Incidence of chromosome aberrations in hothouse workers and the in vitro sensitivity of their lymphocytes to the cytogenetic action of dimatif

The cytogenetical survey of eight persons working in greenhouse and occupationally contacting with a complex of pesticides has shown a significant increase of chromosomal aberration level in comparison with the control group. It is determined that the cytogenetic effect of the greenhouse's workers differed significantly from that of the control group when lymphocytes of the examined persons were treated by the potent mutagen dimatyph in vitro. The occupational group on the whole was more sensitive than control persons to the clastogenic action of dimatyph (43.0 +/- 2.2 and 28.7 +/- 2.4%, respectively). An assumption is advanced on the modifying influence of the pesticide complex on the frequency of induced chromosomal aberrations in vitro.     

Toxic effect of DDT and endosulfan in white shrimp postlarvae Litopenaeus vannamei (Decapoda: Penaeidae) from Chiapas, Mexico

We analized acute toxicity in white shrimp (Litopenaeus vannamei) postlarvae exposed to two chlorinated pesticides, DDT and endosulfan, under laboratory conditions during 168 hours, with controlled temperature (29 +/- 1 degrees C), salinity (3 +/- 1%o) and pH (8 +/- 1). Median lethal concentrations (LC50), "incipient" LC50, median lethal time (LT50) the "maximum acceptable concentration of the toxic compound" (MACT) and "the safety level" (SL) were determined. The concentration of the compounds at which organism growth was reduced by 5 and 50% (EC5 and EC50), as well as changes in oxygen consumption patterns were determined in the surviving postlarvae. They were very sensitive to both compounds and DDT was thrice as toxic as endosulfan. Growth rate decreased 50% and 80% with endosulfan and DDT, respectively, at the experimental pestice concentration. The low resistance of postlarvae to DDT and endosulfan suggests that additional inflow of these pesticides into the aquatic system could affect the rate of shrimp production in the area.     

Metabolic pathways utilized by Phanerochaete chrysosporium for degradation of the cyclodiene pesticide endosulfan.

Recent studies have shown that cultures of white rot fungi not favoring the production of lignin and manganese peroxidases are effective in degrading certain xenobiotics. In this study we have used endosulfan as a model xenobiotic to assess the enzymatic mechanisms of pesticide metabolism under ligninolytic (nutrient-deficient) and nonligninolytic (nutrient-rich) culture conditions. Rapid metabolism of this chlorinated pesticide occurred under each nutrient condition tested. However, the extent of degradation and the nature of the metabolic products differed for nutrient-deficient and nutrient-rich media. The pathways for endosulfan metabolism were characterized by analysis of the fungal metabolites produced. The major endosulfan metabolites were identified by gas chromatography-electron capture detection and gas chromatography-mass spectrometry as endosulfan sulfate, endosulfan diol, endosulfan hydroxyether, and a unknown metabolite tentatively identified as endosulfan dialdehyde. The nature of the metabolites formed indicates that this organism utilizes both oxidative and hydrolytic pathways for metabolism of this pesticide. Piperonyl butoxide, a known cytochrome P-450 inhibitor, significantly inhibited the oxidation of endosulfan to endosulfan sulfate and enhanced hydrolysis of endosulfan to endosulfan diol. We suggest that the metabolism of endosulfan is mediated by two divergent pathways, one hydrolytic and the other oxidative. Judging by the inactivity of extracellular fluid and partially purified lignin peroxidase in metabolizing endosulfan, we conclude that metabolism of this compound does not involve the action of extracellular peroxidases.     

Individual and combined toxicity of common pesticides to teleost Puntius conchonius Hamilton

Individual and combined toxicity of three pesticides, endosulfan, phosphamidon, and aldicarb was evaluated in P. conchonius. The 48 hr LC50 was 21.36 and 446.5 ppm respectively for endosulfan and phosphamidon. When tested jointly, 48 hr LC50 for different ratios of these pesticides were 0.332 (IE:3P), 0.224 (IE:1P), and 0.178 ppm (3E:1P). The cotoxicity coefficients for these combinations were 1793, 3986, and 10009, respectively. An equitoxic mixture of endosulfan, phosphamidon, and aldicarb yielded a 48 hr LC50 of 130.5 ppm. An enhanced toxic impact is indicated when the pesticides are present together rather than as individual compounds.     

