ mRNA expression was significantly up-regulated (P < 0.05). These results suggested that the CPC decreased hybrid grouper growth performance and inflammation function, and different inflammation function responses in PI,MI, and distal intestine (DI) were mediated partly by the TLR-2/MyD88 signaling pathway. According to the analysis of specific growth rate, the dietary optimum replacement level and maximum replacement level were estimated to be 17% and 34%, respectively., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

An 8-week study was conducted to evaluate three different diets supplemented with bovine lactoferrin (LF) at 0 (control), 800, and 1200 mg LF kg -1 diet on somatic growth, hemato-immunological parameters, antioxidant status, and digestive enzyme activities in silvery-black porgy (Sparidentex hasta) juveniles. Fish fed the 800 mg LF kg -1 diet had higher growth performance and feed utilization parameters than the other groups. Hematological and liver antioxidant parameters were not affected by dietary LF supplementation. Fish fed the 800 mg LF kg -1 diet had higher plasma lysozyme activity values than the other groups. Total protease activity was higher in fish fed LF-supplemented diets than the control group. Results indicated that diet supplemented with 800 mg kg -1 for 8 weeks enhanced somatic growth performance, lysozyme activity, and proteolytic digestive enzyme activities in S. hasta, as well as improving feed efficiency parameters like the protein efficiency and feed conversion ratios.

Bacterial ghosts (BGs) can be generated by the controlled expression of the PhiX174 lysis gene E in gram-negative bacteria. They are intriguing vaccine candidates since ghosts retain functional antigenic cellular determinants often lost during traditional inactivation procedures. Here we prepared Edwardsiella tarda ghost (ETG) and tested different concentrations in vaccination trials. The results showed that serum IgM antibody titers were significantly higher in three different concentration immunization groups than control group (P < 0.05), However, there was no significant (P > 0.05) difference between the immunized groups. The phagocytic percentage (PP) was significantly higher (P < 0.05) in ETG immunized groups than in the control group from 3 days post-treatment. The PP continued to rise with time until day 21, when the values of three ETG immunized groups were 45.7%,51.2% and 50.7%, respectively. In addition, phagocytic index (PI) was significantly higher (P < 0.05) in ETG immunized groups than in the control group after 7 days post-treatment. However, there was no significant (P > 0.05) difference of PP or PI between immunized groups. In addition, non-specific immune immunity, such as acid phosphatase, alkaline phosphatase, superoxide dismutase and lysozyme activities displayed a similar pattern in all immunized groups, all immunized fish showed significantly higher activities than control group fish (P < 0.05). Most importantly three ETG immunized groups were all significantly more protected against the E. tarda challenge (19/25, 76% survival), (21/25, 84% survival) and (20/25, 80% survival) respectively, compared to (9/25, 36% survival) survival in the control group, but there was no significant (P > 0.05) difference of survival rate (SR) or relative percent survival (RPS) between immunized groups. All these results suggest that an ETG could stimulate cellular and humoral immunity, and could be used as a vaccine candidate in S.m. In summary, ETG can protect fish from Edwardsiellosis, and there is no significant difference in SR and RPS when three different concentrations of ETG are used, so it can easily be developed as a vaccine for mechanical and artificial operations., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Serum lipid metabolic responses are associated with certain metabolic disorders induced by dietary habits in mammals. However, such associations have not been reported in fish. Lipidomic analyses were performed to investigate fish lipid metabolic responses to a dietary vegetable oil (VO) blend and to elucidate the mechanism of how the dietary VO blend affects serum lipid profiles. Results showed that the dietary VO blend strongly affects serum lipid profiles, especially the ratio of triglyceride/phosphatidylcholine (TAG/PC), via inhibiting hepatic PC biosynthesis and facilitating hepatic and intestinal lipoprotein assembly. Studies in vitro suggested that changes of serum TAG/PC ratio may be partially attributed to altered fatty acid composition in diets. Additionally, the reduction of 16:0/18:1-PC induced by the dietary VO blend may play a role in abnormal lipid deposition through inhibiting PPARA-mediated activation of β-oxidation. These findings suggested that the serum TAG/PC ratio might be a predictive parameter for abnormal lipid metabolism induced by dietary nutrition in fish.

