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Osmoregulatory actions of the GH/IGF axis in non-salmonid teleosts

Salmonid fishes provided the first findings on the influence of the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis on osmoregulation in teleost fishes. Recent studies on non-salmonid species, however, indicate that this physiological action of the GH/IGF-I axis is not restricted to salmonids or anadromous fishes. GH-producing cells in the pituitary of fish acclimated to different salinities show different degrees of activation depending on the species studied. Plasma GH levels either increase or do not change after transfer of fish from freshwater to seawater. Treatment with GH or IGF-I increases salinity tolerance and/or increases gill Na + ,K + -ATPase activity of killifish ( Fundulus heteroclitus ), tilapia ( Oreochromis mossambicus and Oreochromis niloticus ) and striped bass ( Morone saxatilis ). As in salmonids, a positive interaction between GH and cortisol for improving hypoosmoregulatory capacity has been described in tilapia ( O . mossambicus ). Research on the osmoregulatory role of the GH/IGF-I axis is derived from a small number of teleost species. The study of more species with different osmoregulary patterns will be necessary to fully clarify the osmoregulatory role of GH/IGF-I axis in fish. The available data does suggest, however, that the influence of the GH/IGF-I axis on osmoregulation may be a common feature of euryhalinity in teleosts.

Comparative Biochemistry and Physiology, Part B: B

Aryl hydrocarbon receptor function in early vertebrates:Inducibility of cytochrome P450 1A in agnathan and elasmobranch fish

The mammalian aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that controls the expression of cytochrome P450 1A (CYP1A) genes in response to halogenated aromatic hydrocarbons such as 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD). The natural ligand and normal physiologic function of this protein are as yet unknown. One approach to understanding AHR function and significance is to determine the evolutionary history of this receptor and of processes such as CYP1A induction that are controlled by the AHR in mammals. In these studies, AHR function was evaluated in representative cartilaginous fish (little skate, Raja erinacea ) and jawless fish (sea lamprey, Petromyzon marinus and Atlantic hagfish, Myxine glutinosa ), using CYP1A induction as a model AHR-dependent response. Treatment of skate with β -naphthoflavone (BNF) caused an 8-fold increase in hepatic ethoxyresorufin O -deethylase (EROD) activity as well as a 37-fold increase in the content of immunodetectable CYP1A protein. Evidence of CYP1A inducibility was also obtained for another cartilaginous fish, the smooth dogfish Mustelus canis . In contrast, hepatic EROD activity was not detected in untreated lamprey nor in lamprey treated with 3,3′,4,4′-tetrachlorobiphenyl (TCB), a potent AHR agonist in teleosts. A possible CYP1A homolog was detected in lamprey hepatic microsomes by one of three antibodies to teleost CYP1A, but expression of this protein was not altered by TCB treatment. CYP1A protein and catalytic activity were measurable in hagfish, but neither was induced after treatment with TCB. These results suggest that the AHR-CYP1A signal transduction pathway is highly conserved in gnathostomes, but that there may be fundamental differences in AHR signaling or AHR-CYP1A coupling in agnathan fish. Agnathan fish such as hagfish and lamprey may be interesting model species for examining possible ancestral AHR functions not related to CYP1A regulation.

Comparative Biochemistry and Physiology, Part C: P

Possible mechanisms for sensitivity to organophosphorus and carbamate insecticides in eastern screech-owls and American kestrels

Effects of a single dietary exposure to fenthion and carbofuran on the survival, feeding behavior and brain ChE activity of eastern screech-owls, Otus asio and American kestrels, Falco sparverius , were evaluated. Birds were exposed to fenthion (23.6–189.0 ppm) or carbofuran (31.7–253.6 ppm) via meatballs. Carbofuran-exposed owls ate either ≤10% or ≥80% of the meatball whereas all kestrels ate ≤10% of the meatball before exhibiting acute signs of toxicity. Fenthion-exposed owls and kestrels displayed a wide spectrum of meatball consumption (<10–100%). Significant brain ChE inhibition was observed in dead and surviving kestrels exposed to fenthion and carbofuran and dead owls exposed to fenthion ( P <0.0001). Brain ChE activity of owls exposed to carbofuran that survived was not different from that of controls ( P =0.25). Data suggest: (1) slow feeding on a carbamate-contaminated item may provide limited protection from the toxicity of the chemical at certain rates of exposure; (2) the degree of ChE inhibition at neuromuscular junctions may be critical in determining the sensitivity of a species to a carbamate insecticide; (3) sensitivity may be a function of the ChE affinity for the carbamate inhibitor; and (4) the importance of neuromuscular junction ChE depression in determining the sensitivity of an animal may be species-specific.

