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Na+, K+-activated-ATPase inhibition in rainbow trout: A site for organochlorine pesticide toxicity?

1. The Na + , K + -activated, Mg 2+ -dependent-ATPase enzyme system in a heavy microsomal fraction of rainbow trout ( Salmo gairdneri ) brain was inhibited in vitro by chlorinated hydrocarbon pesticides. 2. T 50 (concentration at 50 per cent inhibition) values for dicofol, endosulfan and DDT were 5 × 10 −6 , 3 × 10 −5 and 1 × 10 −4 M respectively. Similar inhibition by these pesticides occurred in kidney and gill ATPase preparations. 3. An unexpected finding was a failure of the classic inhibitor, ouabain, to block the Na + , K + -activated component of ATPase activity in the gill. 4. It is suggested that inhibition of ATPase activity may be a causal factor in the toxic effects of organochlorine pesticides in fishes.

Comparative Biochemistry and Physiology, Part B: C

Biochemical identification of the mallard, Anas platyrhynchos , and black duck, A. rubripes

1. Eleven tissue systems from mallards and black ducks were examined for soluble proteins, lactate dehydrogenases and non-specific esterases through discontinuous polyacrylamide techniques. 2. Biochemical relationships between the black duck and mallard are extremely similar. 3. Hemoglobins and lactate dehydrogenase appear to be common in electrophoretic mobility between the two species. 4. Approximately 89% of the soluble proteins and 58% of the non-specific esterases are common among the two species, indicating both biochemical similarity at the genus level and species-specificity.

Comparative Biochemistry and Physiology, Part B: C

Comparison of techniques for stabilizing hemoglobins of rainbow trout ( Salmo gairdneri ) during frozen storage

1. The stability of hemoglobin of rainbow trout under frozen conditions in oxyform, carboxyform, and cyanometform was examined. 2. Carboxyhemoglobin retained its original electrophoretic banding pattern after 14 days of frozen storage, whereas oxyform and cyanometform hemoglobins did not. 3. Banding patterns changed in some samples in all treatment groups after 21 days of storage.

Comparative Biochemistry and Physiology, Part B: C

Reassociation and hybridization properties of DNAs from several species of fish

Reassociation and hybridization properties from spectrophotometric studies of DNAs from 10 species of fish indicate: 1. Great diversity in the amounts of repeated sequences in the genomes of different species - more specialized fish had less redundancy. 2. Large differences in the complexities of the DNAs - more specialized fish had less information. 3. Little homology between sequences of remotely related species but substantial homology between sequences of closely related species.

Comparative Biochemistry and Physiology, Part B: C

Thymocyte plasma membrane of the rainbow trout, Salmo gairdneri : Associated immunoglobulin and heteroantigens

1. Thymic lymphocytes of the rainbow trout, S. gairdneri were disrupted and a plasma membrane containing fraction isolated by differential and buoyant density centrifugation. 2. Radioiodine introduced into the membrane by the lactoperoxidase catalyzed reaction and immunoglobulin (identified by radioimmunoassay with monoclonal antibody) both copurified in the plasma membrane fraction. 3. Rabbit antibody raised to the plasma membrane fraction showed a strong reaction with trout lymphocytes in immunofluorescence, was mitogenic for trout lymphocytes, and recognized lymphocyte membrane heteroantigens of molecular weight > 70,000 in the thymus and 45,000–95,000 in the head kidney.

Comparative Biochemistry and Physiology, Part B: C

Seasonal variation in diagnostic enzymes and biochemical constituents of captive northern bobwhites and passerines

