Exposure to the herbicide tebuthiuron affects behavior, enzymatic activity, morphology and physiology of the midgut of the stingless bee Partamona helleri.

Partamona helleri is an important pollinator in the Neotropics. However, this bee faces an increased risk of pesticide exposure, potentially affecting both individual bees and entire colonies. Thus, this study aimed to evaluate the effects of the herbicide tebuthiuron on behavior, antioxidant activity, midgut morphology, and signaling pathways related to cell death, cell proliferation and differentiation in P. helleri workers. tebuthiuron significantly reduced locomotor activity and induced morphological changes in the midgut. The activity of the detoxification enzymes superoxide dismutase and glutathione S-transferase increased after exposure, indicating a detoxification mechanism. Furthermore, the herbicide led to alterations in the number of positive cells for signaling-pathway proteins in the midgut of bees, suggesting induction of apoptotic cell death and disruption of midgut epithelial regeneration. Therefore, tebuthiuron may negatively impact the behavior, antioxidant activity, morphology, and physiology of P. helleri workers, potentially posing a threat to the survival of this non-target organism.

Midgut morphology of the predator mosquito Lutzia bigoti (Diptera: Culicidae) and its implications for feeding behavior.

Lutzia mosquitoes (Theobald, 1903) are predaceous during their larval stages, but the adult feeding is not clearly understood, especially in relation to blood feeding. In case these mosquitoes are harmless to humans and related animals, they can be useful in biological control of mosquito vectors of pathogens. Investigating the midgut morphology is a good strategy to understand the feeding behavior of this species. The midgut in Lutzia bigoti Bellardi, 1862 displays two distinct portions, a thin anterior midgut and a more dilated posterior midgut. Digestive cells form a single epithelium in the midgut. These cells have long and packed microvilli at their apex and membrane infoldings at their basal portion, the basal labyrinth. The epithelium is supported by a basal lamina. Regarding their cytoplasm, it is noteworthy the abundance of mitochondria, distributed in an apical-basal fashion, and also a whirl-shaped endoplasmic reticulum in the posterior midgut. Basal cells are also found in the midgut of L. bigoti, resembling regenerative cells. The general organization of L. bigoti's midgut closely resembles that of numerous hematophagous mosquitoes previously documented. However, it diverges due to the presence of a peritrophic matrix even when exclusively fed on sugary solutions. Peculiar aspects of L. bigoti's midgut are discussed and compared to those of other mosquito species.

Exposure to low-concentration fipronil impairs survival, behavior, midgut morphology and physiology of Aedes aegypti larvae.

The Aedes aegypti mosquito is a vector for various arboviruses, including dengue and yellow fever. Insecticides, such as pyrethroids and organophosphates, are widely used to manage and control these insects. However, mosquitoes have developed resistance to these chemicals. Therefore, this study aimed to investigate the effects of the commercial formulation of fipronil (Tuit® Florestal; 80% purity) on the survival, behavior, morphology, and proteins related to signaling pathways of the midgut in A. aegypti larvae under controlled laboratory conditions. Significant reductions in immature survival were observed in all concentrations of fipronil tested. Low insecticide concentration (0.5 ppb) led to decreased locomotor activity in the larvae and caused disorganization of the epithelial tissue in the midgut. Moreover, exposure to the insecticide decreased the activity of detoxifying enzymes such as catalase, superoxide dismutase, and glutathione-S-transferase. On the other hand, the insecticide increased protein oxidation and nitric oxide levels. The detection of LC3, caspase-3, and JNK proteins, related to autophagy and apoptosis, increased after exposure. However, there was a decrease in the positive cells for ERK 1/2. Furthermore, the treatment with fipronil decreased the number of positive cells for the proteins FMRF, Prospero, PH3, Wg, Armadillo, Notch, and Delta, which are related to cell proliferation and differentiation. These findings demonstrate that even at low concentrations, fipronil exerts larvicidal effects on A. aegypti by affecting behavior and enzymatic detoxification, inducing protein oxidation, free radical generation, midgut damage and cell death, and inhibiting cell proliferation and differentiation. Thus, this insecticide may represent a viable alternative for controlling the spread of this vector.

A mixture of mesotrione and atrazine harms adults and larvae of the predatory wasp Polistes satan.

