Immature instars of mayflies are important constituents of the food web in aquatic ecosystems (especially in Neotropical regions) and they are among the most susceptible arthropods to pyrethroid insecticides. These insecticides have been recognized as important stressors of freshwater ecosystems, but their cellular effects in aquatic insects have been neglected. Here, we assessed the susceptibility to deltamethrin (a typical type II pyrethroid) as well as the deltamethrin-mediated cytomorphological changes in the central nervous system and midgut of the mayfly Callibaetis radiatus. While the deltamethrin LC50 for 24h of exposure was of 0.60 (0.46–0.78) μg of a.i/L, the survival of C. radiatus was significantly reduced in deltamethrin concentrations ≥ 0.25 μg a.i/L at 96h of exposure. Sub-lethal deltamethrin exposure severely affected the cytomorphology of C. radiatus midgut (e.g., muscle layer retraction, cytoplasm vacuolation, nucleus and striated border disorganization) and also induced slight cytomorphological changes in the brain (e.g., presence of pyknotic nuclei) and in the thoracic ganglia (e.g., vacuolation of neurons and presence of pyknotic nuclei) of these insects. However, DNA damage was absent in all of these organs, suggesting that the sublethal cellular stress induced by deltamethrin might disrupt physiological processes (e.g., metabolism or electrical signal transmission) rather than cause cell death (e.g., apoptosis) in C. radiatus. Thus, our findings indicated that deltamethrin actions at cellular levels represent a clear indication of sublethal effects on the C. radiatus survival abilities.

Heteroptera have diverse feeding habits with phytophagous, zoophagous, and haematophagous species. This dietary diversity associated with the monophyly of Heteroptera makes these insects a good object for comparative studies of the digestive tract. This work compares the ultrastructure of the middle midgut region in the phytophagous Coptosoma scutellatum (Plataspidae), Graphosoma lineatum (Pentatomidae), Kleidocerys resedae (Lygaeidae), and zoophagous Rhynocoris iracundus (Reduviidae), Nabis rugosus (Nabidae), and Himacerus apterus (Nabidae), to verify if diet affects midgut cells in phylogenetically related insects. The middle region of the midgut was used for comparison because it is the main site for digestion and absorption of the midgut. The digestive cell ultrastructure was similar in the six species, with features of secretory, absorptive, transport, storage, and excretory cells, suggesting a stronger correlation of middle digestive cell ultrastructure with the phylogeny of these species than with the different heteropteran feeding habits.

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In insects, the midgut performs multiple physiologic functions (e.g., digestion and nutrients absorption) and serves as a physical/chemical barrier against pathogens and chemical stressors such as deltamethrin, a pyrethroid insecticide, commonly used in insect control that are agricultural pests and human disease vectors. Here, we described the midgut cell ultrastructure of Callibaetis radiatus nymphs, which are bioindicators of water quality and the ultrastructural alterations in midgut under sublethal exposure to deltamethrin at three different periods (1, 12, 24 h). The digestive cells of deltamethrin-unexposed nymphs had long microvilli, many mitochondria in the apical cytoplasm, a rough endoplasmic reticulum, a basal labyrinth with openings for hemocele, and the midgut peritrophic matrix which is classified as type I. Nymphs exposed to deltamethrin exhibited digestive cells rich in autophagic vacuoles, basal labyrinth loss, and microvilli disorganization since the first hour of contact with deltamethrin. However, these midgut tissues underwent to autophagic cellular recovery along the 24 h of exposure to deltamethrin. Thus, the sublethal exposure to deltamethrin is sufficient to disturb the ultrastructure of C. radiatus midgut, which might reduce the abilities of these insects to survive in aquatic environments contaminated by pyrethroids.