1. A variety of biochemical measurements were taken periodically in captive northern bobwhite ( Colinus virginianus L.), European starlings ( Sturnus vulgaris L.), red-winged blackbirds ( Agelaius phoeniceus L.) and common grackles ( Quiscalus quiscula L.) to determine whether baseline values remain sufficiently stable throughout the year for general clinical use in the absence of concurrent control specimens. 2. Variables included whole blood hemotacrit and hemoglobin, plasma lactate dehydrogenase, α-hydroxybutyrate dehydrogenase, aspartate aminotransferase, alinine aminotransferase, creatine kinase, butyrylcholinesterase, alkaline phosphatase, glucose, albumin, total protein, creatinine, urea nitrogen, uric acid, cholesterol, and triglycerides, and brain acetylcholinesterase. Butyrl- and acetylcholinesterase were included because of their specific uses in toxicology. 3. Significant seasonal differences were detected for each of the variables except brain acetylcholinesterase in at least one of the species. Significant species differences were detected during at least one season for all of the variables measured. 4. All species were maintained outdoors, but only northern bobwhites came into reproductive condition and showed sex-differences in the clinical variables during their normal breeding season. 5. It was concluded that reference values for the 18 clinical variables measured could be calculated from our data for adult specimens of the species studied, and that results for one species cannot be extrapolated with certainty to any other species. 6. Estimated normal bounds for each of the 18 variables measured by commonly used clinical procedures are presented for reproductively quiescent northern bobwhites, European starlings, red-winged blackbirds, and common grackles.

Comparative Biochemistry and Physiology, Part B: C

Lipids and proteins in the Rathke's gland secretions of the North American mud turtle ( Kinosternon subrubrum )

Lipids and proteins in the Rathke's gland secretions of the North American mud turtle ( Kinosternon subrubrum , Kinosternidae) were analyzed by gas chromatography-mass spectrometry (GC-MS) and SDS-polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Analysis by GC-MS indicates 2,3-dihydroxypropanal and C 3 –C 24 free or esterified fatty acids. Analysis by SDS-PAGE indicates a major protein component with an approximate molecular mass of 60 kDa and minor components ranging from ca. 23 to 34 kDa. The major component of K. subrubrum glandular secretions exhibits a mobility that matches that of the Kemp's ridley sea turtle ( Lepidochelys kempi , Cheloniidae), suggesting that these proteins are evolutionarily conserved.

Comparative Biochemistry and Physiology, Part B: C

Biochemical and hematological effects of lead ingestion in nestling American kestrels ( Falco sparverius )

1. One-day old American kestrel ( Faico sparverius ) nestlings were orally dosed daily with 5 μl/g of corn oil (controls), 25, 125 or 625 mg/kg of metallic lead in corn oil for 10 days. 2. Forty per cent of the nestlings receiving 625 mg/kg of lead died after 6 days and growth rates were significantly depressed in the two highest lead dosed groups. At 10 days hematocrit values were significantly lower in the two highest lead treated groups, and hemoglobin content and red blood cell (δ-aminolevulinic acid dehydratase (ALAD) activity was depressed in all lead treated groups. Plasma creatine phosphokinase decreased in the two highest treatment groups. 3. Brain, liver and kidney ALAD activities, brain RNA to protein ratio and liver protein concentration decreased after lead exposure whereas liver DNA, DNA to RNA ratio and DNA to protein ratio increased. Brain monoamine oxidase and ATPase were not significantly altered. 4. Measurements of the ontogeny of hematological variants and enzymes in normal development, using additional untreated nestlings, revealed decreases in red blood cell ALAD, plasma aspartate amino transferase, lactate dehydrogenase, brain DNA and RNA and liver DNA, whereas hematocrit, hemoglobin, plasma alkaline phosphatase, brain monoamine oxidase, brain ALAD and liver ALAD increased during the first 10 days of posthatching development. 5. Biochemical and hematological alterations were more severe than those reported in adult kestrels or precocial young birds exposed to lead. Alterations may be due in part to delayed development.

Comparative Biochemistry and Physiology, Part C: C

Effect on tricaine methanesulfonate (MS-222) on hematocrit values in rainbow trout ( Salmo gairdneri )

1. Anesthesia of rainbow trout ( Salmo gairdneri ) with 70 ppm tricaine methanesulfonate (MS-222) for 3-9 min resulted in a linear increase in hematocrit. 2. Handling of unanesthetized trout caused a higher and more variable hematocrit reading than did exposure to MS-222 for up to 3 min. 3. The range and standard error of hematocrit readings was smallest in trout treated with MS-222 for 1 min.