As amino acids (AAs) are vital molecules in the metabolism of all living organisms and are the building blocks of enzymes, a 6-week feeding trial was conducted for determining the influence of dietary essential amino acid (EAA) deficiencies on pancreatic, plasma, and hepatic enzyme activities in silvery-black porgy (initial weight 4.7 ± 0.01 g) juveniles. Eleven isoproteic (ca. 47%) and isoenergetic (ca. 20.5 MJ kg -1 ) diets were formulated including a control diet, in which 60% of dietary nitrogen were provided by intact protein (fish meal, gelatin, and wheat meal) and 40% by crystalline AA. The other 10 diets were formulated by 40% reduction in each EAA from the control diet. At the end of the experiment, fish fed with threonine-deficient diet showed the lowest survival rate (P < 0.05), whereas growth performance decreased in fish fed all EAA-deficient diets, although the reduction in body growth varied depending on the EAA considered. Pancreatic enzymes (trypsin, lipase, α-amylase, and carboxypeptidase A) activities significantly decreased in fish fed the EAA-deficient diets in comparison with fish fed the control diet (P < 0.05). Fish fed with the arginine-deficient diet had the highest plasma and liver alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels (P < 0.05). Plasma and liver lactate dehydrogenase and superoxide dismutase showed the highest and lowest values, respectively, in fish fed the arginine and lysine-deficient diets (P < 0.05). Plasma metabolites were significantly affected by dietary EAA deficiencies (P < 0.05). The results of this study suggesting dietary EAA deficiencies led to reduction in growth performance as well as pancreatic and liver malfunction. Furthermore, arginine and lysine are the most limited EAA for digestive enzyme activities and liver health in silvery-black porgy.

The immunosuppressive effects of androgens are a key component of the immunocompetence handicap hypothesis (ICHH). Here, we use bluegill sunfish (Lepomis macrochirus) to test two predictions arising from this hypothesis: (1) natural circulating concentrations of the androgen 11-ketotestosterone (11-KT) will be negatively related with measures of immunity, and (2) immune stimulation will lower circulating 11-KT concentration. We found no evidence for a relationship between natural circulating 11-KT concentration and measures of immunity (lymphocyte and granulocyte counts, respiratory burst, cytokine mRNA levels), and an immune stimulation with Vibrio vaccine did not affect circulating 11-KT concentration. We also performed a meta-analysis of immune stimulation studies to help interpret our results, and report evidence suggesting that immune stimulation has weaker effects on androgen levels in fishes compared to other vertebrates. These results suggest that the ICHH may not apply to all vertebrates, although it remains premature to state what factors account for the weaker evidence in fishes that androgens are immunosuppressive., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Epinephelus morio is a large carnivorous species of the Caribbean Sea under reproduction in captivity and nutritional physiology. A diet with raw cornstarch (RCS) was compared to a basal diet without starch (basal) to measure plasma glucose, liver glycogen, and intermediary metabolism. Glucose level did not change (p > 0.05) whereas liver glycogen was significantly higher in fish fed the RCS diet (137.2 ± 14.5 mg g -1 ) than in fish fed the basal diet (87.4 ± 14.5 mg g -1 ). Oral glucose administration (170 mg glucose per 100 g body weight) yielded a slight change; two peaks of plasma glucose were recorded with basal (5.6 mM L -1 ) 2 h after oral administration and at 12 h (6.4 mM L -1 ). After 24 h, with 1.7 mM L -1 , fish returned to initial stage (2.4 mM L -1 ). RCS diet produced the highest level (6.3 mM L -1 ) 2 h after oral administration; lowest level observed at 24 h after oral administration (1.0 mM L -1 ). A significant effect was detected with the presence or absence of dietary carbohydrates (CBH) on hepatic fructose 1,6-bisphosphatase and pyruvate kinase activity. Grouper used two strategies to maintain glucose homeostasis: CBH present in the diet oriented towards gluconeogenesis, whereas no dietary CBH enhanced glycolytic route to liberate glucose and increase liver glycogen.

