Does commercial indoxacarb pose ecotoxicological consequences? Employing a multi-marker approach in the model species Theba pisana.

Indoxacarb is one of the most extensively used oxadiazine insecticides worldwide, but it may exert detrimental effects on ecosystems, population dynamics, and health. Due to the lack of knowledge on the ecotoxicity of indoxacarb, it is still challenging to assess whether this insecticide poses an ecotoxicological impact on terrestrial environments. Therefore, our study aims to provide novel data on the toxic effects of 28-day dietary exposure to commercial grade indoxacarb at two environmentally relevant concentrations, 0.02 µg/mL and tenfold (0.2 µg/mL) on the model species, Theba pisana. Their effects were studied using a multiple biomarker approach by evaluating physiological, biochemical, and histopathological responses. After 28 days of treatment, indoxacarb at both concentrations significantly reduced the food intake and growth of the treated snails. Also, it caused decreases in lipid peroxidation (LPO)  levels after 7 and 14 days of exposure, whereas an opposite effect occurred after 21 and 28 days. All treated snails were found to exhibit a lower content of glutathione (GSH) after all times of exposure. Moreover, catalase (CAT), glutathione-S-transferase (GST), and glutathione peroxidase (GPx) activities, as well as protein content (PC), were elevated in the treated snails after all time intervals. Post exposure to both realistic indoxacarb concentrations, changes in acetylcholinesterase (AChE) activity between a decrease and an increase were observed. Furthermore, indoxacarb caused histo-architectural changes in the hepatopancreas of T. pisana. Our results demonstrate that, at environmentally relevant concentrations, indoxacarb poses negative consequences for T. pisana, indicating its ecotoxicological impacts.

Evaluation of biochemical, hematological, and genotoxic parameters in mice exposed to individual and combined ethoprophos and cadmium.

Environmental contamination by complex mixtures of pesticides and metals is a major health problem in agriculture and industry. In real life scenarios, we are exposed to mixtures of chemicals rather than single chemicals, and therefore it is critical to assess their toxicity. The current work was conducted to assess the toxic effects of a low dose (2% median lethal dose) of ethoprophos (Etho, 0.16 mg kg-1 bw), and cadmium (Cd, 0.63 mg kg-1 bw); each alone or in combination on hematological, biochemical, and genotoxic parameters in male mice for one or four weeks. The tested toxicants resulted in a decline in body and organs weights, the most hematological indices, acetylcholine esterase activity, and the total protein content, while they significantly increased liver and kidney function parameters. Furthermore, they increased the mitotic index (MI), number of abnormal sperms, and chromosomes. In conclusion, Etho and Cd induce deleterious effects on all tested parameters in male mice which reflect more obvious impacts when both combined, particularly after 28 days of exposure. However, further research is needed to confirm toxicokinetic or toxicodynamic interactions between these two toxic compounds in the organisms.

Toxicological assessment of sublethal dose of acetamiprid in male mice and the efficacy of quercetin.

Acetamiprid (ACP) is a neonicotinoid insecticide that is the most effective pesticide for crop protection as well as flea control in agricultural animals and pets in the world. The goal of this study was to look at the in vivo effects of a sublethal dose of ACP on hematotoxicity, oxidative stress, hepatotoxicity, nephrotoxicity, immunotoxicity, and histological alterations, as well as the role of quercetin (QE) in alleviating these effects. Twenty adult male mice were divided into four equal groups orally administered corn oil (control), QE (50 mg kg-1 b.wt.), ACP (1/10 LD50) or ACP plus QE for two weeks. The results showed that ACP significantly lowered the body weight gain, hematological indices, glutathione (GSH), and both cellular and humoral immunity, On the other hand, levels of lipid peroxidation (LPO), glutathione peroxidase (GPx), and liver and kidney marker values were considerably increased in male mice exposed to ACP. In addition, examination under light microscopic showed that ACP induces histological alterations in liver and kidney tissues. The results also revealed that treating intoxicated mice with QE significantly reduced the deleterious effects of ACP. In conclusion, current results show that ACP at the sub lethal dose poses toxic risks to the liver and kidneys, and QE as a natural material enhances antioxidant defenses, which can be used as a potential interventional therapy against negative effects of pesticides like ACP.

Ecotoxicological biomarkers as investigating tools to evaluate the impact of acrylamide on Theba pisana snails.

