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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.)

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.)

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.)

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.)

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.)

Several oxidative stress markers and liver oxygen consumption were measured in different tissues of the marine fish Trichiurus lepturus in late summer and late winter, as well as in juveniles and adult females. Oxygen consumption in liver, superoxide dismutase (SOD) and catalase (CAT) activity in liver, red cells, lens and roe, vitamin E, ubiquinol 10 , β-carotene in liver, red cells, and roe, as well as contents of reduced glutathione (GSH) and lipoperoxidation (TBARS) in red cells were evaluated. Regarding ontogeny, compared to adult fish, juveniles showed significant higher SOD activity in liver and lens, as well as higher liver contents of vitamin E. In contrast, adult females showed higher contents of vitamin E in roe, ubiquinol 10 in liver and roe, and higher GSH levels in red cells, while the other markers remained unchanged. Regarding seasonal changes, no differences were detected in adult females for liver CAT and ubiquinol 10 , CAT in roe, vitamin E in roe and in red cells, liver and red cell ubiquinol 10 , and in GSH in red cells. However, and coinciding with the spawning period of late summer, liver oxygen consumption, SOD and CAT activity and ubiquinol 10 contents in roe and SOD activity in red cells, and red cell TBARS contents were higher compared to late winter. These temporal antioxidant adjustments of Trichiurus lepturus seem to be parallel to the higher oxygen consumption typical of juvenile forms and also to the intense spawning and foraging activities of adult females in late summer., (Copyright © 2017. Published by Elsevier Inc.)

There is no evidence of the effect of 17β-estradiol (E2) administration on stress response and related metabolites after a short-period in chondrostean. In this study, we examined whether E2 is capable of inducing the hypothalamus-pituitary-interrenal axis activity. To accomplish this, sexually immature Acipenser stellatus were injected intraperitoneally with saline or E2 (5mg/kg body mass) and sampled 0, 1, 3, 6, 12, 24, 48, and 72h later. Plasma E2, cortisol, glucose, lactate, hematocrit (Hct), total protein (TP), cholesterol, triglyceride, alkaline phosphatase activity (ALP), calcium (Ca), and phosphorus concentrations were examined. Plasma levels of E2 significantly increased after 1h in E2-injected fish and remained high after 12h. E2 injection enhanced plasma TP and Ca concentrations, but had no effect on other parameters. Injection of either saline or E2 enhanced plasma glucose, lactate, TP, and Ca concentrations, but had no effect on Hct, cholesterol, triglyceride, and ALP. The results demonstrated that acutely elevated level of E2 did not lead to intense changes of stress metabolites and altered biochemical changes compared to the fish in control or saline treatment groups. Therefore, we found no exhibition of E2 on the rate of cortisol synthesis or stress indicators in this species, while E2 could modulate some other related metabolites on vitellogenesis pathway., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The shortnose sturgeon (Acipenser brevirostrum LeSueur, 1818) is a vulnerable species that is found along the eastern coast of North America. Little is known about temperature tolerance in this species and with a rapidly changing global climate, it becomes increasingly important to define the thermal tolerance of this species to better predict population distribution. Using a modified critical thermal maximum test (CTMax), the objectives of this study were to determine the impact of heating rate (0.1, 0.2 and 0.25°Cmin -1 ) on the thermal tolerance, associated hematological responses, and oxygen consumption in juvenile sturgeon. In addition, transcripts associated with physiological stress and heat shock (i.e., heat shock proteins) were also measured. Heating rate did not alter the CTMax values of shortnose sturgeon. Neither heating rate nor thermal stress affected plasma sodium and chloride levels, nor the expression of transcripts that included catalase, glucocorticoid receptor, heat shock protein70 (hsp70), heat shock protein 90α (hsp90α) and cytochrome P450 1a (cyp1a). However, regardless of heating rate, thermal stress increased both plasma potassium and lactate concentrations. Glucose levels were increased at heating rates of 0.2 and 0.25°Cmin -1 , but not at 0.1°Cmin -1 . Overall, oxygen consumption rates increased with thermal stress, but the response patterns were not affected by heating rate. These data support the hypothesis that shortnose sturgeon can tolerate acute heat stress, as many physiological and molecular parameters measured here were non-responsive to the thermal stress., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Prochilodus nigricans, locally known as curimatã, is an Amazonian commercial fish that endures adverse environmental conditions, in particular low dissolved oxygen, during its migration. Poorer environmental conditions are expected in the near future. Prochilodus nigricans overcomes current seasonal and diurnal changes in dissolved oxygen by adjusting erythrocytic levels of ATP and GTP, modulators of Hb-O2 affinity. Will this fish species be endangered under more extreme environmental conditions as hypoxia and acidification tend to occur in a shorter period of time? As P. nigricans does not exhibit any apparent morphological alterations to exploit the air-water interface, it must rely on fast adjustments of blood properties. To investigate this aspect, basic hematology indices, pHe, pHi, plasma lactate, erythrocytic levels of ATP and GTP and functional properties of the hemolysate of P. nigricans were analyzed over a period of 6h in hypoxia and subsequent recovery in normoxia. The levels of erythrocytic GTP were four times higher than ATP and were reduced to ¼ of the original level after 3h under hypoxia. Erythrocytic levels of ATP were unaffected over the experimental period. All other analyzed blood parameters exhibited a time-course change in animals under hypoxia and returned to normoxic levels. Considering the hemolysate functional properties and the ability to regulate the above mentioned blood characteristics, P. nigricans is able to endure short-term changes in dissolved oxygen., (Copyright © 2015 Elsevier Inc. All rights reserved.)

