L'exploitation des terres à des fins agricoles entraîne la présence de nombreux contaminants dans les milieux naturels, ce qui peut nuire à la faune qu'on y retrouve. Nous avons émis l'hypothèse que la présence de contaminants agricoles aurait un impact sur le métabolisme osseux du ouaouaron (Rana catesbeiana), puisque des difformités au niveau des pattes avaient auparavant été observées chez certains individus. Les grenouilles ont été capturées à l'été 2008 en six sites situés aux abords de la rivière Yamaska et choisis selon un gradient d'activité agricole. Elles ont par la suite été disséquées et leurs fémurs et phalanges ont été analysés à l'aide d'une technique d'imagerie numérique, grâce à l'appareil Skyscan 1172. Les phalanges des individus ont été analysées en histologie afin d'estimer leur âge par squelettochronologie. Une analyse sérique des principaux marqueurs du métabolisme osseux a aussi été effectuée. Nos résultats ont démontré que l'activité de la phosphatase alcaline augmentait en fonction du gradient d'exploitation agricole, supposant une atteinte hépatique. Aussi, les ouaouarons provenant du site de plus forte activité agricole présentaient un volume osseux réduit au niveau de la portion trabéculaire de la phalange, ainsi que de la portion corticale du fémur. De plus, les grenouilles capturées à ce site étaient significativement plus jeunes. Nous avons aussi voulu vérifier l'impact de certains pesticides au niveau cellulaire, en utilisant la lignée ostéoblastique murine MC3T3-E1 et la lignée ostéoblastique humaine MG-63. Il s'est avéré que l'atrazine et le métolachlore n'étaient pas cytotoxiques pour les cellules aux concentrations utilisées. Par contre, l'endosulfan, un pesticide organochloré, a été démontré cytotoxique, sa CL50 ayant été mesurée à 42,4µM. Nous avons par la suite tenté de documenter son mécanisme de toxicité par des mesures au cytofluoromètre. Nous avons pu démontrer par un marquage aux sondes PI et Annexine V que l'endosulfan induisait l'apoptose et la nécrose. De plus, le potentiel mitochondrial subit une dépolarisation forte et rapide après traitement à l'endosulfan, ce qui a été mesuré grâce à la sonde JC-1. Bien que l'apoptose soit induite par l'endosulfan et que le potentiel mitochondrial chute, nous n'avons pas été en mesure de mettre en évidence la production de ROS par marquage à la sonde H2DCFDA. Par contre, l'ajout de trois antioxydants différents, soit la vitamine C, la vitamine E et la N-acétyl-cystéine au milieu de culture a significativement réduit la mortalité, ce qui laisse présumer que la production de ROS serait à la base du mécanisme de toxicité de ce pesticide, mais que le marquage au H2DCFDA n' aurait pu le révéler. Le métabolisme osseux est souvent ignoré lors d'études toxicologiques. Ce projet ayant permis de révéler l'impact de contaminants agricoles sur le métabolisme osseux du ouaouaron ainsi que sur des cellules ostéoblastiques indique que cet aspect ne devrait pas être négligé lors de futures études. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : ostéoblaste, apoptose, endosulfan, ouaouaron, pesticides.

