Linking pesticide exposure to neurodegenerative diseases: An in vitro investigation with human neuroblastoma cells.

Although many organochlorine pesticides (OCPs) have been banned or restricted because of their persistence and linkage to neurodegenerative diseases, there is evidence of continued human exposure. In contrast, registered herbicides are reported to have a moderate to low level of toxicity; however, there is little information regarding their toxicity to humans or their combined effects with OCPs. This study aimed to characterize the mechanism of toxicity of banned OCP insecticides (aldrin, dieldrin, heptachlor, and lindane) and registered herbicides (trifluralin, triallate, and clopyralid) detected at a legacy contaminated pesticide manufacturing and packing site using SH-SY5Y cells. Cell viability, LDH release, production of reactive oxygen species (ROS), and caspase 3/7 activity were evaluated following 24 h of exposure to the biocides. In addition, RNASeq was conducted at sublethal concentrations to investigate potential mechanisms involved in cellular toxicity. Our findings suggested that aldrin and heptachlor were the most toxic, while dieldrin, lindane, trifluralin, and triallate exhibited moderate toxicity, and clopyralid was not toxic to SH-SY5Y cells. While aldrin and heptachlor induced their toxicity through damage to the cell membrane, the toxicity of dieldrin was partially attributed to necrosis and apoptosis. Moreover, toxic effects of lindane, trifluralin, and triallate, at least partially, were associated with ROS generation. Gene expression profiles suggested that decreased cell viability induced by most of the tested biocides was related to inhibited cell proliferation. The dysregulation of genes encoding for proteins with anti-apoptotic properties also supported the absence of caspase activation. Identified enriched terms showed that OCP toxicity in SH-SY5Y cells was mediated through pathways associated with the pathogenesis of neurodegenerative diseases. In conclusion, this study provides a basis for elucidating the molecular mechanisms of pesticide-induced neurotoxicity. Moreover, it introduced SH-SY5Y cells as a relevant in vitro model for investigating the neurotoxicity of pesticides in humans.

The effect of DDE and dieldrin and their parental substances (DDT and aldrin) on PGE2 and PGF2α release from bovine endometrial explants collected during 120 to 180 days of the gestational period.

Dieldrin and DDE are environmental metabolites of the organochlorine pesticides aldrin and DDT, respectively. During pregnancy, these chemicals can quickly infiltrate through the placental barrier, accumulate in amniotic fluid and fetus, and act as endocrine disruptors (EDs). The aim of this study was to investigate the effect of DDE and dieldrin and their parental substances at concentrations of 1 and 10 ng/ml on secretion of PGE2 and PGF2α from bovine endometrial explants (120-150 and 151-180 days of pregnancy) after 24 hr of incubation with EDs. The mRNA expression of COX2, PGES and PGFS and the concentrations of PGE2 and PGF2α were measured. EDs did not affect (p>0.05) COX2 gene expression, but DDT and DDE decreased (p⟨0.05) PGES expression and PGE2 secretion in the explants from 120-150 days of pregnancy. Depending on the dose, DDT and DDE increased (p⟨0.05) PGFS expression and PGF2α secretion from the explants from 120-150 days and decreased PGF2α secretion (p⟨0.05) from the explants from 151-180 days of pregnancy. Aldrin and dieldrin decreased (p⟨0.05) PGFS expression and PGF2α secretion from all explants. In summary, EDs disrupt the secretion of PGE2 and PGF2α by influencing the gene expression of PGES and PGFS.

Effects of DDT, DDE, aldrin and dieldrin on prostaglandin, oxytocin and steroid hormone release from smooth chorion explants of cattle.

