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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.)

Many teleosts possess a unique set of respiratory characteristics allowing enhanced oxygen unloading to the tissues during stress. This system comprises three major components: highly pH sensitive haemoglobins (large Bohr and Root effects), rapid red blood cell (RBC) intracellular pH (pHi) protection, and a heterogeneous distribution of membrane-bound plasma-accessible carbonic anhydrase (paCA; absence in the gills). The first two components have received considerable research effort; however, the evolutionary loss of branchial paCA has received little attention. In the current study, we investigated the availability of branchial membrane-bound CA, along with several other CA-related characteristics in species belonging to three basal actinopterygian groups: the Lepisosteiformes, Acipenseriformes and Polypteriformes to assess the earlier hypothesis that Root effect haemoglobins constrain branchial paCA availability. We present the first evidence suggesting branchial membrane-bound CA presence in a basal actinopterygian species: the Senegal bichir (Polypterus senegalus) and show that like the teleosts, white sturgeon (Acipenser transmontanus) and alligator gar (Atractosteus spatula) do not possess branchial membrane-bound CA. We discuss the varying respiratory strategies for these species and propose that branchial paCA may have been lost much earlier than previously thought, likely in relation to the changes in haemoglobin buffer capacity associated with the increasing magnitude of the Bohr effect. The findings described here represent an important advancement in our understanding of the evolution of the unique system of enhanced oxygen unloading thought to be present in most teleosts, a group that encompasses half of all vertebrates., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Starvation has influence on physiology and pharmacokinetic (PK) characteristics of many drugs in land animals. However, similar PK information in fish is lacking. The current study examined the effects of starvation on fish PK, taking florfenicol (FF) in Asian seabass as an example. FF was orally administered at a single dose of 10 mg/kg into 35-day starved fish reared at 25 and 30°C and the serum FF concentration was analyzed by HPLC-FLD. At 30°C, the absorption and elimination half-lives of the starved fish were increased by 30% (from 0.44 to 0.57 h) and 55% (from 7.2 to 11.18 h), respectively. The volume of distribution, clearance, and area under the curve were changed from 1.25 to 0.71 L/kg, 0.120 to 0.044 L/kg/h, and 88 to 228 h·μg/ml, respectively. Similar starvation-induced PK changes were also observed at 25°C. The serum biochemical parameters, mainly the alanine aminotransferase, aspartate aminotransferase, and glucose levels, were significantly reduced in the starvation group. Overall, FF absorption, distribution, and elimination rates were reduced by starvation, resulting in four to five times lower optimal dosage than the non-starved fish. Drug treatment in starved fish should be treated with caution as overdosing and/or tissue residues could perceivably occur., (© 2023 John Wiley & Sons Ltd.)

Lake Shkodra is characterized by a high biodiversity of fish fauna. The high biodiversity that characterized the lake is the result of a good communication with the sea, and of an extensive network of rivers and stream, communicating with the lake. The present study describes different blood parameters of (Cyprinus Carpio, Linnaeus ,1758) population in Shkodra Lake. The samplings were made by catch from the local commercial fishery. The main parameter's measured in fishes blood where: pH, hematocrit, hemoglobin, urea, glucose and total lipid. It was obtained a survey of hematological and biochemical blood parameter's in lake Shkodra fresh water carp. [ABSTRACT FROM AUTHOR]

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 stunning sexual transformation commonly triggered by age, size or social context in some fishes is one of the best examples of phenotypic plasticity thus far described. To date our understanding of this process is dominated by studies on a handful of subtropical and tropical teleosts, often in wild settings. Here we have established the protogynous New Zealand spotty wrasse, Notolabrus celidotus, as a temperate model for the experimental investigation of sex change. Captive fish were induced to change sex using aromatase inhibition or manipulation of social groups. Complete female-to-male transition occurred over 60 days in both cases and time-series sampling was used to quantify changes in hormone production, gene expression and gonadal cellular anatomy. Early-stage decreases in plasma 17β-estradiol (E2) concentrations or gonadal aromatase (cyp19a1a) expression were not detected in spotty wrasse, despite these being commonly associated with the onset of sex change in subtropical and tropical protogynous (female-to-male) hermaphrodites. In contrast, expression of the masculinising factor amh (anti-Müllerian hormone) increased during early sex change, implying a potential role as a proximate trigger for masculinisation. Collectively, these data provide a foundation for the spotty wrasse as a temperate teleost model to study sex change and cell fate in vertebrates., (© 2021. The Author(s).)

Recent studies forecast that many ectothermic animals, especially aquatic stenotherms, may not be able to thrive or even survive predicted climate change. These projections, however, generally do not call much attention to the role of behavior, an essential thermoregulatory mechanism of many ectotherms. Here we characterize species-specific locomotor and respiratory responses to acute ambient warming in two highly stenothermic Antarctic Notothenioid fishes, one of which (Chaenocephalus aceratus) lacks hemoglobin and appears to be less tolerant to thermal stress as compared to the other (Notothenia coriiceps), which expresses hemoglobin. At the onset of ambient warming, both species perform distinct locomotor maneuvers that appear to include avoidance reactions. In response to unavoidable progressive hyperthermia, fishes demonstrate a range of species-specific maneuvers, all of which appear to provide some mitigation of the deleterious effects of obligatory thermoconformation and to compensate for increasing metabolic demand by enhancing the efficacy of branchial respiration. As temperature continues to rise, Chaenocephalus aceratus supplements these behaviors with intensive pectoral fin fanning which may facilitate cutaneous respiration through its scaleless integument, and Notothenia coriiceps manifests respiratory-locomotor coupling during repetitive startle-like maneuvers which may further augment gill ventilation. The latter behaviors, found only in Notothenia coriiceps, have highly stereotyped appearance resembling Fixed Action Pattern sequences. Altogether, this behavioral flexibility could contribute to the reduction of the detrimental effects of acute thermal stress within a limited thermal range. In an ecologically relevant setting, this may enable efficient thermoregulation of fishes by habitat selection, thus facilitating their resilience in persistent environmental change., Competing Interests: The authors have declared that no competing interests exist.

Exosomes are associated with cancer progression, pregnancy, cardiovascular diseases, central nervous system-related diseases, immune responses and viral pathogenicity. However, study on the role of exosomes in the immune response of teleost fish, especially antiviral immunity, is limited. Herein, serum-derived exosomes from mandarin fish were used to investigate the antiviral effect on the exosomes of teleost fish. Exosomes isolated from mandarin fish serum by ultra-centrifugation were internalized by mandarin fish fry cells and were able to inhibit Infectious spleen and kidney necrosis virus (ISKNV) infection. To further investigate the underlying mechanisms of exosomes in inhibiting ISKNV infection, the protein composition of serum-derived exosomes was analyzed by mass spectrometry. It was found that myxovirus resistance 1 (Mx1) was incorporated by exosomes. Furthermore, the mandarin fish Mx1 protein was proven to be transferred into the recipient cells though exosomes. Our results showed that the serum-derived exosomes from mandarin fish could inhibit ISKNV replication, which suggested an underlying mechanism of the exosome antivirus in that it incorporates Mx1 protein and delivery into recipient cells. This study provided evidence for the important antiviral role of exosomes in the immune system of teleost fish.

