Effects of chronic oral exposure to insecticide teflubenzuron on the midgut of the honey bee Apis mellifera workers: histopathological insights into pesticide toxicity.

The honey bee Apis mellifera plays a significant role as a pollinator of native and cultivated plants, by increasing the productivity of several cultures, preserving the flora, and producing forest seeds. However, bee populations are declining worldwide, including A. mellifera, due to Colony Collapse Disorder, mainly resulting from the constant use of pesticides in the crops. Teflubenzuron is a physiological insecticide that belongs to the benzoylurea group, which inhibits chitin synthesis, the main component of the insect integument classified as safe for non-target insects, including bees. However, its effect on non-target organs of insects remains unknown. The midgut is the main organ of the digestive tract, which works in digestion and absorption and may be exposed to pesticides that contaminate food resources. The present work aimed to verify if the insecticide teflubenzuron is toxic and has histopathological effects on the midgut of A. mellifera adult workers. Workers exposed orally and chronically to the field-realistic concentration of teflubenzuron present 81.54% mortality. The epithelium of the midgut of these bees presents high vacuolization, spherocrystals, cell fragments released to the organ lumen, apocrine secretion, nuclear pyknosis, loss of cell-cell contact, and damage to regenerative cell nests and to the peritrophic matrix. These results indicate that the chitin synthesis-inhibiting insecticide teflubenzuron is toxic to A. mellifera after chronic oral exposure, at realistic field concentration, although it is classified as non-toxic to adult and non-target insects.

Resistance of Lepidopteran Pests to Bacillus thuringiensis Toxins: Evidence of Field and Laboratory Evolved Resistance and Cross-Resistance, Mode of Resistance Inheritance, Fitness Costs, Mechanisms Involved and Management Options.

Bacillus thuringiensis (Bt) toxins are potential alternatives to synthetic insecticides for the control of lepidopteran pests. However, the evolution of resistance in some insect pest populations is a threat and can reduce the effectiveness of Bt toxins. In this review, we summarize the results of 161 studies from 20 countries reporting field and laboratory-evolved resistance, cross-resistance, and inheritance, mechanisms, and fitness costs of resistance to different Bt toxins. The studies refer mainly to insects from the United States of America (70), followed by China (31), Brazil (19), India (12), Malaysia (9), Spain (3), and Australia (3). The majority of the studies revealed that most of the pest populations showed susceptibility and a lack of cross-resistance to Bt toxins. Factors that delay resistance include recessive inheritance of resistance, the low initial frequency of resistant alleles, increased fitness costs, abundant refuges of non-Bt, and pyramided Bt crops. The results of field and laboratory resistance, cross-resistance, and inheritance, mechanisms, and fitness cost of resistance are advantageous for predicting the threat of future resistance and making effective strategies to sustain the effectiveness of Bt crops.

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.

Action of dithiocarbimates salts on the honey bee and its pathogen Nosema ceranae.

Apis mellifera, crucial pollinators for both native and cultivated plants, also yield various products such as honey, wax, royal jelly, and propolis, extensively utilized in the food, pharmaceuticals, and cosmetics industries. Nosema ceranae, a prevalent microsporidian worldwide, stands as a significant pathogen for A. mellifera, showing resistance to conventional antibiotics. Consequently, the exploration of novel compounds for N. ceranae control becomes imperative. Dithiocarbimate derivatives emerge as promising antifungal candidates under evaluation for combating various pathogens, particularly those affecting plants. This study assessed the toxicity profile of six dithiocarbimate derivatives on A. mellifera worker survival and N. ceranae pathogen. Among these, four compounds exhibited minimal bee mortality and proceeded to further evaluation against N. ceranae. In vitro assays demonstrated their inhibitory effects on spore germination. Remarkably, the most potent compound suppressed N. ceranae spores by 62% at a concentration of 20 µmol L-1in vivo. Thus, these dithiocarbimate derivatives represent promising new antifungal agents for combatting nosemosis in honey bee populations.

Acute exposure to fungicide fluazinam induces cell death in the midgut, oxidative stress and alters behavior of the stingless bee Partamona helleri (Hymenoptera: Apidae).