Chlorooganic xenobiotics (XBs) such as DDT, DDE, aldrin and dieldrin interfere with release of hormones from chorionic villi that are necessary for sustaining the normal course pregnancy: prostaglandins (PGs), oxytocin (OT), progesterone (P4) and estradiol (E2). Approximately 20 %-40 % of these hormones originate from the smooth chorion. The aim of current studies was to investigate effects of these XBs on synthesis and release of PGE2, PGF2α, OT, P4 and E2 from explants of smooth chorion of cattle, obtained during the120-150 and 151-180 day gestational period. Explants were incubated with DDT, DDE, aldrin or dieldrin at concentrations of 1 and 10 ng/mL for 24 h, and concentrations of PGE2, PGF2α, OT, P4 and E2 in post incubation medium and the relative abundances of COX-2, PTGES, AKR1B1, NP-I/OT, PAM, HSD3B, and CYP19A1 mRNA transcripts in tissue explants were determined. The XBs did not have effects on cell viability in explants (P > 0.05), however, there were effects on prostaglandins, OT and P4 secretion and relative abundance of mRNA transcript for genes encoding the main enzymes involved in synthesis of these hormones (P < 0.05). The XBs that were evaluated did not have effects on E2 synthesis and secretion (P > 0.05). In summary, XBs evaluated in the present study had effects on the pattern of prostaglandin secretion, and can increase OT and P4 release from smooth chorion explants. Because XBs inhibit hormonal action throughout the chorion, there is an increase in risk of abortions or premature births in animals.

Stoichiometric gene-to-reaction associations enhance model-driven analysis performance: Metabolic response to chronic exposure to Aldrin in prostate cancer.

BACKGROUND: Genome-scale metabolic models (GSMM) integrating transcriptomics have been widely used to study cancer metabolism. This integration is achieved through logical rules that describe the association between genes, proteins, and reactions (GPRs). However, current gene-to-reaction formulation lacks the stoichiometry describing the transcript copies necessary to generate an active catalytic unit, which limits our understanding of how genes modulate metabolism. The present work introduces a new state-of-the-art GPR formulation that considers the stoichiometry of the transcripts (S-GPR). As case of concept, this novel gene-to-reaction formulation was applied to investigate the metabolic effects of the chronic exposure to Aldrin, an endocrine disruptor, on DU145 prostate cancer cells. To this aim we integrated the transcriptomic data from Aldrin-exposed and non-exposed DU145 cells through S-GPR or GPR into a human GSMM by applying different constraint-based-methods. RESULTS: Our study revealed a significant improvement of metabolite consumption/production predictions when S-GPRs are implemented. Furthermore, our computational analysis unveiled important alterations in carnitine shuttle and prostaglandine biosynthesis in Aldrin-exposed DU145 cells that is supported by bibliographic evidences of enhanced malignant phenotype. CONCLUSIONS: The method developed in this work enables a more accurate integration of gene expression data into model-driven methods. Thus, the presented approach is conceptually new and paves the way for more in-depth studies of aberrant cancer metabolism and other diseases with strong metabolic component with important environmental and clinical implications.

Do chlorinated insecticides (aldrin and DDT) or products of their transformations (dieldrin and DDE) impair signal transfer from regulators (oxytocin and relaxin) of bovine myometrium motility in vitro?

Aldrin, dieldrin, and DDT are chlorinated insecticides that are unintentionally widespread in the environment. It was previously shown that all of the aforementioned compounds increased secretion of ovarian oxytocin (OT), which is a potent uterotonic agent. However, only DDT and its metabolite (DDE) promoted, while aldrin and dieldrin inhibited basal and OT-stimulated myometrial contractions in cows. Therefore, the aim of this study was to determine the effect of these treatments on the reception and further transmission of the OT-signal for myometrial contractions and on the levels of contractile-associated integral proteins (caveolin; CAV) and gap junction proteins (GAPs). Moreover, their effect on reception of signal for the relaxation of myometrium was also studied. Myometrial strips or cells from non-pregnant (8-12 days of oestrous cycle) or late pregnant (5-8 months) cows were incubated with the studied compounds at environmentally relevant dose (10 ng/ml), which was chosen according to the previous studies. DDT and DDE increased the CAV protein level, while dieldrin decreased the GAPs level. None of the studied compounds affected mRNA expression of the OT receptor and expression of the second messengers (DAG, IP3, PKC, MLCK). Oppositely, DDE and dieldrin decreased mRNA expression of the relaxin (RLX) receptor. Changes in the amount of contractile-associated integral proteins may be involved in the molecular mechanism underlying the adverse effects of the studied insecticides on myometrial motility. Admittedly, none of the studied compounds impaired the reception or further intracellular transmission of the OT signal to promote contractions during the oestrous cycle, while they showed potential to impair the transmission the signal between cells as well as to diminish the effects of one of the primary inhibitor (RLX) of myometrial contractions during gestation.

