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Bioavailability and toxicity of dietborne copper and zinc to fish

To date, most researchers have used dietborne metal concentrations rather than daily doses to define metal exposure and this has resulted in contradictory data within and between fish species. It has also resulted in the impression that high concentrations of dietborne Cu and Zn (e.g.>900 mg kg −1 dry diet) are relatively non-toxic to fish. We re-analyzed existing data using rations and dietborne metal concentrations and used daily dose, species and life stage to define the toxicity of dietborne Cu and Zn to fish. Partly because of insufficient information we were unable to find consistent relationships between metal toxicity in laboratory-prepared diets and any other factor including, supplemented metal compound (e.g. CuSO 4 or CuCl 2 ), duration of metal exposure, diet type (i.e. practical, purified or live diets), or water quality (flow rates, temperature, hardness, pH, alkalinity). For laboratory-prepared diets, dietborne Cu toxicity occurred at daily doses of >1 mg kg −1 body weight d −1 for channel catfish ( Ictalurus punctatus ), 1–15 mg kg −1 body weight d −1 (depending on life stage) for Atlantic salmon ( Salmo salar ) and 35–45 mg kg −1 body weight d −1 for rainbow trout ( Oncorhynchus mykiss ). We found that dietborne Zn toxicity has not yet been demonstrated in rainbow trout or turbot ( Scophthalmus maximus ) probably because these species have been exposed to relatively low doses of metal (<90 mg kg −1 body weight d −1 ) and effects on growth and reproduction have not been analyzed. However, daily doses of 9–12 mg Zn kg −1 body weight d −1 in laboratory-prepared diets were toxic to three other species, carp Cyprinus carpio , Nile tilapia Oreochromis niloticus , and guppy Poecilia reticulata . Limited research indicates that biological incorporation of Cu or Zn into a natural diet can either increase or decrease metal bioavailability, and the relationship between bioavailability and toxicity remains unclear. We have resolved the contradictory data surrounding the effect of organic chelation on metal bioavailability. Increased bioavailability of dietborne Cu and Zn is detectable when the metal is both organically chelated and provided in very low daily doses. We have summarized the information available on the effect of phosphates, phytate and calcium on dietborne Zn bioavailability. We also explored a rationale to understand the relative importance of exposure to waterborne or dietborne Cu and Zn with a view to finding an approach useful to regulatory agencies. Contrary to popular belief, the relative efficiency of Cu uptake from water and diet is very similar when daily doses are compared rather than Cu concentrations in each media. The ratio of dietborne dose:waterborne dose is a good discriminator of the relative importance of exposure to dietborne or waterborne Zn. We discuss gaps in existing data, suggest improvements for experimental design, and indicate directions for future research.

Comparative Biochemistry and Physiology, Part C: T

The oxidation of drugs by fishes

1. Fish liver microsomal systems have been found to catalyze the hydroxylation of aniline and acetanilide, the N-demethylation of aminopyrine and the O-dealkylation of phenacetin. 2. These systems are similar to the corresponding mammalian enzymes and they may be considered to be mixed function oxidase since they require NADPH and oxygen. An absolute requirement for oxygen, however, was difficult to demonstrate for the hepatic phenacetin cleavage system from fish. 3. Microsomal drug metabolizing systems from fish have temperature optima which are considerably lower than those of corresponding mammalian systems

Comparative Biochemistry and Physiology, Part A: M

Stress-induced ascorbic acid depletion and cortisol production in two salmonid fishes

Interrenal ascorbic acid and serum cortisol were measured in non-specificity stressed yearling coho salmon and rainbow trout. Interrenal ascorbate was markedly decreased during stress but increased to normal if adaptation occurred. Serum cortisol was elevated by non-specific stress and remained high after interrenal ascorbate had returned to initial levels.

Comparative Biochemistry and Physiology, Part A: M

Influence of water temperature on gill sodium, potassium-stimulated ATPase activity in juvenile coho salmon (Oncorhynchus kisutch)

1. Gill sodium, potassium-stimulated ATPase activity was determined from December to July in gills of yearling coho salmon ( Oncorhynchus kisutch ) maintained at four temperatures, 6, 10, 15 and 20°C. 2. Compared to fish held at 6°C, elevation in ATPase activity and the associated parr-smolt transformation were accelerated in fish at 10 and 15°C whereas animals at 20°C experienced at best only a transitory elevation in activity. 3. Fish transferred from one temperature to another developed ATPase activities characteristic of fish residing at temperatures to which they were transferred. 4. Cold water (6°C) tended to preserve the elevated ATPase activity while higher temperatures (10 and 15°C) caused decreases after an initial accelerated increase.