Stingless bees (Hymenoptera: Meliponini) are pollinators of both cultivated and wild crop plants in the Neotropical region. However, they are susceptible to pesticide exposure during foraging activities. The fungicide fluazinam is commonly applied in bean and sunflower cultivation during the flowering period, posing a potential risk to the stingless bee Partamona helleri, which serves as a pollinator for these crops. In this study, we investigated the impact of acute oral exposure (24 h) fluazinam on the survival, morphology and cell death signaling pathways in the midgut, oxidative stress and behavior of P. helleri worker bees. Worker bees were exposed for 24 h to fluazinam (field concentrations 0.5, 1.5 and 2.5 mg a.i. mL-1), diluted in 50 % honey aqueous solution. After oral exposure, fluazinam did not harm the survival of worker bees. However, sublethal effects were revealed using the highest concentration of fluazinam (2.5 mg a.i. mL-1), particularly a reduction in food consumption, damage in the midgut epithelium, characterized by degeneration of the brush border, an increase in the number and size of cytoplasm vacuoles, condensation of nuclear chromatin, and an increase in the release of cell fragments into the gut lumen. Bees exposed to fluazinam exhibited an increase in cells undergoing autophagy and apoptosis, indicating cell death in the midgut epithelium. Furthermore, the fungicide induced oxidative stress as evidenced by an increase in total antioxidant and catalase enzyme activities, along with a decrease in glutathione S-transferase activity. And finally, fluazinam altered the walking behavior of bees, which could potentially impede their foraging activities. In conclusion, our findings indicate that fluazinam at field concentrations is not lethal for workers P. helleri. Nevertheless, it has side effects on midgut integrity, oxidative stress and worker bee behavior, pointing to potential risks for this pollinator.

Effects of the insecticide flupyradifurone on Anticarsia gemmatalis caterpillar and its predator Podisus nigrispinus.

The caterpillar Anticarsia gemmatalis (Lepidoptera: Noctuidae) is a prevalent pest in soybean plantations, managed using both natural and synthetic chemical products. However, the emergence of resistance in some populations emphasizes the need to explore alternative insecticides. Flupyradifurone, a neurotoxic insecticide, has not been previously used for controlling A. gemmatalis. This study evaluated the potential of flupyradifurone in the management of A. gemmatalis. Initially, the toxicity and anti-feeding effects, as well as histopathological and cytotoxic impacts, of flupyradifurone on A. gemmatalis were evaluated. Subsequently, the indirect effects of flupyradifurone on the midgut and fat body of the predator Podisus nigrispinus (Hemiptera: Pentatomidae) were verified. The results indicate the susceptibility of caterpillars to flupyradifurone, with an LC50 of 5.10 g L-1. Furthermore, the insecticide adversely affects survival, induces an anti-feeding response, and inflicts damage on the midgut of the caterpillars. However, flupyradifurone also leads to side effects in the predator P. nigrispinus through indirect intoxication of the caterpillars, including midgut and fat body damage. While flupyradifurone demonstrates toxicity to A. gemmatalis, suggesting its potential for the chemical control of this pest, the indirect negative effects on the predator indicate the need for its controlled use in integrated pest management programs with the insecticide and the predator.

First report of resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae) to lambda-cyhalothrin from Pakistan: baseline susceptibility, selection, occurrence of cross-resistance, realized heritability, and inheritance mode of resistance.

