Insecticide resistance by a host-symbiont reciprocal detoxification

Insecticide resistance is one of the most serious problems in contemporary agriculture and public health. Although recent studies revealed that insect gut symbionts contribute to resistance, the symbiont-mediated detoxification process remains unclear. Here we report the in vivo detoxification process of an organophosphorus insecticide, fenitrothion, in the bean bug Riptortus pedestris. Using transcriptomics and reverse genetics, we reveal that gut symbiotic bacteria degrade this insecticide through a horizontally acquired insecticide-degrading enzyme into the non-insecticidal but bactericidal compound 3-methyl-4-nitrophenol, which is subsequently excreted by the host insect. This integrated “host-symbiont reciprocal detoxification relay” enables the simultaneous maintenance of symbiosis and efficient insecticide degradation. We also find that the symbiont-mediated detoxification process is analogous to the insect genome-encoded fenitrothion detoxification system present in other insects. Our findings highlight the capacity of symbiosis, combined with horizontal gene transfer in the environment, as a powerful strategy for an insect to instantly eliminate a toxic chemical compound, which could play a critical role in the human-pest arms race.

A beta-glucosidase of an insect herbivore determines both toxicity and deterrence of a dandelion defense metabolite

Gut enzymes can metabolize plant defense compounds and thereby affect the growth and fitness of insect herbivores. Whether these enzymes also influence feeding preference is largely unknown. We studied the metabolization of taraxinic acid β-D-glucopyranosyl ester (TA-G), a sesquiterpene lactone of the common dandelion (Taraxacum officinale) that deters its major root herbivore, the common cockchafer larva (Melolontha melolontha). We have demonstrated that TA-G is rapidly deglucosylated and conjugated to glutathione in the insect gut. A broad-spectrum M. melolontha β-glucosidase, Mm_bGlc17, is sufficient and necessary for TA-G deglucosylation. Using cross-species RNA interference, we have shown that Mm_bGlc17 reduces TA-G toxicity. Furthermore, Mm_bGlc17 is required for the preference of M. melolontha larvae for TA-G-deficient plants. Thus, herbivore metabolism modulates both the toxicity and deterrence of a plant defense compound. Our work illustrates the multifaceted roles of insect digestive enzymes as mediators of plant-herbivore interactions.

Mechanisms of damage prevention, signalling, and repair impact the ability of Drosophila to tolerate enteric bacterial infection

Many insects thrive on decomposing and decaying organic matter containing a large diversity of both commensal and pathogenic microorganisms. The insect gut is therefore frequently exposed to pathogenic threats and must be able not only to detect and clear these potential infections, but also be able to repair the resulting damage to gut tissues in order to tolerate relatively high microbe loads. In contrast to the mechanisms that eliminate pathogens, we currently know less about the mechanisms of disease tolerance, and most of this knowledge stems from systemic infections. Here we investigated how well-described mechanisms that either prevent, signal, control, or repair tissue damage during infection contribute to the phenotype of disease tolerance during gut infection. We orally infected adult Drosophila melanogaster flies with the bacterial pathogen Pseudomonas entomophila in several loss-of-function mutants lacking epithelial responses including damage preventing dcy (drosocrystallin - a major component of the peritrophic matrix), damage signalling upd3 (unpaired protein, a cytokine-like molecule), damage controlling irc (immune-regulated catalase, a negative regulator of reactive oxygen species) and tissue damage repairing egfr1 (epidermal growth factor receptor). Overall, we detect effects of all these mechanisms on disease tolerance. The deterioration of the peritrophic matrix in dcy mutants resulted in the highest loss of tolerance, while loss of function of either irc or upd3 also reduced tolerance in both sexes. The absence of tissue damage repair signalling (egfr1) resulted in a severe loss in tolerance in male flies but had no substantial effect on the ability of female flies to tolerate P. entomophila infection, despite carrying greater microbe loads than males. Together, our findings provide empirical evidence for the role of damage limitation mechanisms in disease tolerance and highlight how sex differences in these mechanisms could generate sexual dimorphism in immunity.

Isolate locale ale baculovirului entomopatogenic ca o tehnologie de formulare inovatoare, care protejează biopesticidul din degradare a radiației ultraviolete

This paper presents the conceptual conceptual vision a formulation technology for biopesticides in which the active ingredient (baculovirus) is an active coal. Importantly, this indgredient protects the sen-sitive viral DNA from degrading in sunlight, but dissolves in the alkaline insect gut to release the virus, which then infects and kills the pest. We show, using this ingredient, in both laboratory bioassays and field tests, that this can extend the efficacy of the biopesticide well beyond the few hours of existing virus formulations, potentially increasing the spray interval and reducing the need for high application rates. Are presented both theoretical foundations and practical applications and described the results oriented for implementation and functionality of organic agriculture in Republic of Moldova.