Herbicides have been intensively used for weed control, raising concerns about their potentially adverse effects on non-target organisms. Research on the effects of these common agrochemicals on beneficial insects and the ecosystem services they provide (e.g., predation and pollination) is scarce. Therefore, we tested whether a commercial formulation comprising a mixture of mesotrione and atrazine was detrimental to adult females and larvae of the Neotropical predatory social wasp Polistes satan, which is an effective natural enemy of crop pests. Wasps were individually fed syrups contaminated with different concentrations of the herbicide above and below the maximum label rate (MLR = 12 mL/L). Survival was assessed. The locomotor activity, immune response, and midgut morphology of adults as well as the immune response of the larvae were also studied. Herbicide concentrations far above the MLR (12, 40, and 100 times) caused adult mortality, whereas lower concentrations (0.5, 1, and 6 times) did not. Herbicide exposure at 0.5 to 12 times the MLR increased adult activity. Adult exposure at 0.1 or 0.5 times the MLR did not affect melanotic encapsulation of foreign bodies but led to changes in the morphology of the midgut epithelium and peritrophic matrix. In larvae, the ingestion of herbicide at 0.1 or 0.2 times the MLR (corresponding to 9.6 and 19.2 ng of herbicide per individual) did not cause mortality but decreased their melanization-encapsulation response. Increased locomotor activity in herbicide-exposed adults can affect their foraging activity. The altered midgut morphology of adults coupled with the decreased immune response in larvae caused by herbicide exposure at realistic concentrations can increase the susceptibility of wasps to infections. Therefore, herbicides are toxic to predatory wasps.

Exploring honey bee toxicological data as a proxy for assessing dimethoate sensitivity in stingless bees.

The high diversity and distinctive characteristics of stingless bees pose challenges in utilizing toxicity test results for agrochemical registrations. Toxicity assessments were performed on 15 stingless bee species, along with the honey bee, using the insecticide dimethoate, following adapted OECD protocols. Median lethal doses over 24 h (24 h-LD50) were determined for exposure routes (acute oral or contact) and species. Species sensitivity distribution (SSD) curves were constructed and the 5% hazard doses (HD5) were estimated based on 24 h-LD50 values. The SSD curve was adjusted as the body weight and dimethoate response were correlated. Lighter bees (<10 mg) had lower 24 h-LD50 values. Contact exposure for adjusted HD5 suggested insufficient protection for Melipona mondury, whereas the oral exposure HD5 indicated no risks for the other 14 species. Comprehensive risk assessments are crucial for understanding the agrochemical impact on stingless bees, emphasizing the need for a broader species range in formulating conservation strategies.

Toxicological Assessments of Agrochemicals in Stingless Bees in Brazil: a Systematic Review.

The growing concern with the decline of pollinators worldwide is centered on honey bees, due to their wide distribution, economic, and ecological importance. This type of concern remained less evident for stingless bees, which are widely distributed in the Neotropics, until recently. Since exposure to agrochemicals has been identified as one of the potential threats to bees, the present systematic review compiled information from toxicological evaluations in stingless bees in Brazil, home to a considerable portion of the existing species. This systematic review was performed considering species, research institutions, scientific journals, metrics, experimental set ups, and agrochemicals. The first article in this topic was published in 2010. Since then, 93 scientific papers were published, which showed that there are few species of stingless bees used for toxicological evaluations and Brazilian institutions lead these evaluations. Only 1.5% of the stingless bees' species that occur in Brazil were assessed through chronic exposure in the larval stage. The Universidade Federal de Viçosa (UFV) is responsible for 37% of the total publications. The main route of exposure was acute, using adults in laboratory conditions. The main group of agrochemicals studied were insecticides, in particular the neonicotinoids. The current results reveal the advances achieved and point out the gaps that still need to be filled considering toxicological evaluations in stingless bees.

Morphological characterization of antennal sensilla in Toxorhynchites theobaldi, Toxorhynchites violaceus, and Lutzia bigoti adults: a comparative study using scanning electron microscopy.

