Induction of glutathione S-transferase in the freshwater bivalve Sphaerium corneum as a biomarker for short-term toxicity tests?

A study was carried out to examine the use of glutathione S-transferase (GST) in freshwater bivalves as a short-term biochemical marker for organic contaminants. In contrast with earlier reports, induction of GST activity towards 1-Chloro-2,4-dinitrobenzene (CDNB) was not found in any of the experiments performed. These included 10–12 days exposures to sediments spiked with 2,3′,4,4′,5-pentachlorobiphenyl, benzo[a]pyrene or lindane and 7–8 days exposures to dieldrin, lindane or benzo[a]pyrene in a water-only system. A nonsignificant decrease in GST activity (ca. − 20%) was found at the highest sediment concentration of lindane (10,000 μg/kg) and at the highest water concentrations of dieldrin (4.9 and 29 μg/l). Between experiments differences in GST activity were found that were ascribed to seasonal influences and maintenance in the laboratory. It is concluded that under the applied conditions glutathione S-transferase activity of Sphaerium corneum towards CDNB is not a suitable short-term biomarker with respect to the toxicants tested.     

Actions of Insecticides on the Insect Gaba Receptor Complex

Abstract

The actions of insecticides on the insect γ-aminobutyric acid (GABA) receptor were investigated using [³⁵S]t-butylbicyclophosphorothionate ([³⁵S]TBPS) binding and voltage-clamp techniques. Specific binding of [³⁵S]TBPS to a membrane homogenate derived from the brain of Locusta migratoria locusts is characterised by a Kd value of 79.3 ± 2.9 nM and a B_max value of 1770 ± 40 fmol/mg protein. [³⁵S]TBPS binding is inhibited by mM concentrations of barbiturates and benzodiazepines. In contrast dieldrin, ivermectin, lindane, picrotoxin and TBPS are inhibitors of [³⁵S]TBPS binding at the nanomolar range. Bicuculline, baclofen and pyrethroid insecticides have no effect on [³⁵S]TBPS binding. These results are similar to those obtained in electrophysiological studies of the current elicited by GABA in both Locusta and Periplaneta americana central neurones. Noise analysis of the effects of lindane, TBPS, dieldrin and picrotoxin on the cockroach GABA responses reveals that these compounds decrease the variance of the GABA-induced current but have no effect on its mean open time. All these compounds, with the exception of dieldrin, significantly decrease the conductance of GABA-evoked single current.     

Effects of the pesticides atrazine and lindane and of manganese ions on cellular immunity of carp, Cyprinus carpio

Pesticides and metals in the aquatic environment can peturb the fish immune system and, consequently, increase their susceptibility to pathogenic agents. Lindane, an insecticide, was tested for its effects on skin graft rejection after daily administration in the food for 1 month (1000 mg lindane kg⁻¹ food). No difference was observed from the control for the first set of graft rejection times (8 days) or for the second set of graft rejection times (5 days), even if the lymphoid organs were highly contaminated with lindane. Atrazine, lindane and manganese were tested for their effect on in vitro phagocytosis of Yersinia ruckeri and zymosan (yeast extract) by pronephric and splenic macrophages. Neither pesticide had any effect on macrophage phagocytosis at concentrations up to their limit of solubility in water. Manganese ions, in the range 6.25–50 ppm, had a strong enhancing effect on phagocytosis of Y. ruckeri , as did zymosan, but only at a concentration of less than 6.25 ppm.     

Effect of growing Sesamum indicum L. on enhanced dissipation of lindane (1, 2, 3, 4, 5, 6-hexachlorocyclohexane) from soil

The effect of growing Sesamum indicum L. on the dissipation of lindane (gamma-HCH) was studied in spiked soil. For this, S. indicum was grown with four different concentrations of lindane (5, 10, 15, and 20 microg g(-1)). Plant growth, yield, photosynthetic pigments, soluble protein, microbial biomass carbon, lindane uptake, residual lindane concentration in soil and percentage dissipation of lindane from soil were analyzed at 25, 90, and 124 d. The accumulation of lindane in test plants was linearly related to the soil concentration (r2 = 0.897-0.979). At maturity, the accumulation of lindane in S. indicum grown with four spiked concentrations reached up to 7.98, 13.72, 23.71, and 33.29 microg g(-1) dry matter, respectively. There was a marked difference in the dissipation of lindane in vegetated and non-vegetated soils (p < 0.01). After final harvesting, the residual lindane concentrations in four spiked concentrations were reduced by 77.56, 70.12, 62.51, and 58.7%, respectively. Agronomic practice for the onsite application of this species is discussed. Based on the present study, it was calculated that S. indicum could accumulate 2237-2611 mg lindane per acre after 124 d cultivation. S. indicum could thus be used for the phytoremediation of lindane contaminated soil.     