Toxicological studies of pesticides on cytoplasmic streaming in Nitella

In the present study, changes in velocity of cytoplasmic streaming in the giant internodal cells of Nitella for varying concentration of the pesticides, 2,4-D, dieldrin, malathion, methyl parathion and endosulfan, were measured. Marked decrease in the velocity of cytoplasmic streaming was found at the concentrations of 0.01, 0.1, 1, 10 and 100mM. Dieldrin was the most toxic to all the pesticides investigated, followed by methyl parathion, endosulfan, malathion and 2,4-D. Threshold values for dieldrin, methylparathion, endosulfan, malathion and 2,4-D as indicated by the onset of decrease in the normal cytoplasmic streaming velocity were less than 6.25 x 10(-6), 2.5 x 10(-5), 5 x 10(-5), 5 x 10(-5) and 1.25 x 10(-5)M respectively. Cessation of streaming was noticed above 1mM in dieldrin and above 10mM when exposed to methylparathion and endosulfan. Cessation of streaming was not seen up to 100mM concentration of 2,4-D and malathion.     

Evaluation of pesticides against major pests of jute (Corchorus olitorius L.) in West Bengal,India

Among the pesticides evaluated against the jute pests under field conditions, endosulfan 35 EC at 350 g a.i./ha was found to be the most effective insecticide for controlling semilooper, Bihar hairy caterpillar and myllocerus weevil. Endosulfan and fenpropathrin provided good control of yellow mite as well. Even, endosulfan-treated plots gave maximum fibre and stick yield with greater benefit–cost ratio (BCR) over others. Next to endosulfan and chlorpyriphos, neemazal (azadirachtin 50,000 ppm) are useful and safer options against semilooper, Bihar having caterpillar, myllocerus and yellow mite. Methoxyfenocide 10 EC at 200g a.i./ha provided good control (70.60%) of S. obliqua even after 14 days of spraying, but it was not effective against other pests of jute. Further, almost similar efficacy of neemazal and chlorpyriphos was found against the pest complex of jute and based on BCR, neemazal could be an alternative choice over chlorpyriphos.     

Risk Posed by Pesticides to Aquatic Organisms in Rivers of Northern Inland New South Wales,Australia

An ecological assessment was conducted to determine the risk posed by agricultural pesticides to inland rivers of north-west New South Wales (NSW), Australia. A preliminary screening of 30 pesticides provided a short-list of eight for further investigation (atrazine, chlorpyrifos, diuron, endosulfan, fluometuron, metolachlor, profenofos, prometryn). Selection was based on chemical characteristics, toxicity, detection frequencies and environmental concentrations. Hazard quotients were calculated for both spray and non-spray seasons. Where possible, hazard quotients were calculated for both acute and chronic exposures for crustaceans, insects, micro-organisms, molluscs, plants and vertebrates. Chlorpyrifos, endosulfan and profenofos posed a high hazard (HQ > 0.5). A probabilistic risk analysis indicated that chlorpyrifos, endosulfan and profenofos posed a risk from acute exposure during the spray season, while endosulfan also posed a risk from chronic exposure during the spray season. The risks posed by profenofos and chlorpyrifos were characterised by a low probability of detection, but these detections affected a high percentage of species. The risks posed by acute and chronic exposures of endosulfan were characterised by a high probability of detection, but only a limited number of these detections affected a high percentage of species. Risk during the non-spray season was not assessed, as the detection of pesticides was infrequent during this period.     

Screening of soil fungi for in vitro degradation of endosulfan

Intensive use of endosulfan has resulted in contamination of soil and water environments at various sites in Pakistan. This study was conducted to isolate efficient endosulfan-degrading fungal strains from contaminated soils. Sixteen fungal strains were isolated from fifteen specific sites by employing enrichment techniques while using endosulfan as a sole sulfur source, and tested for their potential to degrade endosulfan. Among these fungal strains, Chaetosartorya stromatoides, Aspergillus terricola, and Aspergillus terreus degraded both α- and β-endosulfan upto 75% in addition to ∼20% abiotic degradation of the spiked amount (100 mg l⁻¹) in the broth within 12 days of incubation. Biodegradation of endosulfan by soil fungi was accompanied by a substantial decrease in pH of the broth from 7.0 to 3.2. The major metabolic product was endosulfan diol along with very low concentrations of endosulfan ether. Maximum biodegradation of endosulfan by these selected fungal strains was found at an initial broth pH of 6, incubation temperature of 30°C and under agitation conditions. This study indicates that the isolated strains carried efficient enzyme systems required for bioremediation of endosulfan-contaminated soil and water environments.     