The pharmacokinetics of levamisole were determined in the belugas after single intravascular (IV), and single and multiple-dose oral by feed administrations. Also, the effect of levamisole (LVM) on the stress and immune responses of belugas were assessed. One hundred-fourteen healthy belugas in 4 different groups received single LVM administration at the doses of 50 and 100 mg/kg via IV and oral routes. A separate group of 24 belugas were administered oral LVM at the dose of 100 mg/kg for 5 days. Blood samples were collected at different time points after administrations to measure plasma concentrations of LVM by a validated high-performance liquid chromatography (HPLC) assay. For immunological evaluations, a total of 126 belugas received 50 and 100 mg/kg LVM via medicated feed for 5 days or served as the control without any medication; blood samples were recovered on day 0, 1, 3, 5, 7, 10, and 14 to measure hemolytic activity of the complement system (HAC 50 ), serum lysozyme activity, serum antibacterial activity, glucose, cortisol, total protein, albumin and C3 contents. In the single-dose administration, quantified LVM concentrations were dose-dependent and the oral bioavailability was in the range of 43.2-49.6%. In the multiple-dose administration, the peak plasma concentration at the steady state was 45.2 mg/ml, and accumulation ratio was calculated as 3.6. In the immunological study, LVM especially at the dose of 100 mg/kg increased HAC 50 , lysozyme and antibacterial activity in the sera of treated fish. No significant effect of LVM on glucose and albumin content was observed, but cortisol levels decreased and C 3 content was increased, more significantly by LVM at the dose of 100 mg/kg. Our results indicate that LVM is well absorbed after oral administration and reached to concentrations that can affect stress indicators and improve immune responses in belugas., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This study was done to evaluate the effects of Bisphenol A (BPA) on Siberian sturgeon (Acipenser baerii). As liver is the main organ in the homeostatic adjustments to stress, we used a proteomics method to address molecular response in this tissue. Also, we compared the levels of vitellogenin in plasma and mucus to propose that the last one be a non-invasive method to analyze this biomarker. The fish received 1, 10, and 100 μg g -1  week -1 BPA intraperitoneally for two weeks. The samples were taken on days 0, 7, and 14. Plasma vitellogenin level increased as the highest value was recorded in the group with 100 μg g -1  week -1 of BPA. Changes in the mucus and blood vitellogenin showed a similar pattern, suggesting that mucus could be used for evaluating the changes in blood vitellogenin. Comparative proteomics was used to determine the proteome of the liver of A. baerii in the highest dose of BPA in comparison with the control. Sixteen proteins were identified that their expression changed at least twice between the studied groups. The proteomic results showed that BPA increased the expression of proteins involved in the detoxification and metabolism, activated glycolysis, and produced necrosis in the liver., (Copyright © 2021 Elsevier Inc. All rights reserved.)

A 10-week feeding trial was run to investigate the separate and simultaneous effects of exogenous enzymes (Enz), probiotics (Pro), and Pro-Enz mixtures on the hematology indices, serum biochemical parameters, and innate-immunity status of juvenile Siberian sturgeon. The fish (138.06 ± 3.64 g) were randomly dispersed into 12 tanks (20 individuals per tank) and fed with Enz (Phytase, protease, and xylanase), Pro (Pediococcus pentosaceus and Lactococcus lactis), and Pro-Enz cocktail. At the end of the feeding bioassay, the highest values of red blood cell count, hemoglobin concentration, hematocrit level, and lymphocyte percentage followed by the lowest neutrophil percentage were obtained in Pro-Enz treatment (P < 0.05). Despite a significantly lower level of alkaline phosphatase in the fish fed with Pro supplemented diet (P < 0.05), no significant difference was found in the serum level of alanine aminotransferase and aspartate aminotransferase among the experimental groups (P > 0.05). Total protein content was significantly upregulated in serum and skin mucus samples from those fed with supplemented diets compared to the control group (P < 0.05). In both serum and skin mucus samples, higher immune responses in terms of lysozyme activity, immunoglobulin M, total protein was seen in Pro-Enz treatment compared to the control group followed by the serum complement components (P < 0.05). The results indicate that the combinational supplementation of Siberian sturgeon diet with the exogenous enzymes and probiotics modulates the physiometabolic responses and innate immune system to a higher grade than their individual supplementation., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The size and shape of red blood cells (RBCs) provide key information on life-history strategies in vertebrates. However, little is known about how RBC shape evolved in response to environmental factors, body size and the role of evolutionary rate. Here, we analysed RBC morphometrics in a set of Teleostei (bony fishes) and Elasmobranchii (sharks and rays) species testing the hypothesis that phylogenetic relationship explains species occupation of morphospace. We collected data on cell and nucleus area and volume, nucleus:cytoplasm ratio and shape factor for 65 species belonging to 28 orders. Then, we built phylomorphospaces separately for bony fish and sharks and rays. To test whether phylogenetic relationships predicted phenotypic similarity, we calculated multivariate phylogenetic signal. We also estimated the evolutionary rate of RBC shape for each node and tip using ridge regression. Finally, we tested whether habitat and body size influenced RBC shape using a PGLS. We found a significant phylogenetic signal in RBC shape for bony fish, but not sharks and rays. Saltwater teleost species were more clustered than freshwater ones in the phylomorphospace, suggesting clade disparity. Accordingly, the rate of evolution was highly heterogeneous, with significant decrease in Acanthopterygii. Neither habitat nor body size influenced RBC shape. In conclusion, RBC shape seems to have evolved in fishes in response to multiple selective pressures independent of life-history characters., (© 2021 European Society for Evolutionary Biology. Journal of Evolutionary Biology © 2021 European Society for Evolutionary Biology.)

In the early 20th century, August and Marie Krogh settled one of the most controversial questions in physiology, showing through elegant experiments that oxygen (O 2 ) uptake at the lung is driven by passive diffusion alone. Krogh's later work, on the regulation of local blood flow and capillary recruitment at the tissues, was awarded with the Nobel Prize in 1920. A century later it is still undisputed that O 2 moves across tissues by diffusion, however, animals use active mechanisms to regulate and facilitate the passive process. Teleost fishes have evolved a mechanism by which adrenergic sodium-proton-exchangers (β-NHEs) on the red blood cell (RBC) membrane actively create H + gradients that are short-circuited in the presence of plasma-accessible carbonic anhydrase (CA) at the tissue capillaries. The rapid acidification of the RBC reduces the O 2 affinity of pH-sensitive haemoglobin, which increases the O 2 diffusion gradient to the tissues. When RBCs leave the site of plasma-accessible CA, β-NHE activity recovers RBC pH during venous transit, to promote renewed O 2 loading at the gills. This mechanism allows teleosts to unload more O 2 at their tissues without compromising O 2 diffusion gradients and therefore, to use the available O 2 carrying capacity of the blood to a greater degree. In Atlantic salmon, β-NHE short-circuiting reduces the requirements on the heart by up to 30% during moderate exercise and even at rest, with important ecological implications. Thus, in some teleosts, the RBCs participate in regulating the systemic O 2 flux by actively altering the passive diffusion of O 2 that Krogh discovered., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Reference intervals of blood analytes are important diagnostic tools that clinicians use as a basis for making clinical decisions and that have been documented for many fish species. Although the effects of dietary supplementation on hematology and blood biochemistry variables in clownfish have been shown, there are no published reference intervals to date for routine hematology and blood biochemistry panels for these popular cultured marine fish (including Amphiprion spp.). The objective of this study was to establish de novo reference intervals for selected hemogram data and, using whole-blood analysis, commonly utilized chemistry analytes in two species of aquacultured clownfish: the Tomato Clownfish Amphiprion frenatus and Saddleback Clownfish A. polymnus. This study is based on blood samples from 25 clinically normal Tomato Clownfish and 38 clinically normal Saddleback Clownfish from two clownfish production facilities in Florida. Guidelines by the American Society for Veterinary Clinical Pathology were followed to generate robust reference intervals from the data collected. We report descriptive statistics and reference intervals for several hematology (n = 12) and biochemistry (n = 8) analytes using a small sample size and a small amount of blood per sample. The blood analyte data from this study are comparable with hematology and biochemistry data that have been reported in other teleost species. These results provide baseline health data for clownfish species that can be used in various clinical settings while also serving as a springboard for further research studies., (© 2020 American Fisheries Society.)