Comparative Biochemistry and Physiology, Part C: C

Six different plasma enzymes in bald eagles ( Haliaeetus leucocephalus ) and their usefulness in pathological diagnosis

1. Activities of creatine phosphokinase, glutamic oxalacetic transaminase, glutamic pyruvic transaminase, lactate dehydrogenase, fructose diphosphate aldolase and cholinesterase were measured in plasma of bald eagles. 2. There were no sex differences in the plasma enzyme activities. 3. An acute dieldrin dosage (10 mg/kg) of a female bald eagle resulted in 400% increases in activities of plasma creatine phosphokinase and glutamic oxalacetic transaminase and 250% increases in activities of lactate dehydrogenase and glutamic pyruvic transaminase. 4. At 11 days post-dosage all but one of the plasma enzyme activities had returned to normal; glutamic oxalacetic transaminase activity remained 100% above pre-dosage values. 5. Plasma enzyme assays constitute a non-destrcutive procedure that can be used in valuable wildlife species to screen for the presence and prevalence of environmental contaminants.

Comparative Biochemistry and Physiology, Part C: C

Allantoinase in lake trout ( Salvelinus namaycush ): In vitro effects of PCBs, DDT and metals

1. Allantoinase, an enzyme in the purine-urea cycle, was found in livers of Salvelinus namaycush (Osteichthyes: Salmoniformes). 2. The enzyme was active from pH 6.6 to 8.2 at 37°C and from pH 7.4 to 9.0 at 10°C and had an Arrhenius energy was activation of 11.0 kcal/mol and a temperature quotient of 2.0. The K m of the enzyme homogenate was 8.4 mM allantoin. 3. The concentrations of inorganic metals at which 50% inhibition occurred during in vitro exposure were 6.0 mg/l Cu 2+ , 6.7 mg/l Cd 2+ , 34 mg/l Hg 2+ and 52 mg/l Pb 2+ . The in vitro sensitivity to PCBs, DDT and DDE and kinetics in the presence of metals were determined. 4. Allantoinase activity was negatively correlated with body length for fish from Lake Michigan but not from Lake Superior or the laboratory.

Comparative Biochemistry and Physiology, Part C: C

Effects of petroleum hydrocarbons on hepatic function in the duck

1. The indocyanine green dye clearance test for hepatic function was determined in mallard ducks before and during the chronic ingestion (7 months) of representative paraffinic or aromatic petroleum hydrocarbons (PH). 2. No mortality or visible symptoms of toxicity occurred in any of the tests. Ingestion of 4000 ppm aromatic PH produced significant increases in liver (25%), plasma clearance of indocyanine green (33%) and hepatic blood flow (30%). 3. Although the aromatics elicited a greater hepatic stress response than the paraffins, the ducks tolerated high concentrations of PH for extended periods.

Comparative Biochemistry and Physiology, Part C: C

Plasma corticosterone and thyroxine concentrations during chronic ingestion of crude oil in mallard ducks ( Anas platyrhynchos )

1. Blood samples were collected from mallard ducks after 6, 12, and 18 weeks of dietary exposure to mash containing 0.015%, 0.150%, and 1.500% crude oil. 2. Plasma corticosterone concentrations in ducks fed mash containing 0.150% or 1.500% Alaskan Prudhoe Bay crude oil were uniformly depressed when compared to values in untreated control birds. 3. Plasma thyroxine concentration was not altered in ducks chronically exposed to crude oil. 4. The observed alteration in corticosterone concentration could reduce tolerance to temperature and dietary fluctuations in the environment.

Comparative Biochemistry and Physiology, Part C: C

Organophosphate inhibition of avian salt gland Na, K-ATPase activity

1. Adult black ducks ( Anas rubripes ) were given freshwater or saltwater (1.5% NaCl) for 11 days and half of each group was also given an organophosphate (17 p.p.m. fenthion) in the diet on days 6–11. 2. After 11 days, ducks drinking saltrwater had lost more weight and had higher plasma Na and uric acid concentration and osmolalities than birds drinking freshwater. 3. Saltwater treatment stimulated the salt gland to increased weight and Na, K-ATPase activity. 4. Fenthion generally reduced plasma and brain cholinesterase activity and depressed cholinesterase and Na, K-ATPase activities in salt glands of birds drinking saltwater.