The effects of environmentally relevant concentrations of aldrin and methoxychlor on the testes and sperm of male Clarias gariepinus (Burchell, 1822) after short-term exposure.

The organochlorine pesticides aldrin (0.14 µg/L) and methoxychlor (0.23 µg/L) were both present in the Albasini Dam, Limpopo Province, South Africa, during a field survey in 2014. The use of aldrin has been banned in the USA since 1987 and restricted in South Africa since 1992. The use of methoxychlor, however, remains undefined with little information available about its registration in South Africa despite being banned in Europe (2002) and USA (2003). The aim of this study was to determine the potential effects of environmentally relevant concentrations of aldrin and methoxychlor on the reproductive system of male catfish, Clarias gariepinus. Males were exposed for 96 h to the two pesticides under controlled laboratory conditions. Following exposure, each fish was weighed and measured, and a necropsy performed to determine any macroscopic abnormalities and the general health of the fish. The fish were killed and dissected and the testes removed, weighed and measured to determine the gonadosomatic index (GSI). The right testis of each fish was sectioned for histopathological assessment and to calculate the testes index (IT). The left testis was used for computer-assisted sperm analysis (CASA). The histopathological assessment of the testes showed histopathological changes such as of melano-macrophage centres (MMCs) and vacuolation of spermatogonia and spermatocytes. However, the classification of these changes indicated that the testes tissue structure was normal with slight histological changes. No statistically significant differences (p > 0.05) were found in the CASA parameters between exposure groups. The results of this study showed that the environmentally relevant concentrations of aldrin and methoxychlor did not have a negative effect on the motility of the mature sperm, but adverse effects were noted in the early stages of spermatogenesis, indicating possible effects over longer exposure periods.

Impact of co-exposure of aldrin and titanium dioxide nanoparticles at biochemical and molecular levels in Zebrafish.

Aldrin (ALD), a persistent-organic-pollutant (POP), an organochlorine-cyclodiene-pesticide is highly toxic in nature. Titanium dioxide nanoparticles (TNP) are widely used for various industrial applications. Despite the remarkable research on pesticide toxicity, the work with impact of nanoparticles on POP has been dealt with marginally. Chemicals co-exist in the environment and exhibit interactive effects. An investigation was carried out to evaluate the individual and combined effects of ALD (6 ppm) and TNP (60 ppm) exposure at sub-lethal concentration for 24 h in zebrafish. Significant reversal of lipid peroxidation level in liver and brain tissues and restoration in enhanced catalase activity in all examined tissues were observed in combined group. For other parameters, combined exposure of ALD and TNP does not show significant reversal action on ALD toxicity. Further studies are inline to understand combined effects of both to achieve significant reversal of ALD toxicity by TNP nanoparticles with threshold concentration of aldrin.

Toxicological profile of organochlorines aldrin and dieldrin: an Indian perspective.

Several epidemiological studies have suggested various environmental factors as a possible cause for increased incidence of various abnormalities. Of the various environmental contaminants, the most prevalent and the most discussed are the endocrine disrupting chemicals. Contact of such disruptors with humans has become inevitable today. They are cosmopolitan and present from agriculture to industrial sectors, even in day-to-day consumer products. Aldrin and dieldrin belong to one such class of substances which are known to have a toxic effect on various physiological systems of the human body. Despite an imposed ban on their manufacture and commercial use, these pesticides could still be detected in probable areas of consumption like agriculture. The present review discusses the known possible toxic effects of aldrin and dieldrin and their current existence in the ecosystem across India.

The adverse effects of aldrin and dieldrin on both myometrial contractions and the secretory functions of bovine ovaries and uterus in vitro.