This study examined the effects of implanted cortisol on various aspects of intermediary metabolism of great sturgeon, Huso huso. Prior to experimentation all fish were examined using an endoscope to observe the stage of ovarian development. Subsequently, the 3-year-old female fish in the previtellogenic stage (mean body weight of 6759±53.2g) were intraperitoneally implanted with cocoa butter pellets containing cortisol to mimic the effects of chronic stress. The implant doses were 0 (C0; as control), 5 (C5) and 50 (C50) mg cortisol/kg body weight. Blood samples were taken every seven days during the four weeks of the experiment and analyzed for cortisol, glucose, adrenocorticotropic hormone (ACTH), total protein, total lipid, triiodothyronine (T3), thyroxine (T4), cholesterol and triglyceride content. Growth was reduced in all experimental groups and was not affected by cortisol treatment. Surprisingly, serum cortisol levels were higher in the C5 group than in the C50 throughout the experiment. A significant increase in glucose levels was observed in the cortisol-implanted fish from day 14 onwards. The high dose of cortisol elicited a significant increase in serum T3 and T4 levels. Fish implanted with the high cortisol dose also showed increases in serum ACTH, total lipid and cholesterol levels throughout a 28-day experimental period. The present study reveals that the negative effects of endoscopic surgery remain for at least four weeks and that a sustained-release implant of cortisol to mimic the effects of chronic stress affects metabolic responses. Since the adverse effects of endoscopic surgery on sturgeon welfare can be amplified by cortisol, special attention should be paid to the potential effects of chronic stress on sturgeon in culture., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Selection for growth-related traits in domesticated fishes often results in predictable changes within the growth hormone-insulin-like growth factor (GH-IGF-1) axis. Little is known about the mechanisms controlling changes in growth capacity resulting from fishery-induced evolution. We took advantage of a long-term study where Menidia menidia were selected for size at age over multiple generations to mimic fisheries-induced selection. This selection regime produced three populations with significant differences in intrinsic growth rate. These growth differences partially rebounded, but persisted even after selection was relaxed, resulting in fast, intermediate, and slow-growing lines. Plasma IGF-1 was measured in these populations as a potential target of selection on growth. IGF-1 was significantly correlated with current length and mass, and was positively correlated with growth rate (g d(-1)) in two lines, indicating it may be an appropriate indicator of growth capacity. The slow-growing line exhibited higher overall IGF-1 levels relative to the depressed IGF-1 seen in the fast-growing line, contrary to our prediction. We offer possible explanations for this unusual pattern and argue that somatic growth is likely to be under control of mechanism(s) downstream to IGF-1. IGF-1 provides an interesting basis for understanding endocrine control of growth in response to artificial selection and recovery., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The objective of this study was to examine the effect of cortisol implantations on gonadal development, sex steroid levels, and ovarian cortisol content in cultured great sturgeon Huso huso. Three groups of 5 fish for each treatment were considered. The experimental groups included: control (capsules containing cocoa butter alone), low cortisol (C(5); 5mg cortisol/kg body mass+cocoa butter) and, high cortisol (C(50); 50mg cortisol/kg body mass+cocoa butter). The capsules containing hormones and cocoa butter were intraperitoneally implanted into 3-year-old female fish at pre-vitellogenic stage (mean initial body mass 6809.7 ± 73 g) every 6 weeks over a 6-month period from January to June. The serum levels of cortisol, glucose, cholesterol and sex steroids (testosterone and 17β-estradiol) were determined at the initial time and three weeks after each implantation. Oocyte histological characteristics (the diameter and area of the oocyte, the diameter and area of the nucleus and the ratio of the nucleus area to the oocyte area) were measured at the end of the experiment and compared to those at the initial time. Ovarian cortisol content was measured at the end of the experiment. The results showed that serum cortisol levels varied in a dose-independent manner, so that the highest cortisol concentrations were observed in C(5)-treated fish throughout the experiment. Serum glucose levels were significantly higher in cortisol-treated groups than those in the control group. The high dose of cortisol elicited a significant constant increase in serum cholesterol concentrations. Fish implanted with the high cortisol dose showed significant declines in serum testosterone and 17β-estradiol concentrations throughout the experiment. No significant differences were found in oocyte histological characteristics among experimental groups. The cortisol implants elicited a dose-dependent increase in ovarian cortisol content. At the end of trial, body-growth indices were the lowest in C(50)-implanted fish, while the low cortisol dose had no effect on growth relative to the controls. These results indicated that chronic stress induced by cortisol implantation in great sturgeon suppressed gonadal steroidogenesis and somatic growth but had no effect on ovarian growth and development., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Photoperiod and temperature are known as the main synchronizers of seasonal reproduction in fish. This paper studied the role of photoperiod on the synchronization of F1 Senegal sole reproduction rhythms. Fish were maintained under constant short-photoperiod (9L:15D) from the winter solstice onwards (experimental group) or under naturally-changing photoperiod (control group), and water temperature naturally oscillated in both groups. Blood samples were collected during the reproduction season at pre-spawning (March), spawning (April) and post-spawning (May) to determine the endocrine status. Spawning events and egg quality parameters were also monitored. The results revealed a significant increase in nocturnal melatonin concentration from March to May in the control group, while in the experimental group such seasonal change did not occur. As to plasma levels of vitellogenin, testosterone, estradiol and 11keto-testosterone, differences between groups were found mostly in March, while in April and May levels were often similar. Spawning was observed in both groups, although the experimental group started slightly earlier and also finished earlier than the control group, perhaps as a result of the increase in sex steroids and VTG observed at pre-spawning. Briefly, reproduction rhythms persisted in the absence of the natural lengthening of photoperiod, although photoperiod manipulation altered the seasonal modulation of melatonin, increased sex steroids and vitellogenin at pre-spawning, and slightly advanced the timing of spawning., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Nitric oxide (NO)-mediated angiogenesis may play a role in establishing dense retinal vasculatures of Antarctic hemoglobinless icefishes (suborder: Notothenioidei). We hypothesized that loss of hemoglobin (Hb) leads to elevation in [NO] due to decreased degradation of the compound when the NO-scavenger Hb is absent, thereby inducing vascular growth. We found that total mass of NO metabolites, nitrite plus nitrate (NO(x)), in plasma is greater in icefishes than in red-blooded notothenioids [e.g. C. aceratus (Hb-), 22.7+/-2.9 microM; N. coriiceps (Hb+), 14.7+/-1.7 microM], suggesting a higher NO load in hemoglobinless animals. High NO levels do not appear to be a result of greater NO synthesis; we consistently measured lower activities of the enzyme catalyzing NO production, nitric oxide synthase, in tissues of icefishes than in Hb-expressing notothenioids [e.g. 96+/-10 and 216+/-39 pmol(min g wet wt)(-1) in brain tissue of C. aceratus (Hb-) and G. gibberifrons (Hb+), respectively]. Levels of mRNA for hypoxia-induced (HIF-1alpha and PHD2) and angiogenic genes (VEGF) were similar in red- and white-blooded species, indicating that vascular maintenance in adult animals does not require differences in angiogenic tone. This does not preclude a cause-and-effect relationship between absence of Hb and NO-mediated angiogenesis during earlier ontogenetic stages of icefishes., (Copyright (c) 2010 Elsevier Inc. All rights reserved.)