Fall armyworm (FAW) Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), is a global pest causing damage to several crops. However, its management using chemical control is a challenge due to its capacity to evolve resistance to insecticides. After 6 generations of selection with lambda-cyhalothrin, the LC50 for the insecticide-resistant strain (Lambda-Sel) was 486 ppm, higher than that of the field strain (FAW-MUL) (7.5 ppm), susceptible laboratory strain (Lab-PK) (0.46 ppm), and laboratory unselected strain (UNSEL) (5.26 ppm). Laboratory selection with lambda-cyhalothrin increased resistance from 16.3- to 1056.52-fold and 1.43- to 92.4-fold to lambda-cyhalothrin compared to Lab-PK and UNSEL strains, respectively. The selected strain of S. frugiperda (Lambda-Sel) presented low cross-resistance to chlorpyrifos, moderate to deltamethrin and indoxacarb, very low to spinosad, and no cross-resistance to emamectin benzoate. The realized heritability (h2) of lambda-cyhalothrin resistance in the Lambda-Sel strain was very high (0.88). The reciprocal cross progenies of F1 (Lambda-Sel ♀ × Lab-PK ♂), F1' (Lambda-Sel ♂ × Lab-PK ♀), BC1 (F1 ♀ × Lambda-Sel ♂), and BC2 (F1 ♀ × Lab-PK ♂) showed high resistance ratios of 545.64-, 396.52-, 181.18-, and 146.54-fold, respectively compared to Lab-PK. The degree of dominance values for lambda-cyhalothrin in F1 and F1' indicates incompletely dominant resistance. The difference between observed and expected mortality in backcross populations (BC1 and BC2) revealed a polygenic resistance. In conclusion, the resistance to lambda-cyhalothrin was autosomal, incompletely dominant, and polygenic. These findings provide new insights for insect resistance management strategies to mitigate the occurrence of resistance in this global pest.

Expression and function of the vitellogenin receptor in the hypopharyngeal glands of the honey bee Apis mellifera (Hymenoptera: Apidae) workers.

The vitellogenin receptor (VgR) is essential for the uptake and transport of the yolk precursor, vitellogenin (Vg). Vg is synthesized in the fat body, released in the hemolymph, and absorbed in the ovaries, via receptor-mediated endocytosis. Besides its important role in the reproductive pathway, Vg occurs in nonreproductive worker honey bee, suggesting its participation in other pathways. The objective was to verify if the VgR occurs in the hypopharyngeal glands of Apis mellifera workers and how Vg is internalized by these cells. VgR occurrence in the hypopharyngeal glands was evaluated by qPCR analyses of VgR and immunohistochemistry in workers with different tasks. The VgR gene is expressed in the hypopharyngeal glands of workers with higher transcript levels in nurse honey bees. VgR is more expressed in 11-day-old workers from queenright colonies, compared to orphan ones. Nurse workers with developed hypopharyngeal glands present higher VgR transcripts than those with poorly developed glands. The immunohistochemistry results showed the co-localization of Vg, VgR and clathrin (protein that plays a major role in the formation of coated vesicles in endocytosis) in the hypopharyngeal glands, suggesting receptor-mediated endocytosis. The results demonstrate that VgR performs the transport of Vg to the hypopharyngeal glands, supporting the Ovary Ground Plan Hypothesis and contributing to the understanding of the role of this gland in the social context of honey bees.

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.

Exploring the effects of the acaricide cyflumetofen on the vital organs of the honey bee Apis mellifera (Hymenoptera: Apidae) workers.

Bees are important for maintaining ecosystems, pollinating crops and producing marketable products. In recent years, a decline in bee populations has been reported, with multifactorial causes, including the intensification of pesticide use in agriculture. Among pesticides, cyflumetofen is an insecticide and acaricide used in apple, coffee and citrus crops, whose main pollinator is the honey bee Apis mellifera. Therefore, this bee is a potential target of cyflumetofen during foraging. This study evaluated the histopathological and cytological damage in the midgut, hypopharyngeal glands and fat body of A. mellifera workers exposed to LC50 of cyflumetofen. The midgut epithelium of exposed bees presented cytoplasmic vacuolization, release of vesicles and cell fragments, which indicate autophagy, increased production of digestive enzymes and cell death, respectively. The cytological analysis of the midgut revealed the dilation of the basal labyrinth and the presence of spherocrystals in the digestive cells. The hypopharyngeal glands produced greater amounts of secretion in treated bees, whereas no changes were observed in the fat body. The results indicate that acute exposure to cyflumetofen negatively affect A. mellifera, causing damage to the midgut and changes in the hypopharyngeal glands, which may compromise the survival and foraging of this pollinator.

Interspecific insect relationships on Terminalia argentea (Myrtales: Combretaceae) trees in the Cerrado biome.