The initial experiment of this study was conducted to determine whether sex steroid concentrations varied before, during and after the reproductive season of European grayling (Thymallus thymallus L.) farmed broodstock confined in a recirculation aquaculture system (RAS). The results indicated that the plasma sex steroid (testosterone - T, progesterone - P 4 and estradiol - E 2 ) concentrations varied (P < 0.05) during these reproductive periods. There were greater concentrations of T, P 4 and E 2 before and during the grayling reproductive season indicating there are important functions of these steroid hormones associated with gonadal maturation, gamete production and reproductive behavior during the spawning period. In the second experiment of this study, the effectiveness of European grayling controlled reproduction treatment regimens was analyzed and there were 34.69 % and 68.18 % having ovulations in the 2 and 3-year-old broodfish, respectively. Of the embryos developing to the eyed-egg stage, there were 3.70 % and 6.87 % derived from eggs of 2- and 3-year-old grayling females, respectively. Proportions of embryos developing from the eyed-egg stage to hatching were 38.1 % and 52.1 % from eggs of 2- and 3-year-old grayling females, respectively, when there was culturing in a RAS. The results indicate that with grayling broodfish there is greater efficacy in induction of reproduction when there is imposing of the controlled reproduction treatment regimen on 3- rather than 2-year-old broodfish., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Anabas testudineus (Bloch) was exposed to 0.71 mg/L and 1.42 mg/L (25 and 50% of LC 50 value respectively) naphthalene, a polycyclic aromatic hydrocarbon (PAH), for 21 days. Blood biochemical parameters and erythrocytic morphological alterations were assessed to describe the naphthalene toxicity. Biochemical analysis showed a significant increase in glutamic pyruvic transaminase, GPT (576.7 ± 11.79 and 608.9 ± 12.08 U/L, respectively) and alkaline phosphatase, ALP (12.9 ± 0.69 and 13.4 ± 0.64 U/L, respectively) activities under two doses compared with control. Protein and albumin (ALB) content in blood decreased significantly, in comparison with control value in the tune of 22.67 ± 1.04 and 23.97 ± 1.24 g/dl, respectively and 10.7 ± 0.79 and 11.1 ± 0.67 g/dl, respectively. Erythrocytes showed varied symptomatic morphological changes under naphthalene exposure, which included severe denaturation, swelling in cells, appearance of sickle and tear drop cells, and cellular vacuolation. In particularly, the changes were more prominent under higher naphthalene exposure. Following the results, it has been able to establish that GPT, ALP, protein and ALB, and the morphological manifestations of erythrocytes would be good tools of biomarker in monitoring toxicological paradigm, especially to naphthalene exposure in aquatic bodies., (Copyright © 2020. Published by Elsevier B.V.)

Amazon fish are vulnerable to climate change. Current temperature increases in Amazonian rivers are likely to continue in the coming years. Elevated temperatures impair homeostasis and subject fish to oxidative stress; nevertheless, the effects of elevated temperature on plasma and muscle oxidative stress as well as fillet fatty acid composition in matrinxã (Brycon amazonicus) remain unknown. Therefore, the aims of the present study were to determine whether antioxidant system is able to withstand acute thermal stress to avoid plasma and muscle oxidative damage and to determine the manner in which matrinxã adjust their muscle fatty acid metabolism in a response to acute heat stress. We exposed juvenile matrinxã to four temperatures (28 °C as control and 30, 32, and 34 °C) for 72 h and observed the effects on plasma and muscle oxidant/antioxidant status and on fatty acid composition. Plasma reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in matrinxã exposed to 34 °C compared with the control group, while they were significantly higher in the muscle of matrinxã exposed to 32 °C and 34 °C compared with the control group. Superoxide dismutase (SOD) activity and total antioxidant capacity (TAC) levels were significantly lower in the plasma and muscle of matrinxã exposed to 34 °C compared with the control group. Plasma glutathione peroxidase (GPx) activity was significantly lower in matrinxã exposed to 32 and 34 °C compared with the control group, while it was significantly lower in the muscle of matrinxã exposed to 34 °C. The total content of saturated fatty acid (SFA) was significantly higher in the fillet of matrinxã exposed to 34 °C compared with the control group, while the total content of polyunsaturated fatty acid (PUFA) was significantly lower. Based on this evidence, we conclude that acute heat stress at 32 °C and 34 °C causes plasma and muscular oxidative stress, and the enzymatic and non-enzymatic antioxidant system is unable to prevent oxidative damage. Moreover, increased levels of total SFA and decreased levels of PUFA occur in an attempt to withstand heat stress and maintain membrane fluidity; nevertheless, these responses can represent negative impacts for consumers.

A recombinant giant grouper Luteinizing Hormone (LH) consisting of tethered beta and alpha subunits was produced in a yeast expression system. The giant grouper LH β-subunit was also produced and administered to rabbits for antibody development. The recombinant LH and its antibody were used to develop an Enzyme Linked Immunosorbent Assay (ELISA). This ELISA enabled detection of plasma LH levels in groupers at a sensitivity between 391 pg/ml and 200 ng/ml. Different species of grouper were assayed with this ELISA in conjunction with gonadal histology and body condition data to identify links between circulating LH levels and sexual development. We found that circulating levels of LH decreased when oocytes began to degenerate, and sex-transition gonadal characteristics were apparent when LH levels decreased further. When circulating LH levels were related to body condition (body weight/ body length), transitioning-stage fish had relatively high body condition but low plasma LH levels. This observation was similar across multiple grouper species and indicates that plasma LH levels combined with body condition may be a marker for early male identification in the protogynous hermaphrodite groupers., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Red blood cells (RBCs)-erythrocytes-of Osteichthyes are primarily known for their involvement in the process of gas exchange and respiration. Currently, physiological properties of RCBs in fish should also include their ability to participate in defense processes as part of the innate and adaptive immune mechanisms. In response to viruses, bacteria, and fungi or recombinant nanoparticles, they can modulate expression of genes responsible for immune reactions, influence activity of leukocytes, and produce cytokines, antimicrobial peptides, and paracrine intercellular signaling molecules. Via the complement system (CR1 receptor) and owing to their phagocytic properties (erythrophagocytosis), RBCs of Osteichthyes can eliminate pathogens. In addition, they are probably involved in the immune response as antigen-presenting cells via major histocompatibility complex class II antigens., (Copyright © 2020 Stosik, Tokarz-Deptuła, Deptuła and Deptuła.)

Calcitonin (CT) has a potential function in calcium (Ca) regulation, but there are conflicting observations in fishes. Because of the lack of calcified endoskeleton, sturgeons have low Ca circulating compared with teleost fish and the function of CT on Ca in sturgeon is very less understood. The purpose of this study was to investigate the impact of injection of salmon CT on plasma Ca, magnesium (Mg), phosphate (PHO), and glucose levels of juvenile Siberian sturgeon Acipenser baerii. Sixteen-month-old fish (429.6 ± 12.1 g) were intraperitoneally injected with a single dose of CT (5 μg kg -1 BW) and saline solution as a control group. Thereafter, blood sampling was performed at 0, 1, 2, 4, 6, 12, 24, and 48 h after injection. CT produced marked increases in all variables measured. The highest levels of Ca (6.77 ± 0.53 mg dL -1 ), Mg (9.79 ± 0.16 mg dL -1 ) and PHO (1.74 ± 0.05 mg dL -1 ) were recorded at 2 h after CT injection and showed significant difference compared with control treatment (Ca 4.75 ± 0.12 mg dL -1 ; Mg 5.47 ± 0.16 mg dL -1 and PHO 1.23 ± 0.06 mg dL -1 ). It also likely produced hyperglycemia. However, the differences with the controls were not statistically significant, possibly due to interference with the hyperglycemia induced by the stress of injection. Our results showed that the injection of 5 μg kg -1 BW CT to Siberian sturgeon has an incremental effect on plasma Ca, Mg, and PHO. The increase in plasma Ca level indicated that CT has a potent hypercalcemic effect in sturgeon under laboratory condition, in contrast to the hypocalcemic effects reported for teleosts.

Monthly changes of hepatosomatic index (HSI), gonadosomatic index (GSI), and plasma testosterone (T) levels in the reproductive season of male golden rabbitfish (Siganus guttatus) in captivity were investigated by sampling the liver, testis, and blood. HSI and GSI were estimated as percentage of relative weight of liver and testis to total body weight, respectively. Plasma T levels were measured using enzyme immune assay (EIA). Fragments of the testis were embedded in paraffin, cut to 5 μm thickness, and stained with Harris' hematoxylin and eosin. The results included seasonal changes in HSI, GSI, T, and testicular stages. The plasma T levels were observed high at spermation stage and during February-March and May-June. The values of HSI were monthly fluctuations, but no significant differences between testicular stages were observed. The GSI values continuously exhibited a significant increase from December to April and maintained at high levels until June. GSI continuously increased from stage II (immature testis) to stage V (spawning testis). There was correlation between GSI and testicular stages. Histological sections showed that male golden rabbitfish are multiple spawners and spawning season starts from March to June. These results contribute to further understanding of reproductive biology in golden rabbitfish, which can be implicated for artificial propagation.