Latent effects of insecticide intoxication to marine molluscs

Summary Clams and gastropods survived 96-hour immersion in concentrations up to 25 mg/l of four organophosphorus insecticides (DDVP, dioxathion, methyl parathion, Phosdrin^(R)) or 10 mg/l of four organochloride compounds (p,p-DDT, lindane, endrin, dieldrin) at 24 salinity and 20°C. During a post-treatment observation period of 33 days for gastropods and 133 days for clams, the following was noted: a reduction in number of egg cases deposited by gastropods initially subjected to comparatively high levels of endrin, dieldrin, and lindane; an apparent stimulatory effect on gastropod fecundity by relatively low concentrations of p,p-DDT, endrin, dieldrin, and lindane; and a decrease in survival of clams and gastropods initially subjected to high levels of DDVP or methyl parathion.

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Résumé Des groupes différents de mollusques bivalve, Mercenaria mercenaria, ou gastéropode, Nassa obsoleta, ont survécu des traitements de 25 mg/l d'insecticides organophosphoriques (DDVP, dioxathion, méthyl parathion, Phosdrin^(R)) ou de 10 mg/l de chlorures d'hydrocarbures (p,p-DDT, lindane, endrine, dieldrine). Ces traitements de 96 heures pour chaque groupe exposé à une concentration d'insecticide furent accomplis à une salinité de 24 et à une température de 20°C. Après ce treaement initial les gasteropodes furent placés dans de nouveaux aquariums contenant de l'eau non-contaminée et gardés sous observation pendant 33 jours. Les bivalves, traités de façon identique, furent gardés pendant une periode de 133 jours. La période d'observation révéla les différences suivantes entre les animaux traités et les témoins : a/ réduction du nombre des plaques de ponte pour le gastéropode soumis à de fortes doses d'endrine, de dieldrine, et de lindane; b/ stimulation apparent de la fécundité comme le laisse supposer l'augmentation du nombre des plaques de pontes pour les gastéropodes traités avec de faibles concentrations de p,p-DDT, d'endrine, de dieldrine, et de lindane; c/ une augmentation du taux de mortalité chez les bivalves et les gastéropodes soumis à de fortes doses de DDVP ou de méthyl parathion.

     

Investigations into mechanisms responsible for seizures induced by chlorinated hydrocarbon insecticides: the role of brain ammonia and glutamine in convulsions in the rat and cockerel

Abstract— Seizures were produced in adult female rats and in cockerels (Gallus domesticusL.) after the intracarotid administration of four structurally-unrelated chlorinated hydrocarbon insecticides. The temporal relationship of the course of the convulsions to levels in whole brain of ammonia and glutamine was examined. High levels of brain ammonia occurred in rats and cockerels poisoned acutely with lindane, dieldrin, heptachlor and DDT. The intensity of convulsions and other neurological signs coincided with and were directly correlated with the increases in brain ammonia, whereas interictal periods were associated with decreased levels of ammonia. In both species the order of drug toxicity was lindane > dieldrin > heptachlor > DDT. Significantly elevated levels of brain glutamine were induced in the cockerels after administration of all four insecticides. In contrast, increased levels of glutamine in rats were found only in those intoxicated with lindane and dieldrin. Although the ammonia-binding mechanism (conversion of free ammonia into glutamine) was saturated throughout the experiments in both the rats and cockerels, the latter were more resistant to ammonia toxicity by virtue of being able to convert free ammonia into glutamine more efficiently than rats. These data, together with data reported by others, lead to the conclusion that the four structurally-unrelated chlorinated hydrocarbon insecticides probably induced convulsions in both species by a common mechanism, involving interference with the production and/or utilization of ammonia.     

Comparative biodegradation of alkyl halide insecticides by the white rot fungus, Phanerochaete chrysosporium (BKM-F-1767).

The ability of Phanerochaete chrysosporium to degrade six alkyl halide insecticides (aldrin, dieldrin, heptachlor, chlordane, lindane, and mirex) in liquid and soil-corncob matrices was compared by using 14C-labeled compounds. Of these, only [14C]lindane and [14C]chlordane underwent extensive biodegradation, as evidenced by the fact that 9.4 to 23.4% of these compounds were degraded to 14CO2 in 30 days in liquid cultures and 60 days in soil-corncob cultures inoculated with P. chrysosporium. Although [14C]aldrin, [14C]dieldrin, [14C]heptachlor, and [14D]mirex were poorly mineralized, substantial bioconversion occurred, as determined by substrate disappearance and metabolite formation. Nonbiological disappearance was observed only with chlordane and heptachlor.     