Some mosquitoes, including species of the genus Toxorhynchites, are known for actively preying on other mosquito larvae, making these predators valuable allies in the fight against vector-borne diseases. A comprehensive understanding of the anatomy and physiology of these potential biological control agents is helpful for the development of effective strategies for controlling vector populations. This includes the antennae, a crucial component in the search for hosts, mating, and selection of oviposition sites. This study utilized scanning electron microscopy to characterize the sensilla on the antennae of adult mosquitoes from two species that are exclusively phytophagous, including Toxorhynchites theobaldi and Toxorhynchites violaceus, as well as Lutzia bigoti, which females are allegedly hematophagous. The types of sensilla in each species were compared, and five basic types of antennal sensilla were identified: trichoid, chaetic, coeloconic, basiconic, and ampullacea. The analysis also found that they were morphologically similar across the three species, regardless of feeding habits or sex. The identification and characterization of basic types of antennal sensilla in T. theobaldi, T. violaceus, and L. bigoti suggest that these structures, which play a crucial role in the behavior and ecology, have common functions across different mosquito species, despite differences in feeding habits or sex.

Morphological and morphometrical assessment of spermathecae of Aedes aegypti females

The vectorial capacity of Aedes aegypti is directly influenced by its high reproductive output. Nevertheless, females are restricted to a single mating event, sufficient to acquire enough sperm to fertilize a lifetime supply of eggs. How Ae. aegypti is able to maintain viable spermatozoa remains a mystery. Male spermatozoa are stored within either of two spermathecae that in Ae. aegypti consist of one large and two smaller organs each. In addition, each organ is divided into reservoir, duct and glandular portions. Many aspects of the morphology of the spermatheca in virgin and inseminated Ae. aegypti were investigated here using a combination of light, confocal, electron and scanning microscopes, as well as histochemistry. The abundance of mitochondria and microvilli in spermathecal gland cells is suggestive of a secretory role and results obtained from periodic acid Schiff assays of cell apexes and lumens indicate that gland cells produce and secrete neutral polysaccharides probably related to maintenance of spermatozoa. These new data contribute to our understanding of gamete maintenance in the spermathecae of Ae. aegypti and to an improved general understanding of mosquito reproductive biology.

A comparative study of fat body morphology in five mosquito species

The insect fat body plays major roles in the intermediary metabolism, in the storage and transport of haemolymph compounds and in the innate immunity. Here, the overall structure of the fat body of five species of mosquitoes (Aedes albopictus, Aedes fluviatilis, Culex quinquefasciatus, Anopheles aquasalis and Anopheles darlingi) was compared through light, scanning and transmission electron microscopy. Generally for mosquitoes, the fat body consists of lobes projecting into the haemocoel and is formed by great cell masses consisting of trophocytes and oenocytes. Trophocytes are rich in lipid droplets and protein granules. Interestingly, brown pigment granules, likely ommochromes, were found exclusively in the trophocytes located within the thorax and near the dorsal integument of Anopheles, which is suggestive of the role these cells play in detoxification via ommochrome storage. This study provides a detailed comparative analysis of the fat body in five different mosquito species and represents a significant contribution towards the understanding of the structural-functional relationships associated with this organ.

Insights into the transcriptome of oenocytes from Aedes aegypti pupae

Oenocytes are ectodermic cells present in the fat body of several insect species and these cells are considered to be analogous to the mammalian liver, based on their role in lipid storage, metabolism and secretion. Although oenocytes were identified over a century ago, little is known about their messenger RNA expression profiles. In this study, we investigated the transcriptome of Aedes aegypti oenocytes. We constructed a cDNA library from Ae. aegypti MOYO-R strain oenocytes collected from pupae and randomly sequenced 687 clones. After sequences editing and assembly, 326 high-quality contigs were generated. The most abundant transcripts identified corresponded to the cytochrome P450 superfamily, whose members have roles primarily related to detoxification and lipid metabolism. In addition, we identified 18 other transcripts with putative functions associated with lipid metabolism. One such transcript, a fatty acid synthase, is highly represented in the cDNA library of oenocytes. Moreover, oenocytes expressed several immunity-related genes and the majority of these genes were lysozymes. The transcriptional profile suggests that oenocytes play diverse roles, such as detoxification and lipid metabolism, and increase our understanding of the importance of oenocytes in Ae. aegypti homeostasis and immune competence.