In most vertebrates, red blood cell carbonic anhydrase (RBC CA) plays a critical role in carbon dioxide (CO 2 ) transport and excretion across epithelial tissues. Many early-diverging fishes (e.g., hagfish and chondrichthyans) are unique in possessing plasma-accessible membrane-bound CA-IV in the gills, allowing some CO 2 excretion to occur without involvement from the RBCs. However, implications of this on RBC CA function are unclear. Through homology cloning techniques, we identified the putative protein sequences for RBC CA from nine early-diverging species. In all cases, these sequences contained a modification of the proton shuttle residue His-64, and activity measurements from three early-diverging fish demonstrated significantly reduced CA activity. Site-directed mutagenesis was used to restore the His-64 proton shuttle, which significantly increased RBC CA activity, clearly illustrating the functional significance of His-64 in fish red blood cell CA activity. Bayesian analyses of 55 vertebrate cytoplasmic CA isozymes suggested that independent evolutionary events led to the modification of His-64 and thus reduced CA activity in hagfish and chondrichthyans. Additionally, in early-diverging fish that possess branchial CA-IV, there is an absence of His-64 in RBC CAs and the absence of the Root effect [where a reduction in pH reduces hemoglobin's capacity to bind with oxygen (O 2 )]. Taken together, these data indicate that low-activity RBC CA may be present in all fish with branchial CA-IV, and that the high-activity RBC CA seen in most teleosts may have evolved in conjunction with enhanced hemoglobin pH sensitivity.

Bottlenose dolphins (Tursiops truncatus) are long-lived mammals that can develop chronic aging-associated conditions similar to humans, including metabolic syndrome. Initial studies suggest that these conditions may be attenuated in dolphins using a modified fish diet. Serum metabolomics, fatty acid panels, and blood-based health indices were compared between 20 dolphins on a modified, 50% wild-type diet (50% mullet, 25% capelin, and 25% squid and/or herring) and 10 dolphins on a baseline diet (75% capelin and 25% squid and/or herring). Blood samples were collected at Months 0, 1, 3 and 6. Dolphins on the modified diet had lower insulin (7.5 ± 4.0 and 14.8 ± 14.0 μIU/ml, P = 0.039), lower cholesterol (160 ± 26 and 186 ± 24 mg/dl, P = 0.015) and higher hematocrit (46 ± 3 and 44 ± 3%, P = 0.043) by Month 1 compared to controls. Dolphins with anemia (hemoglobin ≤ 12.5 g/dl, n = 6) or low-normal hemoglobin (12.5-13.5 g/dl, n = 3) before placed on the modified diet had normal hemoglobin concentrations (> 13.5 g/dl) by Month 3. The modified diet caused a significant shift in the metabolome, which included 664 known metabolites. Thirty prioritized metabolites at Months 1 and 3 were 100% predictive of dolphins on the modified diet. Among 25 prioritized lipids, 10 (40%) contained odd-chain saturated fatty acids (OCFAs); C15:0 was the highest-prioritized OCFA. Increased dietary intake of C15:0 (from 1.3 ± 0.4 to 4.5 ± 1.1 g/day) resulted in increased erythrocyte C15:0 concentrations (from 1.5 ± 0.3 to 5.8 ± 0.8 μg/ml, P < 0.0001), which independently predicted raised hemoglobin. Further, increasing age was associated with declining serum C15:0 (R2 = 0.14, P = 0.04). While higher circulating OCFAs have been previously associated with lower risks of cardiometabolic diseases in humans, further studies are warranted to assess potential active roles of OCFAs, including C15:0, in attenuating anemia., Competing Interests: I have read the journal's policy and am the only author of this manuscript with the following competing interests: SV is a shareholder in and employed by Epitracker, Inc. and Seraphina Therapeutics, Inc. SV is an officer of Epitracker, Inc. and Seraphina Therapeutics, Inc. and is named in Epitracker and US Navy patent applications related to relevant products in development. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

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.)

The influence of hypothermia on erythrocyte profile of thermophile teleost species round goby, Neogobius melanostomus (Pallas, 1814), has been studied. Fish were acclimated to temperature 1-2 о С, 15-16 о С and 19-20 о С (control group) and held at given conditions for 5 days. The number of red blood cell precursors (pronormoblasts, basophilic and polychromatophilic normoblasts) in circulating blood has been estimated. Also, the number of abnormal erythrocytes, i.e. cells with micronuclei, nuclei invaginations, red blood cell shades, dacryocytes and cells undergoing amitosis has been determined on smears. The number of immature erythrocytes increased more than two times (p < 0,001) at 1-2 о С. The number of low-differentiated precursors, pronormoblasts and early basophilic normoblasts, increased for the most part. The number of abnormal erythrocytes did not change substantially, The changes in cellular blood composition were accompanied with the increase of plasma lactate concentration, indicating hypoxic state of fish. The results of the present work indicate that hematopoietic tissue remains sensitive to controlling factors at hypothermia, such as hypoxia, and may enhance proliferation and differentiation of erythroid cells., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The present work is designed to compare the chronic toxicity of anthracene [one of the major constituents of polycyclic aromatic hydrocarbons (PAHs)] on Anabas testudineus (Bloch), in an air-breathing carnivorous fish, in laboratory condition under the exposure of two doses of LC 50 value, i.e., 0.0075 mg/l (T1), i.e., 25% and 0.015 mg/l (T2) i.e., 50% for 21 days. A comprehensive comparison was recorded based on biochemical parameters and evaluated the erythrocytic alterations of blood components of the fish. It revealed an enhanced trend of activity of glutamic pyruvic transamin (GPT) 470.7 ± 12.32, 546.6 ± 13.22, 599.4 ± 13.09 U/L and alkaline phosphatase (ALP) 9.2 ± 0.61, 10.4 ± 0.86, 10.9 ± 0.74 U/L in control, T1 and T2 respectively; and reverse trend of protein (PRO) 26.63 ± 1.32, 22.15 ± 1.13, 22.29 ± 1.02 g/dl and albumin (ALB) 11.9 ± 0.71, 9.65 ± 0.91, 10.05 ± 0.94 g/dl in control, T1 and T2 respectively. Under T1 and T2 exposure conditions, it displayed the maximum alterations and appearance of tear drop-like cells (Tr), sickle cells (Sk), swelled cells (Sc) and vacuolated cells (Va) in comparison to control condition. An exclusive experimentation of the present work suggested that biochemical parameters and erythrocytic alterations may be useful tool as biomarkers to monitor the long term toxicological effects, especially to anthracene a constituent of PAHs, in any aquatic environment., (Copyright © 2019. Published by Elsevier B.V.)

The acid-base status is a tightly regulated physiological process, resulting from a balance of ions in the organism relevant to acid-base. The efficiency of the regulatory systems importantly determines the compensatory pH changes for a given disturb. Vertebrates minimize (or compensate) an acid-base disturb by general processes, which include ion transfer and/or PCO 2 changes. Acid-base adjustment in fish is predominantly achieved by branchial exchange of acid-base relevant ions with correlated change in plasma HCO 3 - levels. Conversely, land vertebrates change blood PCO 2 through ventilatory process and hence respiratory control of acid-base regulation plays an important role as a compensatory mechanism. Lungfishes (Dipnoi) have central position on vertebrate's evolution being considered as the sister group to the tetrapods. With an aquatic life mode, lungfish share similarities of respiratory function with tetrapods. This article reviews evidence showing that lungfish's respiratory system regulates acid-base status, like terrestrial ectothermic vertebrates. In the South American lungfish, Lepidosiren paradoxa, the presence of central CO 2 /pH chemoreceptors was unequivocally described. Also, the blood PCO 2 and acid-base status are typical of a terrestrial vertebrate. These aspects are discussed under different environmental conditions that require respiratory acid-base adjustments, such as, exposure to hypercarbia, hypoxia, high temperature and aestivation. Interesting questions regarding the location and cell phenotype of CO 2 /pH central and peripheral chemoreceptors remain an open field to be explored in lungfish., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The fish plasma model (FPM) predicts the fish blood plasma concentration of a pharmaceutical from the water concentration to which the fish is exposed and compares it with the human therapeutic plasma concentration (H ther PC) with the postulate that no adverse toxic effects occur below the H ther PC. The present study provides several lines of evidence supporting the FPM for the beta-adrenergic agonist salbutamol, a small cationic molecule at ambient pH. Salbutamol exhibited very low acute toxicity to early and adult life stages of fish. Biomass reduction in fish early life stages was the most sensitive apical endpoint, with no-observed-effect concentrations (NOECs) in the low mg/L range after continuous exposure for up to 120 d. Given that predicted and measured environmental concentrations are at least 1000-fold lower, the risk of salbutamol in freshwater is deemed very low. Increase in heart beat rate and decrease in total triglyceride content in fish also occurred at the low mg/L range and resembled effects known from humans. This finding supports the FPM assumption of conserved targets in fish with similar functionality. Plasma concentrations measured in adult and juvenile fish exposed to water concentrations at approximately the NOECs exceeded H ther PC and even approached plasma concentrations toxic to humans. This result confirms for salbutamol the FPM hypothesis that no adverse (i.e., population-relevant) toxic effects occur in fish below the H ther PC. Environ Toxicol Chem 2019;38:2509-2519. © 2019 SETAC., (© 2019 SETAC.)