The aquatic air-breathing fish, Trichogaster microlepis, can be found in fresh water and estuaries. We further evaluated the changes in two important osmoregulatory enzymes, Na(+)/K(+)-ATPase (NKA) and vacuolar-type H(+)-ATPase (VHA), in the gills when fish were subjected to deionized water (DW), fresh water (FW), and salinated brackish water (salinity of 10 g/L). Fish were sampled only 4 days after experimental transfer. The mortality, plasma osmolality, and Na(+) concentration were higher in 10 g/L acclimated fish, while their muscle water content decreased with elevated external salinity. The highest NKA protein abundance was found in the fish gills in 10 g/L, and NKA activity was highest in the DW and 10 g/L acclimated fish. The VHA protein levels were highest in 10 g/L, and VHA activity was highest in the DW treatment. From immunohistochemical results, we found three different cell populations: (1) NKA-immunoreactive (NKA-IR) cells, (2) both NKA-IR and HA-IR cells, and (3) HA-IR cells. NKA-IR cells in the lamellar and interlamellar regions significantly increased in DW and 10 g/L treatments. Only HA-IR cells in the lamellar region were significantly increased in DW. In the interlamellar region, there was no difference in the number of HA-IR cells among the three treated. From these results, T. microlepis exhibited osmoregulatory ability in DW and 10 g/L treatments. The cell types involved in ionic regulation were also examined with immunofluorescence staining; three ionocyte types were found which were similar to the zebrafish model., (Copyright 2009 Elsevier B.V. All rights reserved.)

We examined for the first time the hemoglobin components of the blood of the Australian lungfish, Neoceratodus forsteri and their functional responses to pH and the allosteric modulators adenosine triphosphate (ATP), guanosine triphosphate (GTP), 2,3-bisphosphoglyceric acid (BPG) and inositol hexaphosphate (IHP) at 25 degrees C. Lysates prepared from stripped, unfractionated hemolysate produced sigmoidal oxygen equilibrium curves with high oxygen affinity (oxygen partial pressure required for 50% hemoglobin saturation, p(50)=5.3 mmHg) and a Hill coefficient of 1.9 at pH 7.5. p(50) was 8.3 and 4.5 mmHg at pH 6 and 8, respectively, which corresponded to a modest Bohr coefficient (Delta log p(50)/Delta pH) of -0.13. GTP increased the pH sensitivity of oxygen binding more than ATP, such that the Bohr coefficient was -0.77 in the presence of 2 mmol L(-1) GTP. GTP was the most potent regulator of hemoglobin affinity, with concentrations of 5 mmol L(-1) causing an increase in p(50) from 5 to 19 mm Hg at pH 7.5, while the order of potency of the other phosphates was IHP>ATP>BPG. Three hemoglobin isoforms were present and each contained both alpha and beta chains with distinct molecular weights. Oxygen affinity and pH-dependence of isoforms I and II were essentially identical, while isoform III had a lower affinity and increased pH-dependence. The functional properties of the hemoglobin system of Neoceratodus appeared consistent with an active aquatic breather adapted for periodic hypoxic episodes.

The African (Protopterus sp.) and South American lungfish (Lepidosiren paradoxa) inhabit shallow waters, that seasonally dry out, which induces aestivation and cocoon formation in Protopterus. Differently, L. paradoxa has no cocoon, and it aestivates in a simple burrow. In water PaCO(2) is 21.8+/-0.4mmHg (mean values+/-S.E.M.; n=5), whereas aestivation for 20 days increased PaCO(2) to as much as 37.6+/-2.1mmHg, which remained the same after 40 days (35.8+/-3.3mmHg). Concomitantly, the plasma [HCO(3)(-)]-values for animals in water were 22.5+/-0.5mM, which after 20 days increased to 40.2+/-2.3mM and after 40 days to 35.8+/-3.3mM. Initially in water, PaO(2) was 87.7+/-2.0mmHg, but 20 days in aestivation reduced the value to 80.5+/-2.2 and later (40 days) to 77.1+/-3.0mmHg. Meanwhile, aestivation had no effect on pHa and hematocrit. The blood pressures were equal for animals in the water or in the burrow (P(mean) approximately 30mmHg), and cardiac frequency (f(H)) fell from 31beats min(-1) to 22beats min(-1) during 40 days of aestivation. The osmolality (mOsmkgH(2)O(-1)) was elevated after 20 and 40 days of aestivation but declined upon return to water. The transition from activity to aestivation involves new set-points for the variables that determine the acid-base status and PaO(2) of the animals, along with a reduction of cardiac frequency.

Parameters of the antioxidant defense systems of Brycon amazonicus (matrinxã--a neotropical fish) exposed to phenol for 96 h plus the recovery over 1 and 2 weeks were studied in erythrocytes and liver. Hematocrit increase was observed during phenol exposure and recovery for 1 week. Total superoxide dismutases (SOD), glutathione peroxidase (GPx) and reduced glutathione (GSH) did not change during phenol exposure. Erythrocyte glucose-6-phosphate dehydrogenase (G6PDH) increased during that period while catalase (CAT) activity decreased during phenol exposure and recovery for 2 weeks. In the liver, SOD and CAT did not change, whereas GPx increased in the first week of recovery and decreased after 2 weeks. A late response was observed for G6PDH activity which increased only at the second week. Ascorbate concentration in the brain decreased during phenol exposure and increased over recovery. From our results it appears that the oxidative stress was limited in matrinxã exposed to phenol, but seemed to occur during the recovery period.

In this study we measured plasma cortisol, plasma glucose, plasma sodium and potassium, and liver and gill hsp70 levels in juvenile matrinxã (Brycon amazonicus) subjected to a 96 h exposure to phenol (0, 0.2, and 2.0 ppm), and the effect of this exposure on their ability to respond to a subsequent handling stress. Fish were sampled prior to initiation of exposure and 96 h, and at 1, 6, 12, and 24 h post-handling stress. During the 96 h exposure, plasma cortisol and glucose levels remained unchanged in all treatments. While plasma sodium levels were significantly reduced in all groups, plasma potassium levels only decreased in fish exposed to 0 and 0.2 ppm of phenol. Liver hsp70 levels decreased significantly at 96 h in fish exposed to 2.0 ppm of phenol. All groups, except fish exposed to 0.2 ppm of phenol, were able to increase plasma cortisol and glucose levels after handling stress. Fish exposed to 2.0 ppm of phenol showed decreased gill and liver hsp70 levels after the handling stress. Our data suggest that exposure to phenol may compromise the ability of matrinxã to elicit physiological responses to a subsequent stressor.