Several epidemiological studies have suggested a role for environmental chemicals in the etiology of neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Endosulfan (an organochlorine) and zineb (zinc-ethylene-bis-dithiocarbamate) are used as pesticides on a variety of crops worldwide and pose potential health risks to humans and animals. Both endosulfan and zineb are known to affect nervous system. Because the dopaminergic system continues to develop postnatally, we hypothesized that developmental exposure to endosulfan or zineb alone or in combination would result in alteration of nigrostrial neurotransmitters and would render the nigrostrial dopamine system more susceptible to chemical challenge later in life. The objectives of this study were (1) to determine the effects of endosulfan and zineb individually and in combination on dopaminergic or cholinergic pathways in vivo, (2) to investigate the effects of exposure to endosulfan, zineb and their mixtures administered in early life (during brain development) on subsequent exposure to these pesticides on the dopaminergic and cholinergic systems, in vivo, (3) to investigate the mechanism(s) of induction of neuronal cell death caused by these pesticides using human neuroblastoma SH-SY5Y cells in culture, (4) to define the role of oxidative stress in pesticide-induced neuronal cell death in vitro. Male C57Bl/6 mice of 7-9 months old exposed to zineb (50 and 100 mg/kg), endosulfan (1.55, 3.1 and 6.2 mg/kg) and their mixtures every other day over a 2-week period exhibited higher levels of dopamine accumulation in the striatum. Both pesticide-treated groups displayed significantly lower norepinephrine levels in the striatum (Ï ≤ 0.05) than the controls. The developmental exposure to zineb, endosulfan and their combination enhanced the vulnerability to subsequent neurotoxic challenges occurring later in life. Thus, C57BL/6 mice exposed to zineb, endosulfan and their mixtures as juveniles (postnatal days 5 to 19) and re-exposed at 8 months of age showed a significant depletion of striatal dopamine, to 22%, 16%, and 35% of control, respectively. Acetylcholinesterase activity in the cerebral cortex was found to be significantly increased in all pesticide treated groups. Mice given mixtures of pesticides also showed significantly increased levels of normal and aggregated alpha-synuclein, a hallmark of neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. The results of these studies indicate that exposure to these pesticides as neonates and re-exposure as adults could result in neurochemical changes that did not reveal at adulthood when the exposure was at juvenile age only. We further investigated the mechanism(s) of activation of pesticide-induced neuronal cell death in vitro. The characteristic of cell death in SH-SY5Y human neuroblastoma cells was examined. These cells are known to retain catecholaminergic phenotype. Cells were exposed to endosulfan, zineb and mixtures of two pesticides, in concentrations ranging from 50 μM to 400 μM. These exposures caused both apoptotic and necrotic cell death in SH-SY5Y cells as evaluated by lactate dehydrogenase release, 7-aminoactinomycin-D and Annexin-V/PI assays. Exposure to mixtures of the pesticides enhanced both the early apoptosis and late apoptosis/necrosis compared to either chemical alone. Visual evaluation using DNA ladder assay and fluorescence Annexin V/PI assay confirmed the contribution of both apoptotic and necrotic events. Furthermore, endosulfan and zineb alone and in combination altered the caspase-3 activity indicating that both pesticides exposure exert their apoptotic effect via the caspase-3 pathway. Because there has been increasing evidence of the role of reactive oxygen species (ROS) and oxidative stress in pesticide-induced neuronal cell death (apoptosis and necrosis), the levels of ROS and antioxidant enzymes were examined. Cells treated with pesticides were found to enhance the generation of superoxide anion and hydrogen peroxide both in a dose- and time-dependent manner. Mixture of pesticides significantly enhanced the production of these reactive oxygen species compared to cells exposed to individual pesticide. Cells treated with pesticides showed a decrease in superoxide dismutase, glutathione peroxidase, and catalase levels. These pesticides also induced lipid peroxides (thiobarbituric acid reactive products) formation in SH-SY5Y cells. Furthermore, cells exposed to these pesticides were found to have increased in the expression of NFkappaB activity in the nucleus. These data support the hypothesis that oxidative stress was induced in neuronal cells by exposing to these pesticides in vitro. Taken together, the results of this study support the above hypothesis and suggest that the cytotoxicity of endosulfan and zineb and their combinations may, at least in part, be associated with the generation of ROS. Furthermore, mice exposed at early age and re-exposed at adulthood become more susceptible to alteration of neurotransmitter levels compared to mice exposed to these pesticides only as juveniles. These findings could add to the growing body of knowledge on the mechanism of pesticide-induced dopaminergic neuronal cell death and could hold tremendous implication for the future understanding of the possible involvement of environmental risk factors in the pathogenesis of Parkinson's disease. / Ph. D.

