Photorhabdus virulence cassette promotes bacterial invasion into macrophages by activating NF-κB and MAPK signaling pathway / 生物工程学报

Photorhabdus is a Gram-negative bacterium from the family Enterobacteriaceae that lives in a symbiotic association with nematode or insects. In addition to the role of being insect pathogens, one species called Photorhabdus asymbiotica (Pa) causes human infection around the world. Nevertheless, how does this transkingdom infection occur remains elusive. Here we focus on one pathogenic determinant called Photorhabdus virulence cassette (PVC) that is founded in the Pa genome and many other pathogens. The RNA-seq and qPCR data showed that the NF-κB and MAPK pathways were drastically activated in the PVC-treated mammalian macrophages. Western blotting assays using samples treated with various inhibitors of the affected pathways confirmed the results we have observed for MAPK pathway previously. p65 translocation assays validated the NF-κB activation in the macrophages after PVC treatment. Moreover, the bacterial phagocytosis by macrophage was also promoted by PVC at the early stage, and this phagocytosis was inhibited by cytoskeleton inhibitors. Thus, the results indicated that PVC is involved in the bacterial invasion by activating NF-κB and MAPK signaling pathway, providing a new perspective for analyzing the pathogenicity of Pa in human infections.

Inhibition of symbiote fungus of the leaf cutter ant Atta sexdens by secondary metabolites from the bacterium Xenorhabdus szentirmaii associated with entomopathogenic nematodes / Inibição do fungo simbionte da formiga-cortadeira Atta sexdens por metabólitos secundários da bactéria Xenorhabdus szentirmaii associada a nematoides entomopatogênicos

Leaf-cutter ants (Hymenoptera: Formicidae) have evolved as dominant herbivores on the American continent. These social insects remove the leaves of economically important plant species to maintain their colony's food reserves, the symbiotic fungus Leucocoprinus gongylophorus, a basidiomycete. Such fungus can be used for applications of fungicide molecules from metabolites generated by symbiont bacteria (Xenorhabdus and Photorhabdus) from entomopathogenic nematodes (Steinernema and Heterorhabditis). Through isolation and multiplication in tryptic soy broth (TSB) medium of the bacteria Xenorhabdus szentirmaii isolated PAM 25, we conducted laboratorial tests using treatments with 10, 25, and 50% of the metabolites obtained in the sixth day of cultivation. The treatments were centrifuged and filtered to generate a supernatant, which was diluted in potato + dextrose + agar (PDA), to verify the consequences of exposure to the fungus L. gongylophorus in Petri dishes. To confirm metabolite efficiency, the control treatments in PDA only and mixed (PDA+TSB) media were conducted simultaneously for 14 days. We observed total inhibition of the symbiont fungus in both the 25 and 50% dilutions during the first days of the tests. Our results support that these metabolites have inhibitory effect on the development of symbiont fungus of leaf-cutter ants.(AU)

As formigas-cortadeiras (Hymenoptera: Formicidae) evoluíram estabelecendo-se no continente americano como herbívoros dominantes. Esses insetos sociais praticam a desfolha de espécies vegetais de interesse econômico, com a finalidade de manter a reserva alimentar da colônia, o fungo simbionte Leucocoprinus gongylophorus, um basidiomiceto. Tal fungo pode ser alvo de aplicações de moléculas fungicidas encontradas em metabólitos gerados por bactérias simbiontes (Xenorhabdus e Photorhabdus) de nematoides entomopatogênicos (Steinernema e Heterorhabditis). Por meio do isolamento e da multiplicação em meio caldo triptona de soja (TSB) da bactéria Xenorhabdus szentirmaii (isolado PAM 25), foram realizados testes laboratoriais com tratamentos contendo 10, 25 e 50% do metabólito obtido no sexto dia de cultivo. Para tanto, o metabólito foi centrifugado e filtrado, gerando um sobrenadante, o qual foi diluído em batata + dextrose + ágar (BDA), para verificar as consequências da exposição do fungo L. gongylophorus em placas de Petri. Os tratamentos de controle apenas com meio BDA e misto (BDA + TSB) também foram conduzidos simultaneamente por 14 dias, a fim de confirmar a eficiência dos metabólitos. Tanto na diluição de 25 e 50% houve total inibição do fungo simbionte já nos primeiros dias da condução dos testes. Diante dos resultados obtidos, pode-se afirmar que os metabólitos são a causa do efeito inibitório do desenvolvimento do fungo simbionte das formigas-cortadeiras.(AU)