Effect of the Insecticide Chlorpyrifos on Behavioral and Metabolic Aspects of the Spider Polybetes pythagoricus.

The toxicity of pesticides to organisms depends on the total amount of chemical exposure. Toxicity can be minimized if the organism recognizes the pesticide and alters its behavior. Furthermore, the physical barrier of cuticular hydrocarbons can prevent the entrance of the pesticide into the organism. Finally, if the pesticide enters the body, the organism experiences physiological changes favoring detoxification and the maintenance of homeostasis. We analyzed the behavioral and metabolic response of the spider Polybetes pythagoricus at different times of exposure to the organophosphate pesticide chlorpyrifos. First we observed that the individuals are capable of recognizing and avoiding surfaces treated with pesticides based on a behavioral analysis. Subsequently, we characterized cuticular hydrocarbons as a possible barrier against pesticides. Then we observed that the pesticide provoked histological damage, mainly at the level of the midgut diverticula. Finally, we analyzed the activity of several of the spider's enzymes linked to oxidative stress after exposure to chlorpyrifos for different lengths of time (6, 24, and 48 h). We observed that catalase activity was high at the start, whereas the activity of superoxide dismutase and glutathione S-transferase changed significantly at 48 h. Lipid peroxidation became high at 6 h, but decreased at 48 h. In conclusion, although P. pythagoricus can avoid contact with chlorpyrifos, this pesticide causes activation of the antioxidant system when it enters the body. Our results make a significant contribution to the ecotoxicology of spiders. Environ Toxicol Chem 2023;42:1293-1308. © 2023 SETAC.

Sensitivity of histopathological and histochemical parameters in the digestive gland of the apple snail Pomacea canaliculata exposed to cypermethrin.

The aim of this study was to evaluate the toxic effects of the pesticide cypermethrin (CYP) in the digestive gland of the apple snail, Pomacea canaliculata, analysing histological and histochemical alterations. Adult snails were exposed to sublethal CYP concentrations (10, 25, and 100 µg/L) under acute (1 day) and sub-chronic (14 days) conditions. Histological analyses of the morphology of the digestive gland were performed and a histopathological condition index (HI) was calculated. Also, both intracellular accumulation of lipofuscins (LF) and neutral lipids (NL) were evaluated. CYP exposure induced tissue damage to this organ, such as disorganisation of the connective tissue, fibrosis, haemocytic infiltration, atrophy, and necrosis under acute and sub-chronic conditions. These alterations, integrated into a single HI value, revealed notable CYP effects during both acute and sub-chronic exposures. Cell type replacement, measured as VvBAS, was only observed in the sub-chronic treatment. Under acute conditions, the pyrethroid affected NL accumulation at the highest concentration, while in sub-chronic conditions NL accumulation was only observed at the lowest concentrations. P. canaliculata also showed a dose-dependent response of LF under acute CYP exposure conditions. However, under sub-chronic conditions, this parameter was not sensitive to pesticide exposure. All these relevant structural lesions may affect the normal function of the digestive gland, even though the species presented additional mechanisms, as infiltration of hemocyte and basophilic cell hyperplasia, that help it to tolerate the exposure to pollutants. This study showed that some histological and histochemical parameters are sensitive in P. canaliculata at CYP concentrations to which the snail could be exposed in the environments it inhabits.

Behavioral, Histological, and Physiological Evaluation of the Effect of Imidacloprid on the Spider Misumenops maculissparsus.

The aim of the present study was to evaluate the effects of the neonicotinoid insecticide imidacloprid (commercial formulation) on juveniles of the spider Misumenops maculissparsus (Keyserling, 1891). We first analyzed whether spiders recognized the presence of the insecticide on surfaces and in drinking water (in the form of droplets). Next, we investigated if the insecticide generated histologic, physiologic, and/or biochemical alterations. We observed that spiders do not detect the insecticide on a surface (e.g., paper) or in the form of droplets. After the imidacloprid ingestion by droplet intake, most spiders exhibited a paralysis that reverted after 48 h. Consequently, we observed histopathologic damage (i.e., pigment accumulation, necrosis, and cuticle detachment), and an increased catalase (CAT) activity and total-protein concentration in the individuals treated. The activities of glutathione-S-transferase, glutathione peroxidase, glutathione reductase, and superoxide dismutase, however, did not undergo significant variations. The results obtained emphasize the need to consider different classes of biomarkers, such as CAT and other proteins, to identify and evaluate the histologic, biologic, and biochemical effects of imidacloprid, one of the most widely used insecticides. Environ Toxicol Chem 2022;41:2152-2161. © 2022 SETAC.