Pacific tarpon (Megalops cyprinoides) use a modified gas bladder as an air-breathing organ (ABO). We examined changes in cardiac output (V(b)) associated with increases in air-breathing that accompany exercise and aquatic hypoxia. Juvenile (0.49 kg) and adult (1.21 kg) tarpon were allowed to recover in a swim flume at 27 degrees C after being instrumented with a Doppler flow probe around the ventral aorta to monitor V(b) and with a fibre-optic oxygen sensor in the ABO to monitor air-breathing frequency. Under normoxic conditions and in both juveniles and adults, routine air-breathing frequency was 0.03 breaths min(-1) and V(b) was about 15 mL min(-1) kg(-1). Normoxic exercise (swimming at about 1.1 body lengths s(-1)) increased air-breathing frequency by 8-fold in both groups (reaching 0.23 breaths min(-1)) and increased V(b) by 3-fold for juveniles and 2-fold for adults. Hypoxic exposure (2 kPa O2) at rest increased air-breathing frequency 19-fold (to around 0.53 breaths min(-1)) in both groups, and while V(b) again increased 3-fold in resting juvenile fish, V(b) was unchanged in resting adult fish. Exercise in hypoxia increased air-breathing frequency 35-fold (to 0.95 breaths min(-1)) in comparison with resting normoxic fish. While juvenile fish increased V(b) nearly 2-fold with exercise in hypoxia, adult fish maintained the same V(b) irrespective of exercise state and became agitated in comparison. These results imply that air-breathing during exercise and hypoxia can benefit oxygen delivery, but to differing degrees in juvenile and adult tarpon. We discuss this difference in the context of myocardial oxygen supply.

Current understanding of the stress response in fishes has largely come from studies of freshwater-adapted salmonids, with proportionately few comparative studies having examined marine fishes. The current study sought to quantify the magnitude of physiological disturbances, recovery dynamics, and post-exercise behaviour in bonefish (Albula vulpes; a tropical marine fish) exposed to several different exercise and air exposure regimens. Results showed that metabolic disturbances (lactate production, hyperglycemia) increased following exercise and exposure to air, and that the magnitude of metabolic disturbance was proportional to the duration of the stressful event. Fish required between 2-4 h to return to resting values. Exercise and exposure to air also resulted in significant increases in plasma Ca2+, Cl- and Na+, but the magnitude of these ionic changes did not vary with exercise or exposure to air duration and required over 4-h to return to baseline levels. Mortality following exercise was observed only for fish that had been exposed to air for 3 min and not in fish that had been exposed to air for 1 min. Together, results from this study provide a physiological basis for management strategies that can improve the post-release survival of bonefish that have been caught during a catch-and-release angling event.

Host specificity data for gnathiid isopods are scarce because the parasitic stages are difficult to identify and host-parasite contact is often brief. We examined two common nocturnal species, Gnathia falcipenis and Gnathia sp. C, collected in light traps from two locations at Lizard Island on the Great Barrier Reef, Australia. Engorged third stage gnathiids were photographed and permitted to moult into adults to allow identification. We compared approximately 580 bp sequences of 16S mtDNA from blood meals with host sequences available on GenBank using BLASTn. Where homology was <98%, familial identity was investigated with neighbour-joining trees. All blood meal sequences (n=60) and homologous fish sequences (n=87) from GenBank were used in a Bayesian analysis, which identified all but three sequences to family. The host frequency distributions used by each species were significantly different; only four host families were shared. No gnathiids fed on elasmobranchs, blennies or apogonids, and most fed on host families whose representatives are typically large. Gnathia sp. C showed a distinct predilection for nemipterids. Gnathia falcipenis often parasitised sand-dwelling families, and unlike sympatric diurnal gnathiid species, it also frequently parasitised pomacentrids. We conclude that G. falcipenis and Gnathia sp. C operate as generalist micropredators with preferences.

Juvenile matrinxã (Brycon amazonicus) were fed with different dietary vitamin C (ascorbic acid or AA) concentrations for 2 months in order to evaluate its effect on the physiological condition of this species. Five treatments were tested: Reference diet-RD (350 mg AA kg(-1) and 45% crude protein commercial feed); C400, C500, C600 and C800 with supplementation of 400, 500, 600 and 800 mg AA kg(-1). Fish fed 800 mg AA kg(-1) supplemented diets presented greater weight gain and survival. Hematocrit, hemoglobin concentrations and erythrocytes showed significant increase (p<0.05) in C600 and C800 relatively to the others. Total leukocyte values were significantly higher in fish fed vitamin C supplemented diets relatively to those fed RD and leukocytes count of C600 and C800 were the highest among the treatments. Monocytes were the only cells of fish defense mechanism that presented variation, and C600 and C800 presented the lowest values (P<0.05). Glucose level was significantly different relatively to RD only in C400 and the plasmatic total protein values for fish fed vitamin C supplemented diets were higher than for fish fed the RD. Fish plasmatic ions (Na(+) and Cl(-)) were significantly lower for C500, C600 and C800 relatively to C400 and RD. The physiological profile and the growth performance of matrinxã suggest that diet supplemented with 800 mg AA kg(-1) can improve its performance and increase the survival rate.

To investigate the in vivo estrogenic activity of 4-nonylphenol (4-NP) on reproductive function, we have measured the expression level of rockfish vitellogenin (rVTG) mRNA, concentration of plasma VTG and levels of plasma estradiol-17beta(E(2)) and testosterone (T) by radioimmunoassay (RIA). An rVTG mRNA transcript of approximately 4.0 kb was extracted from hepatic tissue for Northern blot analysis, and the effects of 4-NP on rVTG mRNA expression in vivo with both male and female juvenile rockfish were examined. The level of rVTG mRNA expression was increased 48 h after injection with 4-NP of 10 mg/kg body mass and after injection in both male and female rockfish. The level of rVTG mRNA expression was increased 24 h after injection in male rockfish injected with 4-NP of 25 mg/kg body mass, while the level of rVTG mRNA expression was increased 12h after injection in female injected with same dosage. Concentrations of plasma rVTG in female injected with 4-NP of 10 and 25 mg/kg b.w. were increased 72 h after injection and reached 35 and 66 mg/mL, respectively. In male, rVTG concentrations with the dosage of 25 mg/kg body mass were increased 72 h after injection and reached to 14 and 65 mg/mL at 168 h, respectively. Plasma concentrations of T and E(2) in female and male injected with 4-NP were not significantly different between two 4-NP dosages. These results suggest that 4-NP may disrupt the reproductive system of immature rockfish by acting directly on vitellogenesis. Immature females injected with 4-NP were more sensitive than immature males, but VTG in immature males is likely to return to normal conditions slowly. There are obviously large interspecies differences in response of fish to 4-NP.

