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.

Assessing the molluscicidal and biochemical activities of New-Fort®, an inorganic fertilizer against Theba pisana snails.

Terrestrial snails are a significant issue in agricultural production worldwide. The use of nitrogen - phosphorus - potassium (NPK) based fertilizers played an important role in meeting the food demand throughout the world, so its effectiveness against land snails needs to be investigated. This study was conducted to evaluate toxic lethal effect of New-Fort®, an inorganic NPK based fertilizer, in the field for 3, 7 and 10 days and in the laboratory for 24, 48 and 72 h against Theba pisana snails. Also, the impact of its sub-lethal doses (1/10, 1/5, 1/4 and 1/2 of 48 h-LD50) on biochemical parameters were determined under laboratory conditions. The results showed that the snails percent reduction in the field were 21.4, 61.0 and 80.0 % after 10 days' application of quarter, half and one field rate and the values of LD50 in the laboratory were 4.94, 4.56 and 4.24 mg/g b.w at 24, 48 and 72 h, respectively. New-Fort® sub-lethal doses caused a significant inhibition in catalase, γ-glutamyl transferase and acetylcholinesterase activities. It also elicited a significant elevation in glutathione S-transferase activity post exposure to 1/10 and 1/5 of LD50, whereas an opposite effect was occurred after exposure to 1/4 and 1/2 of LD50. Lipid peroxidation level was reduced in snails treated with 1/10 and 1/5 of LD50, whereas it increased in 1/4 and 1/2 of LD50- treated snails. Moreover, a significant inhibition in alkaline phosphatase activity at all tested doses, with the exception of 1/2 of LD50 was observed. An increase in alanine aminotransferase and aspartate aminotransferase activities were occurred after all tested doses exposure. Our findings highlighted on how biochemical changes can be exploited to better understand the mechanisms underlying New-Fort® fertilizer toxicity against the land snail, T. pisana, as well as how to benefit from NPK fertilizers application in snail control.

Bio-molluscicidal potential and biochemical mechanisms of clove oil and its main component eugenol against the land snail, Theba pisana.

The land snail, Theba pisana is a serious pest that adversely affects various crops in sustainable agriculture. Essential oils and their constituents represent an environmentally sound alternative to synthetic pesticides. Our study aimed to investigate the lethal and sub-lethal toxicity of clove oil and its main component eugenol to understand the mechanisms underlying its toxic action against T. pisana. The GC-MS profile of the clove oil composition was characterized. In the laboratory experiment, LD50 of clove oil and eugenol via the contact testing were determined after 48 and 72 h. Moreover, sub-lethal effects of clove oil or eugenol on the survivors following the exposure of snails to the 25 and 50% of the LD50/48 and 72 h were evaluated through using snail tissues for biochemical measurments. The GC-MS analysis showed that eugenol (64.87%) was the major constituent present in the oil. The results also showed that LD50 values at 48 and 72 h were 2006.5 and 1493.5 µg/g b.w for oil and 239.6 and 195.3 µg/g b.w for eugenol, respectively. Compared to control, the sub-lethal effects of clove oil or eugenol at 48 and 72 h showed a significant increase in reduced glutathione (GSH) levels. Catalase (CAT) and glutathione-S-transferase (GST) activities significantly elevated in oil- or eugenol-treated snails, except at low dose after 48 h. After two exposure times, snails exposed to oil or eugenol at both sub-lethal effects had considerably higher γ-glutamyltransferase (γ-GT) and aspartate aminotransferase (AST) activities. Moreover, markedly augmentation in alkaline phosphatase (ALP) and alanine aminotransferase (ALT) activities at all exposure times, with the exception of snails treated with low dose of eugenol after 48 h was observed. Both clove oil and eugenol at the tested doses caused a significant inhibition in acetylcholinesterase (AChE) activity at two exposure times. Our findings highlight the potential of clove oil and eugenol, as an efficient natural molluscicide alternative to its synthetic counterparts for snail control.

Exploring the mechanisms underlying the toxicity of boric acid against the land snail, Theba pisana.

BACKGROUND: The land snail, Theba pisana, is one of the most important threats facing agriculture around the globe. Boric acid (BOA) is currently used as a safe alternative molluscicide to control land snails in sustainable agriculture, but the mechanisms of toxicity have not yet been investigated. The present study characterizes the lethal and sub-lethal (0.5 and 1 mg g-1 ) toxic effects of BOA-contaminated food for 14 days by examining physiological, biochemical and histopathological indicators in T. pisana to understand the mechanisms underlying its toxic action. RESULTS: BOA was found to be lethal against T. pisana with LC50 values of 24.7 and 8.05 mg g-1 after 3 and 7 days of exposure, respectively. BOA sublethal concentrations led to a significant reduction in food consumption and growth of snails after 14 days of exposure. BOA also caused a significant increase in testosterone levels, whereas an opposite effect was observed in estradiol levels. An increase in progesterone levels in snails in the 0.5 mg g-1 BOA group and a decrease in the 1 mg g-1 BOA group were observed after all exposure times. Moreover, the lipid peroxidation level and catalase activity were elevated, whereas acetylcholinesterase activity was inhibited in the treated snails. Alteration in glutathione-S-transferase activity was noticed after exposure to both sublethal concentrations. In addition, BOA induced histopathological alterations in the digestive gland of T. pisana. CONCLUSION: Our findings provide novel insights into how physiological, biochemical and histopathological alterations can be used to explore the mechanisms underlying BOA toxicity against snails. © 2022 Society of Chemical Industry.

