RESUMO
Sugarcane is a crop of major importance used mainly for sugar and biofuel production, and many additional applications of its byproducts are being developed. Sugarcane cultivation is plagued by many insect pests and pathogens that reduce sugarcane yields overall. Recently emerging studies have shown complex multitrophic interactions in cultivated areas, such as the induction of sugarcane defense-related proteins by insect herbivory that function against fungal pathogens that commonly appear after mechanical damage. Fungi and viruses infecting sugarcane also modulate insect behavior, for example, by causing changes in volatile compounds responsible for insect attraction or repelling natural vector enemies via a mechanism that increases pathogen dissemination from infected plants to healthy ones. Interestingly, the fungus Fusarium verticillioides is capable of being vertically transmitted to insect offspring, ensuring its persistence in the field. Understanding multitrophic complexes is important to develop better strategies for controlling pathosystems affecting sugarcane and other important crops and highlights the importance of not only studying binary interactions but also adding as many variables as possible to effectively translate laboratory research to real-life conditions.
RESUMO
During probing and blood feeding, haematophagous mosquitoes inoculate a mixture of salivary molecules into their vertebrate hosts' skin. In addition to the anti-haemostatic and immunomodulatory activities, mosquito saliva also triggers acute inflammatory reactions, especially in sensitized hosts. Here, we characterize the oedema and the cellular infiltrate following Aedes aegypti mosquito bites in the skin of sensitized and non-sensitized BALB/c mice by flow cytometry. Ae. aegypti bites induced an increased oedema in the ears of both non-sensitized and salivary gland extract- (SGE-)sensitized mice, peaking at 6 hr and 24 hr after exposure, respectively. The quantification of the total cell number in the ears revealed that the cellular recruitment was more robust in SGE-sensitized mice than in non-sensitized mice, and the histological evaluation confirmed these findings. The immunophenotyping performed by flow cytometry revealed that mosquito bites were able to produce complex changes in cell populations present in the ears of non-sensitized and SGE-sensitized mice. When compared with steady-state ears, the leucocyte populations significantly recruited to the skin after mosquito bites in non-sensitized and sensitized mice were eosinophils, neutrophils, monocytes, inflammatory monocytes, mast cells, B-cells and CD4+ T-cells, each one with its specific kinetics. The changes in the absolute number of cells suggested two cell recruitment profiles: (i) a saliva-dependent migration; and (ii) a migration dependent on the immune status of the host. These findings suggest that mosquito bites influence the skin microenvironment by inducing differential cell migration, which is dependent on the degree of host sensitization to salivary molecules.
Assuntos
Aedes/imunologia , Quimiotaxia de Leucócito , Edema/imunologia , Mordeduras e Picadas de Insetos/imunologia , Leucócitos/imunologia , Mastócitos/imunologia , Saliva/imunologia , Pele/imunologia , Animais , Microambiente Celular , Modelos Animais de Doenças , Feminino , Cinética , Masculino , Camundongos Endogâmicos BALB C , Infiltração de NeutrófilosRESUMO
BACKGROUND: In tropical sugarcane crops, the fungus Fusarium verticillioides, the agent responsible for the occurrence of the red rot complex, occurs in association with the sugarcane borer Diatraea saccharalis. This fungus, in addition to being transmitted vertically, can manipulate both the insect and the plant for its own dissemination in the field. Due to the complex interaction between F. verticillioides and D. saccharalis, and the high incidence of the fungus in the intestinal region, our objective was to investigate whether F. verticillioides could alter the intestinal structure of the insect. METHODS: We combined analysis of scanning electron microscopy and light microscopy to identify whether the presence of the fungus F. verticillioides, in artificial diets or in sugarcane, could lead to any alteration or regional preference in the insect's intestinal ultrastructure over the course of its development, or its offspring development, analyzing the wall and microvillous structures of the mid-digestive system. RESULTS: Here, we show that the fungus F. verticillioides alters the intestinal morphology of D. saccharalis, promoting an increase of up to 3.3 times in the thickness of the midgut compared to the control. We also observed that the phytopathogen colonizes the intestinal microvilli for reproduction, suggesting that this region can be considered the gateway of the fungus to the insect's reproductive organs. In addition, the colonization of this region promoted the elongation of microvillous structures by up to 180% compared to the control, leading to an increase in the area used for colonization. We also used the fungus Colletotrichum falcatum in the tests, and it did not differ from the control in any test, showing that this interaction is specific between D. saccharalis and F. verticillioides. CONCLUSIONS: The phytopathogenic host F. verticillioides alters the intestinal morphology of the vector insect in favor of its colonization.
RESUMO
The sialotranscriptomes of Aedes aegypti revealed a transcript overexpressed in female salivary glands that codes a mature 7.8 kDa peptide. The peptide, specific to the Aedes genus, has a unique sequence, presents a putative secretory nature and its function is unknown. Here, we confirmed that the peptide is highly expressed in the salivary glands of female mosquitoes when compared to the salivary glands of males, and its secretion in mosquito saliva is able to sensitize the vertebrate host by inducing the production of specific antibodies. The synthetic version of the peptide downmodulated nitric oxide production by activated peritoneal murine macrophages. The fractionation of a Ae. aegypti salivary preparation revealed that the fractions containing the naturally secreted peptide reproduced the nitric oxide downmodulation. The synthetic peptide also selectively interfered with cytokine production by murine macrophages, inhibiting the production of IL-6, IL-12p40 and CCL2 without affecting TNF-α or IL-10 production. Likewise, intracellular proteins associated with macrophage activation were also distinctively modulated: while iNOS and NF-κB p65 expression were diminished, IκBα and p38 MAPK expression did not change in the presence of the peptide. The anti-inflammatory properties of the synthetic peptide were tested in vivo on a dextran sulfate sodium-induced colitis model. The therapeutic administration of the Ae. aegypti peptide reduced the leukocytosis, macrophage activity and nitric oxide levels in the gut, as well as the expression of cytokines associated with the disease, resulting in amelioration of its clinical signs. Given its biological properties in vitro and in vivo, the molecule was termed Aedes-specific MOdulatory PEptide (AeMOPE-1). Thus, AeMOPE-1 is a novel mosquito-derived immunobiologic with potential to treat immune-mediated disorders.