Phenol and its derivatives are xenobiotics present in many industrial wastewaters and in non-specific pesticides. It is a lipophilic compound and, therefore, accumulates along the trophic chain. Phenol is often found in marine and fresh water environments. The aim of this work was to detect metabolic changes induced by phenol in Brycon cephalus juveniles. Several enzymes activities and metabolites were quantified in the liver, white muscle and plasma. Among the enzymes assayed are alanine and aspartate amino transferases (ALAT and ASAT), lactate dehydrogenase (LDH) and malate dehydrogenase (MDH). Glucose, glycogen, lactate, ammonia and pyruvate were also quantified in tissues and plasma (glycogen in tissues only). The liver was the most responsive organ. The activities of the transaminases increased in muscle and liver, followed by an increase in hepatic ammonia. Correlation between ammonia and transaminases points towards phenol-induced consumption of protein. Hepatic glycogen and glucose contents were lower followed exposure to phenol. The same was observed for muscle glucose, suggesting considerable use of carbohydrate stores. The activity of hepatic lactate dehydrogenase increased with negative correlation with muscle lactate. This suggests that hepatic gluconeogenesis supplies tissues like muscle and brain with glucose. These results indicate that phenol intoxication demands metabolic energy and leads to significant changes of the metabolic profile of the fish, inducing to a certain extent a shift from carbohydrate catabolism to protein catabolism and the activation of gluconeogenesis.

A quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) assay was developed to measure transforming growth factor-beta (TGF-beta) in Atlantic menhaden (Brevoortia tyrannus), an estuarine-dependent species plagued by ulcerative skin lesions in the estuaries along the eastern United States. Atlantic menhaden were acclimated in a closed system for two weeks prior to initiation of the study. The synthetic glucocorticoid, triamcinolone acetonide (10mg/kg body weight) was administered by intracoelomic injection and its effect on the splenic mononuclear cell TGF-beta mRNA transcription, liver-somatic index, spleno-somatic index, hematology, and plasma chemistry were compared to untreated fish at 48 and 96h post-treatment. Triamcinolone-treated Atlantic menhaden showed suppression of TGF-beta mRNA production, neutrophilia, monocytosis, lymphopenia, and an increase in blood glucose concentrations. The health indices used in this study may help us interpret some of the changes observed during the development of ulcerative skin lesions in wild-caught menhaden.

Variations of Na(+)/K(+)-ATPase activity and fatty-acid composition in the gills of the sturgeon Acipenser naccarii subjected to progressive acclimation to full seawater (35 ppt) were determined in relation to the hypo-osmoregulatory capacity of this species in the hyperosmotic medium. Blood samples were taken and gills arches were removed at intermediate salinity levels between 0 and 35 ppt and after 20 days at constant salinity (35 ppt). Plasma osmolality and Na(+)/K(+)-ATPase activity increased significantly with growing environmental salinity. Total saturated fatty acids (SFAs) decreased, while total polyunsaturated fatty acids (PUFAs) increased significantly with increasing salinity due mainly to changes in n-3 PUFAs (20:5n-3 and 22:6n-3). The n-3/n-6 ratio increased significantly during the acclimation process. The results show a direct relationship between salinity, increased gill Na(+)/K(+)-ATPase activity and ultrastructural changes of the gill chloride cells. Changes in the fatty-acid composition in gills of A. naccarii during progressive acclimation to full seawater suggest that variations of gill fatty acids may also have a role in osmoregulatory mechanisms.

Salmon catfish and tarpon occur in habitats that periodically become deficient in oxygen resulting in high mortalities of other fish species. The water-breathing catfish, Arius leptaspis, and the facultative air-breathing tarpon, Megalops cyprinoides, both have high haemoglobin and haematocrit, and the oxygen carrying capacity in the air-breather is exceptionally high (15.6+/-1.2 vol%). Iso-pH oxygen equilibria of the red blood cells at 25 degrees C revealed high affinity (P(50)=9 mmHg, pH 7.4) and co-operativity (n(50)>2.2, pH 7.4) in the catfish, and contrasted with low affinity (P(50)=32 mmHg, pH 7.4) and co-operativity (n(50) approximately 1) in the air-breathing tarpon. Oxygen binding was further distinguished by relative pH insensitivity (Bohr factor, Ø=Deltalog P(50)/Deltalog pH=-0.22) in the catfish, compared with a significant Bohr effect in the tarpon (Ø=-0.96). The potential for modulation of haemoglobin-oxygen affinity was indicated by a high ratio of GTP to ATP in the erythrocytes of the catfish, whereas regulation in the tarpon appeared due to ATP alone. Differences in blood respiratory functions between the two species are likely to reflect reduced opportunity for activity under extreme hypoxia in the catfish.

Effects of increasing dietary protein were studied in matrinxa, (Brycon cephalus), an omnivorous teleost from the Amazon Basin in Brazil. Four isocaloric diets were formulated to contain 20%, 27%, 34% and 41% of crude protein (CP). Plasma glucose levels were significantly increased while triacylglycerols were significantly reduced at 41% of CP. Free fatty acids were significantly reduced at each level of rising CP. Plasma amino acids and ammonium followed the dietary CP increase. Liver glycogen and amino acids were reduced; liver glucose and lactate were constant, and ammonium increased with the CP in diets. Muscle glycogen and pyruvate decreased, protein did not change, while lactate and free amino acids increased. Kidney glycogen proportionally rose with the increase of CP from 20-41%. Pyruvate and lactate augmented irregularly from 20-41% CP. A gluconeogenic profile was observed in the kidney; the liver worked as regulator of body glucose. Increase of dietary CP and decrease of carbohydrates pushed muscle and liver catabolism of fat and sugar stores to satisfy energetical demands. CP contents above 34% were not recommended for B. cephalus, if the carbohydrate does not compensate the metabolical demands.

Adult specimens of traira (Hoplias malabaricus Bloch) were subjected to long-term starvation (30 to 240 days) and re-fed for 30 days after 90 and 240 days of food deprivation. Counting of immature erythrocytes in peripheral blood showed that erythropoiesis decreased significantly during the first 30 days of food deprivation. The results suggest that a process of senescence takes place in the pre-existent red blood cells and that the cells are not replaced during starvation. After 240 days of starvation, H. malabaricus had a significantly reduced number of red blood cells, causing changes in hematocrit and blood indices (mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration). Furthermore, during this period, the fish presented leukopenia (lymphocytopenia) and thrombocytopenia. After re-feeding, the number of leukocytes and thrombocytes recovered, but the red blood cell number remained reduced and there was a significant increase in abnormal red cell nuclei.