Insights into the ecotoxicological perturbations induced by the biocide Abamectin in the white snail, Theba pisana.

Abamectin (avermectin B1, ABM) has been widely used as a biocide in agriculture, veterinary and medicine around the world. Yet, there is still a lack of knowledge about the ecotoxicological effects of ABM. In this study, we investigated the acute toxicity and sub-lethal (20% and 60% LD50) biochemical responses of ABM on the non-target land snail, Theba pisana. Mortality of snails increased with the dose increase, resulting 48 h- LD50 value of 1.048 µg/snail. The biochemical results showed a decrease in glycogen content and lipids for two sub-lethal doses after all time intervals, whereas increased the level of total proteins after exposure to 60% LD50 ABM. Overall, the tested sub-lethal doses significantly decreased the total energy reserves. ABM-exposure to snails elevated γ-Glutamyl transferase and Lactate dehydrogenase activities at all-time intervals. A significant increase of Glutathione-S-transferase activity was also recorded in snails exposed to 20% and 60% LD50 after 7 days and all time intervals, respectively. However, ABM inhibited the activity of Aspartate aminotransferase and Alanine aminotransferase after 7 days of exposure. Our investigation provides new insights into the disturbances of energy reserves and enzyme activities in T. pisana that are sensitive and may be used as biomarkers for assessing ABM toxicity.

Emamectin benzoate as a potential molluscicide against white garden snail, Theba pisana in association with biochemical defects.

BACKGROUND: The white garden snail, Theba pisana, is distributed worldwide and is a serious molluscan pest of different crops. Emamectin benzoate (EMB) 'an avermectin derivative' is a novel biorational agent and highly effective pesticide. This study focused on the lethal and in vivo sublethal toxic effect of EMB on the energy reserves (glycogen, lipids and proteins), total energy reserves and activities of glutathione S-transferase (GST), γ-glutamyl transferase (γ-GT), lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the hepatopancreas of T. pisana for up to 7 days of exposure. RESULTS: The median lethal dose (LD50 ) at 48 h of EMB treatment was 5.34 µg g-1 body weight (b.w.). Sublethal doses of 1.07 and 3.20 µg g-1 b.w. (i.e., 20% and 60% of the LD50 ) led to significant dose- and time-dependent decreases in glycogen and lipids; these doses increased the total protein level. Overall, the tested sublethal doses significantly decreased the total energy reserves. Moreover, GST and γ-GT activities were elevated, whereas the activities of AST and ALT were inhibited in the exposed snails. A decrease in LDH activity after 1 and 3 days of exposure and an increase after 7 days of exposure were seen in snails treated with EMB. CONCLUSION: EMB exerted lethal toxicity on T. pisana and consequently caused changes in energy reserve levels and enzyme activities in the animal. © 2022 Society of Chemical Industry.

Contact toxicity and biochemical impact of metaldehyde against the white garden snail Theba pisana (Müller, 1774).

BACKGROUND: Terrestrial snails are one of the most damaging threats to sustainable agriculture. Chemical control using molluscicides is the main approach used to combat these agricultural pests. Metaldehyde is the active ingredient in most snail control products in use. However, its toxicity indices and mode of action have scarcely been investigated. For the first time, we characterized the metaldehyde contact toxicity indices against the white garden snail Theba pisana. The biochemical impact of metaldehyde on acetylcholinesterase (AChE), aspartate aminotransferase (AST) and alanine aminotransferase (ALT), alkaline phosphatase (ALP) and glutathione S-transferase (GST) activities and the lipid peroxidation (LPO) level was investigated. RESULTS: The median lethal dose (LD50 ) values at 24, 48 and 72 h of treatment were 11.33, 8.53, and 6.87 µg g-1 body weight (BW), respectively; while, the median lethal time (LT50 ) values were 88.16, 55.85, and 25.67 h when doses of 6, 8, and 12 µg g-1 BW were applied, respectively. In the snails treated with 2.83 and 5.67 µg g-1 BW (» and ½ LD50 at 24 h of treatment) and 2.13 and 4.27 µg g-1 BW (» and ½ LD50 at 48 h of treatment), higher AChE, GST, AST, ALT, and ALP activities as well as higher levels of LPO were observed compared with that of untreated snails. CONCLUSION: Metaldehyde displayed dose- and time-dependent contact toxicity. The biochemical results suggest that metaldehyde may have neurotoxic and cytotoxic actions in terrestrial snails. Application of metaldehyde in ways that could control pest snails and slugs and reduce its negative impact on the environment are discussed. © 2021 Society of Chemical Industry.

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.