Hundreds of antimicrobial peptides create a selective barrier for insect gut symbionts.

The spatial organization of gut microbiota is crucial for the functioning of the gut ecosystem, although the mechanisms that organize gut bacterial communities in microhabitats are only partially understood. The gut of the insect Riptortus pedestris has a characteristic microbiota biogeography with a multispecies community in the anterior midgut and a monospecific bacterial population in the posterior midgut. We show that the posterior midgut region produces massively hundreds of specific antimicrobial peptides (AMPs), the Crypt-specific Cysteine-Rich peptides (CCRs) that have membrane-damaging antimicrobial activity against diverse bacteria but posterior midgut symbionts have elevated resistance. We determined by transposon-sequencing the genetic repertoire in the symbiont Caballeronia insecticola to manage CCR stress, identifying different independent pathways, including AMP-resistance pathways unrelated to known membrane homeostasis functions as well as cell envelope functions. Mutants in the corresponding genes have reduced capacity to colonize the posterior midgut, demonstrating that CCRs create a selective barrier and resistance is crucial in gut symbionts. Moreover, once established in the gut, the bacteria differentiate into a CCR-sensitive state, suggesting a second function of the CCR peptide arsenal in protecting the gut epithelia or mediating metabolic exchanges between the host and the gut symbionts. Our study highlights the evolution of an extreme diverse AMP family that likely contributes to establish and control the gut microbiota.

Rapid Detection and Identification of Vancomycin-Sensitive Bacteria Using an Electrochemical Apta-Sensor.

In order to combat the complex and diverse infections caused by bacteria, it is essential to develop efficient diagnostic tools. Current techniques for bacterial detection rely on laborious multistep procedures, with high costs and extended time of analysis. To overcome these limitations, we propose here a novel portable electrochemical biosensor for the rapid detection and identification of Gram-positive bacteria that leverages the recognition capabilities of vancomycin and aptamers. A vancomycin-modified screen-printed carbon electrode was used to selectively capture Gram-positive bacteria susceptible to this antibiotic. Electrochemical impedance spectroscopy and scanning electron microscopy demonstrated that capture was achieved in 10 min, with a limit of detection of only 2 CFU/mL. We then tested the device's potential for aptamer-based bacterial identification using Staphylococcus aureus and Bacillus cereus as the test strains. Specifically, electrodes with captured bacteria were exposed to species-specific aptamers, and the resulting changes in current intensity were analyzed using differential pulse voltammetry. When used directly in untreated milk or serum, the system was able to successfully identify a small amount of S. aureus and B. cereus (100 CFU/mL) in less than 45 min. This novel biosensor has the potential to serve as an invaluable tool that could be used, even by inexperienced staff, in a broad range of settings including clinical diagnostics, food safety analysis, environmental monitoring, and security applications.