Comparative biodegradation of alkyl halide insecticides by the white rot fungus, Phanerochaete chrysosporium (BKM-F-1767).

The ability of Phanerochaete chrysosporium to degrade six alkyl halide insecticides (aldrin, dieldrin, heptachlor, chlordane, lindane, and mirex) in liquid and soil-corncob matrices was compared by using 14C-labeled compounds. Of these, only [14C]lindane and [14C]chlordane underwent extensive biodegradation, as evidenced by the fact that 9.4 to 23.4% of these compounds were degraded to 14CO2 in 30 days in liquid cultures and 60 days in soil-corncob cultures inoculated with P. chrysosporium. Although [14C]aldrin, [14C]dieldrin, [14C]heptachlor, and [14D]mirex were poorly mineralized, substantial bioconversion occurred, as determined by substrate disappearance and metabolite formation. Nonbiological disappearance was observed only with chlordane and heptachlor.     

Isolation of four aquatic streptomycetes strains capable of growth on organochlorine pesticides

Ninety-three wild-type isolates identified as actinomycetes were tested against 11 organochlorine pesticides (OPs): aldrin, chlordane, DDD, DDE, DDT, dieldrin, heptachlor, and heptachlor epoxides, lindane, and methoxychlor. Qualitative screening agar assays displayed 62-78% tolerance of strains to OPs. Four strains designed M4, M7, M9 and M15 were selected based on multi-OP-tolerance, and identified as members of the streptomycetes group. Different growth profiles were observed in cultures of the four selected streptomycetes cultured in synthetic medium containing 5-50 microg x l(-1) aldrin or chlordane or lindane. Increase of aldrin removal by the selected microorganisms was concomitant with the 4.8-36.0 microg x l(-1) pesticide concentration range. After 72 h of streptomycete M7 growth in synthetic medium containing 48.0 microg x l(-1) aldrin, the remaining OP concentration in the supernatant was approximately 10% of the initial concentration. Also, in stationary growth phase less than 2.5 microg x l(-1) aldrin residual concentration was detected in the medium.     

Effects of lindane on the glucose metabolism in rat brain cortex cells.

The influence of 0.5 mM gamma-hexachlorocyclohexane (gamma-HCH, lindane) on glucose transport has been investigated using the analog 3-O-methyl-D-(U-14C)glucose. The glucose uptake was lineal for at least 10 sec. Preincubation of dissociated brain cortex cells with lindane decreased the transport of glucose with respect to the controls. The treatment of brain cortex cells with other organochlorine compounds indicated that the alpha-, delta-HCH isomers and dieldrin reproduced the same inhibitory pattern, while beta-HCH and endrin were inactive. The total radioactivity incorporated into CO2 from (U-14C) glucose in the cerebral cortex is also inhibited by lindane in a time dependent manner.     

Lindane effect upon the vasoactive intestinal peptide receptor/effector system in rat enterocytes

Isolated rat enterocytes exposed to the insecticide lindane (the gamma-isomer of hexachlorocyclohexane, HCCH) showed an important decrease in the efficiency of the neuropeptide vasoactive intestinal peptide (VIP) upon the stimulation of cyclic AMP accumulation. The effect of lindane was time- and dose-dependent, optimal conditions being reached after 5 min incubation of cells at 25 degrees C with 0.5 mM of this organochlorine compound. Lindane action exhibited an important degree of specificity since the isomer alpha-HCCH and endrin reproduced the same inhibitory pattern but beta-HCCH and dieldrin were inactive. The inhibition of VIP-induced cyclic AMP accumulation could not be explained by a lindane-dependent reduction in the binding of VIP to its specific receptors. Among various possibilities, the results suggest the modification of membrane fluidity by lindane and/or the activation of Ca2+-dependent protein kinase C by this compound leading to phosphorylation of Gs/adenylate cyclase.     

The formation of chlorobenzene and benzene by the reductive metabolism of lindane in rat liver microsomes

The major metabolite produced by incubating [14C]lindane with rat liver microsomes under anaerobic conditions was determined to be chlorobenzene, with lesser amounts of benzene also being formed. Using relatively high lindane concentrations (250 microM), four nonvolatile metabolites of lindane were also produced anaerobically, the predominant one being identified by mass spectrometry as tetrachlorocyclohexene (TCCH). TCCH, likewise, was reduced to chlorobenzene and benzene in microsomes under anaerobic conditions. Binding of [14C]lindane to microsomal protein occurred under aerobic as well as anaerobic incubation conditions; however, lindane protein binding was greatest in anaerobic incubations compared to those containing an atmosphere of air or 100% oxygen. Hemin reduced by dithionite also readily produced chlorobenzene and benzene from lindane. These results indicate that lindane interacts readily with heme and heme proteins, including cytochrome P-450, in the absence of oxygen to undergo multiple chloride eliminations forming chlorobenzene and benzene as end products.     