In order to better understand how photo-thermal conditions affect oogenesis in captive-bred F1 hāpuku, a wreckfish considered for aquaculture in New Zealand, juvenile (pre-pubertal) fish were assigned to one of two regimes: exposed to a constant temperature of 17°C (CT group) or to seasonally varying temperatures (VT group range: 10-17°C), both under simulated ambient photoperiod, for nearly 2years. Development in females was monitored through repeated gonadal biopsies (histology; target gene mRNA levels) and blood sampling (plasma levels of estradiol-17β; E2). Very little evidence of advancing oogenesis was found in the first year of study, when fish were in their 4th year. In the subsequent year, a proportion of fish reached the pre-spawning stage (fully-grown ovarian follicles); the proportion of females reaching this stage was notably higher in the VT (62%) than the CT (28%) group. Of the few females that did reach maturity in the CT group, significantly lower levels of plasma E2 were observed relative to those in fish from the VT group possibly indicating a temperature-induced endocrine impairment during oogenesis. Interestingly, females that did not reach the pre-spawning stage presented with a small transient, but significant increase in oocyte diameters and plasma E2, suggestive of a dummy run. Clear seasonality was observed for fish under both photo-thermal regimes, and this was reflected in plasma E2 levels and transcript abundances of aromatase, fshr and luteinizing hormone receptor in the ovary; these end points all peaked in maturing females during the late or post-vitellogenic stage. We conclude that captive female F1 hāpuku first mature as five-year-olds and that exposure to a decreased temperature is important for appropriate progression of oogenesis., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The three-spot wrasse, Halichoeres trimaculatus, can change sex from female to male (i.e. protogyny) due to sharp decrease in endogenous estrogen. During the sex change, ovarian tissue degenerates and testicular tissue arises newly. Finally, ovarian tissue disappears completely and replaces into mature testis. In order to predict the molecular mechanisms controlling the processes of sex change, we investigated the expression patterns of four genes (rspo1, figla, sox9b and amh), which have been thought to be associated with ovarian/testicular differentiation in vertebrates. Expression levels of rspo1 and figla, which play important roles for ovarian differentiation in vertebrates, were stable until the middle stage of the sex change, and subsequently down-regulated. Therefore, it was indicated that decrease in rspo1 and figla could result from ovarian degeneration. On the other hand, basis on the expression pattern, it was indicated that sox9b and amh, which are involved in testicular differentiation in vertebrates, were implicated in testicular formation and spermatogenesis during the sex change as well. The present results could be fundamental information for investigating the relationship between these factors and E2 depletion, which is crucial trigger for sex change., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The investigation deals with the effects of Acid Mine Drainage (AMD) of coal mine on fish Channa punctata (Bloch) by examining the incidence of haematological, morphological, histological changes and DNA fragmentation in tissues of C. punctata in laboratory condition. For this study fishes were exposed to 10% of AMD for a period of 30 days. The fusion of the primary and secondary gill lamellae, distortion, loss of alignment, deposition of worn out tissues and mucous on the surface of the lamella in the gills; degeneration of morphological architecture, loss of alignment of tubules, mucous deposition in the kidney; cellular damage, cellular necrosis, extraneous deposition on the surface, pore formation in the liver are some important changes detected by scanning electron microscopy. Fishes of AMD treated group showed gradual significant decrease in TEC, Hb and, increase in TLC and DLC as compared to that of the control. DNA fragmentation observed in kidney of fishes from treated group indicates an intricate pollutant present in the AMD. The high incidence of morphological and histological alterations, haematological changes along with DNA breakage in C. punctata is an evidence of the cytotoxic and genotoxic potential of AMD of coal mines., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Fishes have evolved physiological mechanisms to exhibit stress response, where hormonal signals interact with an array of ion transporters and regulate homeostasis. As major ion transport regulators in fish, cortisol and thyroid hormones have been shown to interact and fine-tune the stress response. Likewise, in fishes many interactions have been identified between stress and immune components, but the physiological basis of such interaction has not yet delineated particularly in air-breathing fish. We, therefore, investigated the responses of thyroid