Exposure of Culex quinquefasciatus to the oomycete Pythium insidiosum: A protocol for in vitro studies.

Pythium insidiosum causes pythiosis, an infection that affects different species of mammals, including humans, and inhabits marshy ecosystems of tropical, subtropical, and temperate regions worldwide. Therefore, this study proposes a protocol to expose Culex quinquefasciatus to P. insidiosum zoospores. Cx. quinquefasciatus immatures (eggs, larvae, and pupae) were exposed to zoospores (8x103 zoospores/mL) of the oomycete for 24 h. The exposure of Cx. quinquefasciatus to the zoospores from L1 to the emergence of adults was evaluated, and P. insidiosum detection was performed by microbiological culture, polymerase chain reaction, and histopathological analysis of stage 4 larvae. The protocol used to produce Cx. quinquefasciatus colonies and adapted for this study proved viable for research on the interaction between P. insidiosum and this Culicidae species. Moreover, P. insidiosum presence was evident in all larval stages of the mosquito, although the presence of the oomycete was not detected in the eggs, pupae, and adults. This study is a pioneer in the development of a protocol to evaluate Cx. quinquefasciatus exposure to P. insidiosum zoospores, and under experimental conditions, P. insidiosum can establish itself in Cx. quinquefasciatus larval stages. The developed protocol is expected to serve as a basis for developing studies to evaluate the interactions of P. insidiosum with these mosquitoes and shed more light on the participation of culicids in expanding the ecological niche of P. insidiosum.

Polymer-based nanostructures loaded with piperine as a platform to improve the larvicidal activity against Aedes aegypti.

Piperine is an alkaloid extracted from the seed of Piper spp., which has demonstrated a larvicidal effect against Ae. aegypti. The incorporation of piperine into nanostructured systems can increase the effectiveness of this natural product in the control of Ae. aegypti larvae. In this study, we evaluated the effectiveness of piperine loaded or not into two nanostructured systems (named NS-A and NS-B) prepared by the nanoprecipitation method. The Ae. aegypti larvae were exposed to different concentrations of piperine loaded or not (2 to 16 ppm) and the mortality was investigated after 24, 48, and 72 hours. The nanostructures prepared were spherical in shape with narrow size distribution and great encapsulation efficiency. The lethal concentration 50 (LC50) for non-loaded piperine were 13.015 ppm (24 hours), 8.098 ppm (48 hours), and 7.248 ppm (72 hours). The LC50 values found for NS-A were 35.378 ppm (24 hours), 12.091 ppm (48 hours), and 8.011 ppm (72 hours), whereas the values found for NS-B were 21.267 ppm (24 hours), 12.091 ppm (48 hours), and 8.011 ppm (72 hours). Collectively, these findings suggested that non-loaded piperine caused higher larval mortality in the first hours of exposure while the nanostructured systems promoted the slow release of piperine and thereby increased the larvicidal activity over time. Therefore, loading piperine into nanostructured systems might be an effective tool to improve the larval control of vector Ae. aegypti.

Reemergence of human malaria in Atlantic Forest of Rio Grande do Sul, Brazil.

Unforeseen Plasmodium infections in the Atlantic Forest of Brazilian Extra-Amazonian region could jeopardise malaria elimination. A human malaria case was registered in Três Forquilhas, in the Atlantic Forest biome of Rio Grande do Sul, after a 45 years' time-lapsed without any malaria autochthonous notification in this southern Brazilian state. This finding represents the expansion of the malaria distribution areas in Brazil and the southernmost human malaria case record in South America in this decade. The coexistence of the bromeliad-breeding vector Anopheles (Kerteszia) cruzii and non-human primates in the Atlantic Forest regularly visited by the patient claimed for the zoonotic origin of this infection. The reemergence of Atlantic Forest human malaria in Rio Grande do Sul was also discussed.

Reemergence of human malaria in Atlantic Forest of Rio Grande do Sul, Brazil

Unforeseen Plasmodium infections in the Atlantic Forest of Brazilian Extra-Amazonian region could jeopardise malaria elimination. A human malaria case was registered in Três Forquilhas, in the Atlantic Forest biome of Rio Grande do Sul, after a 45 years' time-lapsed without any malaria autochthonous notification in this southern Brazilian state. This finding represents the expansion of the malaria distribution areas in Brazil and the southernmost human malaria case record in South America in this decade. The coexistence of the bromeliad-breeding vector Anopheles (Kerteszia) cruzii and non-human primates in the Atlantic Forest regularly visited by the patient claimed for the zoonotic origin of this infection. The reemergence of Atlantic Forest human malaria in Rio Grande do Sul was also discussed.

Microbiota potentialized larvicidal action of imidazolium salts against Aedes aegypti (Diptera: Culicidae).

Mosquitoes are important vectors of pathogens due to their blood feeding behavior. Aedes aegypti (Diptera: Culicidae) transmits arboviruses, such as dengue, Zika, and Chikungunya. This species carries several bacteria that may be beneficial for its biological and physiological development. Therefore, studying the response of its microbiota to chemical products could result in vector control. Recently, imidazolium salts (IS) were identified as effective Ae. aegypti larvicides. Considering the importance of the mosquito microbiota, this study addressed the influence of IS on the bacteria of Ae. aegypti larvae. After exposition of larvae to different IS concentrations, the cultured microbiota was identified through culturomics and mass spectrometry, and the non-cultivated microbiota was characterized by molecular markers. In addition, the influence of the IS on axenic larvae was studied for comparison. There was an alteration in both cultivable species and in their diversity, including modifications in bacterial communities. The axenic larvae were less susceptible to the IS, which was increased after exposing these larvae to bacteria of laboratory breeding water. This highlights the importance of understanding the role of the larval microbiota of Ae. aegypti in the development of imidazolium salt-based larvicides. Such effect of IS towards microbiota of Ae. aegypti larvae, through their antimicrobial action, increases their larvicidal potential.

Aedes-Chikungunya Virus Interaction: Key Role of Vector Midguts Microbiota and Its Saliva in the Host Infection.

Aedes mosquitoes are important vectors for emerging diseases caused by arboviruses, such as chikungunya (CHIKV). These viruses' main transmitting species are Aedes aegypti and Ae. albopictus, which are present in tropical and temperate climatic areas all over the globe. Knowledge of vector characteristics is fundamentally important to the understanding of virus transmission. Only female mosquitoes are able to transmit CHIKV to the vertebrate host since they are hematophagous. In addition, mosquito microbiota is fundamentally important to virus infection in the mosquito. Microorganisms are able to modulate viral transmission in the mosquito, such as bacteria of the Wolbachia genus, which are capable of preventing viral infection, or protozoans of the Ascogregarina species, which are capable of facilitating virus transmission between mosquitoes and larvae. The competence of the mosquito is also important in the transmission of the virus to the vertebrate host, since their saliva has several substances with biological effects, such as immunomodulators and anticoagulants, which are able to modulate the host's response to the virus, interfering in its pathogenicity and virulence. Understanding the Aedes vector-chikungunya interaction is fundamentally important since it can enable the search for new methods of combating the virus' transmission.