Terminalia argentea Mart. (Combretaceae), native to Brazil, is used in habitat restoration programs. Arthropods are bioindicators because their populations reflect changes in the environment. We evaluated the recovery of a degraded area by using ecological indices and analyzing arthropod interactions on T. argentea plants. The richness and diversity of sap-sucking Hemiptera and the abundance of tending ants and Sternorrhyncha predators increased with the number of T. argentea leaves. The correlation of the abundance of tending ants and Sternorrhyncha predators was positive with that of the sap-sucking Hemiptera, and the abundance of Sternorrhyncha predators was negative with that of tending ants and sap-sucking Hemiptera. The positive correlation between the abundance, richness, and diversity of insect groups and numbers of T. argentea leaves is an example of the bottom-up regulation mechanism, with the population dynamics of the lower trophic levels dictating those of higher trophic levels. The contribution of T. argentea, a host plant of many arthropods, to the recovery of ecological relationships between organisms in degraded ecosystems is important.

Morphology of the Female and Male Reproductive Tracts and More Data on the Spermatostyle in the Brazilian Gyretes sp. (Coleoptera, Adephaga, Gyrinidae).

We investigated the male and female reproductive tracts of Gyretes sp. with light and transmission electron microscopies. The male has a pair of testes with a single coiled follicle, followed by short efferent ducts, which have a similar shape and diameter to the testes. Long ducts (epididymides) with differential epithelium open in a pair of long vasa deferentia that lead to the accessory glands. Glycoprotein secretions from the vas deferens epithelium constitute the spermatostyle for spermatozoa aggregation. The female has numerous ovarioles per ovary, a coiled fertilization duct, an accessory gland, and an elongated vagina. Spermatozoa are stored as unaggregated cells in the fertilization duct. In Gyrinidae, the testes and accessory glands show diverse shapes, and the female sperm storage organs vary in shape, size, and type and may play a role in the interaction with sperm aggregates. Testes with a single follicle and vasa deferentia opening in the accessory glands of Gyretes sp. are features shared with other Gyrinidae and other Adephaga. We proposed adding this latter trait to characterize this suborder of beetles. The morphology of the reproductive organs in both sexes contributes to comparative analyses and knowledge of the reproductive biology of Gyretes and may provide additional features for systematics.

Morphology of the Mandibular and Intramandibular Glands of Army Ant Workers of Labidus praedator (Smith 1858) and Labidus coecus (Latreille 1802) (Formicidae: Dorylinae).

Social insects are characterized by having a wide diversity of exocrine glands, with highlights for ants with about 85 glands spreading throughout the body. The mandibular and intramandibular glands are associated with the production of pheromones. The army ants (Dorylinae) play an important role in the structure of the invertebrate community because they are efficient predators and provide suitable conditions for various animals following their invasions in the food search. Labidus coecus (Latreille) is an underground-ameliorating ant and Labidus praedator (Smith) is a generalist surface predator which can deplete invertebrate biomass by up to 75%. This work investigated the morphology of the mandibular and intramandibular glands of L. praedator and L. coecus workers. The glands were analyzed by light microscopy, histochemistry, and scanning electron microscopy. The mandibular and intramandibular glands of the two species were classified as class III glands. The data on the morphology of the mandibular glands has revealed that they have characteristics in common with other subfamilies. The intramandibular glands of the two species of Labidus have similar morphology and chemical composition, which indicates that the components of these glands can have the same function despite their different habits.

Effect of Induced Pulmonary Arterial Hypertension on Testicular Parameters of Wistar Rats Subjected to Resistance Exercise Training.

Pulmonary arterial hypertension (PAH) is characterized by elevated arterial pressure and vascular resistance. PAH may cause alterations in the microcirculation of several organs, including the kidney, liver, brain, and testes. However, it remains unclear whether monocrotaline-induced PAH exerts detrimental effects on animal testes. Thus, we analyzed the impact of PAH on testicular morphology and function. Additionally, we investigated the effect of resistance exercise training (RT) on testicular parameters in PAH rats. Eight healthy Wistar rats and eight PAH rats were subjected to RT training for 30 days; the other PAH and healthy rats (n = 8/group) did not exercise. PAH rats had lower reproductive organ weight, serum testosterone levels, testicular glucose, and nitric oxide (NO) levels, Leydig cell parameters, tubular morphometry, germ cell counts, and daily sperm production than healthy animals did. The practice of RT attenuated the negative impact of PAH on the relative weights of the testes and epididymides, Leydig cell number, nuclear volume, testicular NO levels, and seminiferous epithelium architecture. Moreover, RT positively influenced testosterone levels in PAH animals. We conclude that PAH exerts deleterious effects on testicular histology and function. However, RT can be beneficial to the PAH-affected testicular parameters.

