Diazinon treatment effects on heart and skeletal muscle enzyme activities.

The effect of low levels of diazinon treatment on four marker enzymes in rat heart and skeletal muscle have been investigated. Adult male Wistar rats were treated twice a week with a dose of 0.5 ml X kg-1 X day-1 diazinon for 28 weeks. Diazinon treated rats gained significantly less weight than Sham-treated controls. Typical differences in Succinate dehydrogenase (SDH), Lactate dehydrogenase (LDH), Phosphofructo kinase (PFK) and Hexokinase (HK) activities were observed between heart and skeletal muscles. Diazinon feeding had no effect on heart, soleus, gastrocnemius and plantaris SDH, LDH and PFK enzyme activities after 28 weeks. HK activity was significantly increased in sham-control soleus and plantaris muscles after 28 weeks. Diazinon feeding inhibited HK activity in plantaris muscle after 28 weeks treatment. These results demonstrate that chronic low levels of diazinon have little effect on the glycolytic and oxidative activity in heart and skeletal muscle.

Effects of chronic intake of diazinon on blood and brain monoamines and amino acids.

Male rats were treated bi-weekly by gavage with the equivalent of 0.5 mg X kg-1 X day-1 technical diazinon for up to 28 weeks. The animals were sacrificed at specific time intervals (7, 14 and 28 weeks) and compared with age matched controls. Blood and brain tissues were analysed for cholinesterase activity and for concentrations of catecholamines and amino acids. Only Plasma cholinesterase was significantly reduced by the low level pesticide treatment. Erythrocyte acetyl cholinesterase and brain acetyl cholinesterase were unchanged while during the same period several putative brain neurotransmitters aspartate, glutamate (excitatory) and taurine as well as GABA (inhibitory) were significantly reduced in experimental vs control animals whereas no significant changes occurred between weeks in similarly fed animals. Blood serotonin was significantly elevated but no other blood or brain monoamine was significantly altered. Overt manifestations of brain toxicity observed were not apparent in experimental compared with control animals save for a significant decrease in growth observed in experimental animals. It was concluded that oral administration of low doses of diazinon exerts significant effects other than as an anticholinesterase on important brain neurotransmitters even at the low dose levels administered in this study.

Studies on the toxicity, metabolism, and anticholinesterase properties of acephate and methamidophos.

The toxicity of acephate to four species of aquatic insects, as well as the metabolism and cholinesterase-inhibiting properties of the chemical in the rat were studied. The results indicated that mayfly larvae were very sensitive to the toxic effects of acephate, whereas larvae of the stonefly, damselfly and mosquito were much less sensitive. In the rat, orally-administered acephate was rapidly absorbed from the intestines and severely inhibited the cholinesterases in the blood and brain. The enzymes began to recover after 24 hours, while the chemical was completely eliminated within three days. The amount of methamidophos observed in the liver was extremely low. The cholinesterase-inhibiting properties of acephate and methamidophos were compared in vitro to that of paraoxon, a known strong anticholinesterase. Enzymes from four vertebrates were used. In all cases, except one, acephate was found to be six orders of magnitude weaker than paraoxon, whereas methamidophos was three orders weaker. Trout brain cholinesterase was the exception; it was as sensitive to paraoxon as it was to methamidophos. Finally, four cholinesterases were inhibited with methamidophos, and their ability to reactivate spontaneously or to recover by induction with pyridine aldoxime methiodide (PAM) in vitro were determined. The results suggested that methamidophos-inhibited cholinesterases did not reactivate spontaneously; instead the enzymes remained inhibited either in a phosphorylated or an aged state. The significance of these results are discussed in relation to the use of acephate for forest insect pests.

Hydrolytic and metabolic products of acephate in water and mouse liver.

Acephate was incubated in distilled water of three different pH's at 37 degrees C for 7 days. Three hydrolytic products were formed: methamidophos, O,S-dimethyl phosphorothiolate (DMPT), and O-methylacetyl phosphoramidothiolate (OMPT). A single dose of acephate was also fed to mice, and their livers were excised and analyzed for metabolic products up to 30 hours. Three products were detected: methamidophos, DMPT, and S-methylacetyl phosphoramidothiolate (SMPT). The anticholinesterase properties of acephate, methamidophos, DMPT, SMPT, and OMPT were determined. Only acephate and methamidophos had measurable inhibitory effects on the mouse erythrocyte enzyme, methamidophos being about ten times more effective than acephate. The amount of methamidophos formed in the water and mouse liver was too low to have any direct effect on the toxicity of acephate. Acephate toxicity to aquatic insects would depend on its persistence in water, its uptake by the insects, its conversion to methamidophos, and the combined inhibitory effect of acephate and methamidophos on the cholinesterase enzyme. The toxicity of acephate to mammals would depend on the direct anticholinesterase effect of the chemical and to a small extent on methamidophos.