hormones and cortisol, ion transporter functions and non-specific immune response of an obligate air-breathing fish Anabas testudineus Bloch to zymosan treatment or hypoxia stress or both, to understand how immune challenge modifies the pattern of stress response in this fish. Induction of experimental peritonitis in these fish by zymosan treatment (200ngg -1 ) for 24h produced rise in respiratory burst and lysozomal activities in head kidney phagocytes. In contrast, hypoxia stress for 30min in immune-challenged fish reversed these non-specific responses of head kidney phagocytes. The decline in plasma cortisol in zymosan-treated fish and its further suppression by hypoxia stress indicate that immune challenge suppresses the cortisol-driven stress response of this fish. Likewise, the decline in plasma T 3 and T 4 after zymosan-treatment and the rise in plasma T 4 after hypoxia stress in immune-challenged fish indicate a critical role for thyroid hormone in immune-stress response due to its differential sensitivity to both immune and stress challenges. Further, analysis of the activity pattern of ion-dependent ATPases viz. Na + /K + -ATPase, H + /K + -ATPase and Na + /NH 4 + -ATPase indicates a functional interaction of ion transport system with the immune response as evident in its differential and spatial modifications after hypoxia stress in immune-challenged fish. The immune-challenge that produced differential pattern of mRNA expression of Na + /K + -ATPase α-subunit isoforms; nkaα1a, nkaα1b and nkaα1c and the shift in nkaα1a and nkaα1b isoforms expression after hypoxia stress in immune-challenged fish, presents transcriptomic evidence for a modified Na + /K + ion transporter system in these fish. Collectively, our data thus provide evidence for an interactive immune-stress response in an air-breathing fish, where the patterns of cortisol-thyroid hormone interaction, the ion transporter functions and the non-specific immune responses are reversed by hypoxia stress in immune-challenged fish., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Juvenile sablefish, Anoplopoma fimbria (mean length 17.1±2.4cm, and mean weight 75.6±5.7g) were used to evaluate toxic effects on antioxidant systems, immune responses, and stress indicators by ammonia exposure (0, 0.25, 0.75, and 1.25mg/L) at different water temperature (12 and 17°C) in 1 and 2 months. In antioxidant responses, superoxide dismutase (SOD) and catalase (CAT) were significantly increased by ammonia exposure, whereas glutathione (GSH) was decreased. In immune responses, lysozyme and phagocytosis activity were significantly increased by ammonia exposure. In stress indicators, plasma glucose, heat shock protein 70 (HSP 70), and cortisol were significantly increased. At high water temperature (17°C), alterations by ammonia exposure were more distinctly. The results of this study indicated that ammonia exposure can induce toxic effects in the sablefish, and high water temperature can affect the ammonia exposure toxicity., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The sex identity of fish can be easily manipulated by exogenous hormones. Treatment with 17-methyltestosterone (MT) has been widely used to induce a male fate, but the molecular and cellular processes underlying sex changes induced by MT treatments and the withdrawal of MT are not well studied. In this study, we systematically investigated gonadal histology, gene expression profiles, sex steroid hormone levels, and cellular changes during sex changes induced by MT-feeding and MT-feeding withdrawal in the protogynous orange-spotted grouper, Epinephelus coioides. Based on gonadal histology, we demonstrated that MT-feeding-induced sex reversal can be divided into early and late phases: in the early phase, male and female germ cells coexist, and MT-feeding withdrawal leads to a female fate; in the late phase, only male germ cells are observed, and MT-feeding withdrawal does not reverse the process, leading to a male fate. In both the early and late phases, cytochrome P450 family19 subfamily A member 1 (cyp19a1a) gene expression increased in response to MT-feeding withdrawal. Finally, by tracing doublesex- and Mab-3-related transcription factor 1 (dmrt1)-expressing cells, we found that gonia-like cells in the germinal epithelium might be the major germ cell sources for developing testes during sex reversal. Collectively, our findings provide insights into the molecular and cellular mechanisms underlying sex changes induced by exogenous hormones., (© The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Hypothalamic peptides, gonadotropin-releasing hormone (GnRH) and gonadotropin inhibitory hormone (GnIH), play pivotal roles in the control of reproduction and gonadal maturation in fish. In the present study we tested the possibility that stress-mediated reproductive dysfunction in teleost may involve changes in GnRH and GnIH activity. We studied expression of brain GnIH, GnIH-R, seabream GnRH (sbGnRH), as well as circulating levels of follicle stimulating