Mancozeb Research Articles

Pre-imaginal exposure to mancozeb induces morphological and behavioral deficits and oxidative damage in Drosophila melanogaster

Mancozeb (MZ), a manganese/zinc containing ethylene-bis-dithiocarbamate, is a broad-spectrum fungicide. Chronic exposure to MZ has been related to several organisms’ neurological, hormonal, and developmental disorders. However, little is known about the post-natal effects of developmental exposure to MZ. In this study, Drosophila melanogaster was subjected to a pre-imaginal (eggs-larvae-pupae stage) model of exposure to MZ at 0.1 and 0.5 mg/mL. The emergence rate, body size, locomotor performance, sleep patterns, and molecular and biochemical parameters were evaluated in post-emerged flies. Results demonstrate that pre-imaginal exposure to MZ significantly impacted early emerged flies. Additionally, reduced progeny viability, smaller body size and delaying in emergence period, locomotor impairment, and prolonged sleep time were observed. Content of glucose, proteins, and triglycerides were altered, and the bioenergetics efficiency and oxidative phosphorylation at complex I were inhibited. mRNA stade state levels of genes responsive to stress, metabolism, and regulation of circadian cycle (Nrf2, p38, Hsp83, Akt1, GPDH, tor, per, tim, dILP2, and dILP6) were augmented, pointing out to stimulation of antioxidant defenses, insulin-dependent signaling pathway activation, and disruption of sleep regulation. These data were followed by increased lipid peroxidation and lower glutathione levels. In addition, the activity of catalase and glutathione-S-transferase were induced, whereas superoxide dismutase was inhibited. Together, these results demonstrate that developmental exposure to MZ formulation led to phenotype and behavioral alterations in young flies, possibly related to disruption of energetic metabolism, oxidative stress, and deregulation of genes implied in growth, sleep, and metabolism.

Effect of Nelumbo nucifera Petals Extract on Antioxidant Activity and Sperm Quality in Charolais Cattle Sperm Induced by Mancozeb.

The white Nelumbo nucifera petals aqueous extraction (NAE) was prominent in phytochemical content, antioxidant activity, and enhanced rat sperm viability induced by FeSO4, a heavy metal. Mancozeb (MZ) contains heavy metals and is widely used for fungal control in agriculture and industry. It induces oxidative stress and causes of spermatogenesis and reproductive organs’ abnormalities in both humans and animals. The aims of the present study were to investigate the effects of white Nelumbo nucifera petals aqueous extraction (WNAE) on sperm quality in cattle sperm induced by MZ. Moreover, this study investigated phytochemical compounds by liquid chromatography-mass spectrometry. A protein profile related to sperm quality with SDS-page and sperm energy preservation for each treatment was determined. The results found nine phytochemical compounds, in which quercetin-3-O-arabinoglycoside was a major flavonoid that was found in the WNAE. MZ induced free radicals in cells, leading to LPO and protein oxidation, while decreasing sperm motility, sperm viability, acrosome integrity, and normal sperm morphology. The cattle sperm found four proteins related to sperm quality including MWs of 17, 31, 34, and 55 kDa. The WNAE effectively increased energy preservation, sperm motility, sperm viability, acrosome integrity, and normal sperm morphology. The WNAE enhanced sperm qualities by reducing oxidative stress. It might be suggested that WNAE has benefits for sperm preservation which may be used to guard against toxicity in animals or humans exposed to MZ contaminants.