Aldrin and dieldrin are chloroorganic insecticides which are recognised as endocrine disruptors. The aim of the study was to investigate their effect on the secretory functions of the uterus and ovary and on myometrial contractions. Myometrial strips and uterine and ovarian cells from nonpregnant cows were incubated with the xenobiotics (0.1, 1 or 10 ng/ml) for 24 or 72 h. Next, their effect on viability of myometrial, endometrial, granulosa and luteal cells, myometrial strip contractions, the synthesis and secretion of prostaglandins (PGs: PGF2α and PGE2) from uterine cells, the secretion of oestradiol (E2), testosterone (T) and oxytocin (OT) from granulosa cells and the secretion of progesterone (P4) and OT from luteal cells were determined. Neither of the xenobiotics (10 ng/ml) affected (P>0.05) the viability of the ovarian and uterine cells, while both (0.1-10 ng/ml) decreased (P<0.05) the basal and OT-stimulated myometrial contractions. In spite of these effects, neither of the insecticides affected (P>0.05) the synthesis and the secretion of PGs from the myometrial cells. Although they also did not impair the secretion of the PGs from the endometrial cells, they abolished (P<0.05) the stimulatory effect of OT (P<0.05) on the secretion of the PGs and stimulated (P<0.05) the secretion of OT from the granulosa and luteal cells. Moreover, aldrin and dieldrin stimulated secretion of E2 and T from the granulosa cells, while only dieldrin increased (P<0.05) the secretion of P4 from luteal cells. The data show that aldrin and dieldrin stimulated the secretory function of the cultured granulosa and luteal cells and inhibited the myometrial contractions of cows in vitro, which may affect on natural parturition.

Role of a γ-aminobutryic acid (GABA) receptor mutation in the evolution and spread of Diabrotica virgifera virgifera resistance to cyclodiene insecticides.

The western corn rootworm, Diabrotica virgifera virgifera, is a damaging pest of cultivated corn that was controlled by applications of cyclodiene insecticides from the late 1940s until resistance evolved ∼10 years later. Range expansion from the western plains into eastern USA coincides with resistance development. An alanine to serine amino acid substitution within the Rdl subunit of the gamma-aminobutyric acid (GABA) receptor confers resistance to cyclodiene insecticides in many species. We found that the non-synonymous single nucleotide polymorphism (SNP) G/T at the GABA receptor cDNA position 838 (G/T(838)) of D. v. virgifera resulted in the alanine to serine change, and the codominant SNP allele T(838) was genetically linked to survival of beetles in aldrin bioassays. A phenotypic gradient of decreasing susceptibility from west to east was correlated with higher frequencies of the resistance-conferring T(838) allele in the eastern-most populations. This pattern exists in opposition to perceived selective pressures since the more eastern and most resistant populations probably experienced reduced exposure. The reasons for the observed distribution are uncertain, but historical records of the range expansion combined with the distribution of susceptible and resistant phenotypes and genotypes provide an opportunity to better understand factors affecting the species' range expansion.

Organochlorine pesticides and polychlorinated biphenyls in sediments and fish from freshwater cultured fish ponds in different agricultural contexts in north-eastern France.

Organochlorine pesticides (HCB, HCH with α-, ß-, and γ isomers, heptachlor, cis-heptachlor epoxyde, trans-heptachlor epoxyde, endosulfan with α- and ß isomers, sulfate endosulfan, o,p'-DDT, p,p'-DDT, o,p'-DDE, p,p'-DDE, o,p'-DDD, p,p'-DDD, chlorothalonil, alachlor, aldrin, dieldrin, methoxychlor, oxychlordane, chlordane with α- and γ isomers, p,p'-dicofol and o,p'-dicofol) and indicators PCBs (IUPAC nos. 28, 52, 101, 118, 138, 153, and 180) were studied both in sediments and muscles of farmed fish species (Cyprinus carpio and Perca fluviatilis). Samples were collected from fish ponds located in the hydrographic basin of the Moselle River (Lorraine Region, France). OCPs and PCBs were present at low concentrations both in sediments and fish muscles. Concerning sediments, ∑DDTs revealed concentrations ranging from 0.2 to 2.30 ng g(-1) dw and ∑PCBs ranged from 0.3 to 3.5 ng g(-1) dw. Concerning fish muscles, the highest concentrations in OCPs were those of p,p'-DDE, with average concentrations of 0.57±0.44 ng g(-1) ww for carp and 0.58±0.29 ng g(-1) ww for perch. The contamination profiles proved to be different depending on the fish species. Indeed, HCH-isomers, HCB, and dieldrin were detected only for the carp and always at low concentrations. For example, the highest concentration of HCHs was observed for ß-HCH with a mean value of 0.64±0.15 ng g(-1) ww for carp. As for PCBs, the levels of ∑PCBs ranged from 0.3 to 6.4 ng g(-1) ww in carp muscles and from 0.90 to 5.60 ng g(-1) ww in perch muscles.