L'influence du vétiver sur le devenir des polluants métalliques (Cu et Cd) et organique (insecticide : endosulfan) dans le système sol-eau-vétiver a été étudiée dans deux sols représentatifs du Burkina Faso, un sol vertique et un sol ferrugineux. Des études d'adsorption en batch, des essais en vase de végétation en conditions contrôlées et en lysimètres en conditions naturelles ont été réalisées. Les sols plantés au vétiver montrent des coefficients d'adsorption (Kf) de l'endosulfan significativement plus élevés comparés à ceux des sols non plantés. En vases de végétation, le temps de demi-vie estimé de l'endosulfan dans les sols plantés est 1,1 fois moins important comparé aux sols non plantés, en relation avec l'augmentation significative de la microflore dégradant l'endosulfan en présence du vétiver. Par ailleurs, les teneurs en Cu et Cd dans le vétiver sont significativement plus élevées pour le sol ferrugineux que pour le sol vertique 6 mois après traitement. Le coefficient d'absorption biologique (CAB) de Cu et Cd et leurs facteurs de bioconcentration (FBC) et de translocation (FT) supérieurs à 1 dans le vétiver démontrent son efficacité à absorber et à accumuler Cu et Cd. En conditions extérieures, 6 mois après le traitement, les quantités lessivées de Cu sont très faibles pour l'ensemble des colonnes. Pour les sols plantés au vétiver, les quantités lessivées d'endosulfan et de Cd sont plus de deux fois inférieures à celles des sols non plantés, en relation avec leur adsorption plus élevée et la dégradation favorisée de l'endosulfan dans les sols plantés. / The influence of vetiver on the fate of two heavy metals (Cu and Cd) and an insecticide (endosulfan) in the soil-water-vetiver system was studied in two representative soils of Burkina Faso, a vertisol and a lixisol. Batch adsorption coefficient measurements, pot and leaching experiments, under phytotronic and outdoor conditions, respectively, were conducted. For both soils, adsorption coefficient values (Kf) of endosulfan, Cu and Cd were higher on planted soils than on unplanted soils. In the pot experiments, the estimated endosulfan half-life (DT50) in planted soils was 1.1-fold lower compared to unplanted soils, in accordance with the significant increase of the microflora degrading endosulfan in planted soils. Moreover, the Cu and Cd contents in vetiver grass were significantly higher for the lixisol than for the vertisol. The biological absorption coefficient (BAC), the bioconcentration and translocation factors (BCF and TF, respectively) of Cu and Cd were greater than 1 in vetiver, that showed the efficiency of vetiver to absorb and accumulate Cu and Cd. In outdoor conditions, 6 months after treatment, the leached Cu amounts were very low for all columns. In the presence of vetiver, the Cd and endosulfan leached amounts were more than 2-fold lower than those of unplanted soils, related to their higher adsorption and the faster degradation of endosulfan in planted soils.

Juvenile male blue crabs move into the tidal, freshwater James River during warmer months to feed and grow by undergoing molting. In crustaceans, growth and molting are hormonally controlled. The physiological effects of a multiple-stressor environment are determined by comparing the blue crab’s oxygen uptake after exposure to pure sand, James River sediment, or endosulfan-spiked sand. The effect of multiple stressors on molting is measured by the activity level of N-acetyl-ß-glucosaminidase (NAG), an enzyme in epidermal tissue important to molting. The oxygen uptake was decreased by exposure to James River sediment but not for exposure to endosulfan for seven days. Exposure to James River sediments over two days caused a similar suppression of epidermal NAG activity as exposure to endosulfan. These results indicate that the blue crab’s exposure to James River sediments and moderate hypoxia has the potential to cause short-term effects on physiology and long-term effects on growth.