Ball-milled synthesis of maize biochar-ZnO nanocomposite (MB-ZnO) and estimation of its photocatalytic ability against different organic and inorganic pollutants

Different industrial and agricultural practices release a variety of dyes and pesticides in soil and water. Degradation of these pollutants is very important to avoid health and environmental issues. In this study, the photocatalysis technique has been optimized and adopted to degrade organic and inorganic pollutants. First of all, biochar with distinctive physicochemical properties, like high specific surface, highly carbonaceous property, and the electron-conductive nature was prepared in a vacuum furnace from maize straw. These properties depicted the effective absorbance ability of prepared biochar. Using a solvent-free ball-milling method, ZnO loaded maize biochar nanocomposite (MB-ZnO) was synthesized from this biochar and used as a photocatalyst to degrade aqueous pollutants, under different light sources. The adsorption and photocatalytic activity of MB-ZnO was assessed against Safranin O (Saf) and Mancozeb (MC) within a closed system using different light conditions including dark, UV and visible light. To understand the mechanism of Saf and MC removal from aqueous solution, reaction kinetics was calculated according to the pseudo-first-order kinetic model. MB-ZnOcomposite exhibited variable photocatalytic performances to degrade Saf under visible light (83.5%), UV radiations (81.0%) and dark conditions (78%) in 60 min. Similarly, maximum MC degradation by MB-ZnO nanocomposite was exhibited in visible light (56.5%), followed by UV radiations (27.5 %) and dark conditions (25.2%). The findings of this study concluded that MB-ZnO nanocomposite can be used as an excellent catalyst to remove aqueous pollutants, under both light and dark conditions.

Combination of Potassium Phosphite and Reduced Doses of Fungicides Encourages Protection against Phytophthora infestans in Potatoes

Late blight caused by the oomycete Phytophthora infestans is considered the biggest threat to potato farming worldwide. For susceptible cultivars, the disease is often managed by frequent applications of fungicides to reduce yield loss. The use of bio-based compounds that interfere with biologically active systems is an innovative strategy for improving disease management. In the present work, the control of P. infestans infection on potatoes by potassium phosphite (KPhi) combined with recommended and reduced doses of active ingredients (Ais) from different fungicides was evaluated. The protective effects of different combinations were initially assessed in vivo and subsequently compared with a greenhouse screening. The active ingredients cyazofamid (CFD) and mancozeb (MCB), used at recommended and reduced doses, were less effective at reducing P. infestans infections than when combined with KPhi. In greenhouse trials, CFD, mandipropamid (MPD) and MCB at recommended doses were the most effective treatments when combined with KPhi; meanwhile, the combination of KPhi with azoxystrobin (AZ), benthiavalicarb-isopropyl/mancozeb (ISO/MCB), and CFD at reduced doses exhibited strong protective activity compared to other similar combinations. This decreased the severity of infection by P. infestans up to ~89%. Greenhouse experiments also demonstrated that a combination of KPhi and CFD at both doses caused the highest reduction in disease severity (up to ~90%) within 35 days of infection. In microplot experiments, KPhi delayed the progression of late blight in susceptible potato varieties; therefore, in the combined treatments AUDPC values were significantly lower than those obtained after applications with CFD doses, providing sufficient protection against late blight. Our data suggest that optimizing the formulation with addition of KPhi could result in a lower recommended dose. This would result in a reduction of the active compounds of the fungicides in potato farming. Furthermore, the impact of KPhi on late blight development makes it a potential component for incorporation into an integrated pest management system.

Cytotoxicity of mancozeb on Sertoli–germ cell co-culture system: Role of MAPK signaling pathway

Mancozeb (MZB) is a worldwide fungicide for the management of fungal diseases in agriculture and industrial contexts. Human exposure occurs by consuming contaminated plants, drinking water, and occupational exposure. There are reports on MZB's reprotoxicity such as testicular structure damage, sperm abnormalities, and decrease in sperm parameters (number, viability, and motility), but its molecular mechanism on apoptosis in testis remains limited. To investigate the molecular mechanisms involved in male reprotoxicity induced by MZB, we used primary cultures of mouse Sertoli-germ cells. Cells were exposed to MZB (1.5, 2.5, and 3.5μM) for 3h to evaluate viability by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, reactive oxygen species (ROS) generation, and oxidative stress parameters (lipid peroxidation). Cell death and mitogen-activated protein kinase (MAPK) signaling were measured in these cells using flow cytometry and western blotting. In addition, some groups were exposed to N-acetylcysteine (NAC, 5mM) in the form of co-treatment with MZB. Mancozeb reduced viability and increased the level of intracellular ROS, p38 and c-Jun N-terminal kinases (JNK) MAPK proteins phosphorylation, and apoptotic cell death, which could be blocked by NAC as an inhibitor of oxidative stress. The present study indicated for the first time the toxic manifestations of MZB on the Sertoli-germ cell co-culture. Redox imbalance and p38 and JNK signaling pathway activation might play critical roles in MZB-induced apoptosis in the male reproductive system.