Acrylamide (ACR) is a widespread industrial chemical with recognized adverse effects not only to humans but to other organisms in the environment as well. In the present study, the ecotoxicological effects of dietary exposure to sublethal concentration (1/20 LC50) of ACR on the land snail, Theba pisana after 2 weeks of exposure and 1-week recovery with respect to oxidative stress parameters; lipid peroxidation (LPO), reduced glutathione (GSH), catalase (CAT), and glutathione-S-transferase (GST), cytogenetic parameter; deoxyribonucleic acid (DNA) content, as well as immunological parameters; cell death, phagocytosis, lysosomal membrane stability (LMS), lectins, superoxide anion (O2-) generation, phenoloxidase (PO), peroxidase (POD), and hemocyanin (Hc) were examined. The results showed that ACR significantly increased LPO level and the activity of CAT and GST, cell death, and Hc level, whereas a significant decline in DNA and GSH contents, phagocytic activity, LMS, lectins, O2- generation, POD, and PO activities compared to the controls after 2-week exposure was observed. After 1-week recovery, most of the tested parameters in exposed snails were permanent and not reversible to the control levels. This study suggests that the tested multiple parameters of T. pisana species may be used as biomarkers of ACR exposure. Besides, T. pisana snails could be used as a good sentinel organism for ACR exposure in pollution monitoring studies.

Physiological traits of land snails Theba pisana as simple endpoints to assess the exposure to some pollutants.

In the current study, the toxicity bioassay of three pollutants abamectin (ABM), thiamethoxam (TMX), and acrylamide (ACR) against land snails Theba pisana was measured. Also, the ecotoxicological effects of dietary exposure to sublethal concentration (1/20 LC50) of these pollutants for 2-week exposure and 1-week recovery on some physiological endpoints evaluated as feeding activity, growth response, and carbonic anhydrase activity as a marker in charge of shell formation and seromucoid level as a marker in charge of mucus synthesis of the snails were studied. The results exhibited that the 48-h LC50 values were 0.91, 313.8, and 45.7 µg/g dry food for ABM, TMX, and ACR, respectively. The sublethal concentrations of these pollutants in the diet after 2-week exposure were found to reduce the food consumption and inhibit growth rate of the snails. Also, the data illustrated that carbonic anhydrase activity was significantly decreased. On the other hand, there was a significant increase in the seromucoid level as a marker responsible for mucus synthesis in ABM- and TMX-exposed snails, while ACR showed significantly decreased level when compared to control. After 1-week recovery, the tested endpoints of treated snails were slightly repaired but still less than that of the untreated animals. The overall outcome of this investigation suggests the utility of this animal as a good bioindicator organism for ABM, TMX, and ACR exposure in pollution monitoring studies.

Preparation of antibodies and development of an enzyme immunoassay for determination of atrazine in environmental samples.

An indirect competitive enzyme-linked immunosorbent assay (ELISA) has been developed and optimized for atrazine determination in soil at different depths (0-10, 10-20, and 20-30 cm) before and after 48 h of application, corn shoot and cow milk samples collected from Dina farm, Egypt. This assay was based on a specific polyclonal antibodies (PAb) raised by immunizing New Zealand rabbits with an immunogen prepared by coupling 3-{4-(ethylamino)-6-(isopropylamino)-1,3,5-triazine-2-yl} thiopropanoic acid to bovine serum albumin (BSA) via N-hydroxysuccinimide (NHS) active ester method. The sensitivity (estimated as IC50 value) was 17.5 µg mL⁻¹ with a detection limit of 0.1 ng mL⁻¹. The maximum atrazine concentration was found in soil especially in the deepest layer (325 and 890 µg kg⁻¹ before and after application, respectively). Atrazine concentration in corn shoot was 333.28, µg kg⁻¹ dry plant, while there was no detectable amount in milk. All samples screened by ELISA were validated by gas chromatography mass spectrometer procedure (GC/MS). Good correlation was achieved between the two methods (r = 0.997 for soil and 0.9814 for plant). This study demonstrates the utility and convenience of the simple, practical and cost-effective ELISA method in the laboratory for analysis of environmental samples. The method is ideal for the rapid screening of large numbers of samples in laboratories where access to GC/MS facilities, is limited or lacking.

The role of vitamin C as antioxidant in protection of oxidative stress induced by imidacloprid.

Pesticides may induce oxidative stress leading to generate free radicals and alternate antioxidant or oxygen free radical scavenging enzyme system. This study was conducted to investigate the acute toxicity of imidacloprid toward male mice and the oxidative stress of the sublethal dose (1/10 LD(50)) on the lipid peroxidation level (LPO), reduced glutathione content (GSH) and activities of the antioxidant enzymes; catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glucose-6-phosphate dehydrogenase (G6PD), and glutathione-s-transferase (GST). Also, the protective effect of vitamin C (200mg/kg bw) 30 min before or after administration of imidacloprid were investigated. The results demonstrated that the median lethal dose (LD(50)) of imidacloprid after 24h was 149.76 mg/kg bw. The oral administration of 14.976 mg/kg imidacloprid significantly caused elevation in LPO level and the activities of antioxidant enzymes including CAT, SOD, GPx and GST. However, G6PD activity remained unchanged, while the level of GSH content was decreased. In addition, the results showed that vitamin C might ameliorate imidacloprid-induced oxidative damage by decreasing LPO and altering antioxidant defense system in liver. The protective effect of the pre-treatment with vitamin C against imidacloprid-induced oxidative stress in liver mice is better than the post-treatment.