Effects of the insecticide imidacloprid on the post-embryonic development of the honey bee Apis mellifera (Hymenoptera: Apidae).

The widespread use of pesticides in agriculture has been linked to declines in bee populations worldwide. Imidacloprid is a widely used systemic insecticide that can be found in the pollen and nectar of plants and has the potential to negatively impact the development of bee larvae. We investigated the effects of oral exposure to a realistic field concentration (20.5 ng g-1) of imidacloprid on the midgut and fat body of Apis mellifera worker larvae. Our results showed that larvae exposed to imidacloprid exhibited changes in the midgut epithelium, including disorganization of the brush border, nuclear chromatin condensation, cytoplasm vacuolization, and release of cell fragments indication cell death. Additionally, histochemical analysis revealed that the midgut brush border glycocalyx was disorganized in exposed larvae. The fat body cells of imidacloprid-exposed larvae had a decrease in the size of lipid droplets from 50 to 8 µm and increase of 100 % of protein content, suggesting possible responses to the stress caused by the insecticide. However, the expression of de cdc20 gene, which plays a role in cell proliferation, was not affected in the midgut and fat body of treated larvae. These results suggest that imidacloprid negatively affects non-target organs during the larval development of A. mellifera potentially impacting this important pollinator species.

Hemocyte morphology of worker subcastes of the leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae).

Hemocytes are cells present in the hemolymph of insects that play a role in combating invasive pathogens, ensuring defense by the immune system in these organisms. While the types of hemocytes are well known in some insect representatives, data on these cells in Hymenoptera are limited to certain bees and wasps, with little information available for ants. Among ants, the genus Atta has environmental and economic importance, forming highly organized colonies consisting of the queen and workers, with the latter subdivided into subcastes: gardeners, waste removers, foragers, and soldiers, which are exposed to different pathogens. This study describes the morphology of hemocytes in the worker subcastes of Atta sexdens rubropilosa. Hemolymph samples from the ant were submitted to light, confocal, and scanning electron microscopy analyses. Five types of hemocytes were identified in the hemolymph of all ant subcastes, including prohemocytes, oenocytoids, spherulocytes, plasmatocytes, and granulocytes. They exhibited nuclei with a predominance of decondensed chromatin. The granulocytes were the most abundant cell type in the subcastes, followed by prohemocytes, plasmatocytes, oenocytoids, and spherulocytes. Phagocytosis assays reveal that plasmatocytes and granulocytes are the main phagocytic cells in all castes evaluated. This study fills an important gap in understanding the immune response in this ant species.

Midgut Cell Damage and Oxidative Stress in Partamona helleri (Hymenoptera: Apidae) Workers Caused by the Insecticide Lambda-Cyhalothrin.

The stingless bee Partamona helleri plays a role in pollinating both native and cultivated plants in the Neotropics. However, its populations can be reduced by the pyrethroid insecticide lambda-cyhalothrin. This compound may cross the intestinal barrier and circulate through the hemolymph, affecting various non-target bee organs. The aim of the present study was to assess the extent of cellular damage in the midgut and the resulting oxidative stress caused by lambda-cyhalothrin in P. helleri workers. Bees were orally exposed to lambda-cyhalothrin. The lethal concentration at which 50% of the bees died (LC50) was 0.043 mg a.i. L-1. The P. helleri workers were fed this concentration of lambda-cyhalothrin and their midguts were evaluated. The results revealed signs of damage in the midgut epithelium, including pyknotic nuclei, cytoplasm vacuolization, changes in the striated border, and the release of cell fragments, indicating that the midgut was compromised. Furthermore, the ingestion of lambda-cyhalothrin led to an increase in the activity of the detoxification enzyme superoxide dismutase and the levels of the NO2/NO3 markers, indicating oxidative stress. Conversely, the activities of the catalase and glutathione S-transferase enzymes decreased, supporting the occurrence of oxidative stress. In conclusion, the ingestion of lambda-cyhalothrin by P. helleri workers resulted in damage to their midguts and induced oxidative stress.