Comparative Biochemistry and Physiology, Part A: P

Indocyanine green: A test of hepatic function and a measure of plasma volume in the duck

1. The exponential removal of ICG from the plasma by the mallard duck liver made possible the measurement of fractional dye clearance (K), plasma volume (PV) and plasma clearance (PC). 2. Values obtained for K (14.9%/min), PV (39.2 ml/kg) and PC (5.8 ml/min per kg) agreed with those obtained by other techniques used in a number of species. 3. Sex did not affect the removal of ICG by the liver. However, increases in K, PV and PC were noted in hen mallards in laying condition. 4. The data should prove useful as baseline values for physiological and pathological studies on the avian liver

Comparative Biochemistry and Physiology, Part A: P

The influence of hunger on meal to pellet intervals in barred owls

1. Barred owls fed at a sub-maintenance (SM) level had significantly (P < 0.01) longer meal to pellet intervals (MPI)/g eaten/kg body weight (BW) than those fed at an above maintenance (AM) level; MPI/g per kg for owls fed at a maintenance (M) level was intermediate but significantly (P < 0.01) different from both SM and AM. 2. During SM feeding, MPI/g per kg gradually increased. 3. The proportion of a meal occurring in a pellet was less in “hungry” owls whether losing weight (SM) or gaining (AM) as compared to owls maintaining their normal body weight (M). 4. SM fed owls appear to be able to increase digestion time as well as thoroughness of digestion.

Comparative Biochemistry and Physiology, Part A: P

The bile acid composition of crane gallbladder bile

1. The biliary bile acids of the whooping crane ( Grus americana ) and the Florida sandhill crane ( G. canadensis pratensis ) have been examined. 2. Cholic acid (CA), chenodeoxycholic acid (CDOCA) and lithocholic acid were found in bile from both species of these North American cranes. 3. CDOCA and CA were the primary bile acids in both species, together constituting 70% or more of the bile acids by weight. 4. The primary bile acids of cranes appear to be the same as those that have been identified in other avian species.

Comparative Biochemistry and Physiology, Part A: P

Composition and energy contents of mature inshore spawning capelin ( Mallotus villosus ): Implications for seabird predators

1. Lipid levels of capelin are highest in late fall and lowest during the summer spawning season; protein levels are constant at 13–14% body wt throughout the year. 2. Ovid females contained significantly more lipid and protein and less water and had higher energy densities than males and spent females. 3. Surgically-removed egg masses made up 34.2 ± 10.3% female body wt and were very similar in composition and energy density to gravid females, differing from spent females and males in similar respects. Owing to the ovarian development of females, sexes differ in energy density only during the spawning season. 4. Sexes were similar in amino acid composition. Analysis of capelin and three other seabird forage species revealed that isoleucine levels were lower than minimum avian maintenance and growth requirements. 5. Implications for the foraging behaviour and food preferences of diving seabird predators (murres, puffins) are discussed

Comparative Biochemistry and Physiology, Part A: M

Effects of dietary excesses of branched-chain amino acids on the metabolism and tissue composition of lake trout ( Salvelinus namaycush )

1. Excesses of either leucine, isoleucine or valine were fed in separate experiments to determine if the branched-chain amino acid antagonism reported in other animals occur in trout ( Salvelinus namaycush ). 2. Parameters measured include growth rate, feed utilization efficiency, plasma and muscle-free amino acids, carcass composition and branched-chain amino acid aminotransferase. 3. Dietary excesses of leucine or isoleucine caused an increase in the valine requirement. 4. The inability of leucine and isoleucine supplementations to ameliorate the effects of excess dietary valine are interpreted as a valine toxicity rather than an antagonism.

Comparative Biochemistry and Physiology, Part A: P

Response of rainbow trout to source and level of supplemental dietary methionine

1. Methionine and total sulfur amino acid (TSAA) requirements of rainbow trout ( Salmo gairdneri ) were investigated by feeding graded isosulfurous levels of l - and dl -methionine, l -cystine, and the free acid and calcium forms of methionine hydroxy analog (MHA). 2. Added cystine did not promote growth, survival or prevent cataracts. 3. l -methionine produced fastest growth, followed by dl -methionine, CaMHA and free acid MHA. 4. Trout fed CaMHA gained 85.7 and 92.3% as much as those fed l -methionine and dl -methionine. 5. Within each experiment, the level of L-methionine isomer that prevented cataracts was constant (1.86 g/100g protein in experiment (1), 1.45 in experiment (2) and was lower than for maximum growth (2.89 and 2.15 g) regardless of methionine source.