Gut Microbiota Dysbiosis Influences Metabolic Homeostasis in Spodoptera frugiperda

Insect gut microbiota plays important roles in acquiring nutrition, preventing pathogens infection, modulating immune responses, and communicating with environment. Gut microbiota can be affected by external factors such as foods and antibiotics. Spodoptera frugiperda (Lepidoptera: Noctuidae) is an important destructive pest of grain crops worldwide. The function of gut microbiota in S. frugiperda remains to be investigated. In this study, we fed S. frugiperda larvae with artificial diet with antibiotic mixture (penicillin, gentamicin, rifampicin, and streptomycin) to perturb gut microbiota, and then examined the effect of gut microbiota dysbiosis on S. frugiperda gene expression by RNA sequencing. Firmicutes, Proteobacteria, Bacteroidetes, and Actinobacteria were the most dominant phyla in S. frugiperda. We found that the composition and diversity of gut bacterial community were changed in S. frugiperda after antibiotics treatment. Firmicutes was decreased, and abundance of Enterococcus and Weissella genera was dramatically reduced. Transcriptome analysis showed that 1,394 differentially expressed transcripts (DETs) were found between the control and antibiotics-treated group. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) results showed that antibiotics-induced dysbiosis affected many biological processes, such as energy production, metabolism, and the autophagy–lysosome signal pathway. Our results indicated that dysbiosis of gut microbiota by antibiotics exposure affects energy and metabolic homeostasis in S. frugiperda, which help better understand the role of gut microbiota in insects.

Evaluation of Anti-Malaria Potency of Wild and Genetically Modified Enterobacter Cloacae Expressing Effector Proteins in Anopheles Stephensi

Background: Malaria is one of the most lethal infectious diseases in tropical and subtropical areas of the world. To fight the disease, paratransgenesis using symbiotic bacteria offers a sustainable and environmentally-friendly strategy. Here we evaluated the disruption of malaria transmission in the Anopheles stephensi-Plasmodium berghei assemblage, using wild and modified insect gut bacterium, Enterobacter cloacae.Methods: The assay was carried out using E. cloacae dissolvens wild-type (WT) and its three engineered strains expressing GFP-defensin (GFP-D), scorpine-HasA (S-HasA), and HasA, The 3-5 day-old female mosquitoes were supplemented overnight with the studied bacteria [1×109cells/mL of 5% (wt/vol), fructose and red dye (1/50 ml)] soaked on cotton-wool. Each group of sugar-fed mosquitoes was then starved for 4-6 hours and fed on a P. berghei–infected mouse for 20 min in the dark at 17-20°C. The blood-fed mosquitoes were kept at 19±1°C and RH 80±5, and parasite infection was measured by midgut dissection and oocyst counting 10 days post-infection (dpi). Results: Both wild-type and genetically modified bacterial strains significantly (P< 0.0001) disrupted the P. berghei development in the An. stephensi midgut, in comparison with the control group. The mean parasite inhibition of E. cloacaeWT, E. cloacaeHasA, E. cloacaeS-HasA, and E. cloacaeGFP-D was measured as 72, 86, 92.5 and 92.8 respectively. Conclusions: The wild and modified E. cloacae might abolish oocyst development by providing a physical barrier or by excretion of intrinsic effector molecules. These findings reinforce the case for the use of either wild or genetically modified E. cloacae bacteria as a powerful tool to combat malaria.

Parasitoid Causes Cascading Effects on Plant-Induced Defenses Mediated Through the Gut Bacteria of Host Caterpillars

Koinobiont endoparasitoid wasps whose larvae develop inside a host insect alter several important facets of host physiology, potentially causing cascading effects across multiple trophic levels. For instance, the hijacking of the host immune responses may have effects on how insects interact with host plants and microbial associates. However, the parasitoid regulation of insect–plant–microbiome interactions is still understudied. In this study, we used the fall armyworm (FAW), Spodoptera frugiperda, and the braconid parasitoid Cotesia marginiventris to evaluate impacts of parasitism on the gut microbiome of FAW larvae, and respective maize plant defense responses. The level of reactive oxygen species and the microbial community in larval gut underwent significant changes in response to parasitism, leading to a significant reduction of Enterococcus, while elevating the relative abundance of Pseudomonas. FAW with parasitism had lower glucose oxidase (GOX) activity in salivary glands and triggered lower defense responses in maize plants. These changes corresponded to effects on plants, as Pseudomonas inoculated larvae had lower activity of salivary GOX and triggered lower defense responses in maize plants. Our results demonstrated that parasitism had cascading effects on microbial associates across trophic levels and also highlighted that insect gut bacteria may contribute to complex interrelationships among parasitoids, herbivores, and plants.

Species Identity, Life History, and Geographic Distance Influence Gut Bacterial Communities in Lab-Reared and European Field-Collected Culicoides Biting midges

Bacteria are part of the insect gut system and influence many physiological traits of their host. Gut bacteria may even reduce or block the transmission of arboviruses in several species of arthropod vectors. Culicoides biting midges are important arboviral vectors of several livestock and wildlife diseases, yet limited information is available on their gut bacterial communities. Addressing this gap will help inform how these communities can be manipulated and ultimately used as novel tools to control pathogens. To assess how bacterial communities change during the life stages of lab-reared C. nubeculosus and C. sonorensis, endosymbiotic bacteria were identified using Illumina sequencing of 16S rRNA and taxonomically characterised. Analyses were conducted to determine how gut bacterial communities in adults are influenced by species identity and geographic distance among biting midge populations. Communities of the two lab-reared Culicoides species significantly changed after pupation and with maturation into 6-day-old adults. Pseudomonas, Burkholderiaceae and Leucobacter bacteria were part of a core community that was trans-stadially transmitted and found throughout their life cycle. Among field-collected biting midges, the bacterial communities were unique for almost each species. Cardinium, Rickettsia and Wolbachia were some of the most abundant bacteria in midges collected from wetlands. Only Pseudomonas was present in high relative abundance in all field-collected species. In this study, species identity, as well as geographic distance, influenced the gut bacterial communities and may partly explain known inter- and intra-species variability in vector competence. Additionally, stably associated bacterial species could be candidates for paratransgenic strategies to control vector-borne pathogens.