Cardiac developmental toxicity and transcriptome analyses of zebrafish (Danio rerio) embryos exposed to Mancozeb

Mancozeb (MZ), an antibacterial pesticide, has been linked to reproductive toxicity, neurotoxicity, and endocrine disruption. However, whether MZ has cardiactoxicity is unclear. In this study, the cardiotoxic effects of exposure to environment-related MZ concentrations ranging from 1.88 μM to 7.52 μM were evaluated at the larval stage of zebrafish. Transcriptome sequencing predicted the mechanism of MZ-induced cardiac developmental toxicity in zebrafish by enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO). Consistent with morphological changes, the osm, pfkfb3, foxh1, stc1, and nrarpb genes may effect normal development of zebrafish heart by activating NOTCH signaling pathways, resulting in pericardial edema, myocardial fibrosis, and congestion in the heart area. Moreover, differential gene expression analysis indicated that cyp-related genes (cyp1c2 and cyp3c3) were significantly upregulated after MZ treatment, which may be related to apoptosis of myocardial cells. These results were verified by real-time quantitative RT-qPCR and acridine orange staining. Our findings suggest that MZ-mediated cardiotoxic development of zebrafish larvae may be related to the activation of Notch and apoptosis-related signaling pathways.

Toxicological Comparison of Mancozeb and Zoxamide Fungicides at Environmentally Relevant Concentrations by an In Vitro Approach.

Mancozeb (MZ) and zoxamide (ZOX) are fungicides commonly used in pest control programs to protect vineyards. Their toxic and genotoxic potential were investigated in vitro on HepG2 and A549 cell lines at environmentally relevant concentrations. Cytotoxicity, apoptosis, necrosis and intracellular reactive oxygen species (ROS), comet assay and a micronucleus test with CREST immunofluorescence were used. The expression of a panel of genes involved in apoptosis/necrosis (BAX/BCL2), oxidative stress (NRF2), drug metabolism (CYP1A1) and DNA repair (ERCC1/OGG1) was evaluated by real-time PCR. Both fungicides were cytotoxic at the highest tested concentrations (295.7 and 463.4 µM, respectively); MZ induced necrosis, ZOX did not increase apoptosis but modulated BAX and BCL2 expression, suggesting a different mechanism. Both compounds did not increase ROS, but the induction of CYP1A1 and NRF2 expression supported a pro-oxidant mechanism. The comet assay evidenced MZ genotoxicity, whereas no DNA damage due to ZOX treatment was observed. Positive micronuclei were increased in both cell lines treated with MZ and ZOX, supporting their aneugenic potential. ERCC1 and OGG1 were differently modulated, indicating the efficient activation of the nucleotide excision repair system by both fungicides and the inhibition of the base excision repair system by MZ. Overall, MZ confirmed its toxicity and new ZOX-relevant effects were highlighted.

Effects of mancozeb on heat Shock protein 70 (HSP70) and its relationship with the thermal physiology of Physalaemus henselii (Peters, 1872) tadpoles (Anura: Leptodactylidae).