Le vétiver a été choisi pour former des haies autour des parcelles cotonnières et maraîchères du Burkina Faso, afin de limiter la dispersion des polluants vers les plantes, les eaux de surface et souterraines. L'influence du vétiver sur le devenir des polluants métalliques (Cu et Cd) et organique (insecticide : endosulfan) dans le système sol-eau-vétiver a été étudiée dans deux sols représentatifs du Burkina Faso, un sol vertique et un sol ferrugineux. Des études d'adsorption en batch, des essais en vase de végétation en conditions contrôlées et en lysimètres en conditions naturelles ont été réalisées. Les sols plantés au vétiver montrent des coefficients d'adsorption de l'endosulfan (Kf = 6,5 - 9,7 mg1-n Ln kg-1), de Cu (3162,6 - 4445,1 mg1-n Ln kg-1) et de Cd (76,8 - 280,2 L kg-1) significativement plus élevés comparés à ceux des sols non plantés (Kf = 6,3 - 7,2, 328,1 - 2921,9 et 37,2 - 206,1 mg1-n Ln kg-1, respectivement). De plus, le sol vertique adsorbe plus ces micropolluants que le sol ferrugineux, en relation avec sa CEC plus importante résultant de ses plus fortes teneurs en carbone organique et en argiles. En vases de végétation, 6 mois après traitement, l'endosulfan n'est détecté ni dans les parties aériennes et racinaires du vétiver, ni dans les sols. La non-détection de l'endosulfan pourrait s'expliquer par sa dégradation, sa minéralisation, sa volatilisation et la formation de résidus non extractibles. Dans nos conditions, le temps de demi-vie estimé de l'endosulfan (DT50) est supérieur dans le sol ferrugineux (43,1 jours) comparé à celui du sol vertique (40,6 jours). Dans les sols plantés, le DT50 de l'endosulfan est 1,1 fois moins important comparé aux sols non plantés, en relation avec l'augmentation significative de la microflore dégradant l'endosulfan en présence du vétiver. Par ailleurs, les teneurs en Cu et Cd dans le vétiver sont significativement plus élevées pour le sol ferrugineux que pour le sol vertique 6 mois après traitement, en relation avec les teneurs en CuMgCl2 et CdMgCl2 supérieures dans le sol ferrugineux comparé au sol vertique. Le coefficient d'absorption biologique (CAB) de Cu et Cd et leur facteur de bioconcentration (FBC) supérieurs à 1, ainsi que leur facteur de translocation (FT) inférieure à 1 dans le vétiver démontrent son efficacité à absorber et à accumuler Cu et Cd. En conditions extérieures, 6 mois après le traitement, les quantités lessivées de Cu sont très faibles pour l'ensemble des colonnes (0,001 - 0,010%). Pour les deux autres micropolluants, les pertes par lessivage à travers le sol ferrugineux (0,07% d'endosulfan et 1,48% de Cd) sont supérieures à celles du sol vertique (0,01% d'endosulfan et 0,04% de Cd). En présence du vétiver, les quantités lessivées d'endosulfan et de Cd sont plus de deux fois inférieures à celles des sols non plantés, en relation avec leur adsorption plus élevée et la dégradation favorisée de l'endosulfan dans les sols plantés. L'efficacité du vétiver à réduire le transfert des micropolluants vers les eaux de surface et souterraines nécessiterait d'être validée à l'échelle de la parcelle avant d'encourager la plantation de haies autour des parcelles cotonnières et maraîchères du Burkina Faso.

The focus of this thesis was to investigate developmental neurotoxic effects of different persistent and non-persistent environmental pollutants, alone or in binary mixtures, when exposure occurs during a critical period of brain development, in mice. The compounds investigated included a perfluorinated compound, perfluorohexane sulphonate (PFHxS), and four different pesticides, endosulfan, cypermethrin, chlorpyrifos and carbaryl. Both persistent and non-persistent pollutants are detected in the environment and in humans, which shows that exposure to these compounds is occurring in real life. Humans can therefore be exposed to various pollutants during their whole lifetime, starting from the gestational period to adulthood. Furthermore, exposure to environmental pollutants is rarely exclusive to a single compound, but rather occurs through combinations of various pollutants present in the environment. Exposure to environmental pollutants during human brain development have been suggested to be a possible cause for neuropsychiatric disorders, such as autism spectrum disorder (ASD) and attention deficit hyperactivity disorder (ADHD). Previous studies have shown that chemicals can induce irreversible disorders in brain function when exposure to these chemicals occurs during a critical defined period of the brain development known as the brain growth spurt (BGS). The BGS is characterized by a rapid growth and development of the immature brain. In humans, and mice, this period also overlaps the lactation period indicating that newborns and toddlers can be exposed via mothers’ milk as well. This thesis has shown that a single oral exposure to PFHxS, endosulfan, cypermethrin, chlorpyrifos or carbaryl can induce developmental neurotoxic effects in mice, when exposure occurs during a critical period of brain development. These effects are manifested as persistent altered adult spontaneous behavior in a novel home environment, modified habituation, altered susceptibility of the cholinergic system and changed levels of neuroproteins in the mouse brain. Furthermore, a single neonatal co-exposure to a binary mixture of carbaryl/chlorpyrifos or PFHxS/endosulfan can interact and exacerbate the adult behavioral effects. These effects were seen at dosages were the single compound did not elicit a response or induced a much weaker behavioral effect. This indicates that risk assessments conducted on single compounds might underestimate interaction effects of mixtures when co-exposed.