Residues of lindane in adipose tissue of lambs and sheep from islands Krk and Cres

Concentrations of lindane from adipose tissue i.e. omentum of sheep from homesteads of island Krk that were treated with lindane, were compared with the concentrations of lindane measured in adipose tissue of non treated sheep from husbandry cooperative on island Cres. Evaluation of food safety for this samples were estimated by comparing obtained results and maximum tolerance concentration (MTC) for lindane (2.0 and 0.2 mg/kg). In treated lambs from Krk measured concentrations of lindane were in range from 0.0038 to 1.8644 mg/kg, and in treated sheep from 0.0094 to 1.646 mg/kg. In control group of lambs and sheep concentration of lindane was < or = 0.0051 mg/kg. Estimation of food safety based on the results from this research are discussable, regarding the fact that MTC prescribed before year 2007 was 2.0 mg/kg. When the new law Regulations of MTC for residues of pesticides in food and animal feed (NN 119/07) came into the force prescribed MTC was 0.02 mg/kg. If food safety of sheep meat from Krk would be estimated based on latter, then 73.4% of samples of lamb meat and 80% of sheep meat could be proclaimed unsafe for human consumption. It is encouraging that after completion of using lindane in year 2005, levels of lindane in adipose tissue of lamb from Krk in year 2006 were under the concentration set by new legislate (< 0.02 mg/kg).     

Comparative toxicity and biotransformation of lindane in C57BL/6 and DBA/2 mice

DBA/2 mice, previously identified as "unresponsive" to aromatic hydrocarbons which induce microsomal enzymes in C57BL/6 mice, are more vulnerable to the convulsant effect of repeated doses of lindane than similarly treated C57BL/6 mice. Death in convulsions and higher blood and brain lindane concentrations indicate that less efficient disposition of lindane itself accounts for the greater vulnerability of the DBA/2 mice. The same two principal chlorophenolic metabolites of lindane were identified in the blood and tissues of both strains, but the time-courses of blood concentrations in response to repeated lindane dosing were different.     

Activation of gamma-aminobutyric acid insensitive chloride channels in mouse brain synaptic vesicles by avermectin B1a.

The interaction of avermectin B1a (AVMB1a) with mouse brain chloride channels was characterized using a radiochloride efflux assay. The loss of intravesicular chloride from synaptoneurosomes preloaded with 36Cl involved an initial rapid phase followed by a slower phase that approached equilibrium within 10 min. AVMB1a stimulated a 30% loss of intravesicular chloride within the first 2 s of exposure; however, AVMB1a had no effect on the rate of the slower phase of chloride loss. Experiments with lysed synaptoneurosomes showed that both chloride loading and basal and AVMB1a-stimulated chloride release required the presence of intact vesicles. The efflux of 36Cl from mouse brain synaptosomes and the stimulation of efflux by AVMB1a were qualitatively similar to the results obtained with synaptoneurosomes but involved much lower overall levels of chloride loading and release. AVMB1a produced half-maximal stimulation of chloride efflux from synaptoneurosomes at a concentration of 2.1 +/- 0.3 microM and a 35.4 +/- 1.4% maximal loss of intravesicular chloride at saturating concentrations. gamma-Aminobutyric acid (GABA), bicuculline, or the chloride channel blockers picrotoxinin, t-butylbicyclophosphorothionate (TBPS) 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), and anthracene 9-carboxylic acid (9-CA) had little or no effect on the loss of chloride from synaptoneurosomes either in the presence or the absence of AVMB1a. However, the chlorinated cycloalkane insecticides dieldrin and lindane were equally effective as inhibitors of GABA-dependent chloride uptake and AVMB1a-stimulated chloride efflux. These data demonstrate that AVMB1a-stimulated chloride efflux from mouse brain synaptic vesicles results from the activation of GABA-insensitive chloride channels and that this action is distinct from their previously documented effects on GABA-gated chloride channels in mouse brain preparations. Our findings imply that both GABA-gated and GABA-insensitive chloride channels may be toxicologically significant targets for the action of avermectins.     