Comparative Biochemistry and Physiology, Part A: P

Whole body and tissue blood volumes of two strains of rainbow trout ( Oncorhynchus mykiss )

1. Estimates of apparent packed cell, plasma and total blood volumes for the whole body and for 13 selected tissues were compared between Kamloops and Wytheville strains of rainbow trout ( Oncorhynchus mykiss ) by the simultaneous injection of two vascular tracers, radiolabeled trout erythrocytes ( 51 Cr-RBC) and radioiodated bovine serum albumin ( 125 I-BSA). 2. Whole body total blood volume, plasma volume and packed cell volume were slightly, but not significantly greater in the Wytheville trout, whereas, the apparent plasma volumes and total blood volumes in 4 of 13 tissues were significantly greater in the Kamloops strain. 3. Differences were most pronounced in highly perfused organs, such as the liver and kidney and in organs of digestion such as the stomach and intestines. 4. Differences in blood volumes between the two strains may be related to the greater permeability of the vascular membranes in the Kamloops strain fish.

Comparative Biochemistry and Physiology, Part A: M

Male sex steroids and hormonal control of male courtship behavior in the yellow-bellied slider turtle, Trachemys scripta

Survey of androgens and estrogens in serum, liver and testes of male yellow-bellied slider turtles, Trachemys (= Pseudemys ) scripta , a species exhibiting dissociated gametogenesis age-dependent melanism, revealed the presence of numerous androgen precursors, androgens, androgen metabolites, and estrogens in quantities varying with season, tissue, and male coloration. The most commonly found and abundant androgens in all males were dehydroepiandrosterone (DHEA) and androstenedione, which were present at significantly higher levels in testes than in serum or liver. Epitestosterone was found in the serum and liver of only melanistic males; testosterone was only rarely recovered. Behavioral testing of castrated male turtles implanted with various androgens, as well as intact and sham-operated controls, revealed that melanistic males courted more frequently than nonmelanistic males, and that exogenous testosterone was most effective in inducing courtship behavior in castrated males.

Comparative Biochemistry and Physiology, Part A: M

Effects of salinity on chloride cells and Na+ K+-ATPase activity in the teleost Gillchthys mirabilis

1. Longjawed mudsuckers, Gillichthys mirabilis , in 30ppt seawater (SW) were transferred to 1.5, 30 and 60ppt SW. 2. In the first 1–3 days after transfer, plasma chloride level and plasma osmolarity rose in the 60ppt SW fish, and decreased in the 1.5ppt SW fish. 3. By day 21, however, plasma chloride and osmolarity were at or near the levels seen in the controls (30ppt). 4. Branchial and jawskin Na + , K + -ATPase activities were high in all salinities, and did not differ significantly among treatments. 5. The vital fluorescent stains DASPEI and anthroylouabain were used to detect mitochondria and Na + , K + -ATPase, respectively, in chloride cells. 6. Both stains indicated that jawskin chloride cell density did not differ among treatment groups. 7. In contrast, chloride cell size increased significantly with increasing salinity. 8. The chloride cells of fish in 60 ppt SW were noticeably angular in outline, whereas those of both the 1.5 and 30ppt SW fish were circular. 9. The results are discussed in relation to the ion transport requirements encountered in the intertidal habitat of the mudsucker.

Comparative Biochemistry and Physiology, Part A: P

Protein retention and liver aminotransferase activities in Atlantic salmon fed diets containing different energy sources

Atlantic salmon ( Salmo salar ) fingerlings (14.4 g) were fed diets containing either glucose, dextrin, raw corn starch and lipid, or a high protein U.S. Fish and Wildlife Service open-formula diet (ASD2-30) for 12 weeks. Significant differences in weight gain and feed: gain ratio were not observed among salmon fed the diets containing glucose, dextrin or ASD2-30. Diets containing dextrin and glucose supported greater protein retention and reduction in alanine aminotransferase activity than the other diets. Activity of aspartate aminotransferase was not affected by the dietary treatment. Protein retention correlated highly with alanine aminotransferase activity.