Negative impacts on amphibians have been reported due to contamination by agrochemicals. However, until now, no study has tested the effect of the fungicide mancozeb (MZ) on thermal tolerance and its relationship with the expression of heat shock proteins (HSPs). MZ is the best-selling broad-spectrum fungicide in the world, which negatively affects non-target organisms. Here, we tested for the first time the effects of MZ on critical thermal maximum (CTmax) and its relationship to the expression of heat shock protein 70 (HSP70) in tadpoles of Physalameus henselii, a colder-adapted species in southernmost of the Neotropical region. A sublethal concentration of 2mg/L was used. We found that the CTmax of the MZ-treated group was lower than that of the control group. In addition, there was an increase in HSP70 expression in tadpoles exposed to MZ and in tadpoles that underwent heat treatment. However, tadpoles subjected to MZ and heat treatment showed no induced HSP70 protein expression. Our results demonstrated that sublethal doses of the fungicide MZ negatively affected the thermal physiology and heat shock protein expression in tadpoles of P. henselii by inducing an increase in HSP70 concentration and by reducing the critical CTmax supported by tadpoles. It is important to understand the relationship between environmental contamination and physiological thermal limits in our current scenario of high rates of habitat conversion associated with unrestricted use of agrochemicals, as well as the challenging environmental changes induced by global warming.

Épocas de aplicação de azoxistrobina + mancozebe no controle de mancha branca do milho

Um dos principais métodos de controle da mancha branca do milho, o controle químico com estrobilurinas e fungicidas protetores demonstraram efeito no controle da doença, no entanto, condicionada ao momento de sua realização em estádios fenológicos da cultura. O objetivo deste trabalho foi avaliar o controle e efeito no progresso temporal da mancha branca do milho com a associação dos fungicidas mancozebe e azoxistrobina em diferentes épocas de aplicação, dois cultivares, duas localidades de plantio e o efeito na produtividade da cultura. Os experimentos foram conduzidos nos municípios de Marechal Cândido Rondon e Sertanópolis, PR. Foram utilizados dois híbridos suscetíveis, DKB230PRO3 e DKB290PRO3. O delineamento experimental utilizado foi de blocos casualizados com 4 repetições. As avaliações foram efetuadas 10 dias após cada aplicação, além de uma avaliação no estádio de maturidade fisiológica. A severidade da doença foi avaliada através da quantificação da área foliar lesionada em 10 plantas por parcela e utilizadas no cálculo da Área Abaixo da Curva de Progresso da Doença, submetidos a análise de variância e o teste de médias. As curvas de progresso da doença foram ajustadas aos modelos logístico e exponencial. A produtividade foi estimada após a colheita manual, com o peso de mil grãos e produtividade (kg ha-1). Aplicações dos fungicidas em estádios V8 e V4+V8+VT proporcionaram melhor controle da mancha branca. Aplicações no estádio V4 não proporcionaram severidades diferentes da testemunha sem aplicação. Não foram observados ganhos significativos em produtividade em função da aplicação dos fungicidas.

Disruption of mitochondrial complexes, cytotoxicity, and apoptosis results from Mancozeb exposure in transformed human colon cells

Ethylene bisdithiocarbamate pesticides, including Mancozeb (MZ), are used as fungicides. Effects of MZ on apoptosis induction and mitochondrial activity of HT-29 colon cells were investigated. MZ exposed cells exhibited blebbing and cellular membrane disruption in scanning electron micrographs. Positive fluorescent staining with Annexin V at doses of 60–140 μM supports apoptosis as the mechanism of cell death. Activity of all electron transport chain complexes were evaluated. Mitochondrial Complex I activity was decreased in 100 μM treated cells. Mitochondrial Complex III activity was decreased in 60 and 100 μM MZ treated cells. Mitochondrial Complex II and Complex IV activities were decreased in cells treated with 60, 100, and 140 μM. Cells treated with 60 μM exhibited a decrease in Complex V enzyme activity. It is concluded that MZ exposure inhibits all mitochondrial complexes of HT-29 cells and that positive fluorescent microscopy and blebbing support previous studies of cell death via apoptosis.