Kinetics of Endosulfan Biodegradation by Stenotrophomonas maltophilia EN-1 Isolated From Pesticide-Contaminated Soil

Endosulfan is one of the persistent organochlorinated pesticides used extensively throughout the world, particularly in the developing countries. Its microbial metabolic transformation product endosulfan sulphate is more toxic and persistent than the parent compound itself. In order to completely mineralize endosulfan, augmentation of soil microbial community with efficient endosulfan degradation properties could a potentially viable option. In the present study, endosulfan degrading bacterium was isolated from the agriculture-contaminated soil of Shujaabad, Multan, Pakistan by using enrichment technique. The isolated bacterial strain EN-1 (Endosulfan-1) was identified as S. maltophilia by 16S rRNA sequencing and biochemical analysis. EN-1 has demonstrated the ability to utilize endosulfan as sole sulfur source. Kinetics of endosulfan degradation was studied at various initial concentrations ranges from 5 mg/L to 100 mg/L by growth dependent and growth independent kinetic models. Biodegradation kinetics revealed that the bacterium was highly efficient in endosulfan degradation. The average values of kinetic constants i.e. K_s, and µ_max were 13.73 mg/L and 0.210 h^?1 respectively, while µ_max/K_s ratio was 0.015. Addition of sulfur decreased the rate of degradation as the µ_max/K_s was observed to reduce. GC-MS analysis revealed that the bacterium metabolised the endosulfan into non-toxic metabolite i.e. endosulfan diol. The study instigates a complete elucidation of degradation process for commercial applications.     

Optimization of environmental parameters for biodegradation of alpha and beta endosulfan in soil slurry by Pseudomonas aeruginosa

Aim: To determine optimal environmental conditions for achieving biodegradation of α‐ and β‐endosulfan in soil slurries following inoculation with an endosulfan degrading strain of Pseudomonas aeruginosa. Methods and Results: Parameters that were investigated included soil texture, soil slurry: water ratios, initial inoculum size, pH, incubation temperature, aeration, and the use of exogenous sources of organic and amino acids. The results showed that endosulfan degradation was most effectively achieved at an initial inoculum size of 600 μl (OD = 0·86), incubation temperature of 30°C, in aerated slurries at pH 8, in loam soil. Under these conditions, the bacterium removed more than 85% of spiked α‐ and β‐endosulfan (100 mg l^?1) after 16 days. Abiotic degradation in noninoculated control medium within same incubation period was about 16%. Biodegradation of endosulfan varied in different textured soils, being more rapid in course textured soil than in fine textured soil. Increasing the soil contents in the slurry above 15% resulted in less biodegradation of endosulfan. Exogenous application of organic acids (citric acid and acetic acid) and amino acids (l ‐methionine and l ‐cystein) had stimulatory and inhibitory effects, respectively, on biodegradation of endosulfan. Conclusion: The results of this study demonstrated that biodegradation of endosulfan by Ps. aeruginosa in soil sediments enhanced significantly under optimized environmental conditions. Significance and Impact of the Study: Endosulfan is a commonly used pesticide that can contaminate soil, wetlands and groundwater. Our study demonstrates that bioaugmentation of contaminated soils with an endosulfan degrading bacterium under optimized conditions provides an effective bioremediation strategy.     

Dynamic genome-scale modeling of Saccharomyces cerevisiae unravels mechanisms for ester formation during alcoholic fermentation