Morphology of the digestive gland of the marine panpulmonate limpet Siphonaria lessonii: A cytological, histochemical, and ultrastructural description.

The molluskan digestive gland has been widely studied and its structural and ultrastructural descriptions have allowed the understanding of its several functions. Despite siphonarids are broadly distributed around the world, morphological studies on their digestive system are poorly represented. The panpulmonate limpet Siphonaria lessonii is the most abundant gastropod and the dominant herbivore in the rocky intertidal coast of Buenos Aires. The aim of this study was to describe the morphology, histology, ultrastructure, and histochemistry of the digestive gland of this gastropod as well as the cycle of activity of digestion. For that, different histochemical techniques along with light microscopy, transmission electron microscopy, and scanning electron microscopy were employed. This study revealed a complex epithelium, composed of a simple layer with five cell types. Digestive cells and vacuolated cells are responsible for intracellular digestion and energy accumulation; basophilic cells, secrete substances that would be involved in extracellular digestion; pigmented cells might have an excretory function and thin cells would correspond to undifferentiated cells. In addition, the tubules present a changing morphology according to the digestive activity that they undergo. As S. lessonii is a grazer that feeds continuously, the cycle of activity of the digestive gland seems to be daily.

Multiple-biomarker approach in the assessment of the health status of a novel sentinel mussel Brachidontes rodriguezii in a harbor area.

The objective of this study is to analyze whether a combination of biomarkers at different levels of biological complexity could be used to assess the health status of a population of Brachidontes rodriguezii associated to a harbor area in Mar del Plata, Argentina. A battery of biomarkers of general stress was measured in mussels collected from impacted and non-impacted areas. This included: condition index; shell analysis; histopathological alterations; atrophy and integrity of the digestive gland tissue; changes in cell type composition of the digestive gland, and glycogen accumulation. The studied biomarkers were integrated into the Integrative Biological Response (IBR/n) index. Overall, the IBR/n indicated a higher level of stress in mussels located in the harbor area. Thus, this biomarker index is a sensitive analytical tool that could be used to classify the ecotoxicological risk in coastal sites.

Histopathological effects of cypermethrin and Bacillus thuringiensis var. israelensis on midgut of Chironomus calligraphus larvae (Diptera: Chironomidae).

Pesticides are extensively used for the control of agricultural pests and disease vectors, but they also affect non-target organisms. Cypermethrin (CYP) is a synthetic pyrethroid used worldwide. Otherwise, bioinsecticides like Bacillus thuringiensis var. israelensis (Bti) have received great attention as an environmentally benign and desirable alternative. In order to evaluate the toxicity of those pesticides, Chironomus calligraphus was selected due to its high sensitivity to some toxicants. Third and fourth instars larvae were exposed to serial dilutions of CYP and Bti to determine LC50 values. In order to evaluate the potentially histopathological alterations as biomarkers, after 96-h of exposure, live larvae were fixed for histological analysis of the mid region of digestive tract. The 96-h LC50 values were 0.52 and 1.506µg/L for CYP and Bti, respectively. Midgut histological structure of the control group showed a single layer of cubical cells with microvilli in their apical surface and a big central nucleus. The midgut epithelium of larvae exposed to a low concentration of CYP (0.037µg/L) showed secretion activity and vacuolization while at high concentration (0.3µg/L) cells showed a greater disorganization and a more developed fat body. On the other hand, Bti caused progressive histological damage in this tissue. Chironomus calligraphus is sensitive to Bti and CYP toxicity like other Chironomus species. The histopathological alterations could be a valuable tool to assess toxicity mechanism of different pesticides.