Fate and toxicity of acephate (Orthene) added to a coastal B. C. stream.

Acephate was added to a small, coastal B. C. stream to yield a concentration of 1100-1200 ppb for a 5-h period. It was rapidly taken up by fish, sediments, insect nymphs and larvae. No fish or insect mortality was noted although the more toxic methamidophos was found in both groups of animals. Acephate and methamidophos residues in animals and sediments declined to trace or non-detectable levels in 24 h and to these levels in water by 96 h. The impact of acephate on the stream and its fauna was limited and localized.

Detection of incipient effects of anticholinesterase insecticides in rats and humans by electromyography and cholinesterase assay.

Rats, fed low levels of diazinon (0.5 and 5.0 mg/kg) and parathion (0.1, 0.5 and 1.0 mg/kg) daily for 26 weeks, and agricultural workers chronically exposed to anticholinesterase insecticides, were monitored by electromyographic (EMG) and blood cholinesterase determinations. The two diazinon-fed and the highest parathion-fed groups of rats gained less weight, whereas the lowest parathion-fed group gained more weight than the control; rats fed 0.5 mg/kg parathion daily did not show any difference in weight gain when compared to the control. In addition, the two diazinon-fed groups of rats gained significantly less weight than the three parathion-fed groups. Both erythrocyte acetylcholinesterase (AChE) and plasma cholinesterase (ChE) activities were severely inhibited in the treated groups. However, parathion inhibited ChE much less than diazinon, and slight recovery of both enzymes was evident in the parathion-treated groups. No changes in amplitudes of EMGs were observed in the treated rats. Agricultural workers were examined monthly during one growing season. Although there was high variation in the data, EMG amplitudes and AChE levels were lower in the workers than in persons not exposed to pesticides; ChE levels were statistically different only once. These results have demonstrated that the rat is not suitable for investigating the use of EMG as a method for monitoring exposure to anticholinesterase insecticides, but that EMG (or AChE) may be used to monitor such exposures in humans.

Image analysis of hepatocyte nuclei from chlordane-treated rats.

The treatment of rats with hepatic dysfunction with chlordane at below the "no-effect" dose produces an increase in the number of hepatocyte nuclei with doubled DNA content. More important, though, it can be shown that even cells with unaltered DNA contents undergo statistically significant changes in their chromatin distribution pattern. Thus, image analysis techniques are capable here of detecting and substantiating toxic effects not apparent by visual inspection.

Residues in cutthroat trout (Salmo clarki) and California newts (Tarichia torosa) from a lake treated with technical chlordane.

The water of a lake was sprayed with technical chlordane to give a concentration of approximately 10 ppb. Resident California newts (Tarichia torosa) and cutthroat trout (Salmo clarki) were subsequently collected and analyzed for residue concentrations. The newts were collected at 14, 279, 451, and 1,036 days; the trout at 93, 279, 421, and 1,014 days. At first, the concentrations of residues were very high in both species and their composition, except for heptachlor, resembled that of technical chlordane. The heptachlor was quantitatively epoxidized in news after 14 days but in trout, some heptachlor was still present after 93 days. Concentrations of total chlordane in body tissues of both species declined more than 98% with time. Of seven individually determined constituents of technical chlordane, trans-nonachlor was by far the most persistent, accounting for 49 to 55% of total chlordane in specimens collected last. Both animals produced oxychlordane, probably from gamma-chlordane. Heptachlor, heptachlor epoxide and gamma-chlordeine were non-detectable in both species after 279 days. The results strongly suggest that the California newt metabolizes and eliminates chlordane residues more effectively than the cutthroat trout.

The fate of acephate and carbaryl in water.

Acephate was resistent to hydrolysis in distilled, buffered water at pH 4.0 to 6.9, but not at pH 8.2, held for 20 days at 20 or 30 degrees C. The maximum conversion to methamidophos was 4.5% of the added acephate at pH 8.2 and 20 degrees C. The persistence of acephate in two natural waters, held at 9 degrees C for up to 42 and 50 days varied: 80% were recovered from pond water after 42 days, and 45% from creek water after 50 days. Rates of acephate degradation increased greatly when treated water samples were incubated in the presence of sediments, but not if water and sediment were autoclaved prior to treatment and incubation. The greatest conversion to methamidophos, 1.3% of the added acephate, had occurred after 42 days in pond water without sediment. Under the same conditions, carbaryl was less persistent than acephate in the natural waters: 18 to 20% were recovered from pond water after 42 days, and 37 to 40% from creek water after 50 days. The presence of sediment did not affect its degradation significantly. But more than 55% were recovered after 50 days if water and sediment were autoclaved prior to treatment and incubation. Neither acephate, methamidophos, nor carbaryl could be shown to escape from water into the atmosphere.