Comparative Biochemistry and Physiology, Part C: P

Identification of in vitro cytochrome P450 modulators to detect induction by prototype inducers in the mallard duckling ( Anas platyrhynchos

Seven modulators of mammalian monooxygenase activity were screened for their ability to selectively stimulate or inhibit in vitro monooxygenase activities of hepatic microsomes from mallard ducklings treated with phenobarbital, β -naphthoflavone, 3,3′,4,4′,5-pentachlorobiphenyl or vehicle. Microsomes were assayed fluorometrically for four monooxygenases: benzyloxy-, ethoxy-, methoxy-, and pentoxyresorufin- O -dealkylase, in combination with each of the seven modulators. Four combinations: α -naphthoflavone and 2-methylbenzimidazole with benzyloxyresorufin, and Proadifen with methoxy- and ethoxyresorufin, respectively, were evaluated further. β -Naphthoflavone-treated groups were clearly distinguished from the corn oil vehicle control group by all of the assays and by the effects of the modulators in three of the four assay/modulator combinations. Enzyme activities of the phenobarbital and saline groups were statistically similar ( P ≥0.05) when assayed without modulator added, but each assay/modulator combination distinguished between these groups. The PCB-treated group was distinguished from the corn oil vehicle control group only for BROD activity, with or without the presence of modulator. Graphing of per cent modulation of BROD activity versus initial BROD activity provided the clearest distinction between all of the study groups. Identification of these selective in vitro modulators may improve detection and measurement of low level cytochrome P450 induction in avian species. Also, both the monooxygenase activities induced and the impacts of the modulators indicated differences between mammalian and avian cytochromes P450.

Comparative Biochemistry and Physiology, Part C: P

The oral bioavailability and toxicokinetics of methylmercury in common loon ( Gavia immer ) chicks

We compared the toxicokinetics of methylmercury in captive common loon chicks during two time intervals to assess the impact of feather growth on the kinetics of mercury. We also determined the oral bioavailability of methylmercury during these trials to test for age-related changes. The blood concentration-time curves for individuals dosed during feather development (initiated 35 days post hatch) were best described by a one-compartment toxicokinetic model with an elimination half-life of 3 days. The data for birds dosed following completion of feather growth (84 days post hatch) were best fitted by a two-compartment elimination model that includes an initial rapid distribution phase with a half-life of 0.9 days, followed by a slow elimination phase with a half-life of 116 days. We determined the oral bioavailability of methylmercury during the first dosing interval by comparing the ratios of the area under the blood concentration-time curves (AUC 0&rarr;&infin; ) for orally and intravenously dosed chicks. The oral bioavailability of methylmercury during the first dosing period was 0.83. We also determined bioavailability during both dosing periods using a second measure because of irregularities with intravenous results in the second period. This second bioavailability measure estimated the percentage of the dose that was deposited in the blood volume (f), and the results show that there was no difference in bioavailability among dosing periods. The results of this study highlight the importance of feather growth on the toxicokinetics of methylmercury.

Comparative Biochemistry and Physiology, Part A: M

Immune function and hematology of male cotton rats ( Sigmodon hispidus ) in response to food supplementation and methionine

We examined effects of supplementation of food quantity and quality (=enhanced methionine) on hematologic and immunologic parameters of wild, but enclosed, adult male cotton rats ( Sigmodon hispidus ) in north-central Oklahoma. Sheet metal enclosures were stocked with a high density of wild-caught cotton rats (160 animals/ha) and randomly assigned a treatment of no supplementation, mixed-ration supplementation or methionine-enhanced supplementation. Aside from small increases in counts of red blood cells and hematocrit levels, most indices of erythrocytic characteristics were not affected by supplementation with the mixed-ration or enhanced methionine. In contrast, platelet counts were highest in mixed-ration and methionine treatments and counts of total white blood cells were highest with methionine supplementation, albeit relative proportions of different leukocytes did not differ among treatments. Immunologically, neither delayed-type hypersensitivity response nor hemolytic-complement activity differed among treatments. Supplementation of food quantity and quality did not broadly affect hematologic parameters and immune function of male cotton rats, but enhanced platelet and leukocyte counts may confer advantages to overall health. Clarification of the role of such effects on population limitation or regulation requires additional research.