Species-dependent effects of dietary lindane and/or zineb on the activation of aflatoxin B1 into mutagenic derivatives

Sprague-Dawley rats and Swiss mice were given diets containing lindane, 125 ppm, or zineb, 5200 ppm, or a mixture of both at the above-mentioned concentrations for 2 and 4 weeks. The effect of pesticide ingestion on the ability of liver S9 to metabolize aflatoxin B1 (AFB1) into mutagenic derivatives was tested by the Salmonella (TA100)/microsome test according to Ames. Control mouse-liver S9 was less efficient (13%) than the corresponding preparation from control rat liver. The ingestion of lindane produced a similar increase in the activities of both rat- (68%) and mouse-liver S9 (62%). Pretreatment with zineb inhibited (46%) rat-liver S9 but caused a marked increase (400%) in the activity of mouse-liver S9. Concomitant exposure to both pesticides showed that lindane released the inhibitory action of zineb on rat-liver S9 and reduced the stimulatory effect of zineb on mouse-liver S9. The inducing action of zineb in mice was a function of the dietary concentration of the pesticide. No effect was observed at dietary concentrations of zineb up to and including 500 ppm.     

Lindane degradation by root epiphytic bacterium Achromobacter sp. strain A3 from Acorus calamus and characterization of associated proteins

Abstract

Lindane degrading root epiphytic bacteria were isolated from wetland plant Acorus calamus. Bacterial strain A3 identified as Achromobacter sp. A3, showed maximum degradation potential of 88.7?±?1.24% for 50?mg?l^?1 lindane. Lindane biodegradation was followed by decrease in pH as well as increase in concentration of chloride ions in the culture medium. Lindane degradation potential of Achromobacter sp. A3 was also studied at different concentrations of lindane. Maximum degradation was at 10?mg l^?1 followed by 50?mg l^?1 and 100?mg l^?1 lindane. Also, lindane induced proteins were studied using SDS-PAGE. The induced proteins were identified as alpha/beta hydrolase fold-3 domain-containing protein, involved in lindane hydrolysis and extracellular solute-binding family protein having role in transmembrane transport of lindane for utilization of lindane by bacteria. The appearance of unique polypeptides in lane corresponding to media supplemented with lindane showed that the exposure of bacterial cells to lindane has resulted in regulative expression of certain proteins. So far as known, this is the first report to isolate and study lindane degrading root epiphytic bacteria from A. calamus.     

Gene mapping and cross-resistance in cyclodiene insecticide-resistant Drosophila melanogaster (Mg.)

Resistance to the cyclodiene insecticide dieldrin maps to a single gene (Rdl) on the left arm of chromosome III in Drosophila melanogaster (Meigen). The gene was further mapped by the use of chromosomal deficiencies to a single letter sub-region, 66F, on the polytene chromosome. The cross-resistance spectrum of a backcrossed strain lacking elevated mixed function oxidase activity, a common resistance mechanism, was examined. Levels of resistance similar to those found in other insects were found to dieldrin, aldrin, endrin, lindane, and picrotoxinin. Strong similarity of this single major gene with that found in other cyclodiene resistant insects is suggested by its cross-resistance spectrum and chromosomal location, via homology with other Diptera. The significance of major genes in insecticide resistance is discussed.     

Developmental and Lactational Exposure to Dieldrin Alters Mammary Tumorigenesis in Her2/neu Transgenic Mice

Breast cancer is the most common cancer in Western women and while its precise etiology is unknown, environmental factors are thought to play a role. The organochlorine pesticide dieldrin is a persistent environmental toxicant thought to increase the risk of breast cancer and reduce survival in the human population. The objective of this study was to define the effect of developmental exposure to environmentally relevant concentrations of dieldrin, on mammary tumor development in the offspring. Sexually mature FVB-MMTV/neu female mice were treated with vehicle (corn oil), or dieldrin (0.45, 2.25, and 4.5 µg/g body weight) daily by gavage for 5 days prior to mating and then once weekly throughout gestation and lactation until weaning. Dieldrin concentrations were selected to produce serum levels representative of human background body burdens, occupational exposure, and overt toxicity. Treatment had no effect on litter size, birth weight or the number of pups surviving to weaning. The highest dose of dieldrin significantly increased the total tumor burden and the volume and number of tumors found in the thoracic mammary glands. Increased mRNA and protein expression of the neurotrophin BDNF and its receptor TrkB was increased in tumors from the offspring of dieldrin treated dams. This study indicates that developmental exposure to the environmental contaminant dieldrin causes increased tumor burden in genetically predisposed mice. Dieldrin exposure also altered the expression of BNDF and TrkB, novel modulators of cancer pathogenesis.