Integrated management of fusarium wilt of chickpea

Chickpea (Cicer arietinum L.) is one of the most important pulse crop grown all over India. Chickpea wilt caused by Fusarium oxysporum f. sp. ciceri is one of the major disease on chickpea in Northern Karnataka, which is soil and seed borne. Heavy inoculum in the soil and favorable environment condition results in the death of infected plant and therefore total yield loss. In this study, integrated management was conducted in both pot culture and in field with two antagonists, one botanical and one fungicide were studied against Fusarium oxysporum f. sp. ciceri causing chickpea wilt. Pot culture studies found that seed treatment with Carbendazim 12% + Mancozeb 63% @ 2 g/kg of seeds + Trichoderma harzianum 4 g/kg of seeds + Curcuma longa rhizome extract (Turmeric) @ 150 ml/ kg of seeds gave minimum wilt incidence (1.46%), maximum wilt incidence was noticed in untreated control with 100 per cent wilted plants. Field studies found that seed treatment with Carbendazim 12% + Mancozeb 63% @ 2 g/kg of seeds + Trichoderma harzianum 4 g/kg of seeds + Curcuma longa rhizome extract (Turmeric) @ 150 ml/ kg of seeds gave minimum wilt incidence (1.46%) and maximum yield (10.34 q/ha) followed by seed treatment with Carbendazim 12% + Mancozeb 63% @ 2 g/kg of seeds. Bio agents Trichoderma harzianum and Bacillus sp. @ 4 g/ kg of seeds and aqueous plant extract of Curcuma longa (Turmeric) rhizome extract @ 150 ml/ kg of seeds were also significant in reducing wilt incidence and more yield as compared to control.

Endocrine-disrupting activity of mancozeb.

The objective of the present study was to review the existing data on the mechanisms involved in the endocrine disrupting activity of mancozeb (MCZ) in its main targets, including thyroid and gonads, as well as other endocrine tissues that may be potentially affected by MCZ. MCZ exposure was shown to interfere with thyroid functioning through impairment of thyroid hormone synthesis due to inhibition of sodium-iodine symporter (NIS) and thyroid peroxidase (TPO) activity, as well as thyroglobulin expression. Direct thyrotoxic effect may also contribute to thyroid pathology upon MCZ exposure. Gonadal effects of MCZ involve inhibition of sex steroid synthesis due to inhibition of P450scc (CYP11A1), as well as 3β-HSD and 17β-HSD. In parallel with altered hormone synthesis, MCZ was shown to down-regulate androgen and estrogen receptor signaling. Taken together, these gonad-specific effects result in development of both male and female reproductive dysfunction. In parallel with clearly estimated targets for MCZ endocrine disturbing activity, namely thyroid and gonads, other endocrine tissues may be also involved. Specifically, the fungicide was shown to affect cortisol synthesis that may be mediated by modulation of CYP11B1 activity. Moreover, MCZ exposure was shown to interfere with PPARγ signaling, being a key regulator of adipogenesis. The existing data also propose that endocrine-disrupting effects of MCZ exposure may be mediated by modulation of hypothalamus-pituitary-target axis. It is proposed that MCZ neurotoxicity may at least partially affect central mechanisms of endocrine system functioning. However, further studies are required to unravel the mechanisms of MCZ endocrine disrupting activity and overall toxicity.

Mancozeb impairs mitochondrial and bioenergetic activity in Drosophila melanogaster

Mancozeb (MZ) is a broad-spectrum fungicide used worldwide in several crops. Neurological disorders in humans and animals have been associated with exposure to this compound by mechanisms still not fully understood. Drosophila melanogaster represents a reliable model in toxicological studies, presenting genetic and biochemical similarities with mammals. In this study, D. melanogaster flies were exposed for 15 days to MZ through the food (5 and 10 mg/mL). After that period, the efficiency of mitochondrial respiration complexes and metabolic markers were analyzed and evaluated. Flies presented weight loss, lower glucose, trehalose, and glycogen levels, and augmented levels of triglycerides concerning control (non-treated group). Acetyl-CoA Synthetase (ACeCS-1) and Acyl-Coenzyme Synthetase (ACSL1) contents were unchanged by MZ treatment. Mitochondrial respiration of flies was targeted by MZ treatment, evidenced by a decrease in oxygen consumption and bioenergetics rate and inhibition in mitochondrial complexes I/II. These results suppose that an impairment in mitochondrial respiration jointly with reduced levels of energetic substrates might be a mechanism involved in MZ deleterious effects, possibly by the limitation of ATP's availability, necessary for essential cellular processes.