Comparative Biochemistry and Physiology, Part C: C

Avian species differences in the intestinal absorption of xenobiotics (PCB, dieldrin, Hg 2+ )

1. Intestinal absorption of a polychlorinated biphenyl, dieldrin, and mercury (from HgCl 2 ) was measured in adult Northern bobwhites, Eastern screech owls, American kestrels, black-crowned night-herons and mallards in vivo by an in situ luminal perfusion technique. 2. Bobwhites, screech owls and kestrels absorbed much more of each xenobiotic than black-crowned night-herons and mallards. 3. Mallards absorbed less dieldrin and mercury than black-crowned night-herons. 4. Mercury absorption by kestrels was more than twice that in screech owls and eight times that observed in mallards. 5. Pronounced differences in xenobiotic absorption rates between bobwhites, screech owls and kestrels on the one hand, and black-crowned night-herons and mallards on the other, raise the possibility that absorptive ability may be associated with the phylogenetic classification of birds.

Comparative Biochemistry and Physiology, Part C: C

Depression of plasma luteinizing hormone concentration in quail by the anticholinesterase insecticide parathion

1. To examine the effects of parathion on basal plasma luteinizing hormone (LH) concentration, male Japanese quail ( Coturnix japonica ) were orally intubated with 0, 5 or 10 mg/kg parathion and sacrificed after 4, 8 and 24 hr. 2. At the 5 mg/kg dose, plasma LH levels were reduced at 4 and 8 hr, but returned to control values by 24 hr. Brain acetylcholinesterase activity was substantially reduced by 10 mg/kg parathion (52, 75 and 37% inhibition at 4, 8 and 24 hr, respectively) and plasma LH concentration remained depressed through the 24-hr period. 3. These findings suggest that the organophosphorus insecticide parathion may alter plasma LH concentration in a manner which might impair reproductive activity, and provide indirect evidence for a cholinergic component in the regulation of LH secretion in quail.

Comparative Biochemistry and Physiology, Part C: C

Comparative cardiovascular effects of four fishery anesthetics in spinally transected rainbow trout, oncorhynchus mykiss

1. We compared the effects of four anesthetics on heart rate, dorsal and ventral aortic blood pressure, and electrocardiograms of rainbow trout ( Oncorhynchus mykiss ). 2. Exposure to the local anesthetics tricaine methanesulfonate (MS-222) and benzocaine hydrochloride (BZH) produced minimal cardiovascular alterations. Mean dorsal aortic pressure (DAP) decreased during exposure to MS-222, and mean DAP and mean ventral aortic pressure (VAP) increased 15% during recovery from BZH. 3. Exposure to the general anesthetic 2-phenoxyethanol (2-PE) or the hypnotic agent etomidate (ET) dramatically decreased heart rate and blood pressures and altered EKG patterns. 4. During recovery, VAP and DAP increased above baseline for an extended period. Heart rate and EKG patterns rapidly returned to normal.

Comparative Biochemistry and Physiology, Part C: C

Relation of fatty acid composition in lead-exposed mallards to fat mobilization, lipid peroxidation and alkaline phosphatase activity

The increase of n-6 polyunsaturated fatty acids (PUFA) in animal tissues has been proposed as a mechanism of lead (Pb) poisoning through lipid peroxidation or altered eicosanoids metabolism. We have studied fatty acid (FA) composition in liver and brain of mallards ( Anas platyrhynchos ) feeding for 3 weeks on diets containing combinations of low or high levels of vitamin E (20 or 200 UI/kg) and Pb (0 or 2 g/kg). Saturated FA, n-6 PUFA and total concentrations of FA were higher in livers of Pb-exposed mallards, but not in their brains. The percentage of n-6 PUFA in liver and brain was slightly higher in Pb-exposed mallards. The increase of n-6 PUFA in liver was associated with decreased triglycerides and increased cholesterol in plasma, thus could be in part attributed to feed refusal and fat mobilization. The hepatic ratios between adrenic acid (22:4 n-6) and arachidonic acid (20:4 n-6) or between adrenic acid and linoleic acid (18:2 n-6) were higher in Pb exposed birds, supporting the existing hypothesis of increased fatty acid elongation by Pb. Among the possible consequences of increased n-6 PUFA concentration in tissues, we found increased lipid peroxidation in liver without important histopathological changes, and decreased plasma alkaline phosphatase activity that may reflect altered bone metabolism in birds.

Comparative Biochemistry and Physiology, Part C: T