We examined the estrogenic effect of 2,2'4,4'5,5'-hexachlorobiphenyl (PCB 153) on the rockfish, Sebastes schlegeli. We measured levels of plasma estradiol-17beta (E(2)) and testosterone (T) by radioimmunoassay (RIA). Plasma concentrations of T and 17alpha-hydroxyprogesterone in female and male fish injected with PCB 153 using two dosages (0.16 mg/kg body weight. and 0.57 mg/kg) did not differ significantly between sexes or from sham-injected controls of the same sex. Plasma concentrations of E(2) in females injected with PCB 153 (both levels) increased at 12 and 24 h. Concentrations of plasma vitellogenin (VTG) in females increased 72 h after injection with PCB 153 and reached 0.38 and 1.25 mg/mL, respectively. No VTG was detected in males injected with the same dosage. These results suggest that PCB 153 may lead to the production VTG in female rockfish through a synergistic effect with E(2), resulting in indirect disruption of the aromatization process.

In mammals the pentraxins, C-reactive protein (CRP) and serum amyloid P component (SAP) are important components of the immune response. Although pentraxins have been isolated from a number of fish species few studies detail their functional immunological role. In this paper we report the establishment of a flow cytometry based assay to measure the phagocytic activity of isolated snapper head kidney leukocytes (HKLs). This assay was then used to examine the ability of a pentraxin-like protein isolated from the serum of snapper (P. auratus) (Sn-PLP) to act as an opsonin. Incubation of snapper head kidney leukocytes (HKL) with FITC-labelled beads resulted in uptake of these particles by approximately 35% of HKLs. Incubation of beads with Sn-PLP enhanced phagocytosis by snapper HKLs in a dose-dependant manner. Enhanced phagocytosis could be inhibited by addition of a rabbit anti-Sn-PLP antibody suggesting that Sn-PLP may act as a ligand for a HKL cell surface receptor. This study provides further evidence toward a functional role for pentraxins in the host defence repertoire of fish.

Pentraxin-like molecules have been isolated from a number of fish species. However, little is known about the function of these proteins in the teleosts. In this study we report the isolation and characterization of a pentraxin-like molecule from the serum of snapper (Pagrus auratus) that has the ability to activate complement. This pentraxin-like protein was isolated from serum by calcium-dependent binding to agarose. SDS-PAGE analysis demonstrated an oligomeric protein of approximately 200k Da consisting of non-covalently bound subunits of 26 and 23 kDa. Protein sequencing revealed significant (50%) sequence identity with pentraxins from both Atlantic salmon (S. salar) and rainbow trout (O. mykiss). However, polyclonal antibodies raised against snapper pentraxin did not recognise salmon or trout pentraxin in Western blot analysis. Following LPS injection, snapper pentraxin levels increased 2-fold before gradually returning to basal levels. Most significantly, the isolated pentraxin initiated complement-mediated lysis of ligand-coated sheep erythrocytes in a dose-dependent fashion. In view of the similarity between the known fish pentraxins, and their similarity to mammalian serum amyloid P-components we conclude that the isolated protein may be a snapper pentraxin homologue.

The effects of time of day and water temperature on the acute physiological stress response were investigated in young-of-the-year green sturgeon (Acipenser medirostris). The response to a 1-min air-emersion stressor was assessed during the day (08.00 h) and at night (20.00 h), as well as after acclimation to either 11 degrees C or 19 degrees C. Blood samples were collected prior to stress and at several times after exposure to the stressor, and plasma concentrations of cortisol, lactate, and glucose were determined. The magnitudes of cortisol (19.1 ng ml(-1) vs. 4.9 ng ml(-1)) and lactate (190.6 mg l(-1) vs. 166.7 mg l(-1)) were significantly higher in fish stressed at night when compared with the day. There were no significant differences in glucose levels between time periods. Although, acclimation temperature did not affect peak cortisol concentrations (56.7 and 50.3 ng ml(-1) at 11 degrees C and 19 degrees C, respectively), the duration of the response was significantly extended at 11 degrees C. Post-stressor lactate increases were similar between temperature groups, but at 11 degrees C post-stressor glucose levels were significantly increased through 6 h, suggesting stressor-induced glycogenolysis and gluconeogenesis or decreased glucose utilization. These data demonstrate that the physiological stress response in green sturgeon is modified by both time of day and temperature.

Increased vitellogenin (vtg) levels in the blood of male fish are frequently used as an indicator of estrogenic exposure. Similar responses are expected for mussels, where the concentration of vtg-like proteins has been reported to depend on estrogens. To verify the role of hemolymph during vitellogenesis of mussels, the saltwater mussel Mytilus edulis and the freshwater mussel Anodonta cygnea were exposed to 17beta-estradiol (E2) and wastewater treatment plant effluents, known for their estrogenic potential. Gel electrophoresis did not reveal any significant induction (or repression) of plasma proteins compared to control plasma. Our results do not support the hypothesis that mussel hemolymph is a carrier of estrogen-dependent major egg-yolk precursors (vtg-like proteins). However, additional information on a 35+/-2-kDa hemolymph protein, previously reported to bind heavy metals, was obtained by high-resolution two-dimensional electrophoresis. It was resolved in a cluster of single proteins with properties that match the characteristics of a previously reported histidine-rich glycoprotein.

Following a 7.5-h transport haul, juvenile pallid sturgeon (Scaphirhynchus albus) showed a small but significant increase in plasma cortisol to 4.7 ng ml(-1) but similar increases did not occur after fish were handled in a net held in the air for 30 s. Subsequent experiments on yearling pallid sturgeon and hybrid pallidxshovelnose (S. albusxplatorynchus) sturgeon using the same 30-s handling stressor failed to evoke increases in plasma cortisol, lactate or glucose. Plasma cortisol increased significantly from about 2 to 13-14 ng ml(-1) in both pallid and hybrid sturgeon during a 6-h severe confinement stressor with handling. Plasma cortisol in 2-year-old pallid sturgeon subjected to the same stressor demonstrated a linear pattern of increase during the initial 1 h. Plasma lactate increased from 1.11 to about 2.11 mmol l(-1) in hybrid sturgeon during the first hour of severe confinement but did not change throughout the entire confinement period in pallid sturgeon. A significant increase in plasma cortisol to 5.4 ng ml(-1) in 2-year-old pallid sturgeon 1 h after being subjected to 30 s handling at 19:00 h but not at 07:00 or 13:00 h suggests that a small diurnal variation in their stress response may exist. Although both pallid and hybrid sturgeons were responsive to stress, they exhibited very low physiological responses compared with those following equivalent stressors in most teleostean fishes or another chondrostean, the paddlefish (Polyodon spathula). Reasons for the apparent low responses to handling and confinement in scaphirhynchid sturgeons are not known but may relate to their evolutionary history, neuroendocrine mechanisms involved in their corticosteroid responses, or anatomy of their interrenal tissue structure.