Comparative Biochemistry and Physiology, Part A: M

Electrocardiographic consequences of a peripatetic lifestyle in gray wolves ( Canis lupus )

Cardiac chamber enlargement and hypertrophy are normal physiologic responses to repetitive endurance exercise activity in human beings and domestic dogs. Whether similar changes occur in wild animals as a consequence of increased activity is unknown. We found that free-ranging gray wolves ( Canis lupus , n =11), the archetypical endurance athlete, have electrocardiographic evidence of cardiac chamber enlargement and hypertrophy relative to sedentary captive gray wolves ( n =20), as demonstrated by significant increases in QRS duration, QT interval, and QT interval corrected for heart rate, a tendency towards increased Q, R, and S wave voltages in all leads, and a significant decrease in heart rate. We conclude that exercise activity level and therefore lifestyle affects physiologic variables in wild animals. An immediate consequence of this finding is that physiologic measurements obtained from a captive wild-animal population with reduced exercise activity level may not accurately reflect the normal physiologic state for free-ranging members of the same species.

Comparative Biochemistry and Physiology, Part A: M

Maternal steroids and contaminants in common tern eggs: A mechanism of endocrine disruption?

We looked for evidence for the hypothesis that exposure of female birds to polychlorinated biphenyls (PCBs) results in alteration of blood steroid hormone concentrations and alters subsequent hormone transfer of steroids to eggs. Eggs of three-egg clutches were collected from a PCB-exposed common tern ( Sterna hirundo ) colony (Ram Island, Buzzards Bay, MA, USA) and from a relatively clean colony (Bodkin Island, Chesapeake Bay, MD, USA), and were analyzed for concentrations of organochlorine contaminants and steroid hormones (17β-estradiol, 5α-dihydrotestosterone, testosterone and androstenedione). There was no relationship between total PCBs and steroid concentrations considering all eggs together, considering eggs of different laying order or considering differences between sequentially laid eggs in a clutch. Similarly, concentrations of di- and tri-chlorinated biphenyls and steroids in eggs were not related. The concentrations of PCBs, mercury and selenium were below estimated thresholds for toxicity to embryos. Maternal steroids, except estradiol, were present in yolk of all eggs, with increasing concentrations in the second and third eggs laid. Our data provided no evidence for a maternal toxicological event that might alter the amount of maternal steroid hormone transferred to eggs.

Comparative Biochemistry and Physiology, Part C: T

Fasting modifies Aroclor 1254 impact on plasma cortisol, glucose and lactate responses to a handling disturbance in Arctic charr

Integrated effects of polychlorinated biphenyl (PCB) and nutritional status on responses to handling disturbance were investigated in the Arctic charr ( Salvelinus alpinus ). The fish were orally contaminated with Aroclor 1254 and held either with or without food for 5 months before they were subjected to a 10-min handling disturbance. Food-deprived fish were given 0, 1, 10 or 100 mg PCB kg −1 and the fed fish 0 or 100 mg PCB kg −1 . Plasma cortisol, glucose and lactate levels were measured at 0 (pre-handling), 1, 3, 6 and 23 h after the handling disturbance. Food-deprived control fish had elevated plasma cortisol levels compared with fed fish before handling. These basal cortisol levels were suppressed by PCB in food-deprived fish, and elevated by PCB in fed fish. The immediate cortisol and glucose responses to handling disturbance were suppressed by PCB in a dose-dependent way in food-deprived fish. Although these responses were also lowered by PCB in the fed fish, the effect was much less pronounced than in food-deprived fish. There were only minor effects on plasma lactate responses. Our findings suggest that the stress responses of the Arctic charr are compromised by PCB and that the long-term fasting, typical of high-latitude fish, makes these species particularly sensitive to organochlorines such as PCB.

