山本 大介

Anopheles mosquitoes spread malaria, which is caused by Plasmodium parasite infection. In nature, both male and female mosquitoes typically ingest sugars from floral nectar; however, few studies have investigated the effects of sugars as an insecticide for mosquitoes. During our previous work, the lifespan of An. stephensi was shortened by the feeding of high concentrations of lactose. The mechanism by which lactose can shorten the lifespan of mosquitoes is still unidentified. The present study aims to evaluate the potential of lactose as an insecticide against mosquitoes. A sublethal concentration of lactose decreased the lifespan of An. stephensi but did not affect their blood-feeding ratio or the number of eggs they laid. Female Anopheles mosquitoes refused to ingest lactose-containing sugars when a lactose-free sugar is available; however, male mosquitoes ingest them. Both sexes excreted lactose from their bodies after feeding, while other sugars were completely absorbed and digested. An ingestion volume assay using food dye revealed that lactose reduces the volume of the solution in the mosquito body. Female mosquitoes forced to ingest lactose for 3 wk showed a slight change in the composition of their midgut microbiota but not in their relative quantity. These findings suggest that lactose feeding causes osmotic diarrhea-like symptoms in Anopheles mosquitoes. The use of attractive toxic sugar baits with lactose as an insecticide is expected to control male Anopheles mosquitoes.

Phlebotomine sand flies are vectors of the protozoan parasite Leishmania spp. Although the intestinal microbiota is involved in a wide range of biological and physiological processes and has the potential to alter vector competence, little is known about the impact of host species and environment on the gut microbiome. To address this issue, a comparative analysis of the microbiota of sand fly vector populations of Leishmania major and L. tropica in a mixed focus of cutaneous leishmaniasis in Tunisia was performed. Bacterial 16S rRNA gene amplification and Illumina MiSeq sequencing were used to characterize and compare the overall bacterial and fungal composition of field-collected sand flies: Phlebotomus papatasi, Ph. perniciosus, Ph. riouxi, and Ph. sergenti. Thirty-eight bacterial genera belonging to five phyla were identified in 117 female specimens. The similarities and differences between the microbiome data from different samples collected from three collections were determined using principal coordinate analysis (PCoA). Substantial variations in the bacterial composition were found between geographically distinct populations of the same sand fly species, but not between different species at the same location, suggesting that the microbiota content was structured according to environmental factors rather than host species. These findings suggest that host phylogeny may play a minor role in determining the insect gut microbiota, and its potential to affect the transmission of the Leishmania parasite appear to be very low. These results highlight the need for further studies to decode sand fly Leishmania-microbiota interactions, as even the same bacterial species, such as Enterococcus faecalis, can exert completely opposite effects when confronted with different pathogens within various host insects and vice versa.

Resilin is an elastic protein that is vital to insects’ vigorous movement. Canonical resilin proteins possess the R&R Consensus, a chitin-binding domain conserved in a family of cuticular proteins, and highly repetitive sequences conferring elastic properties. In the malaria vector mosquito, Anopheles gambiae, however, a cuticular protein has been found that has an R&R Consensus resembling that of resilin but lacks the repetitive sequences (here, we call it resilin-related or resilin-r). The relationship between resilin-r and resilin was unclear. It was also unknown whether resilin-r is conserved in mosquitoes. In this paper, phylogenetic and structural analyses were performed to reveal the relationship of resilin homologous proteins from holometabolous insects. Their chitin-binding abilities were also assessed. A resilin-r was found in each mosquito species, and these proteins constitute a clade with resilin from other insects based on the R&R Consensus sequences, indicating an evolutionary relationship between resilin-r and resilin. The resilin-r showed chitin-binding activity as same as resilin, but had distinct structural features from resilin, suggesting that it plays specialized roles in the mosquito cuticle. Another resilin-like protein was found to exist in each holometabolous insect that possesses resilin-like repetitive sequences but lacks the R&R Consensus. These results suggest that similar evolutionary events occurred to create resilin-r and resilin-like proteins.

BACKGROUND: Blood-sucking phlebotomine sand flies are vectors of the protozoan parasites Leishmania spp. Although the intestinal microbiota is involved in a wide range of biological and physiological processes and has the potential to alter vector competence, little is known about the factors that modify the gut microbiota composition of sand flies. As a key step toward addressing this issue, we investigated the impact of host species on the gut bacterial composition in Phlebotomus and Lutzomyia sand flies reared under the same conditions. METHODS: Bacterial 16S rRNA gene amplification and Illumina MiSeq sequencing were used to characterize the overall bacterial composition of three laboratory-reared sandflies: Phlebotomus papatasi, Ph. duboscqi, and Lutzomyia longipalpis. RESULTS: Our results showed that the larvae of the three sand fly species harbored almost the same microbes but had different relative abundances. Adult Ph. papatasi and Ph. duboscqi revealed similar microbiome compositions, which were distinct from that of adult Lu. longipalpis. Furthermore, we showed that Ph. papatasi and Ph. duboscqi are hosts for different bacterial genera. The experiment was repeated twice to improve accuracy and increase reliability of the data, and the same results were obtained even when a distinct composition of the microbiome among the same species was identified probably because of the use of different larvae food batch. CONCLUSIONS: The present study provides key insights into the role of host species in the gut microbial content of different sand fly species reared under the same conditions, which may influence their susceptibility to Leishmania infection.

A rare sugar, allose, was reported to inhibit the development of Plasmodium parasites in Anopheles mosquitoes; however, the mechanism remains unknown. The present study addressed the inhibitory mechanism of allose on the development of the Plasmodium parasite by connecting it with bacteria involvement in the midgut. In addition, further inhibitory sugars against Plasmodium infection in mosquitoes were explored. Antibiotic-treated and antibiotic-untreated Anopheles stephensi were fed fructose with or without allose. The mosquitoes were infected with luciferase-expressing Plasmodium berghei, and parasite development was evaluated by luciferase activity. Bacterial composition analysis in gut of their mosquitoes was performed with comprehensive 16S ribosomal RNA sequencing. As the result, allose inhibited the development of oocysts in mosquitoes regardless of prior antibiotic treatment. Microbiome analysis showed that the midgut bacterial composition in mosquitoes before and after blood feeding was not affected by allose. Although allose inhibited transient growth of the midgut microbiota of mosquitoes after blood feeding, neither toxic nor inhibitory effects of allose on the dominant midgut bacteria were observed. Ookinete development in the mosquito midgut was also not affected by allose feeding. Additional 15 sugars including six monosaccharides, four polyols, and five polysaccharides were tested; however, no inhibitory effect against Plasmodium development in mosquitoes was observed. These results indicated that allose inhibits parasite development in midgut stage of the mosquito independently of midgut microbiota. Although further studies are needed, our results suggest that allose may be a useful material for the vector control of malaria as a "transmission-blocking sugar."