hormone (FSH), and luteinizing hormone (LH) in the cinnamon clownfish, Amphiprion melanopus. Treatment with cortisol increased GnIH mRNA level, but reduced sbGnRH mRNA and circulating levels of LH and FSH in cinnamon clownfish. Using double immunofluorescence staining, we found expression of both GnIH and GnRH in the diencephalon region of cinnamon clownfish brain. These findings support the hypothesis that cortisol, an indicator of stress, affects reproduction, in part, by increasing GnIH in cinnamon clownfish which contributes to hypothalamic suppression of reproductive function in A. melanopus, a protandrous hermaphroditic fish., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Biologically active recombinant yellowtail kingfish follicle stimulating hormone (rytkFsh) was produced in yeast Pichia pastoris and its biological activity was demonstrated by both in-vitro and in-vivo bioassays. Incubation of ovarian and testicular fragments with the recombinant hormone stimulated E 2 and 11-KT secretion, respectively. In-vivo trial in immature female YTK resulted in a significant increase of plasma E 2 levels and development of oocytes. In males at the early stages of puberty, advancement of spermatogenesis was observed, however plasma 11-KT levels were reduced when administered with rytkFsh., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The use of cleaner fish in Norwegian aquaculture has to a large extent been based on wild catches, but breeding of lumpfish and ballan wrasse is currently increasing. Due to disease problems and required vaccine development, tools to study immune responses and a better understanding of the immune system in these species is demanded. The present study comprises lumpfish (Cyclopterus lumpus) and five species of wrasses: Ballan wrasse (Labrus bergylta), rock cook (Centrolabrus exoletus), cuckoo wrasse (Labrus mixtus), corkwing wrasse (Symphodus melops), and goldsinny wrasse (Ctenolabrus rupestris). We present a comparison of the IgM sequences, phylogenetic relationship to other teleosts and characteristic features of IgM in the species studied. The lumpfish IgM heavy chain sequence was assembled from high throughput cDNA sequencing whereas the wrasse sequences were determined by molecular cloning. The secreted form of the IgM heavy chain from all species consisted of four constant Ig domains. IgM was purified from lumpfish and ballan wrasse sera by gel filtration followed by anion exchange chromatography, and polyclonal sera were produced against these proteins. Antisera against ballan wrasse IgM showed cross-reactivity to all analyzed species of wrasses, some cross-reactivity to lumpfish, very low reaction to salmon, and no reaction to cod. Anti- IgM sera against lumpfish cross-reacted to the light chain of all species studied. Wrasses and lumpfish IgM showed high binding affinities for protein A. IgM concentration in adult ballan wrasse (700-800 g) was measured by single radial immunodiffusion assay and found to be 13.4 mg/ml which is about 36% of the total protein concentration. The IgM concentration in lumpfish (600-3600 g) was estimated to 1-2.6 mg/ml, which corresponds to approximately 3% of the total protein concentration., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Neutrophils are a short-lived, terminally differentiated, innate immune cell, that are critical first responders during infection. Research into neutrophil-pathogen interactions in fish has primarily employed cells derived from the pro-nephros and nephros. Since these sites are also the location of neutrophil and other immune cell development, there may be some ambiguity in maturation and functional ability of these cells, and difficulty in differentiating the effects of neutrophils from those of macrophages and monocytes. In contrast, peripheral blood circulating neutrophils are mature and ready to respond, thus it may be more physiologically relevant to use these cells for immune studies when evaluating interactions with blood-borne pathogens. The enrichment of tropical, euryhaline fish blood cells cannot follow classic mammalian enrichment methods for several reasons: Fish have nucleated red blood cells (RBC's), a high number of reticulocytes, a very low number of granulocytic leukocytes and an osmotic tolerance, rendering techniques such as water lysis ineffective. Enrichment of neutrophils, while minimizing RBC contamination, is imperative for studies where luminescence or fluorescence signals may be confounded by background from an overabundance of RBC's. We have optimized a method for enriching neutrophils from peripheral blood, with an initial settlement step employing 6% dextran (Mr 450,000-650,000), for 30-60 min at room temperature, followed by density separation on an 8-step Percoll density gradient. This method provides a cell suspension comprising 20-50% neutrophils, free of contamination from reticulocytes. These are then suitable for luminometric or fluorometric downstream analyses., (Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.)