Fish are dependent on aerobic metabolism. They respond to changes in oxygen availability by a wide spectrum of compensatory and respiratory adjustments to safeguard tissue oxygenation. Such adjustments are directed to facilitate both oxygen uptake at the gas exchange surfaces and oxygen unloading to tissues. The importance of erythrocytic organic phosphates as regards oxygen transfer has been recognised since 1967 when the 'dramatic' effect of 2,3DPG on human haemoglobin was first reported. The present review examines the appearance of all the major erythrocytic organic phosphates during the evolutionary radiation of fish. In addition, it provides examples illustrating qualitative and quantitative ontogenetic changes of organic phosphates in the red blood cell of several fish species and describes their effects on oxygen affinities. The interaction of the organic phosphates with haemoglobins and divalent cations are also examined. Of particular interest is the regulation of erythrocytic organic phosphates according to both environmental and physiological conditions.

Exposure of a facultative air breather, Hoplosternum littorale, to 12.5, 25, and 37.5% of the water soluble fraction (WSF) of Urucu crude oil, resulted in a rapid increase in air-breathing frequency (ABF) sustained over the 45 min period of exposure. Following 4 h exposure to a graded increase in WSF up to 50%, there was no significant affect on haematocrit, or plasma [Na+] and [K+]. Crude oil ingestion resulted in some degree of ion regulatory impairment, however results were variable. A single oral dose of 3.0 ml/kg of Urucu crude elevated net whole body Na+ efflux and resulted in a 7% reduction in plasma [Na+] 72 h following ingestion. A single oral dose of 3.0 ml/kg resulted in a significant net whole body K+ efflux and a reduction in plasma [K+] 24 h after ingestion. No mortalities were observed in any exposure regime in this study. An oral dose of Urucu crude oil at 3.0 ml/kg also resulted in a 24% reduction in ATP:Hb ratio (from 0.206 to 0.157) and a 31% reduction in GTP:Hb ratio (0.455 to 0.315) 24 h following ingestion indicating that these fish may be hypoxemic. Taken together, these results indicate that exposure of H. littorale to Urucu crude oil affects gas exchange and ion regulation.

The blood oxygen affinity of vertebrates is regulated, in part, through changes in red cell phosphate levels and increased oxygen affinity during reductions in inspired oxygen and is a well-described and common feature. However, during anaemia, when oxygen delivery is compromised by a reduction in blood oxygen carrying capacity, a lowering of blood oxygen affinity will facilitate oxygen unloading in the tissues, while oxygen loading at the gas exchange organ is not impaired. The present study investigated the effects of artificially induced anaemia in vivo on the blood oxygen affinity and red cell nucleoside triphosphate (NTP) concentrations in the turtle, Chrysemys picta. Blood was obtained from conscious animals through an arterial catheter and oxygen equilibrium curves were determined using the Tucker method while NTP concentrations were analyzed spectrophotometrically. Before induction of anaemia haematocrit averaged 23% and P50 was 18.5 +/- 0.7 with a NTP/Hb of 0.20 +/- 0.01 (mmol/mmol). After the haematocrit had been reduced to approximately 10% by bleeding (48-96 h) (blood volume was maintained by re-infusion of plasma and Ringer) there were no effects on P50 or red cell NTP concentrations. Thus, in contrast to fish and mammals, turtles do not exhibit a change in blood oxygen affinity during anaemia.

The concentrations of ammonia in the plasma of the mudskipper Boleophthalmus boddaerti exposed to cyanide for 1-6 days were significantly greater than the respective values of the controls. This was due to an increase in the production of NH3 in the muscle and an increase in the retention of NH3 and/or NH4+ in the blood of the cyanide-exposed fish when compared to controls. Cyanide exposure significantly increased the specific activity of muscle AMP deaminase. Since adenylosuccinate synthetase and lyase were also present in the muscle, exposure to cyanide might increase the production of NH3 from the catabolism of purine nucleotides. B. boddaerti exposed to cyanide excreted significantly less ammonia than the control fish. Results indicate changes in the permeability of the epithelial surfaces of the cyanide-exposed fish to NH3 and/or NH4+. Since the tissues and organs needed time to activate the inducible cyanide detoxification mechanisms, the increase in the production of NH3 might be an important defensive mechanism for B. boddaerti during the early phase of cyanide exposure.

The physiological role of the catecholamines (CA), adrenaline and noradrenaline in fish has been frequently reviewed, but the metabolic consequences of these hormones have received less attention. The purpose of this review is to examine the recent literature dealing with CA actions on whole fish and tissue metabolism. The CA increase glucose production both in vivo and in vitro, at least in isolated hepatocytes. Although the data are less clear, lipid mobilization is also a consequence of elevated circulating CA. The difficulty with using the whole fish for such studies is that CA may alter other circulating hormone levels, CA turnover in the circulation quickly, and it is difficult to define precisely the tissue being affected. Much of our understanding is derived, therefore, from the study of isolated tissues, and especially the hepatocyte. Catecholamines stimulate both glycogenolysis and gluconeogenesis in hepatocytes isolated from a large number of fish species. This review examines the steps involved in the signal transduction system, from the binding of CA to alpha- and beta-adrenoceptors to the ultimate effects of specific enzyme phosphorylation. Recent literature demonstrates that the complexity of the adrenoceptor system noted for mammals, also is expressed in fish. Adrenoceptor subtypes are specific to species, to tissues and to function of the tissues, and these issues are discussed especially as they are related to external and to internal stressors. Future research will pursue better definitions of the adrenoceptor systems, molecular biology of the components of these receptor systems and development of alternative cell models. There still remains a poor explanation of the reason for the diversity of adrenoceptor systems, and there are a number of fish systems that may provide unique opportunities to understand this question.

Vitellogenin was isolated by gel filtration and ion-exchange chromatography from plasma of estradiol-treated male Zoarces viviparus. The purification of vitellogenin was followed through each step by native polyacrylamide gelelectrophoresis (PAGE). The purified vitellogenin was used to raise anti-vitellogenin antibodies in rabbits. The specificity of the affinity purified antibody raised against vitellogenin was assessed by Western blot analysis. No crossreactivity was observed with plasma from non-induced control males. A direct enzyme-linked immunosorbent assay (ELISA) was developed by use of the raised anti-vitellogenin. The detection limit for the purified standard vitellogenin by the ELISA method was 5 ng ml-1 Vitellogenin levels were quantified in plasma of pregnant female Zoarces viviparus, which were held in aquaria with different concentrations of the estrogenic compound 4-nonylphenol dissolved in the ambient seawater. A marked effect of exposure to ambient 4-nonylphenol was observed by significant dose-dependent increases in the level of plasma vitellogenin of the pregnant fish as well as of embryos exposed in vitro.