Organisms with a parasitic lifestyle comprise a high proportion of biodiversity in aquatic and terrestrial environments. However, there is considerable variation in the ways in which they acquire nutrients. Hematophagy is a common consumption strategy utilized by some terrestrial, aquatic, and marine organisms whereby the parasite removes and digests blood from a host. Gnathiid isopods are marine hematophagous parasites that live in benthic substrates from the intertidal to the abyss. Although ecologically similar to ticks and mosquitoes, they feed only during each of 3 juvenile stages and adults do not feed. They have long been considered as generalist fish parasites and to date, there have been no reports of their successfully feeding on invertebrates. Based on observations of gnathiids attached to soft-bodied invertebrates collected from light traps, we conducted a laboratory experiment in which we collected and individually housed various common Caribbean invertebrates and placed them in containers with gnathiids to see if the gnathiids would feed on them. All fed gnathiids were subsequently removed from containers and given the opportunity to metamorphose to the next developmental stage. In total, 10 out of the 260 gnathiids that were presented with 1 of 4 species of potential invertebrate hosts had fed by the next morning. Specifically, 9 of a possible 120 gnathiids fed on lettuce sea slugs ( Elysia crispata ), and 1 of a possible 20 fed on a bearded fireworm ( Hermodice carunculata ). Eight of these 10 fed gnathiids metamorphosed to the next stage (5 to adult male, 2 to adult female, and 1 to third-stage juvenile). Even though feeding rates on invertebrates were considerably lower than observed for laboratory studies on fishes, this study provides the first documented case of gnathiids' feeding on and metamorphosing from invertebrate meals. These findings suggest that when fish hosts are not readily available, gnathiids could switch to soft-bodied invertebrates. They further provide insights into the evolution of feeding on fluids from live hosts in members of this family.

In the present study, we aimed to screen the potential probiotic Bacillus subtilis isolated from the gut of healthy fish using in vitro assays and to evaluate its effect on Dabry's sturgeon (Acipenser dabryanus) using in vivo feeding experiments. Among the isolates, B. subtilis BSth-5 and BSth-19 exhibited antimicrobial effect against four sturgeon-pathogenic bacteria, including Aeromonas hydrophila, A. veronii, A. media, and Streptococcus iniae. The cell number of B. subtilis BSth-5 and BSth-19 changed little after 2 h of exposure to pH 3.0 or fresh Dabry's sturgeon bile at 2.5% and 5.0%. Meanwhile, B. subtilis BSth-5 and BSth-19 produced extracellular protease, cellulose, and lipase. And it was proved that B. subtilis BSth-5 and BSth-19 were harmless after injection of Dabry's sturgeon. One group of Dabry's sturgeon was fed a control diet and two groups were fed experimental diets containing 2.0 × 10 8  CFU/g BSth-5 (T1 group) or BSth-19 (T2 group) for 8 weeks. No significant differences in final weight, weight gain rate, and special growth rate were observed in the T1 and T2 groups compared to the control group (P > 0.05), but a significant improvement in survival rate was detected after 4 and 8 weeks of feeding (P < 0.05). After 8 weeks, serum total antioxidant capacity, total superoxide dismutase activity, and IgM levels were significantly higher in the T1 and T2 groups compared to the control group (P < 0.05). Moreover, serum lysozyme activity was significantly higher in the T1 group relative to the control group during the whole experiment period (P < 0.05); however, the differences were not significant between the T2 and control groups (P > 0.05). Serum malondialdehyde levels in the T1 and T2 groups were significantly lower than those in the control group after 4 weeks (P < 0.05). Sturgeons in the T1 and T2 groups showed a higher survival rate after Aeromonas hydrophila infection. To summarize, dietary supplementation with BSth-5 and BSth-19 could enhance the survival rate, antioxidant activity, serum immunity, and disease resistance in A. dabryanus., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Non-competitive, enzyme-linked immunosorbent assays (ELISAs) for three distinct sole vitellogenins (VtgAa, VtgAb and VtgC) were designed using their purified lipovitellin (Lv) products and corresponding digoxigenin-labeled, anti-Lv polyclonal antibodies, primarily for employment in monitoring estrogenic pollution of the environment. The working range of the ELISAs was from 0.97 to 1,000 ng/mL for all Vtg subtypes. Each ELISA appeared to be specific to the targeted Vtg subtype. Intra- and inter-assay coefficients of variation in the developed ELISAs were lower than 10%. Three Vtg subtypes were induced in serum of immature fish by estradiol-17β (E2) injection (0.5 mg/kg body weight). All Vtg subtypes were induced one day after the injection, reaching peak levels (Lv equivalents) within three days, as follows: 39.1 ± 28.9 μg/mL (VtgAa), 57.9 ± 30.7 μg/mL (VtgAb) and 12.6 ± 4.8 μg/mL (VtgC). In wild-caught males, VtgAa, VtgAb and VtgC were detected in ranges from 0.26 to 1.21, 0.19 to 8.69, and 0.17 to 53.50 μg/mL, respectively, over various sampling periods. In vitellogenic females sampled in January, the average level of VtgAb (8,744.43 ± 733.93 μg/mL) was significantly higher than for VtgAa (150.33 ± 22.35 μg/mL) or VtgC (57.08 ± 6.00 μg/mL); thus VtgAb appeared to be the most dominant Vtg subtype. The present study entails the first report on development of subtype-specific Vtg ELISAs in marbled sole, which empowers us to detect and monitor estrogenic contamination in aquatic environments inhabited by this species., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Nowadays, use of the natural plant derivatives as an immunostimulant in aquaculture is becoming more useful than antibiotics which are destructive to target animals, consumers, and the environment. The present study aimed to evaluate the efficacy of dietary substitution of wheat flour at 2, 5, 7.5, and 10% of olive pomace (OP 2 , OP 5 , OP 7.5 , and OP 10 ) in some immune-physiological and antioxidant variables of yearling Siberian sturgeon (Acipenser baerii) weighing 165 ± 6.8 g at 19.1 ± 1.52 °C for 56 days. Fifteen fish per tank were randomly accommodated in triplicates per treatment and the control group was also included. No significant difference was seen in hematological indices including red blood cells (RBS) and immunocompetent cell population sizes among the treated groups (P > 0.05). Also, malondialdehyde (MDA) activity and superoxide dismutase (SOD) in serum remained unaffected in different treatments (P > 0.05). Total protein (TP), high-density lipoprotein (HDL), catalase (CT), and lysozyme were affected by dietary OP levels (P < 0.05), and the highest value was observed at the OP 5 diet, but no influence was seen in serum IgM by OP. These data show that application of olive pomace as a partial substitute of wheat flour in the diet of Siberian sturgeon is feasible with stimulatory and antioxidant functions.