Background: The microbiota plays an essential role in host health, particularly through competition with opportunistic pathogens. Changes in total bacterial load and microbiota structure can indicate early stages of disease, and information on the composition of bacterial communities is essential to understanding fish health. Although Red Snapper (Lutjanus campechanus) is an economically important species in recreational fisheries and a primary aquaculture candidate, no information is available on the microbial communities of this species. The aim of this study was to survey the microbiota of apparently healthy, wild-caught Red Snapper from the Gulf of Mexico. Sampled Red Snapper showed no physical signs of disease. Tissues that are either primary entry routes for pathogens (feces, gill) or essential to disease diagnosis (blood) were sampled. Bacteria were enumerated using culture-based techniques and characterized by pyrosequencing., Results: Aerobic counts of feces and gill samples were 10 7 and 10 4  CFU g -1 , respectively. All individuals had positive blood cultures with counts up to 23 CFU g -1 . Gammaproteobacteria dominated the microbiota of all sample types, including the genera Pseudoalteromonas and Photobacterium in feces and Pseudomonas in blood and gill. Gill samples were also dominated by Vibrio while blood samples had high abundances of Nevskia. High variability in microbiota composition was observed between individuals, with percent differences in community composition ranging from 6 to 76 % in feces, 10 to 58 % in gill, and 52 to 64 % in blood., Conclusions: This study provides the first characterization of the microbiota of the economically significant Red Snapper via pyrosequencing. Its role in fish health highlights the importance of understanding microbiota composition for future work on disease prevention using microbial manipulation.

This study confirmed that observations of blue-green colouration in plasma fractions of the ballan wrasse Labrus bergylta were caused by the linear tetra-pyrrole biliverdin and that the molecule was of the physiologically relevant IXα isomer. Accumulation appears driven by chromogenic association with an unknown protein moiety which precludes enzymatic reduction and would suggest active management. It was demonstrated that the pigment did not fluctuate relative to ontogeny, or indeed binary gender in the species of interest, but mobilisation and depletion in the subset of individuals undergoing sex change at the time of study supports a potential association with gender inversion processes. It is of note that although biliverdin does have some effect on external colouration, the evidence is indicative that crypsis is a supplementary function thus other factors must be considered., (© 2016 The Fisheries Society of the British Isles.)

The present study was undertaken to develop a comprehensive understanding of how environmental cues and sex steroids relate with cyclic changes in spermatogenesis in freshwater spotted snakehead Channa punctatus that is nutritious and economically important. The seasonal histological changes in testis and annual profile of gonadosomatic index (GSI) of C. punctatus delineated the testicular cycle into four phases: regressed (December-March), preparatory (April-June), spawning (July and August) and postspawning (September-November). Among environmental variables, correlation and regression analyses exhibited an important relationship between photoperiod and testicular weight while role of rainfall was seen confined to spawning. The seasonal profile of plasma sex steroids when correlated with cyclic changes in spermatogenesis in spotted snakehead, testosterone (T) seems to be involved in controlling the major events of spermatogenesis from renewal of stem cells to spawning of spermatozoa. Another important androgen prevalent in teleosts, 11-ketotestosterone (11-KT), was high during preparatory phase, suggesting that 11-KT in addition to T plays an important role in progression of spermatogenesis and spermiation in C. punctatus. However, 11-KT was not seen to be associated with milt production and release of spermatozoa during spawning. Plasma profile of estradiol-17β (E2) during different reproductive phases revealed the involvement of E2 in repopulation of stem cells during postspawning phase and in maintaining quiescence of testis during regressed phase.

The aim of this study was to evaluate the lead toxic effects on the stress parameters and immune responses of Sebastes schlegelii. Juvenile rockfish, S. schlegelii (mean length 14.2±1.9cm, and mean weight 57.3±5.2g) were exposed for 4 weeks with the different levels of dietary lead (Pb(2+)) at 0, 30, 60, 120 and 240mg/L. The plasma cortisol and heat shock protein 70 was evaluated as stress indicators. The plasma cortisol of S. schlegelii was significantly increased in response to the dietary lead exposure over 60mg/kg at 2 weeks. After 4 weeks, the significant increase in the plasma cortisol was observed at 30 and 60mg/kg, but the level was decreased over 120mg/kg. The heat shock protein 70 of S. schlegelii was also notably elevated over 60mg/kg for 4 weeks. In the immune response, the immunoglobulin M of S. schlegelii was considerably increased over 120mg/kg for 4 weeks. A significant increase was observed in lysozyme activity. The plasma lysozyme activity of S. schlegelii was elevated over 120mg/kg after 2 weeks and 60mg/kg after 4 weeks, and kidney lysozyme activity was also increased at 240mg/kg after 2 weeks and over 120mg/kg after 4 weeks. The results indicate that dietary Pb exposure can cause a significant stress and immune stimulation of S. schlegelii., (Copyright © 2016 Elsevier B.V. All rights reserved.)