Comparative Biochemistry and Physiology, Part A: P

Kangaroo rat bone compared to white rat bone after short-term disuse and exercise

Kangaroo rats ( Dipodomys ordii ) were used to study the effects of confinement on mechanical properties of bone with a long range objective of proposing an alternative to the white rat model for the study of disuse osteoporosis. Kangaroo rats exhibit bipedal locomotion, which subjects their limbs to substantial accelerative forces in addition to the normal stress of weight bearing. We subjected groups of kangaroo rats and white rats ( Rattus norvegicus ) to one of two confinement treatments or to an exercise regime; animals were exercised at a rate calculated to replicate their (respective) daily exercise patterns. White laboratory rats were used as the comparison because they are currently the accepted model used in the study of disuse osteoporosis. After 6 weeks of treatment, rats were killed and the long bones of their hind limbs were tested mechanically and examined for histomorphometric changes. We found that kangaroo rats held in confinement had less ash content in their hind limbs than exercised kangaroo rats. In general, treated kangaroo rats showed morphometric and mechanical bone deterioration compared to controls and exercised kangaroo rats appeared to have slightly “stronger” bones than confined animals. White rats exhibited no significant differences between treatments. These preliminary results suggest that kangaroo rats may be an effective model in the study of disuse osteoporosis.

Comparative Biochemistry and Physiology, Part A: P

A review of lead poisoning in swans

Nearly 10,000 swans of six species or subspecies from 14 countries have died from poisoning caused by lead that originated from ingestion of fishing weights, shotgun pellets (shot), or contaminated vegetation or sediments associated with mining and smelting wastes. Lead contamination in mute swans in England caused local population declines during the late 1970s and 1980s. More tundra swans died from lead poisoning than any other species. The extreme record involved an estimated 7200 tundra swans that died over five winters at one locality in North Carolina. The recent legislation to ban lead fishing weights in most of England and Wales and recent replacement of lead shot with steel shot for waterfowl hunting in the United States and a few areas of Europe, including Denmark, are expected to reduce the incidence of lead poisoning in swans.

Comparative Biochemistry and Physiology, Part C: P

Regional cholinesterase activity in white-throated sparrow brain is differentially affected by acephate (Orthene®)

Effects of a 14-day dietary exposure to an organophosphorus pesticide, acephate (acetylphosphoramidothioic acid O,S-dimethyl ester), were determined on cholinesterase activity in three regions (basal ganglia, hippocampus, and hypothalamus) of the white-throated sparrow, Zonotrichia albicollis , brain. All three regions experienced depressed cholinesterase activity between 0.5–2 ppm acephate. The regions exhibited cholinesterase recovery at 2–16 ppm acephate; however, cholinesterase activity dropped and showed no recovery at higher dietary levels (>16 ppm acephate). Evidence indicates that the recovery is initiated by the magnitude of depression, not the duration. In general, as acephate concentration increased, differences in ChE activity among brain regions decreased. Three terms are introduced to describe ChE response to acephate exposure: 1) ChE resistance threshold, 2) ChE compensation threshold, and 3) ChE depression threshold. It is hypothesized that adverse effects to birds in the field may occur at pesticide exposure levels customarily considered negligible.

Comparative Biochemistry and Physiology, Part C: P

Conservation of Toll-like receptor signaling pathways in teleost fish

In mammals, toll-like receptors (TLR) recognize ligands, including pathogen-associated molecular patterns (PAMPs), and respond with ligand-specific induction of genes. In this study, we establish evolutionary conservation in teleost fish of key components of the TLR-signaling pathway that act as switches for differential gene induction, including MYD88, TIRAP, TRIF, TRAF6, IRF3, and IRF7. We further explore this conservation with a molecular phylogenetic analysis of MYD88. To the extent that current genomic analysis can establish, each vertebrate has one ortholog to each of these genes. For molecular tree construction and phylogeny inference, we demonstrate a methodology for including genes with only partial primary sequences without disrupting the topology provided by the high-confidence full-length sequences. Conservation of the TLR-signaling molecules suggests that the basic program of gene regulation by the TLR-signaling pathway is conserved across vertebrates. To test this hypothesis, leukocytes from a model fish, rainbow trout ( Oncorhynchus mykiss ), were stimulated with known mammalian TLR agonists including: diacylated and triacylated forms of lipoprotein, flagellin, two forms of LPS, synthetic double-stranded RNA, and two imidazoquinoline compounds (loxoribine and R848). Trout leukocytes responded in vitro to a number of these agonists with distinct patterns of cytokine expression that correspond to mammalian responses. Our results support the key prediction from our phylogenetic analyses that strong selective pressure of pathogenic microbes has preserved both TLR recognition and signaling functions during vertebrate evolution.

Comparative Biochemistry and Physiology, Part D: G