Characterization of the isomers perfluorooctane sulfonamide (PFOSA), a key intermediate of the perfluorooctane sulfonate (PFOS) precursors, is a prerequisite to understand the contribution of precursors to PFOS in the environmental and biological matrices. However, the lack of commercial standards makes quantification of the PFOSA isomers in complex matrices a big challenge. For the first time, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established to identify and quantify PFOSA isomers. Improvement on chromatographic separation with selected reaction monitoring allowed full resolution of six monomethyl branched isomers of PFOSA. The isomers were identified and quantified using a series of characteristic fragment ions and the specific product ions of molecular anion m/z 498: m/z 78 (n-), m/z 169 (iso-), m/z 419 (1m-), m/z 164 (2m-), m/z 259 (3m-), m/z 269 (4m-) and m/z 219 (5m-PFOSA). With the aid of 19 F nuclear magnetic resonance to define one technical product as standard, the developed method was used to quantify PFOSA isomers in other technical products and spiked fish blood samples. The results indicated that the developed method displayed strong application prospect for measuring trace level of PFOSA isomers in complex samples. The method detection limits for all isomers were in the range of 0.1-1 pg/g ww for blood samples., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Eight nuclear abnormalities of genotoxicity and cytotoxicity were studied in peripheral blood erythrocytes of herring (Clupea harengus membras), flounder (Platichthys flesus), and Atlantic cod (Gadus morhua) sampled (2010-2017) from the Polish and the Lithuanian Exclusive Economic Zones (EEZ) in the Baltic Sea. At all study stations, total genotoxicity (∑Gentox) was found to be higher than total cytotoxicity (∑Cytox). A significant time-related decrease in genotoxicity was detected in the Lithuanian EEZ (2015-2017), while in the Polish EEZ (2014-2016), the opposite tendency was revealed. The highest ∑Gentox and ∑Cytox values recorded in fish sampled at the study stations located relatively close to each other clearly indicate an increased environmental genotoxicity and cytotoxicity pressure for fish in these areas. Exceptionally high and high-level genotoxicity risks to herring followed by those to flounder and cod were determined at a higher percentage of the stations studied.

Sources of lead exposure of many bird species are poorly understood. We analyzed blood lead concentrations from osprey (n = 244; Pandion haliaetus) and bald eagles (n = 68; Haliaeetus leucocephalus) and documented potential sources of lead they may encounter. Adult bald eagles had higher blood lead concentrations than did adult osprey. However, blood lead concentrations of nestlings were similar for both species. Although 62% of osprey had detectable lead concentrations ( x ¯  = 1.99 ± 4.02 μg/dL, mean ± standard deviation [SD]), there was no difference in the detection frequency or lead concentrations between osprey adults and nestlings. Likewise, we found no differences in the detection frequency or lead concentrations in osprey adults and nestlings from high- and low-salinity areas. Of the bald eagle samples tested, 55% had detectable lead levels ( x ¯  = 6.23 ± 10.74 μg/dL). Adult bald eagles had more detectable and higher lead concentrations than did nestlings or pre-adults. Among environmental samples, paint had the highest lead concentrations, followed by sediment, blue catfish (Ictalurus furcatus), and gizzard shad (Dorosoma cepedianum). There was no correlation between blood lead concentrations of osprey adults and their offspring. Our results indicate that, in the Chesapeake Bay region of Maryland and Virginia (USA), there are multiple sources by which piscivorous raptors may be exposed to lead. Environ Toxicol Chem 2019;38:862-871. © Published 2019 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America., (© Published 2019 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.)

In an effort to overcome the palatability issues currently constraining the effective delivery of praziquantel (PZQ) via feed to treat monogenean parasites in yellowtail kingfish, this study compared the bioavailability and palatability of PZQ in hydrogenated castor oil (HCO) solid lipid nanoparticles (SLN) against pure PZQ in this species. Improving bioavailability would facilitate lower dietary inclusion levels to achieve the same therapeutic dose and therefore reduce the bitterness of feeds containing PZQ. Bioavailability was determined by co-administering feed with either pure PZQ, HCO-SLN or HCO-SLN coated with chitosan via intubation and quantifying the pharmacokinetics response. In contrast to studies with mammals, the results demonstrated that PZQ in HCO-SLN had equal bioavailability to pure PZQ in yellowtail kingfish, including when HCO-SLN were coated with chitosan. We hypothesise that the lack of improvement in bioavailability may be due to the lack of M cells and Peyer's patches in fish and the subsequent inability of fish to take nanoparticles directly into the lymphatic system. Furthermore, palatability of the feeds medicated with PZQ was not improved when the PZQ was incorporated into HCO-SLN, possibly due to the low loading rate of PZQ within the HCO-SLN and the subsequent thick coating of nanoparticles that was required on the surface of the feed pellets. Combined, these data demonstrate that the SLN used in the current study are not capable of delivering the benefits required to enable effective in-feed treatment of PZQ against monogenean parasites in yellowtail kingfish., (Crown Copyright © 2018. Published by Elsevier Inc. All rights reserved.)

An optical communication type biosensor system has been developed which can measure blood glucose concentration, which is a stress indicator of fish, in real-time while fish swimming freely. However, this system is hard to make instant acknowledgment of fish stress level which has to contain an unavoidable delay in the judgment. In this research, we aimed to develop a novel stress visualization system which can quickly judge the levels for fish stress response instantly based on a color changeable LED while another LED was designed to send data. The present system is based on the principle of converting the output current value measured by the glucose biosensor corresponding to the stress response into a voltage value. Then, the color and stress switching points of the LED (Red, Yellow, Green) were decided based on the voltage value gained from the biosensor which mentioned above. Furthermore, we attempted to use our biosensor system to make real-time monitoring of fish stress in vivo. As results, the proposed sensor can make real-time measurement of glucose and shows a great response to those of actual fish sample in the range from 35.36 to 300 mg dl -1 (R = 0.9899). When the glucose concentration in the collected sample was switched to the concentration pre-sett, it was successful to switch the LED color according to the gained voltage value both in vitro and in vivo. Furthermore, when monitoring the stress responses of the fish in vivo, color switching corresponding to the sensor output current value was observed successfully., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The objective of this study was to evaluate toxicogenetic potential of surface water samples from rivers of center-west Brazil and analyze the influence of land use and cover and physicochemical parameters in genetic damage. Samples were collected during winter (June) and summer (November) at sampling sites from Dourados and Brilhante Rivers (Mato Grosso do Sul/Brazil). The toxicogenetic variables, including chromosomal alterations, micronuclei, and mitotic index, were analyzed in meristematic cells of Allium cepa; and micronuclei, nuclear abnormalities, and DNA strand breaks (arbitrary units, AU T ) were analyzed in erythrocytes of Astyanax lacustris. The rivers presented physicochemical values outside the Brazilian laws, which can be a characteristic of human pollution (domestic sewage and local agriculture). The results of A. cepa test suggest that the water samples from Dourados and Brilhante rivers exerted significant (p < 0.05) cytotoxic and genotoxic effects, in both periods of collection, especially alterations in mitotic index. In blood cells of A. lacustris, genotoxic effect of the water samples from the rivers also was observed as significant nuclear abnormalities, DNA breaks (UA T ), in both sampling periods, compared with the negative control. Spearman correlation analyses revealed that data of land use and cover and physicochemical parameters were statistically correlated with DNA damages in bioassays. This study demonstrates toxicogenetic potential of water samples from Dourados and Brilhante rivers; furthermore, the type of land use and land cover and physicochemical parameters were revealed to have influence on toxicogenetic damage.

