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Divergent effects of postmortem ambient temperature on organophosphorus- and carbamate-inhibited brain cholinesterase activity in birds

Time- and temperature-dependent postmortem changes in inhibited brain cholinesterase (ChE) activity may confound diagnosis of field poisoning of wildlife by anticholinesterase pesticide. Carbamate-inhibited ChE activity may return to normal within 1 to 2 days of exposure of intact carcass to moderate ambient temperature (18-32C). Organophosphorus-inhibited ChE activity becomes more depressed over the same time. Uninhibited ChE activity was resilient to above freezing temperature to 32C for 1 day and 25C for 3 days. Carbamate- and organophosphorus-inhibited ChE can be separated by incubation of homogenate for 1 hour at physiological temperatures; carbamylated ChE can be readily reactivated while phosphorylated ChE cannot.

Pesticide Biochemistry and Physiology

DDE increased the toxicity of parathion to coturnix quail

Adult male Japanese quail (Coturnix coturnix) were exposed to DDE or chlordane in the diet and subsequently dosed with parathion or paraoxon. Pretreatment with 5 or 50 ppm DDE in the diet for 12 weeks resulted in increased cholinesterase (ChE) activity in plasma, but not in the brain. Dietary concentrations of 5 and 50 ppm DDE caused increased susceptibility of quail that were challenged with parathion or paraoxon. The increased mortality resulting from DDE pretreatment was reflected in brain ChE inhibition. The synergistic action of DDE was apparent after 3 days of exposure to 50 ppm DDE and 1 week of exposure to 5 ppm DDE. Birds exposed for 3 weeks to 5 or 50 ppm DDE retained their DDE-potentiated sensitivity to parathion after 2 weeks on clean diet. Chlordane pretreatment resulted in decreased susceptibility (antagonism) to parathion, but not to paraoxon dosage. Implications of differing responses in ChE and mortality among controls, DDE-, and chlordane-pretreated birds after parathion or paraoxon dosage are discussed.

Pesticide Biochemistry and Physiology

Bone development in black ducks as affected by dietary toxaphene

Black ducks, Anas rubripes, were exposed to dietary toxaphene concentrations of 0, 10, or 50 μg/g of food for 90 days prior to laying and through the reproductive season. Toxaphene did not affect reproduction or survival, but reduced growth and impaired backbone development in ducklings. Collagen, the organic matrix of bone, was decreased significantly in cervical vertebrae of ducklings fed 50 μg/g, and calcium conentrations increased in vertebrae of ducklings fed 10 or 50 μg/g. The effects of toxaphene were observed only in female ducklings. In contrast to effects on vertebrae, toxaphene exposure did not alter tibia development. Toxaphene residues in carcasses of these ducklings averaged slightly less than the dietary levels.

Pesticide Biochemistry and Physiology

Recovery of cholinesterase activity in five avian species exposed to dicrotophos, an organophosphorus pesticide

The responses of brain and plasma cholinesterase (ChE) activities were examined in mallard ducks, bobwhite quail, barn owls, starlings, and common grackles given oral doses of dicrotophos, an organophosphorus insecticide. Up to an eightfold difference in response of brain ChE activity to dicrotophos was found among these species. Brain ChE activity recovered to within 2 SD of normal within 26 days after being depressed 55 to 64%. Recovery of brain ChE activity was similar among species and followed the model Y = a + b (log 10 X ).

Pesticide Biochemistry and Physiology

Hormonal responses and tolerance to cold of female quail following parathion ingestion

Thirty-week-old female bobwhite quail (Colinus virginianus), maintained at 26 + 1?C, were provided diets containing 0,25, or 100 ppm parathion ad libitum. After 10 days, birds were exposed to mild cold (6 + 1?C) for 4,8, 12, 24, or 48 hr. Brain acetylcholinesterase activity was inhibited in a dose-dependent manner in birds receiving 25 and 100 ppm parathion. Body weight, egg production, and plasma luteinizing hormone and progesterone concentrations were reduced in birds receiving 100 ppm parathion compared with other groups. Cold exposure did not alter plasma corticosterone levels in the 0- and 25-ppm parathion groups, but a two- to five fold elevation of plasma corticosterone was observed in birds fed 100 ppm parathion. These findings indicate that (i) short-term ingestion of parathion can impair reproduction possibly by altering gonadotropin or steroid secretion, and (ii) tolerance to cold may be reduced following ingestion of this organophosphate.