Enantioselective behavior of alpha-HCH in mouse and quail tissues.

alpha-HCH (hexachlorocyclohexane) is chiral and can still be detected in almost all environmental media. In this study, the enantioselective behavior of alpha-HCH in mice (CD1) and quail (Coturnix japonica) was investigated and compared after a single dose of exposure. The primary nerve cell culture was conducted to evaluate the enantioselective metabolic capacity of nerve cells of mouse and quail for alpha-HCH. In various tissues of the mice and quail, the alpha-HCH concentrations showed a typical pattern of first-order dynamics after exposure. The enantiomeric fractions (EFs) in nonbrain tissues of mice decreased substantially, indicating continuous depletion of (+)-alpha-HCH in mice. Tissue-specific EF trends in quail and enantioselective degradation of (-)-alpha-HCH in quail liver were observed. These observations indicated that the dynamic changes of EFs in mice and quail were independent of concentration changes in the same tissues. In brain tissues, the enantioenrichment of (+)-enantiomer was totally independent of their concentrations in blood. The in vitro metabolism of alpha-HCH in the primary nerve cells were negligible, and the slight EF changes in primary nerve cells demonstrated that metabolism, uptake, and excretion in the brain cells would not lead to the observed dramatic enantioenrichment of (+)-alpha-HCH in the brain tissues of the two animals. The enantioselective transport across the blood-brain barrier was the primary cause for the enantioenrichment of (+)-alpha-HCH in the brain tissues.

Empirical and parametric likelihood interval estimation for populations with many zero values: application for assessing environmental chemical concentrations and reproductive health.

BACKGROUND: Understanding the health effects associated with environmental chemicals is challenging when individuals have concentrations at or below the laboratory limits of detection as well as when the values may round to zero or are presented in the form of 0 to substitute for missing values, which may result in many zeros in the database. Comparison of mean concentrations between individuals with and without disease necessitates estimation procedures that allow for data with many zero values. The main aim of this article is to propose and examine parametric and distribution-free methods for comparing data sets containing many zero observations. An important application of this approach is related to assessing environmental chemical concentrations and reproductive health. METHODS: We extended the empirical likelihood technique for estimating confidence intervals (CIs) in data sets with many zeros. We examined the proposed empirical likelihood interval estimations via a broad Monte Carlo study that compares the proposed method with parametric techniques. Certain characteristics of Monte Carlo simulations were chosen to be close to parameters of the real data set. We applied the method to a cohort study comprising 84 women aged 18-40 years who had undergone laparoscopy between 1999 and 2000 in whom serum concentrations of 2 organochlorine pesticides--Aldrin and beta-Benzene hexachloride (ß-BHC) were measured using gas chromatography with electron capture. RESULTS: When applied to the cohort study, the method produced efficient 95% CIs, allowing for the comparison of mean serum Aldrin concentrations for women with and without endometriosis (0.000338, 0.003561) and (0.000803, 0.004211), respectively. Mean ß-BHC concentrations also could be compared (0.000493, 0.005869) and (0.000680, 0.003807) based on individuals with and without the disease, respectively. Differences in mean concentrations for Aldrin and ß-BHC could be estimated (-0.001563, 0.003025) and (-0.003522, 0.002890), respectively. CONCLUSIONS: We found the empirical likelihood method for estimating CIs robust when data sets contain many zeros. In so doing, mean concentrations of Aldrin or ß-BHC did not differ by endometriosis diagnosis.

A whole sample toxicity assessment to evaluate the sub-lethal toxicity of water and sediment elutriates from a lake exposed to diffuse pollution.