Pesticide use in vegetable production in rural Uganda - A case study of Kabale District, South western Uganda

A study to investigate commonly grown vegetables, commonly used pesticides, and pesticide use practices was conducted in Kabale District, in South-western Uganda. This is because indiscriminate pesticide use and poor application practices can leave pesticide residues in food rendering it unsafe for consumption. The study revealed extensive pesticide application in Brassica oleracea; var. capitata (cabbage), Brassica oleracea; var. botrytis (cauliflower), Solanum lycopersicum, (tomato) and Beta vulgaris (beet root). Information obtained using interviews revealed that 16.5% of the traders in Kabale Municipality sold pesticides and 70% of the farmers in the major vegetable growing subcounties of Kaharo, Kyanamira and Kamuganguzi sprayed their vegetables with pesticides. Only 18% of the interviewed farmers could interpret instructions on pesticide container or bag labels correctly. All farmers (100%) had never attended any training on pesticide use. Cypermethrin, dimethoate, dichlorvos, metalaxyl, profenofos, malathion and mancozeb were mentioned as commonly used pesticides in vegetables grown in the district. Some of the farmers (42%) used mixed different pesticides in the vegetables. Limited knowledge about pesticide application, inability to interpret instructions, non-observance of pre-harvest intervals, mixing pesticides and lack of training on pesticide use contribute to pesticide use malpractices which may put farmers’ health at risk and reduce food quality. Therefore, there is need to address the identified knowledge gaps on safer pesticide application in order to attain safe agricultural productivity for sustainable food security, safeguarding human health and community development in Kabale District, Uganda. Key words: Pesticides, vegetables, pesticide use practices, Kabale District, Uganda.

Behavioral changes occur earlier than redox alterations in developing zebrafish exposed to Mancozeb.

As agriculture expands to provide food and wellbeing to the world's growing population, there is a simultaneous increasing concern about the use of agrochemicals, which can harm non-target organisms, mainly in the aquatic environment. The fungicide Mancozeb (MZ) has been used on a large-scale and is a potent inducer of oxidative stress. Therefore, there is an urgent need for the development of more sensitive biomarkers designed to earlier biomonitoring of this compound. Here we tested the hypothesis that behavioral changes induced by sublethal MZ concentrations would occur first as compared to biochemical oxidative stress markers. Embryos at 4h post-fertilization (hpf) were exposed to Mancozeb at 5, 10 and 20μg/L. Controls were kept in embryo water only. Behavioral and biochemical parameters were evaluated at 24, 28, 72, and 168 hpf after MZ exposure. The results showed that MZ significantly altered spontaneous movement, escape responses, swimming capacity, and exploratory behavior at all exposure times. However, changes in ROS steady-stead levels and the activity of antioxidant enzymes were observable only at 72 and 168 hpf. In conclusion, behavioral changes occurred earlier than biochemical alterations in zebrafish embryos exposed to MZ, highlighting the potential of behavioral biomarkers as sensitive tools for biomonitoring programs.

Manganese-induced neurotoxicity in cerebellar granule neurons due to perturbation of cell network pathways with potential implications for neurodegenerative disorders.