Some heavy metals related to human activities were measured in the water of Chascomús lake. The maximum concentrations were: 0.23 μg/L for Cd, 4.28 μg/L for Cr, 22.09 μg/L for Cu, 2.49 μg/L for Ni, 3.24 μg/L for Pb and 210.76 μg/L for Zn. The values of Cd, Cr, Cr, Pb and Zn were above the Argentine National Guidelines for the Protection of the Aquatic life. The analysis of gonadal condition of pejerrey fish (Odontesthes bonariensis) from this lake did not revealed any reproductive damages. However, exposures with environmental concentrations of Cd, Cr, Cu and Zn under laboratory conditions of pejerrey males (14 days), caused a significant increase of the expression of the three variants of gnrh in the brain (within Cd exposure) and a decrease in cyp19a1b mRNA (within Cu exposure). Furthermore, at pituitary level, a decrease in fshb transcript levels was observed in the fish exposed to Cd and Cr and a decrease in the expression of both gonadotropin receptors at gonadal level in Zn exposure. Moreover, the gonads of the fish exposed to all the tested metals suffered structural damages showing shortness of the spermatic lobules, fibrosis, testis ova and the presence of piknotic cells. All these findings alert that heavy metals pollution affects the expression of key reproductive genes and gonadal structure of fish species that represent the predominant group of organisms and are considered sentinel species in the aquatic ecosystems., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Low concentrations of nonylphenol (NP) in aquatic environment can induce drastic effects on the endocrine system in animals. In this study, we examined the modulatory effects of NP on reproductive and physiological parameters in juveniles of the red seabream and black rockfish following waterborne NP exposure (0, 1, 10, and 50 μg L -1 ) for 60 days. In red seabream exposed to 50 μg L -1 NP, plasma levels of 17β-estradiol (E2) and 11-ketotestosterone (11-KT) were significantly lower at 30 and 60 days, while E2 levels were slightly higher in 10 μg L -1 -exposed individuals at day 30. Similarly, significantly lower levels of E2 and 11-KT were observed in 10 and 50 μg L -1 -exposed black rockfish at 60 days, whereas the E2 level was higher in 1 μg L -1 -exposed individuals at day 30. After exposure to NP, plasma and mRNA levels of vitellogenin (VTG) were significantly higher in both species at 30 and 60 days, similar to the inducible effects from synthetic estrogen. Plasma cortisol levels were significantly elevated by relatively higher concentrations of NP (10 and 50 μg L -1 ) at 30 and 60 days. Finally, 60 days of exposure of 50 μg L -1 NP significantly decreased the gonadosomatic index (GSI) and increased the hepatosomatic index (HSI) in both species. The results obtained from this study provide an evidence of the endocrine disrupting potential of waterborne NP on early stages of economically important marine fish. The NP-triggered endocrine modulation can induce effects on the development of reproductive and metabolic organs in fish species., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Only a small proportion of pharmaceuticals available for commercial use have been monitored in the aquatic environment, and even less is known about the effects on organisms. With thousands of pharmaceuticals in use, it is not feasible to monitor or assess the effects of all of these compounds. Prioritisation schemes allow the ranking of pharmaceuticals based on their potential as environmental contaminants, allowing resources to be appropriately used on those which are most likely to enter the environment and cause greatest harm. Many different types of prioritisation schemes exist in the literature and those utilising predicted environmental concentrations (PECs), the fish plasma model (FPM), critical environmental concentrations (CECs) and acute ecotoxicological data were assessed in the current study using the 50 most prescribed drugs in the UK. PECs were found to be overestimates of mean measured environmental concentrations but mainly underestimations of maximum concentrations. Acute ecological data identified different compounds of concern to the other effects assessments although the FPM and CECs methods were more conservative. These schemes highlighted antidepressants, lipid regulators, antibiotics, antihypertensive compounds and ibuprofen as priority compounds for further study and regulation.

Substituted diphenylamine antioxidants (SDPAs) and benzotriazole UV stabilizers (BZT-UVs) are additives used in industrial and commercial applications to prevent degradation by oxidation and are contaminants of emerging environmental concern. Little is known about the fate of these contaminants in wildlife, particularly in reptiles, birds and marine mammals. Nine SDPAs and six BZT-UVs were measured in blood plasma of seven fish species, snapping turtles (Chelydra serpentina), double-crested cormorants (Phalacrocorax auritus), and bottlenose dolphins (Tursiops truncatus) from various locations in North America. Plasma SDPAs were more frequently (90-100%) detected and with higher concentrations (median: 25-270 pg g -1 , wet weight (ww)) in organisms from urban areas than rural locations (median: -1 ). The concentrations of most SDPAs generally followed the order of fish ≥ snapping turtles > double-crested cormorants > bottlenose dolphins. Of the three quantifiable BZT-UVs, 2-(2H-benzotriazol-2-yl)-4,6-di-tert-pentylphenol (UV328) showed higher detection frequency in most species of fish, bird and turtle (range of 0-67%), indicating the widespread distribution of UV328 in the aquatic environment of lower Great Lakes region., (Crown Copyright © 2018. Published by Elsevier B.V. All rights reserved.)

In order to understand pharmacokinetics in vivo and tissue residues of florfenicol in Hybrid sturgeon cultured in cold water, changes in concentration of florfenicol in plasma, muscle, liver and kidney of sturgeon after administration were measured by high performance liquid chromatography mass spectrometry. The results show that after 15.0 mg/kg dose administration, the concentration change in plasma and liver, kidney and muscle of sturgeon are suitable for the one compartment open model. Florfenicol pharmacokinetic parameters in plasma, liver, kidney and sturgeon muscle showed that the peak concentration (C max ) were 549.7, 551.8, 958.2 and 405.4 μg/kg, respectively. The peak time (T max ) was 17.5, 14.2, 14.0 and 20.8 hr, respectively. The elimination half-life (T 1/2 ) was 75.1, 99.8, 68.8 and 75.5 hr, respectively. Area under the curve were 70,048.9, 63,850.6, 109,579.9 and 53,437.3 μg/kg hr, respectively. Mean residence time was 100.6, 96.4, 96.5 and 104.6 hr, respectively. The absorption speed and elimination of florfenicol in cold water cultured sturgeon is slow, and the peak concentration is low, but the florfenicol was widely distributed in the body of sturgeon. The results provide a theoretical basis for formulation of medication and withdrawal period of florfenicol used in sturgeon cultured in cold water., (© 2018 John Wiley & Sons Ltd.)

The effect of various feeding strategies was evaluated on growth performance and biochemical parameters in two sizes of Siberian sturgeon (465.75 ± 11.18 and 250.40 ± 12 g) during 45 days. Fish were distributed into six experimental treatments including large fish with satiation feeding (LA), small fish with satiation feeding (SA), large fish with 50% satiation feeding (LR), small fish with 50% satiation feeding (SR), large starved fish (LS), and small starved fish (SS). Differences in final weight between LA and LR treatments were not noticeable, whereas SA and SR treatments showed significant differences. Growth parameters were more affected in small fish. In condition factor and weight gain in starved treatments, a considerable reduction occurred by interaction between feeding strategies and fish size, so the lowest values were obtained in SS treatment. Glucose levels significantly decreased in small fish during the starvation. Interaction between feeding strategy and fish size indicated the highest and lowest albumin level in SA and SS treatment, respectively. Cholesterol, triglyceride, total protein, and globulin showed no significant differences. It can be deduced that small fish are more sensitive to starvation than the large fish. Since glucose and albumin showed significant decrease in starved small fish, these parameters can help to monitor nutritional status and feeding practices. It was indicated that in both sizes of Siberian sturgeon, feeding 50% satiation reduced the food cost without negative impact on physiological condition, and it can be considered as an appropriate strategy to face unfavorable circumstances.

Here, we investigated the early development of two closely related myxozoan parasites, the highly pathogenic Myxobolus cerebralis, the causative agent of the whirling disease in salmonids, and Myxobolus pseudodispar, a common, non-pathogenic parasite of cyprinids. The aim of our study was to examine under in vivo laboratory conditions whether fish blood is involved in the intrapiscine development of the two parasite species and investigate if there is dissimilarity between the parasite infection intensity in blood and if it varies in terms of host susceptibility and parasite pathogenicity. Highly susceptible, less susceptible and non-susceptible hosts were involved. Blood samples were taken 1 day, 1 week and 1 month post exposure to M. cerebralis and M. pseudodispar, respectively. The prevalence and infection intensity was estimated by parasite-specific quantitative real-time PCR. Although previous findings assumed that M. cerebralis might escape from host immune system by migrating via peripheral nerves, our experimental results demonstrated that M. cerebralis is present in blood during the early stage of intrapiscine development. For the non-pathogenic M. pseudodispar, the highest infection prevalence was found in the original host, common roach Rutilus rutilus, whereas the highest infection intensity was detected in rudd Scardinius erythrophthalmus, a "dead-end" host of the parasite. The presence of M. pseudodispar developmental stages in the blood of both susceptible and non-susceptible cyprinids suggests that the susceptibility differences remain hidden during the early stage of infection. Our findings supply further evidence that host specificity is not determined during the early, intrapiscine development involving the vascular system. Furthermore, we found remarkable differences in the infection dynamics of the two parasite species examined, possibly due to their distinct pathogenicity or variations in adaptive capabilities to immune components in host blood., Competing Interests: The authors have declared that no competing interests exist.