Comparative Biochemistry and Physiology, Part C: T

Comparative health assessment of western Pacific leatherback turtles (Dermochelys coriacea) foraging off the coast of California, 2005-2007

Leatherback turtles ( Dermochelys coriacea ) are critically endangered, primarily threatened by the overharvesting of eggs, fisheries entanglement, and coastal development. The Pacific leatherback population has experienced a catastrophic decline over the past two decades. Leatherbacks foraging off the coast of California are part of a distinct Western Pacific breeding stock that nests on beaches in Indonesia, Papua New Guinea, and the Solomon Islands. Although it has been proposed that the rapid decline of Pacific leatherback turtles is due to increased adult mortality, little is known about the health of this population. Health assessments in leatherbacks have examined females on nesting beaches, which provides valuable biological information, but might have limited applicability to the population as a whole. During September 2005 and 2007, we conducted physical examinations on 19 foraging Pacific leatherback turtles and measured normal physiologic parameters, baseline hematologic and plasma biochemistry values, and exposure to heavy metals (cadmium, lead, and mercury), organochlorine contaminants, and domoic acid. We compared hematologic values of foraging Pacific leatherbacks with their nesting counterparts in Papua New Guinea ( n =11) and with other nesting populations in the Eastern Pacific in Costa Rica ( n =8) and in the Atlantic in St. Croix ( n =12). This study provides the most comprehensive assessment to date of the health status of leatherbacks in the Pacific. We found significant differences in blood values between foraging and nesting leatherbacks, which suggests that health assessment studies conducted only on nesting females might not accurately represent the whole population. The establishment of baseline physiologic data and blood values for healthy foraging leatherback turtles, including males, provides valuable data for long-term health monitoring and comparative studies of this endangered population.

California

Mercury and drought along the Lower Carson River, Nevada: III. effects on blood and organ biochemistry and histopathology of snowy egrets and black-crowned night-herons on Lahontan Reservoir, 2002-2006

A 10-year study (1997-2006) was conducted to evaluate reproduction and health of aquatic birds in the Carson River Basin of northwestern Nevada (on the U.S. Environmental Protection Agency Natural Priorities List) due to high mercury (Hg) concentrations from past mining activities. This part of the study evaluated physiological associations with blood Hg in young snowy egrets (Egretta thula) and black-crowned night-herons (Nycticorax nycticorax), and organ biochemistry and histopathological effects in snowy egrets on Lahontan Reservoir (LR) from the period 2002-2006. LR snowy egret geometric mean total Hg concentrations (&mu;g/g ww) ranged from 1.5 to 4.8 for blood, 2.4 to 3.1 liver, 1.8 to 2.5 kidneys, 1.7 to 2.4 brain, and 20.5 to 36.4 feathers over these years. For night-herons, mean Hg for blood ranged from 1.6 to 7.4. Significant positive correlations were found between total Hg in blood and five plasma enzyme activities of snowy egrets suggesting hepatic stress. Histopathological findings revealed vacuolar changes in hepatocytes in LR snowy egrets as well as correlation of increased liver inflammation with increasing blood and tissue Hg. Hepatic oxidative effects were manifested by decreased hepatic total thiol concentration and glutathione reductase activity and elevated hepatic thiobarbituric acid-reactive substances (TBARS), a measure of lipid peroxidation. However, other hepatic changes indicated compensatory mechanisms in response to oxidative stress, including decreased oxidized glutathione (GSSG) concentration and decreased ratio of GSSG to reduced glutathione. In young black-crowned night-herons, fewer correlations were apparent. In both species, positive correlations between blood total Hg and plasma uric acid and inorganic phosphorus were suggestive of renal stress, which was supported by histopathological findings. Both oxidative effects and adaptive responses to oxidative stress were apparent in kidneys and brain. Vacuolar change and inflammation in peripheral nerves were found to correlate with blood and tissue Hg. Hg-associated effects related to the immune system included alterations in specific white blood cells and lymphoid depletion in the bursa that were correlated with blood and tissue Hg. When the number of plasma variables that differed between young snowy egrets from the LR site and the reference site were compared between wet and drought years, over twice as many variables were affected during drought years. This resulted in many more variables correlating with blood total Hg during dry than during wet years, suggesting the combination of drought and Hg was more stressful than Hg alone. Drought may have exacerbated Hg-related effects as reported previously for overall productivity. This relationship was not evident in black-crowned night-herons, although data were more limited.

Nevada