The impact of diffuse pollution in aquatic systems is of great concern due to the difficult to measure and regulate it. As part of an ecological risk assessment (ERA), this study aims to use a whole sample toxicity assessment to evaluate the toxicity of water and sediment from Lake Vela, a lake that has been exposed to diffuse pollution. In this way, standard (algae: Pseudokirchneriella subcapitata; cladoceran: Daphnia magna) and local species (algae: Aphanizomenon flos-aquae; cladoceran: Daphnia longispina) were exposed to surface water, and sediment elutriates were collected seasonally from two sites at Lake Vela: one near the east bank (ES), surrounded by agricultural lands; and the other near the west bank (WS), surrounded by a forest. The results confirmed the seasonal contamination of both environmental compartments by pesticides, including organochlorine pesticides, and the presence of high concentrations of nutrients. Although both sites were contaminated, higher levels of pesticides and nutrients were detected in ES, particularly in the sediments. Bioassays showed that water samples (100% concentration) collected in summer and autumn significantly affected the growth rate of P. subcapitata, which could be attributed to the presence of pesticides. Likewise, they revealed an apparent toxicity of elutriates for P. subcapitata and for both daphnids, in summer and autumn. In fact, although pesticides were not detected in elutriates, high levels of un-ionized ammonia were recorded, which is considered highly toxic to aquatic life. By comparing the several species, P. subcapitata was revealed to be the most sensitive one, followed by the daphnids, and then by A. flos-aquae. Results obtained in this study underlined the importance of whole samples toxicity assessment for characterizing the ecological effects of complex mixtures from diffuse inputs, in the ERA processes.

Discriminating the molecular basis of hepatotoxicity using the large-scale characteristic molecular signatures of toxicants by expression profiling analysis.

Predicting the potential human health risk posed by chemical stressors has long been a major challenge for toxicologists, and the use of microarrays to measure responses to toxicologically relevant genes, and to identify selective, sensitive biomarkers of toxicity is a major application of predictive and discovery toxicology. To investigate this possibility, we investigated whether carcinogens (at doses known to induce liver tumors in chronic exposure bioassays) deregulate characteristic sets of genes in mice. Male C3H/He mice were dosed with two hepatocarcinogens (vinyl chloride (VC, 50-25 mg/kg), aldrin (AD, 0.8-0.4 mg/kg)), or two non-hepatocarcinogens (copper sulfate (CS, 150-60 mg/kg), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T, 150-60 mg/kg)). Large-scale molecular changes elicited by these four hepatotoxicants in liver tissues were analyzed using DNA microarray. Three days after administration, no significant phenotypic changes were induced by these four different hepatotoxicants in terms of histological examination or blood biochemical assay. However, unsupervised hierarchical analysis of gene expressional changes induced by hepatotoxicants resulted in two major gene subclusters on dendrogram, i.e., a carcinogen (VN, AD) and non-carcinogen group (CS, 2,4,5-T), and also revealed that distinct molecular signatures exist. These signatures were founded on well-defined functional gene categories and may differentiate genotoxic and non-genotoxic carcinogens. Furthermore, Venn diagram analysis allowed us to identify carcinogen and non-carcinogen-associated molecular signatures. Using statistical methods, we analyzed outlier genes for four different classes (genotoxic-, non-genotoxic-carcinogen, genotoxic-, non-genotoxic non-carcinogen) in terms of their potential to predict different modes-of-action. In conclusion, the identification of large-scale molecular changes in different hepatocarcinogen exposure models revealed that different types of hepatotoxicants are associated with different epigenetic changes and molecular pathways and that these large-scale characteristic molecular changes could be used as predictable toxicity markers.

Effect of aldrin on life history characteristics of rotifer Brachionus calyciflorus pallas.

The organochlorine insecticide aldrin is commonly used in intensive agriculture, and demonstrates estrogenic activity. Rotifers such as Brachionus calyciflorus are favored test animals in aquatic toxicology because of their more sensitivity to most toxicants. In the tested concentration range of 0.04-1.28 mg/L, aldrin shortened significantly the durations of embryonic development. Lower concentrations of aldrin had an intriguing effect on the reproduction of the rotifers and are beneficial to their survival. Different endpoints of both development and reproduction had different sensitivity to aldrin. The reproductive endpoint of the rotifers is more sensitive to aldrin than the developmental endpoint.

The toxic potential of aldrin and heptachlor on Danio rerio juveniles (Cypriniformes, Cyprinidae).