Manganese (Mn) is essential for living organisms, playing an important role in nervous system function. Nevertheless, chronic and/or acute exposure to this metal, especially during early life stages, can lead to neurotoxicity and dementia by unclear mechanisms. Thus, based on previous works of our group with yeast and zebrafish, we hypothesized that the mechanisms mediating manganese-induced neurotoxicity can be associated with the alteration of protein metabolism. These mechanisms may also depend on the chemical speciation of manganese. Therefore, the current study aimed at investigating the mechanisms mediating the toxic effects of manganese in primary cultures of cerebellar granule neurons (CGNs). By exposing cultured CGNs to different chemical species of manganese ([[2-[(dithiocarboxy)amino]ethyl]carbamodithioato]](2-)-kS,kS']manganese, named maneb (MB), and [[1,2-ethanediylbis[carbamodithioato]](2-)]manganese mixture with [[1,2-ethanediylbis[carbamodithioato]](2-)]zinc, named mancozeb (MZ), and manganese chloride (MnCl2)), and using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, we observed that both MB and MZ induced similar cytotoxicity (LC50∼ 7-9 μM), which was higher than that of MnCl2 (LC50∼ 27 μM). Subsequently, we applied systems biology approaches, including metallomics, proteomics, gene expression and bioinformatics, and revealed that independent of chemical speciation, for non-cytotoxic concentrations (0.3-3 μM), Mn-induced neurotoxicity in CGNs is associated with metal dyshomeostasis and impaired protein metabolism. In this way, we verified that MB induced more post-translational alterations than MnCl2, which can be a plausible explanation for cytotoxic differences between both chemical species. The metabolism of proteins is one of the most energy consuming cellular processes and its impairment appears to be a key event of some cellular stress processes reported separately in other studies such as cell cycle arrest, energy impairment, cell signaling, excitotoxicity, immune response, potential protein accumulation and apoptosis. Interestingly, we verified that Mn-induced neurotoxicity shares pathways associated with the development of Alzheimer's disease, Amyotrophic Lateral Sclerosis, Huntington's disease, and Parkinson's disease. This has been observed in baker's yeast and zebrafish suggesting that the mode of action of Mn may be evolutionarily conserved.

Study on the protective effect of hydroalcoholic Olive Leaf extract (oleuropein) on the testis and sperm parameters in adult male NMRI mice exposed to Mancozeb

This study aimed to assess the protective effects of the Olive Leaf Extract (OLE) against Mancozeb (MZB)-induced reproductive damage in male NMRI mice.In this experimental study, NMRI mice were randomly divided into 7 groups (n = 8) including 1: control, 2: MZB, 3: MZB + OLE (200 mg/kg), 4: MZB + OLE (400 mg/kg), 5: MZB + atropine (0.25 mg/kg), 6: OLE (200 mg/kg) and 7: OLE (400 mg/kg). Mice were sacrificed after 5 weeks, and blood samples were drawn to measure serum testosterone level. Sperm samples were collected and analyzed according to standard criteria. The left testis was removed to assess testicular parameters using H&E staining. The Protamine level and DNA fragmentation were determined by chromomycin A3 (CMA3) and SCD assays, respectively.Sperm viability decreased in MZB- induced groups. Also, the sperm's protamine deficiency (higher than 30%) and DNA fragmentation increased within MZB induced. There were significantly higher structural deformities of sperm's head, neck, and tail in MZB-treated groups as well (p < 0.05).Within treated groups with OLE, these sperm factors could repair defects versus MZB group. Furthermore, there were significant differences comparing sperm motility (type A, B, and D); protamine level, DNA fragmentation (SCD1, SCD3, and SCD4), and testosterone levels compared to the groups (p < 0.05). Sperm motility, seminiferous tubules diameter, the height of the germinal epithelium, and testosterone level within OLE 200 mg/kg was higher than other groups, while these factors in OLE 400 mg/kg was less than OLE 200 mg/kg.Results showed that oral administration of OLE could be useful in ameliorating MZB induced toxicity in the reproductive system of male mice within 200 mg/kg dosage; while OLE 400 mg/kg probably could exacerbate the harmful effects.

Green Synthesized Ag Nanoparticles for Bio-Sensing and Photocatalytic Applications.