Neurotoxic effects of leptophos (PhosvelR) in chickens and rats following chronic low-level feeding.

Leptophos (O-[4-bromo-2,5 dichlorophenyl] O-methyl phenylphosphonothioate) (PhosvelR) was administered orally to chickens and rats in doses of 0.5 and 5.0 mg/kg/day for 26 weeks. Hens fed 5.0 mg/kg, except one, showed ataxia and became paralysed in the legs at varying times from 8 to 19 weeks. A fifth hen showed ataxia early in the experiment but recovered fully for the remainder of the experiment. Rats fed both doses and chickens fed 0.5 mg/kg showed no signs of delayed neurotoxicity. All hens fed 5.0 mg/kg stopped laying by about the third week. Animals of both species fed 5.0 mg/kg either lost weight (chickens) or gained less weight (rats) than the others. Erythrocyte acetylcholinesterase (AChE) of the chickens given both doses was significantly depressed at first, then increased, and later dropped to control levels. AChE of rats fed 0.5 mg/kg was significantly inhibited but soon recovered to within control levels. On the other hand, the AChE of rats fed 5.0 mg/kg was inhibited throughout the experiment. Plasma cholinesterase (ChE) of both species was first inhibited and then recovered erratically for both insecticide concentrations. Histological alterations in the spinal cord of paralysed hens included axon and myelin degeneration in the ventral, lateral and posterior columns. In the paralysed hens, 79% of the neurotoxic esterase in the brain were inhibited, whereas in the non-paralysed hens (including the one non-paralysed hen receiving 5.0 mg/kg/day) and all rats only about half as much was inhibited.

Effects of sub-chronic low-level dietary intake of chlordane on rats with cirrhosis of the liver.

Male rats, 60 days old, were treated with chlordane during or after induction of liver cirrhosis with carbon tetrachloride to determine the effect of treatment with chlordane on the response of the rats to the disease. When liver cirrhosis was induced simultaneously with chlordane treatment the disease symptoms were aggravated; the lipid content of the tissue was lowered significantly, growth rate was significantly lower than controls and there was no apparent replacement of damaged liver tissue by liver growth. The cytochrome P450 content of the liver was similar after both treatments. Continuation of the chlordane treatment after termination of the carbon tetrachloride treatment brought about a more rapid recovery from the induced cirrhotic condition. All these responses were to a dose range one tenth the recommended "no effect" level for healthy animals.

Persistence of residues in water and sediment of a fresh-water lake after surface application of technical chlordane.

A fresh-water lake, free from detectable pesticide residues before this study, was treated with a commercial formulation of technical chlordane. Water and sediment samples were analysed for chlordane residues 7, 24, 52, 279. and 421 days after treatment. Residues moved rapidly from the water to the lake bottom, supporting earlier results of a laboratory study with other organochlorines. In water, alpha- and gamma-chlordane concentrations remained proportional to total chlordane concentrations, as determined by total-peak area measurements of gas-liquid chromatogrammes. But in bottom sediments, alpha- and gamma-chlordane were more persistent than other constituents of technical chlordane, supporting recent evidence that quantification of technical chlordane residues on the basis of gamma-chlordane quantification only (or that of alpha- and gamma-chlordane only), can lead to incorrect results and that this method, although simple and fast, should no longer be recommended.

Chlorinated hydrocarbon residues in fish, crabs, and shellfish of the Lower Fraser River, its estuary, and selected locations in Georgia Strait, British Columbia--1972-73.

Between August 1972 and September 1973, fish, crabs, and shellfish were collected from the lower Fraser River, its estuary, and selected areas of Georgia Strait in British Columbia. Samples were analyzed for aldrin, dieldrin, alpha- and gamma-chlordane, p,p'-DDT, p,p'-DDE, p,p'-DDD, heptachlor, heptachlor epoxide, lindane, and polychlorinated biphenyls (PCB's). Of these, p,p'-DDT, p,p'-DDE, p,p'-DDD, heptachlor epoxide, and one PCB, Aroclor 1254, were detected in samples of many fish, crabs, and shellfish from the lower Fraser River and its estuary. Generally, compounds found in decreasing order of magnitude in samples from the Fraser River and its estuary were: PCB's, p,p'-DDE, heptachlor epoxide, p,p'-DDT, and p,p'-DDD. Greatest concentrations of these compounds occurred in biota from the waters adjacent to the City of Vancouver. With one exception, animals from Georgia Strait and those away from the immediate influence of Fraser River water contained no detectable levels of chlorinated hydrocarbons.