Thesis (MTech (Chemical Engineering))--Peninsula Technikon, Cape Town, 2003 / The monitoring of pesticide contamination in surface and groundwater is an essential aspect of an assessment of the potential environmental and health impacts of widespread pesticide use. Previous research in three Western Cape farming areas found consistent (37% to 69% of samples) pesticide contamination of rural water sources. However, despite the need, monitoring of pesticides in water is not done due to lack of analytical capacity and the cost of analysis in South Africa. The Solid Phase Microextraction (SPME) sampling method has been developed over the last decade as a replacement for solvent-based analyte extraction procedures. The method utilizes a short, thin, solid rod of fused silica coated with an absorbent polymer. The fibre is exposed to the pesticide contaminated water sample under vigorous agitation. The pesticide is absorbed into the polymer coating; the mass absorbed depends on the partition coefficient of the pesticide between the sample phase and the polymeric coating, the exposure time and factors such as agitation rate, the diffusivity of the analyte in water and the polymeric coating, and the volume and thickness of the coating. After absorption, the fibre is directly inserted into the Gas Chromatograph (GC) injection port for analysis. For extraction from a stirred solution a fibre will have a boundary region where the solution moves slowly near the fibre surface and faster further away until the analyte is practically perfectly mixed in the bulk solution by convection. The boundary region may be modelled as a layer of stationary solution surrounded by perfectly mixed solution.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-09-16T15:27:12Z No. of bitstreams: 2 Dissertação - Marcia Felipe Mendes - 2016.pdf: 3733938 bytes, checksum: a83e0262d1671e990c617cd0061f8d2f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-09-16T15:27:41Z (GMT) No. of bitstreams: 2 Dissertação - Marcia Felipe Mendes - 2016.pdf: 3733938 bytes, checksum: a83e0262d1671e990c617cd0061f8d2f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-09-16T15:27:41Z (GMT). No. of bitstreams: 2 Dissertação - Marcia Felipe Mendes - 2016.pdf: 3733938 bytes, checksum: a83e0262d1671e990c617cd0061f8d2f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-02-26 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Because of the toxicological relevance of pesticides introduced into the environment through agricultural practices, it is necessary to develop simple methodologies with low cost, enabling remove these pollutants from the environment, in particular of water intended for human consumption. The adsorption process is an efficient and low cost technique that have wide application. The adsorbent capacity of the leaves of Typha angustifolia L. herbaceous perennial plant belonging to the family Typhaceae, in Brazil it is also popularly known as taboa, was investigated for the removal of pesticides trifluralin, chlorpyrifos and α-endosulfan from aqueous medium. The the analytical method using solid phase extraction (SPE) and chromatography with electron capture detector (GC/ECD) was optimizates and validate. The validation criterions were reached and the analytical methos was considered efficient for trifluralin, chlorpyrifos and α-endosulfan quantification in an aqueous medium, providing credibility to the results obtained in the adsorption experiments. The adsorption capacity of T. angustifolia was evaluated under different parameters: treatment of the material, amount of material, contact time between the plant material and the pesticides solution and concentration of the pesticides solution. The results showed that treatment of the material with distilled water and was satisfactory and, for all doses evaluated the pesticide removal was efficient, with best results obtained at a dose of 3.0 g L-1 , equivalent to 60 mg of adsorbent in 20.0 mL of solution, in which the pesticide removal were between 73 and 