O corpo gorduroso de Aedes (Stegomyia) aegypti (Diptera; Nematocera) (Linnaeus, 1762): estudo morfológico do órgão em diferentes idades e condições alimentares, isolamento, cultivo primário e transcriptoma dos enócitos

O corpo gorduroso é o principal órgão do metabolismo intermediário dos insetos e a principal fonte de componentes da hemolinfa. Dois tipos celulares estão presentes no corpo gorduroso dos dípteros: os trofócitos e os enócitos. O presente trabalho teve como objetivos testar as seguintes hipóteses: (a) o corpo gorduroso de A. aegypti é capaz de se reorganizar do ponto de vista ultra-estrutural, histoquímico e morfométrico de acordo com a idade e com o tipo de alimentação; (b) os enócitos de A. aegypti podem ser cultivados e (c) os mesmos expressam transcritos relacionados com os processos de desintoxicação, síntese de lipídios e imunidade inata. Para tanto, foram utilizadas fêmeas recém-emergidas, fêmeas com 18d de idade, sendo um grupo 18h e outro quatro dias após a alimentação sangüínea e fêmeas com 18d alimentadas com açúcar. Nas fêmeas alimentadas com açúcar, o corpo gorduroso está mais desenvolvido do que nas pós-mergidas e nas alimentadas com sangue. Nessas mesmas fêmeas, os trofócitos apresentam o citoplasma preenchido basicamente por gotículas de lipídio devido à lipogênese. Após a alimentação sangüínea, os lóbulos do corpo gorduroso estão achatados, o que pode ser explicado pelo esvaziamento do órgão que exporta nutrientes para os ovários. As alterações dos trofócitos após a alimentação sangüínea incluem o aumento do diâmetro nuclear, a diminuição das gotículas de lipídios, o alargamento das mitocôndrias e a presença de lisossomos nos trofócitos. Os enócitos puderam ser mantidos viáveis por 57 dias em cultivo primário e o citoplasma dessas células é quase todo preenchido pelo retículo endoplasmático liso (REL). Os transcritos mais abundantes encontrados nos enócitos correspondem à citocromo P450, responsável pela desintoxicação e às outras proteínas relacionadas com o metabolismo de lipídios.

Também são encontrados transcritos. relacionados com o reconhecimento e de destruição de patógenos. Provavelmente, a presença do REL e a localização periférica dos enócitos no corpo gorduroso de A. aegypti facilita o processo de desintoxicação, a produção e a secreção de lipídios para a hemolinfa, além do reconhecimento de patógenos e da secreção de componentes antimicrobianos. A presença do REL desenvolvido e a expressão de várias enzimas comprometidas com a desintoxicação e o metabolismo de lipídios sugerem que os enócitos participam da homeostasia e síntese lipídica em A. aegypti. Além disso, nossos resultados confirmam o corpo gorduroso de A. aegypti como um órgão capaz de reorganizar sua microanatomia, componentes citoplasmáticos e aspectos subcelulares de acordo com a idade e o tipo de dieta. Também é possível cultivar os enócitos e sua purificação, cultivo primário e transcriptoma constituem importantes ferramentas para que futuros estudos possam investigar a atuação dessas células na fisiologia e na imunidade inata de mosquitos

O corpo gorduroso de Aedes (Stegomyia) aegypti (Diptera; Nematocera) (Linnaeus, 1762): estudo morfológico do órgão em diferentes idades e condições alimentares, isolamento, cultivo primário e transcriptoma dos enócitos / The fat body Aedes (Stegomyia) aegypti (Dipetra; Nematocera) (Linnaeus, 1762): Morphological study of the organ in different ages and diets, isolation, primary culture and the transcriptome of the oenocytes