In this work, sensing and photocatalytic activities of green synthesized silver nanoparticles (Ag NPs) are investigated. Ag NPs have been synthesized by the reduction of silver nitrate (AgNO3) using different leaf extracts. An optimum surface plasmon resonance (SPR) behavior is obtained for neem leaf extracts because of the presence of a high concentration of diterpenoids, as evidenced from gas chromatography mass spectroscopy results. The underlying mechanism for the formation of Ag NPs is highlighted. The Ag NPs are in spherical shape and exhibit the hexagonal crystal phase and also show a good stability. The biosensing property of the Ag NPs is evaluated using mancozeb (MCZ) agro-fungicide, and the SPR peak position exhibited a linear response with MCZ concentration. The sensitivity is found to be 39.1 nm/mM. Further, the photocatalytic activity of Ag NPs is tested using 0.5 mM MCZ solution as a model under UV–visible illumination. It is observed that photocatalytic activity is caused by the formation of reactive oxygen species. Therefore, the green synthesized Ag NPs are potential candidates for biosensing and photocatalytic applications.

Feasibility of Vis/NIR spectroscopy to detect and estimate fungicide residues on intact lettuces

Pesticides are applied repeatedly to grain, fruit, and vegetable crops for protection against pathogens, pests, and weeds, in short periods of time before harvest. The effective and fast monitoring of chemical residues in agricultural products is important for the assurance of healthy food. This study was accomplished to evaluate the feasibility to detect and estimate the concentration of dithiocarbamate fungicide (mancozeb) residues on intact lettuce leaves based on Vis/NIR spectral reflectance measurements and multivariate data analysis. In the pre-harvest interval, a high initial rate of decline of dithiocarbamate residues was observed between one and seven days after pesticide spraying (decrease of 90.3%), while a slower decline was verified from seventh to fourteenth day (decrease of 8.7%). The usefulness of this spectrometric method has been evidenced by determination of dithiocarbamate residues at concentrations between 0.23 and 10.3 mg CS2 kg-1, with detection and quantitation limits of 0.49 and 1.41 mg CS2 kg-1, respectively. Vis/NIR spectral reflectance combined to the partial least square analysis have potential to be applied for estimating dithiocarbamate concentrations on intact lettuce leaves, presenting advantages such as real-time measurements and the possibility to be built into the industrial processing lines.

Toxic effects of a mancozeb-containing commercial formulation at environmental relevant concentrations on zebrafish embryonic development.

The toxicological knowledge of mancozeb (MZ)-containing commercial formulations on non-target species is scarce and limited. Therefore, the objective of this work was to represent a realistic application scenario by evaluating the toxicity of environmental relevant and higher concentrations of a commercial formulation of MZ using zebrafish embryos. Following determination of the 96-h LC50 value, the embryos at the blastula stage (~ 2h post-fertilisation, hpf) were exposed to 0.5, 5, and 50μgL-1 of the active ingredient (~ 40× lower than the 96-h LC50). During the exposure period (96h), lethal, sublethal, and teratogenic parameters, as well as behaviour analysis, at 120hpf, were assayed. Biochemical parameters such as oxidative stress-linked enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR)), reactive oxygen species (ROS) levels, and glutathione levels (GSH and GSSG), as well as the activity of degradation (glutathione S-transferase (GST) and carboxylesterase (CarE)), neurotransmission (acetylcholinesterase (AChE)), and anaerobic respiration (lactate dehydrogenase (LDH))-related enzymes, were analysed at the end of the exposure period. Exposed embryos showed a marked decrease in the hatching rate and many malformations (cardiac and yolk sac oedema and spinal torsions), with a higher prevalence at the highest concentration. A dose-dependent decreased locomotor activity and a response to an aversive stimulus, as well as a light-dark transition decline, were observed at environmental relevant concentrations. Furthermore, the activities of SOD and GR increased while the activity of GST, AChE, and MDA contents decreased. Taken together, the involvement of mancozeb metabolites and the generation of ROS are suggested as responsible for the developmental phenotypes. While further studies are needed to fully support the hypothesis presented, the potential cumulative effects of mancozeb-containing formulations and its metabolites could represent an environmental risk which should not be disregarded.