Midgut microbiota can participate in the detoxification and metabolism processes in insects, but there are few reports on the relationship between midgut microbiota and insecticide resistance in mosquitoes. In this study, we performed metagenomic sequencing on a susceptible strain (SS), a field-collected Hainan strain (HN), and a deltamethrin-resistant strain (RR) of Culex pipiens quinquefasciatus to understand the diversity and functions of their midgut microbiota. The results revealed differences in midgut microbiota among the three strains of Cx. pipiens quinquefasciatus. At the phylum level, Proteobacteria was the most prominent, accounting for nearly 70% of their midgut microbes. At the genus level, Aeromonas made up the highest proportion. In addition, Aeromonas, Morganella, Elizabethkingia, Enterobacter, Cedecea, and Thorsellia showed significant differences between strains. At the species level, Bacillus cereus, Enterobacter cloacae complex sp. 4DZ3-17B2, Streptomyces sp. CNQ329, and some species of Pseudomonas and Wolbachia were more abundant in the two resistant strains. Principal component analysis (PCA) showed that the SS strain had significantly different metagenomic functions than the two deltamethrin-resistant strains (HN and RR strain). The HN and RR strains differed from the SS strain in more than 10 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The analysis of species abundance and functional diversity can provide directions for future studies.

The beneficial effects of the probiotic strain Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) are based on its adherence and colonization ability in the gut. However, little is known about the migration and long-term gut colonization of the strain. This study evaluated the gut colonization modes of Lactiplantibacillus plantarum RS-09 to identify the strain with long-term gut colonization potential. We established CFDA/SE-labeled RS-09 to study the temporal and spatial distribution of RS-09 in the intestine as well as to analyze its persistence in different parts of the intestine by flow cytometry. This study has shown that the RS-09 strain maintains strong adhesion abilities under acid (pH 2.5) and base (pH 8.5) conditions. In addition, CFDA/SE can be used as an indicator for the labeling of L. plantarum RS-09 in the intestinal tract in vivo. We established a growth kinetics model of RS-09 to elucidate its persistence in the intestine. In vivo persistence experiments showed that the persistence rate of RS-09 was the highest in the cecum (69.5%) and the lowest in the duodenum (12.8%) at 96 h. After 20 days, RS-09 was predominantly localized in the cecum and colon steadily. These studies provide new insights into the long-term persistence of L. plantarum in the gastrointestinal tract. The CFDA/SE label system may be used to study the in vivo colonization dynamics of other probiotic strains.

Lactobacillus plantarum can interact with macrophages against bacterial enteropathy due to its potential ability to modulate macrophage polarization. However, this mechanism is not completely understood. TLR2 can recognize microbial components and trigger macrophage cytokine responses to different gram-positive strains. The aim of this study was to investigate whether probiotic Lactobacillus plantarum RS-09 can induce macrophage polarization against Salmonella Typhimurium infection via TLR2 signalling. BALB/c mice were preadministered RS-09 continuously for 7 days and then infected with Salmonella Typhimurium ATCC14028. Mouse RAW264.7 mononuclear macrophages were stimulated with RS-09 and coincubated with ATCC14028 or PBS controls. The results of the in vivo study indicated that RS-09 could relieve S. Typhimurium-induced splenomegaly, body weight loss and death rate. RS-09 also limited the colonization and translocation of S. Typhimurium in the gastrointestinal tract and thereby protected against infection. We also observed that RS-09 upregulated the production of M1 macrophage characteristics (e.g., CD11c and IL-6) against S. Typhimurium. Furthermore, RS-09 induced the expression of TLR2 in macrophages. In an in vitro study, treatment of RAW264.7 cells with RS-09 either concurrently with or before S. Typhimurium challenge enhanced the secretion of Reactive oxygen species and Nitric oxide. This effect was related to TLR2 and NF-κB activation. Based on these findings, Lactobacillus plantarum RS-09 was shown to modulate M1 macrophage polarization and induce TLR2-linked NF-κB signalling activity in the innate immune response to S. Typhimurium infection.

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