O corpo gorduroso é o principal órgão do metabolismo intermediário dos insetos e a principal fonte de componentes da hemolinfa. Dois tipos celulares estão presentes no corpo gorduroso dos dípteros: os trofócitos e os enócitos. O presente trabalho teve como objetivos testar as seguintes hipóteses: (a) o corpo gorduroso de A. aegypti é capaz de se reorganizar do ponto de vista ultra-estrutural, histoquímico e morfométrico de acordo com a idade e com o tipo de alimentação; (b) os enócitos de A. aegypti podem ser cultivados e (c) os mesmos expressam transcritos relacionados com os processos de desintoxicação, síntese de lipídios e imunidade inata. Para tanto, foram utilizadas fêmeas recém-emergidas, fêmeas com 18d de idade, sendo um grupo 18h e outro quatro dias após a alimentação sangüínea e fêmeas com 18d alimentadas com açúcar. Nas fêmeas alimentadas com açúcar, o corpo gorduroso está mais desenvolvido do que nas pós-mergidas e nas alimentadas com sangue. Nessas mesmas fêmeas, os trofócitos apresentam o citoplasma preenchido basicamente por gotículas de lipídio devido à lipogênese. Após a alimentação sangüínea, os lóbulos do corpo gorduroso estão achatados, o que pode ser explicado pelo esvaziamento do órgão que exporta nutrientes para os ovários. As alterações dos trofócitos após a alimentação sangüínea incluem o aumento do diâmetro nuclear, a diminuição das gotículas de lipídios, o alargamento das mitocôndrias e a presença de lisossomos nos trofócitos. Os enócitos puderam ser mantidos viáveis por 57 dias em cultivo primário e o citoplasma dessas células é quase todo preenchido pelo retículo endoplasmático liso (REL). Os transcritos mais abundantes encontrados nos enócitos correspondem à citocromo P450, responsável pela desintoxicação e às outras proteínas relacionadas com o metabolismo de lipídios.

Também são encontrados transcritos. relacionados com o reconhecimento e de destruição de patógenos. Provavelmente, a presença do REL e a localização periférica dos enócitos no corpo gorduroso de A. aegypti facilita o processo de desintoxicação, a produção e a secreção de lipídios para a hemolinfa, além do reconhecimento de patógenos e da secreção de componentes antimicrobianos. A presença do REL desenvolvido e a expressão de várias enzimas comprometidas com a desintoxicação e o metabolismo de lipídios sugerem que os enócitos participam da homeostasia e síntese lipídica em A. aegypti. Além disso, nossos resultados confirmam o corpo gorduroso de A. aegypti como um órgão capaz de reorganizar sua microanatomia, componentes citoplasmáticos e aspectos subcelulares de acordo com a idade e o tipo de dieta. Também é possível cultivar os enócitos e sua purificação, cultivo primário e transcriptoma constituem importantes ferramentas para que futuros estudos possam investigar a atuação dessas células na fisiologia e na imunidade inata de mosquitos

Biochemical and morphological aspects of salivary glands of the predator Brontocoris tabidus (Heteroptera: Pentatomidae)

The aim of this work was to study the feeding habits of the predator Brontocoris tabidus (Heteroptera: Pentatomidae) analyzing the salivary gland complex (SGC). The SGC was dissected and subjected to histological analyses and biochemical assays. Results showed that a pair of bilobed principal salivary glands and one pair of tubular accessory salivary glands form the SGC and different parts of salivary gland synthesizes similar substances. Lipases, a-amylase and trypsin-like enzymes were detected at both lobes of the principal salivary gland. These data together with observations related to the predator's diet suggested that it could be considered an obligate zoophytophagous.

Esta pesquisa contribui para a compreensão dos hábitos alimentares do percevejo predador Brontocoris tabidus (Heteroptera: Pentatomidae) por meio de análises morfológicas e bioquímicas de suas glândulas salivares. Espécimes tiveram o sistema salivar dissecado, e parte das amostras obtidas foi submetida a análises histológicas. Secções foram coradas com hematoxilina e eosina ou submetidas aos testes histoquímicos azul de bromofenol, PAS e azul do Nilo. A outra parte das amostras dissecadas teve seu conteúdo analisado em SDS-PAGE e em ensaios de atividade enzimática. Os resultados mostram que o sistema salivar de B. tabidus é formado por um par de glândulas bilobadas principais e um par de glândulas tubulares acessórias, e que suas diferentes regiões sintetizam substâncias semelhantes. Os ensaios enzimáticos detectaram atividade para tripsina, amilase e lipase, que juntamente com observações de laboratório relativas à dieta do predador indicam que o mesmo pode ser considerado um zoofitófago obrigatório.