80%. The adsorption kinetics of the pesticide by T. angustifolia showed that the adsorption equilibrium was reached in 40 minutes for all pesticides, removing up to 90% and using a very small amount of plant material (3 g L-1 ). The experimental data were evaluated using the kinetic models of pseudo-first order, pseudo-second order and Avrami. One of the limitations encountered in the concentration influence study is the limited solubility of the agrochemical in aqueous medium (between 0.33 and 1.00 mg L-1 ), resulting in the study of a small concentration range in the adsorption experiments. To study the pesticides adsorption mechanism by T. angustifolia material, the experimental data were applied to Langmuir, Freundlich and Sips non-linear isotherms models. Additional tests showed that the adsorption capacity is unaffected when using natural water in the adsorption experiments. The results indicate that the material obtained from of T. angustifolia leaves, has a great potential to be used as an alternative adsorbent material in pesticide contaminated aqueous solution treatment. / Devido à relevância toxicológica dos agrotóxicos introduzidos no ambiente através de práticas agrícolas, torna-se necessário o desenvolvimento de metodologias simples e de baixo custo, que permitam remover esses poluentes do meio ambiente, em especial, das águas destinadas a consumo humano. O processo de adsorção é uma técnica que possui grande aplicação, por ser eficiente e de baixo custo. A capacidade adsorvente das folhas de Typha angustifolia L., planta herbácea perene pertencente à família Typhaceae, no Brasil também conhecida popularmente como taboa, foi investigada para a remoção dos agrotóxicos trifluralina, clorpirifós e α-endossulfam de meio aquoso. Foi realizada a otimização e validação da metodologia analítica de extração em fase sólida (SPE) e determinação por cromatografia gasosa utilizando-se o detector por captura de elétrons (GC/ECD). A metodologia (SPEGC/ECD) atendeu aos critérios de validação estabelecidos e foi eficiente para a quantificação dos agrotóxicos em meio aquoso, fornecendo credibilidade aos resultados obtidos nos ensaios de adsorção. A capacidade de adsorção da T. angustifolia foi avaliada sob diferentes parâmetros: tratamento do material, dose de material, tempo de contato entre o material vegetal e a solução dos agrotóxicos e concentração dos agrotóxicos na solução. Os resultados obtidos mostraram que o tratamento do material com água destilada foi satisfatório e que para todas as doses de adsorvente avaliadas a remoção dos agrotóxicos foi eficiente, sendo obtido um melhor resultado para a dose de 3,0 g L-1 , equivalente a 60 mg de adsorvente para 20,00 mL de solução, onde se obteve remoção entre 73 e 80% dos agrotóxicos. O estudo cinético do processo de adsorção dos agrotóxicos pela T. angustifolia mostrou que o equilíbrio de adsorção foi alcançado em 40 minutos, para todos os agrotóxicos, com remoção de até 90%, utilizando uma dose muito pequena de material vegetal (3 g L-1 ). Os dados experimentais foram avaliados utilizando-se os modelos cinéticos de pseudo-primeira ordem, pseudo-segunda ordem e Avrami. Uma das limitações encontradas no estudo da influência da concentração foi a solubilidade limitada dos agrotóxicos em meio aquoso (entre 0,33 e 1,00 mg L-1 ), que fez com que os ensaios de adsorção fossem avaliados para uma pequena faixa de concentração. Para estudar o mecanismo de adsorção dos agrotóxicos pelo material T. angustifolia, os dados experimentais foram aplicados aos modelos não lineares de Langmuir, Freundlich e Sips. Ensaios adicionais mostraram que a capacidade de adsorção é pouco afetada quando se utiliza água natural obtida em represas nos ensaios de adsorção. Os resultados obtidos indicam que o material vegetal, obtido a partir das folhas de T. angustifolia, possui um grande potencial para ser utilizado como material adsorvente alternativo no tratamento de meio aquoso contaminado com os agrotóxicos trifluralina, clorpirifós e α-endossulfam.