Fermentation employing Saccharomyces cerevisiae has produced alcoholic beverages and bread for millennia. More recently, S. cerevisiae has been used to manufacture specific metabolites for the food, pharmaceutical, and cosmetic industries. Among the most important of these metabolites are compounds associated with desirable aromas and flavors, including higher alcohols and esters. Although the physiology of yeast has been well-studied, its metabolic modulation leading to aroma production in relevant industrial scenarios such as winemaking is still unclear. Here we ask what are the underlying metabolic mechanisms that explain the conserved and varying behavior of different yeasts regarding aroma formation under enological conditions? We employed dynamic flux balance analysis (dFBA) to answer this key question using the latest genome-scale metabolic model (GEM) of S. cerevisiae. The model revealed several conserved mechanisms among wine yeasts, for example, acetate ester formation is dependent on intracellular metabolic acetyl-CoA/CoA levels, and the formation of ethyl esters facilitates the removal of toxic fatty acids from cells using CoA. Species-specific mechanisms were also found, such as a preference for the shikimate pathway leading to more 2-phenylethanol production in the Opale strain as well as strain behavior varying notably during the carbohydrate accumulation phase and carbohydrate accumulation inducing redox restrictions during a later cell growth phase for strain Uvaferm. In conclusion, our new metabolic model of yeast under enological conditions revealed key metabolic mechanisms in wine yeasts, which will aid future research strategies to optimize their behavior in industrial settings.     

Changes in gill structure induced by BHC, lindane and endosulfan in the fresh-water teleost Puntius ticto (Ham.).

The effect of three organochlorinated pesticides on the gill structure of Puntius ticto, a freshwater teleost, was investigated. Fish exposed to sublethal concentrations of BHC (0.17 ppm), lindane (0.19 ppm) and endosulfan (0.20 ppm) were studied. The pesticides were detected qualitatively in the gill tissue by thin-layer chromatography (TLC). The results showed that they could be detected after 15 days' exposure, but not after 96 h exposure. Histopathological examination revealed several structural and functional changes in the gills. Exposure to BHC was followed by an inflammatory reaction and complete dystrophy of the lamellar structure of the gills. Lindane-treated fish showed disruption of the epithelial covering of the gills and excessive haemorrhage in the blood vessels. In exposure to endosulfan the gill lamellae shrank and became thinner.     

Reactive oxygen species in in vitro pesticide-induced neuronal cell (SH-SY5Y) cytotoxicity: role of NFkappaB and caspase-3

Oxidative stress has been implicated in pesticide-induced neurotoxicity, based on its role in the cascade of biochemical changes that lead to dopaminergic neuronal cell death. We have, therefore, examined the role of oxidative stress caused by the pesticides endosulfan and zineb in human neuroblastoma cells (SH-SY5Y) in culture. Upon treatment with 50-200 microM concentrations of either of these pesticides, SH-SY5Y cells generated both superoxide anion and hydrogen peroxide in a dose-and time-dependent manner. Mixtures of the pesticides significantly enhanced the production of these reactive oxygen species compared to individual pesticide exposures. Pesticide treatment decreased superoxide dismutase, glutathione peroxidase, and catalase activities in SH-SY5Y cells. Additionally, these pesticides induced lipid peroxide (thiobarbituric acid reactive products) formation in these cells. While both pesticides individually (at 100 microM) increased caspase-3 activity, cells exposed to a mixture of the pesticides exhibited significantly low levels of this enzyme, probably due to excessive necrotic cell death. Furthermore, exposure to these pesticides increased nuclear NFkappaB activity. Taken together, these findings suggest that the cytotoxicity of endosulfan and zineb, both individually and in mixtures may, at least in part, be associated with the generation of reactive oxygen species with concomitant increased expression of NFkappaB.     

Evaluation of the mutagenic potential of endosulfan using the Salmonella/mammalian microsome assay

The mutagenic potential of endosulfan, a polychlorinated insecticide, was assessed using the highly sensitive Salmonella tester strains TA97(a), TA98, TA100 and TA102. It exhibited a toxic effect at dose levels of 50 micrograms/plate and higher. Plate incorporation studies did not show mutagenic response with any of the tester strains used. A modification of the assay using a preincubation procedure showed mutagenic activity with and without metabolic activation with TA97(a) only. Increased toxicity was observed after addition of phenobarbital-induced S9 mix.     