Pesticide Biochemistry and Physiology

Osmoregulatory function in ducks following ingestion of the organophosphorus insecticide fenthion

Salt gland function and osmoregulation in aquatic birds drinking hyperosmotic water has been suggested to be impaired by organophosphorus insecticides. To test this hypothesis, adult black ducks (Anas rubripes) were provided various regimens of fresh or salt (1.5% NaCl) water before, during, and after ingestion of mash containing 21 ppm fenthion. Ducks were bled by jugular venipuncture after I, 7. and 12 days of treatment, and were then killed. Brain and salt gland acetylcholinesterase activities were substantially inhibited (44-61% and 14-36%) by fenthion. However, salt gland weight and Na + -K + -ATPase activity, and plasma Na + , CI- , and osmolality, were uniformly elevated in all groups receiving salt water including those ingesting fenthion. In a second study, salt gland Na + -K + -ATPase activity in mallards (A. platyrhynchos) was not affected after in vitro incubation with either fenthion or fenthion oxon at concentrations ranging from 0.04 to 400 ?M, but was reduced in the presence of 40 and 400 ?M DDE (positive control). These findings suggest that environmentally realistic concentrations of organophosphorus insecticides do not markedly affect osmoregulatory function in adult black ducks.

Pesticide Biochemistry and Physiology

Enhancement of parathion toxicity to quail by heat and cold exposure

Effects of ambient temperature on the acute oral toxicity of parathion were investigated in Japanese quail (Coturnix japonica) maintained at thermoneutral temperature (26.degree. C) or exposed to elevated (37.degree. C) or reduced (4.degree. C) temperatures commonly encountered by free-ranging wild birds. Based upon estimates of the median lethal dosage, there was up to a two-fold enhancement of parathion toxicity in birds chronically exposed to heat or cold. Twenty-four hours after administration of a low dosage (4 mg/kg body wt, po), there was markedly greater cholinesterase inhibition in surviving heat-exposed quail compared with those reared at 26.degree. C (e.g., brain acetylcholinesterase depression of 42% versus 12%). There were no differences in hepatic activities of parathion oxidase, paraoxonase, or paraoxon deethylase which could account for greater toxicity to chronically heat-exposed birds. In contrast, 4 mg parathion/kg wt elicited less plasma cholinesterase inhibition in cold-exposed quail compared to thermoneutral controls (e.g., < 10% versus 48% depression after 24 hr). Increased liver weight and a doubling of paraoxonase activity may have been associated with greater tolerance to sublethal doses of parathion in chronically cold-exposed quail. These findings, together with limited field observations, indicate that the hazard associated with anticholinesterase exposure of wild birds is substantially influenced by environmental temperature.

Pesticide Biochemistry and Physiology

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

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

Dopaminergic and anti-estrogenic responses in juvenile steelhead (Oncorhynchus mykiss) exposed to bifenthrin

The frequency of detection and concentrations of bifenthrin, a pyrethroid insecticide, in the waterways inhabited by the endangered species, steelhead trout ( Oncorhynchus mykiss ), has become a significant concern for regulatory agencies. Endocrine disruption has been observed with estrogenic and anti-estrogenic responses in fish species at different life stages. Since several studies have indicated alterations in dopaminergic signaling associated with endocrine responses, juvenile steelhead were exposed to environmentally relevant concentrations of 60 or 120 ng/L bifenthrin for two weeks. Fish brains were assessed for dopamine levels and the expression of genes involved in dopaminergic and estrogenic processes, such as catechol- o -methyltransferase ( comt ) and monoamine oxidase ( mao ). Vitellogenin ( vtg ) and estrogenic receptors ( ERα1 , ERβ1 , and ERβ2 ) were also evaluated in livers of the animals. Dopamine concentrations were significantly higher in fish brains following bifenthrin exposure. Consistent with a reduction in dopamine clearance, there was a significant decrease in the mRNA expression of comt with increased bifenthrin concentration. Hepatic expression of ERα1 and ERβ2 mRNA was significantly decreased with increased bifenthrin concentration. These data support the possible mechanism of bifenthrin altering the dopaminergic pathway at low ng/L concentrations, in juvenile steelhead, which could interfere with endocrine feedback loops. These findings support the need for and importance of identifying species and life stage differences in pesticide modes of action to reduce uncertainties in risk assessments.

Comparative Biochemistry and Physiology Part C: To

Percutaneous absorption of several chemicals, some pesticides included, in the red-winged blackbird

Percutaneous absorption in vivo through the skin of the feet of the red-winged blackbird (Agelaius phoeniceus) has been investigated. Absorption after 18-24 hours exposure to 0.01 M solutions of salicylic acid, caffeine, urea, 2,4-D, dieldrin, diethylstilbesterol, and DDT was measured. Of these, only DDT and diethylstilbesterol were not absorbed to a measurable degree. The solvents ethanol, dimethylsulfoxide (DMSO), and vegetable oil were compared with water in their effects on the absorption ofcaffeine, urea, and salicylic acid. Ethanol, DMSO,and oil each decreased percutaneous absorption of salicylic acid. DMSO increased absorption of caffeine, and ethanol had no effect on it. Neither DMSO nor ethanol affected penetration of urea. Partition coefficients (K) (epidermis/water) were determined for all seven penetrants. Compounds with higher values of K showed lower percutaneous absorption. These findings suggest that K may be useful to predict percutaneous absorption in vivo. It appears unlikely that percutaneous absorption contributes greatly to the body burden of 2,4-D and dieldrin in A. phoeniceus.

Environmental Physiology and Biochemistry