Carbonic anhydrase and proton ATPase are co-distributed, being restricted to the apical regions of the gill epithelium of freshwater teleosts. Carbonic anhydrase supplies protons to the apical proton ATPase. Carbonic anhydrase is absent from the basal regions of the gill epithelium. Plasma flowing through the gills has no available carbonic anhydrase activity and plasma CO2/bicarbonate reactions are uncatalyzed. Thus, bicarbonate dehydration in plasma is negligible, and catalyzed bicarbonate dehydration occurs in erythrocytes in blood flowing through the gills. This results in tight coupling of carbon dioxide excretion to oxygen uptake and the evolution of hemoglobins with large Haldane effects but low buffering capacities, typical of many freshwater teleosts. Tight coupling of carbon dioxide and oxygen transfer in these fish also ensures that the Root shift does not impair oxygen uptake at the gills. Under these conditions, there is a selective advantage for hemoglobins with a Root shift. The presence of a Root shift augments oxygen transfer to the tissues in general and the eye and swimbladder in particular.

Several membrane ion transporters playing a role in gas transport and exchanges, cell volume regulation and intracellular acid-base regulation have been identified in fish red blood cells (RBCs). This short review focuses on Na+/K+ATPase and its role in establishing the ionic gradients across the membrane, on the Cl-/HCO3- exchanger and its key role in respiration and possibly in inducing a chloride conductance, on the Na+/H+ exchanger and the recent advances on its molecular mechanisms of activation and regulation, on the different types of K-Cl cotransports, the different hypotheses and suggested models and their role in cell volume regulation. There is no evidence in the literature for ionic channels in fish RBCs. We present original data obtained with the patch-clamp technique that shows for the first time the existence of a DIDS-sensitive chloride anionic conductance measured in whole cell configuration and the presence of a stretch-activated nonselective cationic channel recorded in cell-attached and excised inside-out configuration. The part played by these ionic conductances is discussed in relation with their possible involvement in volume regulation.

Vitellogenin of matrotrophic viviparous eelpout (Zoarces elongatus) was purified from estradiol-17 beta (E2) treated immature male sera by gel chromatography and anion exchange chromatography. Isolated vitellogenin has a molecular weight of 540 kDa estimated by gel chromatography. Serum levels of vitellogenin in females were measured during oocyte development and gestation by single radial immunodiffusion. Serum vitellogenin level was low (less than 0.2 mg/ml) during the early vitellogenic period, increased in the late vitellogenic period to a peak level (6.4 +/- 2.1 mg/ml) at the beginning of gestation. After that it rapidly decreased to a low level (0.1 +/- 0.1 mg/ml) during the early gestation period. Levels of vitellogenin remained low throughout the gestation period. Serum E2 levels in females showed increased from 1.3 to 3.0 ng/ml during the late vitellogenic period, and declined to 0.4 ng/ml during the early gestation period. Serum levels of E2 showed good correlation with serum vitellogenin levels, suggesting that the vitellogenin synthesis is controlled by E2 in this species. These results combined with the matrotrophic growth of embryo during gestation suggest that there is a shift in the synthesis of maternal nutritional products for embryos from the yolk to other nutrients.

Glucose-6-phosphate dehydrogenase (G6PD) and L-glutamate dehydrogenase (GDH) from Antarctic fish were isolated and characterized. G6PD was purified from the erythrocytes of red-blooded Dissostichus mawsoni and from the colorless blood of the icefish Chionodraco hamatus. Structural and functional characterization showed that the two enzymes do not differ significantly from each other. GDH was purified from the liver of the icefish Chaenocephalus aceratus. As in other fish ODHs, it showed a marked preference for NAD-. The amino acid sequence of the active-site peptide is virtually identical to that of other fish and vertebrate counterparts. Although the basic structural features of the Antarctic enzymes are similar to those of mesophilic organisms, some catalytic and thermodynamic properties make the Antarctic enzymes more suited to cold-adapted organisms.

Radiolabelled vitellogenin produced by juvenile grouper following injection of 3H-leucine, 32P-orthophosphate and estradiol-stimulation was purified by anion-exchange and gel filtration chromatography and characterized by electrophoresis and chemical analysis. The radiolabelled product was found to exist in two heterogeneous molecular weight forms by electrophoresis under nondenaturing conditions on polyacrylamide gel with M(r), of 260,000 and 525,000. It showed two protein monomers (M(r) 113,000 and 140,000) on electrophoresis under denaturing condition on a sodium dodecyl sulphate-polyacrylamide gel. The purified material contained 17.2% total lipid with 1.3% cholesterol and 2.1% triglycerides, 8-21 micrograms/mg protein total carbohydrate and 6.8 micrograms phosphate/mg protein; amino acid analysis showed a profile comparable to that found for vitellogenins isolated from the grouper, medaka, goldfish and rainbow trout. The study showed grouper vitellogenin to be a glycophospholipoprotein similar in composition to vitellogenins from other teleosts and demonstrated that vitellogenins can be induced in juveniles by injection with estradiol-17 beta.

1. Relative to human HbA, opossum (Didelphis virginiana) hemoglobin was found to be more susceptible to autoxidation. While the initial rate of autoxidation of spot (Leiostomus xanthurus) hemoglobin is close to that of HbA, complete oxidation occurs in 50 hr. 2. Direct addition of hydrogen peroxide (H2O2) induced oxidation of hemoglobins in a definite order: spot Hb > HbA > opossum Hb. Excess H2O2 led to heme degradation and precipitation that occurred much faster for spot Hb than the case with other proteins. 3. Exposure of hemoglobins to a continuous flux of H2O2, generated by the glucose/glucose oxidase system, induced the formation of heterogeneous protein-associated oxidation products. 4. Differential reactivity among these hemoglobins under the same or different oxidative conditions, with respect to methemoglobin formation and stability of the ferric form, may reflect the differences in the local heme environment of these proteins.

1. Tilapia sparrmanii was exposed to 0.098 mg/l potassium dichromate. 2. The effect of chromium on blood coagulation was measured with a thrombelastograph at a pH of 5, 7.4 and 9 over 96 hr, as well as an uncontrolled pH after 2 weeks of exposure. 3. The prolonged kinetic times and reduced maximal amplitudes at a pH of 7, 4 and 9, differed significantly from the control values. 4. Thrombelastograms reflected a decrease in the clotting ability of fish blood, with an increase in pH, which is typical of thrombocytopenia. 5. It was evident that the exposure of fish to chromium led to clotting defects that caused internal bleeding.