Comparison of colorimetric and chemiluminescent enzyme‐linked immunosorbent assay for the detection of endosulfan in food samples

Pesticides have become part of food protection since their inception. Endosulfan, an organochlorine insecticide, has been used against insect pests such as whiteflies, aphids, red spiders and mites. Methods of immunochemical assays have been devised for the determination and analysis of pesticides and commonly used for the analysis of contaminants in food, water, soil and body fluids. Chicken IgY antibodies raised against endosulfan haptens were used for the detection of endosulfan. We have compared colorimetric (CO) and chemiluminescence (CL) enzyme‐linked immunosorbent assay (ELISA) techniques for the detection of endosulfan isomers in a food matrix. CL ELISA assay was found to be more sensitive than CO assay. The mean recovery was 81.2–95.6% for α‐ and β‐endosulfan‐spiked food samples with 2.8–4.6% relative standard deviation. The detection of the endosulfan isomers was linear in the range 100 µg/mL–5 fg/mL, with a limit of detection at 100 µg/mL and 5 fg/mL for the CL ELISA method and 100 µg/mL and 1 ng/mL for the CO ELISA method respectively. These methods can be used for the rapid and reliable detection of organochlorine pesticide endosulfan. Copyright © 2015 John Wiley & Sons, Ltd.     

Metabolic activation of cytochrome P-450/P-448 in the yeast Saccharomyces cerevisiae

The strains D6 and JD1 of the yeast Saccharomyces cerevisiae were used to assay the genetic activity of several compounds, benzo[a]pyrene, 15,16-dihydro-11-methyl-cyclopenta[a]phenanthren-17-one, 2-naphthylamine and cyclophosphamide, which require metabolic activation by cytochromes P-450 and P-448 to produce genetically active chemical species. Cells from both strains were harvested from cultures grown in low concentrations of glucose and switched to growth in high glucose containing media. Treatments under these conditions resulted in increased sensitivity of the test systems without the presence of an exogenous S9 mix and the presence of S9 was found not to enhance this sensitivity. The yeasts used under these treatment conditions showed a P-450/P-448 type metabolism.     

Frequencies of chromosomal aberrations in smokers exposed to pesticides in cotton fields

Blood samples were collected from 50 smokers who were exposed to the pesticides DDT, BHC, endosulfan, malathion, methyl parathion, monocrotophos, quinolphos, dimethoate, phosphomidon, cypermethrin and fenvelrate. Samples were also collected from 20 non-smokers (control I) and 27 smokers (control II) who were unexposed to pesticides. Control II showed a significant increase in chromosomal aberrations when compared to control I. There was a significant increase in total chromosomal aberrations in smokers exposed to pesticides when compared to unexposed populations.     

Mutagenic and recombinogenic action of pesticides in Aspergillus nidulans.

Thirteen pesticides, aminotriazole, benomyl, captafol, captan, dalapon-Na, dichlorvos, dinobuton, dodine, ioxynil, mecoprop, neburon, picloram and tordon were tested for ability to induce (1) point mutations to 8-azaguanine resistance, (2) mitotic crossing-over, and (3) mitotic non-disjunction and haploidization in Aspergillus nidulans. Tests were performed at three different pHs, i.e. 4.5, 7, 8.2. Three of the pesticides, captan , captafol and dichlorvos induced point mutations; dichlorvos also induced a high frequency of mitotic crossing-over and non-disjunction; benomyl induced a very high frequency of non-disjunction whereas aminotriazole induced weakly both types of somatic segregation.     

Regulation of Hepatic Drug Transporter Activity and Expression by Organochlorine Pesticides

Organochlorine (OC) pesticides constitute a major class of persistent and toxic organic pollutants, known to modulate drug‐detoxifying enzymes. In the present study, OCs were demonstrated to also alter the activity and expression of human hepatic drug transporters. Activity of the sinusoidal influx transporter OCT1 (organic cation transporter 1) was thus inhibited by endosulfan, chlordane, heptachlor, lindane, and dieldrine, but not by dichlorodiphenyltrichloroethane isomers, whereas those of the canalicular efflux pumps MRP2 (multidrug resistance‐associated protein 2) and BCRP (breast cancer resistance protein) were blocked by endosulfan, chlordane, heptachlor, and chlordecone; this latter OC additionally inhibited the multidrug resistance gene 1 (MDR1)/P‐glycoprotein (P‐gp) activity. OCs, except endosulfan, were next found to induce MDR1/P‐gp and MRP2 mRNA expressions in hepatoma HepaRG cells; some of them also upregulated BCRP. By contrast, expression of sinusoidal transporters was not impaired (organic anion‐transporting polypeptide (OATP) 1B1 and OATP2B1) or was downregulated (sodium taurocholate co‐transporting polypeptide (NTCP) and OCT1). Such regulations of drug transporter activity and expression, depending on the respective nature of OCs and transporters, may contribute to the toxicity of OC pesticides.