Morphology and Morphometry of the Reproductive Tract of the Cotton Boll Weevil after Prolonged Feeding on Alternative Diets.

Anthonomus grandis Boheman (Coleoptera: Curculionidae) survives on alternative diets; however, this induces reproductive diapause. The objective was to evaluate the morphology and morphometry of the reproductive tract of this weevil after feeding on alternative diets. The experimental design was completely randomized with 160 replications and treatments arranged in a factorial design 3 × 3, represented by A. grandis adults fed on 3 food types (fragments of banana (T1) or orange (T2) endocarp, or with cotton squares of cultivar BRS 286 (T3, control)) and three evaluation periods (30, 60, and 90 days) and after each of these periods they were fed with cotton squares for 10 days. The reproductive tract of 100% of A. grandis females fed banana endocarp, orange endocarp, and cotton squares for 30 and 60 days and then cotton squares were morphologically adequate for reproduction, and after 90 days, only 50% of those fed cotton squares were in this condition. The length of the ovarioles and the width of the mature oocyte were greater for A. grandis fed on cotton squares and smaller in those with banana and orange endocarps. Histological sections reveal that male testes even with strong degenerative signals are already producing spermatozoa. On the other hand, females displayed ovaries with nurse cells in the tropharium and some maturing oocytes in the vitellarium. The body length was longer and the testis area and diameter smaller in males fed on cotton squares than in those with banana and orange endocarp. Anthonomus grandis females fed for ≥90 days with alternative food sources do not recover the functionality of their reproductive tract, even after consuming, for 10 days, a diet that favors reproduction. On the other hand, the males remain with their reproductive organs functional with this condition.

A beetle for prosecution: exogenous DNA detection from larval and adult gut of a Neotropical carrion beetle.

The use of recovered DNA ingested by necrophagous or hematophagous insects has increased in forensic sciences, mainly with representatives of flies. However, some beetles are also important for medico-legal forensic entomology because they feed on carcasses until advanced decomposition. This study evaluated whether the Neotropical carrion beetle Oxelytrum discicolle (Silphidae) has the potential for the detection of exogenous DNA into the gut. The whole gut or the gut contents were extracted from O. discicolle larvae and adult previously fed on pig carcass. The pig DNA recovery rate was 33.3% in larvae and 25% in adults, indicating that the carrion beetle's gut may be useful for DNA identification of ingested food. Samples with the whole gut or only gut contents showed the same DNA recovery rate. Exogenous DNA from the whole gut was recovered from samples stored in ethanol at -20 ºC for 11 days, showing that samples of O. discicolle can be stored in the forensic laboratory without loss in DNA recovery rate.

From the dual cyclone harvest performance of single conidium powder to the effect of Metarhizium anisopliae on the management of Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae).

Insect pests introduced in eucalyptus plantations in Brazil are mostly of Australian origin, but native microorganisms have potential for their management. High quality biopesticide production based on entomopathogenic fungi depends on adequate technologies. The objective of this study was to evaluate Mycoharvester® equipment to harvest and separating particles to obtain pure Metarhizium anisopliae conidia to manage Thaumastocoris peregrinus Carpintero & Dellapé, 2006 (Hemiptera: Thaumastocoridae). The Mycoharvester® version 5b harvested and separated M. anisopliae spores. The pure conidia were suspended in Tween 80® (0.1%) and calibrated to the concentrations of 1 x 106, 107, 108 and 109 conidia/ml to evaluate the pathogenicity, lethal concentration 50 and 90 (LC50, LC90) and lethal time 50 and 90 (LT50, LT90) of this fungus to T. peregrinus. This equipment harvested 85% of the conidia from rice, with production of 4.8 ± 0.38 x 109 conidia/g dry mass of substrate + fungus. The water content of 6.36% of the single spore powder (pure conidia) separated by the Mycoharvester® was lower than that of the agglomerated product. The product harvested at the concentrations of 108 and 109 conidia/ml caused high mortality to T. peregrinus third instar nymphs and adults. The separation of conidia produced by solid-state fermentation with the Mycoharvester® is an important step toward optimizing the fungal production system of pure conidia, and to formulate biopesticides for insect pest management.