The relationship between plasma glucose concentration and intracellular glucose (liver, heart, brain, gill, gonad, intestine, kidney, spleen, white muscle) was determined in fish species with a range in plasma glucose (Atlantic cod, 5.06 mM; cunner, 3.8 mM; rainbow trout, 3.7 mM; lumpfish, 0.9 mM; short-horned sculpin, 0.6 mM; and winter flounder, 0.6 mM). The ratio of intracellular glucose/plasma glucose was always higher than one in liver for all species consistent with a diffusion gradient from the tissue to the plasma. In all other tissues in Atlantic cod, cunner, and rainbow trout the diffusion gradient was from the plasma to the intracellular space. In short-horned sculpin, the mean ratio in heart and white muscle exceeded one and in winter flounder the ratio was significantly greater than one at 5.97 and 2.92 for heart and muscle, respectively. The presence of an active glucose 6-phosphatase in white muscle could account for elevated amounts of free glucose. The white muscle of all species displayed phosphoenolpyruvate carboxykinase and in winter flounder the activity was as high in white muscle as in liver suggesting that gluconeogenesis may be associated with a relatively high-muscle glucose content. The glycogen content was highest in liver followed by heart with lower amounts in all other tissues. There was an inverse correlation between heart glycogen content and plasma glucose concentration when all species were included. It is contended that in species with low plasma glucose, heart glycogen is accumulated at a slow rate under normoxia, to be called upon under hypoxic conditions when the gradient for inward diffusion is unfavourable for high rates of glucose metabolism.

An eleven weeks feeding trial was conducted to determine the effects of different levels of dietary soybean lecithin (SBL) on growth performance, blood chemistry and immunity in juvenile stellate sturgeon (Acipenser stellatus). Fish were fed seven isoproteic (44% crude protein) and isolipidic (17% crude fat) diets containing graded levels of SBL: 0 (control), 1, 2, 4, 6, 8 and 10%. Results showed that dietary SBL supplementation significantly improved the final body weight (BW) and weight gain (WG). Fish fed 6% SBL showed the highest BW and WG values in comparison to fish fed the control diet (P < 0.05), whereas increasing SBL levels above 6% had little practical benefit in terms of somatic growth performance. The inclusion of SBL in diets significantly improved the immune response as data from lysozyme, total Ig levels, alternative complement, phagocytic and bactericidal activities indicated (P < 0.05). The broken-line regression analysis of immunological variable revealed that depending on the parameter considered, the optimal SBL levels in diets for stellate sturgeon juveniles varied. In particular, dietary SBL levels requirements in stellate sturgeon when considering the phagocytic activity rate were determined at 3.3%, whereas 4.1-4.2% were recommended when considering data from lysozyme, alternative complement and bactericidal activities. In contrast, the highest minimum dietary SBL content was estimated at 6.9% when data from total Ig levels were considered. These results indicated that dietary PLs are required for boosting innate immunity in stellate sturgeon, although their minimal level changed depending on the immunological parameter considered. Therefore, we assume that SBL levels comprised between 3.3 and 6.9% may be used as a prophylactic measure to improve the health status in stellate sturgeon. Red blood cell count, hemoglobin and hematocrit levels increased with increasing dietary SBL levels, especially in those sturgeons fed the diet with 6% SBL (P < 0.05). In addition, white blood cell counts significantly increased as dietary SBL levels increased from 4 to 8% in comparison to the control group. Blood biochemistry was also affected by different dietary SBL levels. In particular, significantly higher levels of glucose, cholesterol, HDL and triglycerides were detected in fish fed >6%, >4%, >2% and 2% SBL, respectively (P < 0.05). Based on somatic growth parameters, blood chemistry and systemic immunity parameters, diets containing ca. 6% SBL are recommended for juvenile stellate sturgeon., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

One of the challenges of sturgeon aquaculture is that sturgeon takes an extended amount of time to reach sexual maturity. The pattern of the protein expression in relation to the late maturity of sturgeon can help to better understand changes in sexual maturity. 17β-estradiol (E2), testosterone (T) and vitellogenin (Vtg) levels were examined at all stages of sexual maturation in Sterlet sturgeon (Acipenser ruthenus). Two-dimensional gel electrophoresis and mass spectrometry analysis were used to show the pattern of the ovarian proteins. The T levels increased from the previtellogenic to the postvitellogenic stages (P < 0.05) and Vtg showed a decremental pattern in pre- and postvitellogenic, and atresia (not significantly). The analysis showed 900 protein spots, 19 of which were successfully identified and had significant differences between the previtellogenic and the vitellogenic groups (P < 0.05). Among the identified proteins, 40% involved in cell defense (heat shock protein, Glutathione peroxidase, natural killer enhancing factor, peroxiredoxin-2), 30% in transcription and translation (constitutive photomorphogenesis 9 and Ybx2), 20% in metabolism and energy production (triose-phosphate isomerase (TPI)) and 10% in transport (glycolipid transfer protein). In the vitellogenic stage, the proteins were related to metabolism and energy production (TPI, ES1, creatin kinase, enolase, nucleoside diphosphate kinase, 50%), cell defense (thioredoxin and dislophid isomerase, 20%) and transport (fatty acid binding protein, 10%). Our findings show changes in protein expression pattern from cell defense to metabolism during egg development., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Administration of probiotic candidates in fish has generally been shown as a useful strategy to improve growth performance, survival, digestive enzyme activity, and gut microbiota. Unfortunately, the sero-immunological responses of different fish to different probiotic candidates are poorly understood. The present study assessed the effect of Pediococcus acidilactici as a probiotic on the biochemical and immunological parameters of beluga. Fish (248.32 ± 10.21 g) were fed a control diet (without P. acidilactici( and three different doses of P. acidilactici-supplemented diets (10 7 , 10 8 , and 10 9  CFUg -1 diets) for 8 weeks. On week 8, blood and serum were sampled. Dose-dependent increase of immunological parameters (respiratory burst activity, lysozyme content, serum antibacterial activity, and total immunoglobulin) and biochemical parameters (total protein and albumin levels) was observed. However, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were significantly increased in the juvenile beluga fed by 10 9  CFUg -1 P. acidilactici-supplemented diet compared to the other groups. Based on the results of this evaluation, it is reasonable to conclude that the inclusion of P. acidilactici as probiotic in diets for juvenile beluga improves the sero-immunological parameters of the fish and should be considered by farmers as a strategy to improve fish health.

Field surveys of the impact of environmental estrogen (EE) pollution in aquatic wildlife have been conducted using vitellogenin (VTG) as a biomarker to evaluate the influence of EE. However, a standard baseline of VTG level that can be used to evaluate EE pollution has not been fully determined. To the best of our knowledge, the present study is the first to determine the standard baseline VTG level for evaluating the biological effects of EE pollution using the Japanese common goby (Acanthogobius flavimanus) as the target model fish. Plasma VTG and estradiol-17β (E 2 ) levels associated with the reproductive cycle of wild goby inhabiting an unpolluted environment were measured. Mean plasma VTG and E 2 levels exhibited similar changes, increasing in the yolk vesicle stage and peaking in the tertiary yolk stage in females. However, plasma VTG and E 2 levels showed no significant changes in males, remaining at low levels throughout the reproductive cycle. The highest VTG levels in females and males were 1.6 mg ml -1 and 124.87 ng ml -1 , respectively. These results indicate that the baseline level (normal level) in males was approximately 130 ng ml -1 at most. We concluded that the threshold between normal and abnormal levels with a 10% risk rate was 150 ng ml -1 in the wild male goby. Plasma VTG levels in males captured from Nagasaki Harbor were higher than the threshold in each reproductive developmental stage, indicating the possibility of EE pollution at this site. The biological standard baseline for VTG established in this study is useful for assessing EE pollution in natural waters., (Copyright © 2018 John Wiley & Sons, Ltd.)