With the objective of evaluating the effects of organochlorine pesticides (aldrin and heptachlor) on the survival, growth and gill morphology of juvenile zebrafish (Danio rerio), four partial chronic toxicity bioassays were conducted (seven days' duration) with both compounds in a semi-static system with renewal every 24 h. The results did not show any effects on the fish's survival, but did on their growth and gill morphology. Aldrin was more toxic than heptachlor, since the chronic value (CV) for growth was nearly 8.7 times less, a result confirmed by analyzing the histology of the gills, in which the changes detected were considered more severe for the former substance. In general, they were found branchial lesions of first stage, in other words, cell proliferation between the secondary lamellae, hyperplasia, lifting of respiratory epithelial cells; fusion of several secondary lamellae and dilation of blood vessels. Aneurysms (alterations of second stage) they were verified in the exposed organisms to the aldrin, which also presented about 10% of reduction in the standard length and 30% of reduction in the total weight in relation to the exposed organisms to the control. For heptachlor these values were of 8% and 25%, respectively. The intensity of the gill lesions and growth of the fish did not depend on the pesticide concentration, suggesting different modes of action of the products.

Biomarkers of oxidative stress in Wallago attu (Bl. and Sch.) during and after a fish-kill episode at Panipat, India.

The present investigation was carried out by sampling water, sediment and fish during a fish-kill episode at Panipat (Haryana, India), and again sampling at the same site was conducted after a gap of two months. During the second sampling no fish-kill was observed and the water was relatively less turbid and clear. Antioxidant profile and lipid peroxidation (LPO) in fish tissues were studied. Analysis was also carried out on the physico-chemical characteristics of water samples along with heavy metal and pesticide analysis in water and sediment samples during and after the episode. Dissolved oxygen level was substantially low during the fish-kill episode. Heavy metals (copper and chromium) and pesticides like BHC (Benzene hexachloride), DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane) were also detected during the episode in water and sediment samples. Various oxidative stress biomarkers in liver, kidney and gill tissues in the Indian freshwater fish Wallago attu (Bl. and Sch.) collected from the site were investigated. The levels of reduced glutathione and non-protein thiol were significantly (P<0.001) higher in the liver of Wallago attu collected from Panipat after the fish-kill episode. Ascorbic acid levels in all the tissues did not change significantly after the episode. The LPO in liver, kidney and gills was significantly low (P<0.01-0.001) in all tissues of fish collected after the fish-kill episode. The protein carbonyl levels were significantly low (P<0.05-0.01) in all the fish organs sampled after the fish-kill episode. The findings suggest that industrial effluent may result in the massive loss of a commercial commodity. The simultaneous measurement of the physicochemical parameters of the water samples showed a good correlation between the biomarkers responses and the environmental chemical stress conditions.

Determination of oxidative metabolism in Collembolan Proisotoma minuta (Tullberg).

An oxidative metabolism of Collembolan Proisotoma minuta was determined with a model compound of aldrin and dieldrin in this paper. The seven-day LD(50) values for aldrin, dieldrin, and piperonyl butoxide in salt solution were 0.496, 0.367, and 8.346 mg L(-1), respectively. When P. minuta were exposed to aldrin, dieldrin was the sole metabolite. The conversion of aldrin to dieldrin was known to be catalyzed by P450 monooxygenases system. It has been shown that the synergist piperonyl butoxide inhibited the metabolism of aldrin in P. minuta.

Cyclodiene organochlorine insecticide-induced alterations in the sulfur-redox cycle in CHO-K1 cells.

The effect of the cyclodiene organochlorine pesticides aldrin, dieldrin and endosulfan was assessed on CHO-K1 cultures at fractions of their lethal doses, determined by the neutral red (NRI) incorporation assay (NRI6.25, NRI12.5 and NRI25). Glutathione peroxidase, reductase and S-transferase, and total and oxidised glutathione were evaluated along the standard growth curve of the cultures. After a 24-h incubation with each insecticide, glutathione peroxidase incurred a large increase, while glutathione reductase and S-transferase activities were slightly higher than untreated controls. Unlike oxidised glutathione, the content of total glutathione declined significantly after exposure to cyclodiene insecticides. Changes in cell membrane integrity were assessed by the lactate dehydrogenase (LDH) release assay and lipid peroxidation for a wide range of pesticide concentrations. Membrane leakage and peroxide production were significantly enhanced at concentrations of aldrin and as low as 12.5 microg/ml, whereas dieldrin and endosulfan increased membrane fragility at much higher concentrations.