Maintenance of genomic integrity with high fidelity is of prime importance to any organism. An insult which may result in compromised genome integrity is prevented or its consequences are monitored by advanced cellular networks, collectively called the DNA damage response (DDR). Various DNA repair pathways, which are part of DDR, constantly correct the genome in the event of any undesirable change in the genetic material and prevent the transmission of any impairment to daughter cells. Non homologous DNA end joining (NHEJ) is the predominant DNA repair pathway associated with DDR in higher eukaryotes, correcting double-strand breaks (DSBs). Microhomology mediated end joining (MMEJ), an alternate mechanism to NHEJ also exists in cells, which is associated with erroneous joining of broken DNA, leading to mutagenesis. DDR is of paramount importance in cellular viability and therefore, any defects in DDR or the imbalance of repair pathways contribute to mutations, cellular transformations and various neurodegenerative and congenital abnormalities. Here, we investigate the DDR via NHEJ and MMEJ pathways during embryonic development in mice as well as in presence of an environmental pollutant, Endosulfan, in order to understand how physiological and exogenous factors condition the balance of repair pathways. Among various classes of pesticides known to cause side effects, organochlorine pesticides (OCPs) lead the list, possessing high transport potential, and a variety of toxic and untoward health effects. Endosulfan is a widely used organochlorine pesticide and is speculated to be detrimental to human health. However, very little is known about mechanism of its genotoxicity. Using in vivo and ex vivo model systems, we showed that exposure to Endosulfan induced reactive oxygen species (ROS) in a concentration dependent manner. Using an array of assays and equivalents of sub-lethal concentrations comparable to the detected level of Endosulfan in humans living in active areas of exposure, we demonstrated that ROS production by Endosulfan resulted in DNA double-strand breaks in mice, rats and human cells. In mice, the DNA damage was predominantly detected in type II pneumocytes of lung tissue; spermatogonial mother cells and primary spermatids of testes. Importantly, Endosulfan-induced DNA damage evoked DDR, which further resulted in elevated levels of classical NHEJ. However, sequence analyses of NHEJ junctions revealed that Endosulfan treatment resulted in extensive processing of broken DNA, culminating in increased and long junctional deletions, thereby favouring erroneous repair. We also find that exposure to Endosulfan led to significantly increased levels of MMEJ, which is a LIGASE III dependent, alternative, non classical repair pathway, encompassing long deletions and processing of DNA. Further, we show that the differential expression of proteins following exposure to Endosulfan correlated with activation of alternative DNA repair. At the physiological level, using mouse model system, we showed that exposure to Endosulfan affected physiology and cellular architecture of organs and tissues. Among all organs, damage to testes was extensive and it resulted in death of different testicular cell populations. We also found that the damage in testes resulted in qualitative and quantitative defects during spermatogenesis in a time dependent manner, increasing epididymal ROS levels and affecting sperm chromatin integrity. This further culminated in reduced number of epididymal sperms and actively motile sperms, which finally resulted in reduced fertility in male but not in female mice. Repair of DSBs is important for maintaining genomic integrity during the successful development of a fertilized egg into a whole organism. To date, the mechanism of DSB repair in post implantation embryos has been largely unknown except for the differential requirement of DNA repair genes in the course of development. These studies relied on null mutation analysis of animal phenotypes and therefore a quantitative understanding of repair pathways was absent. In the present study, using a cell free repair system derived from different embryonic stages of mice, we found that canonical NHEJ is predominant at 14.5 day of embryonic development. Interestingly, all types of DSBs tested were repaired by LIGASE IV/XRCC4 and Ku-dependent classical NHEJ. Characterization of end-joined junctions and expression studies further showed evidence for C-NHEJ. Strikingly, we observed non canonical end joining accompanied by DSB resection, dependent on microhomology and LIGASE III in 18.5-day embryos. Further we observed an elevated expression of CtIP, MRE11, and NBS1 at this stage, suggesting that it could act as a switch between classical and microhomology-mediated end joining at later stages of embryonic development. Keeping these observations in mind, we wondered if Endosulfan affected the differential regulation of DDR during development, similar to mice tissues. Upon analysing the effect of endosulfan on NHEJ/MMEJ at above mentioned stages of mouse embryonic development, we found that C-NHEJ efficiency remained low or unaltered while the efficiency of MMEJ was upregulated significantly, perturbing the repair balance during embryo development and hence facilitating mutagenic repair. Thus, our results establish the existence of both classical and non classical NHEJ pathways during the post implantation stages of mammalian embryonic development. Our studies also provide deeper insights into physiological and molecular events leading to male infertility upon Endosulfan exposure and its impact on impairing the differential regulation of DNA repair during embryonic development. Our findings suggest the plasticity of DNA repair pathways in physiological and pathological conditions and provide insights into mechanism of genome instability due to DNA repair imbalance, when exposed to environmental mutagens.