Metagenomic Strain-Typing Combined with Isolate Sequencing Provides Increased Resolution of the Genetic Diversity of <i>Campylobacter jejuni</i> Carriage in Wild Birds

oleh: Malte Herold, Louise Hock, Christian Penny, Cécile Walczak, Fatu Djabi, Henry-Michel Cauchie, Catherine Ragimbeau

Format: Article
Diterbitkan: MDPI AG 2023-01-01

Deskripsi

As the world’s leading cause of human gastro-enteritis, the food- and waterborne pathogen <i>Campylobacter</i> needs to be intensively monitored through a One Health approach. Particularly, wild birds have been hypothesized to contribute to the spread of human clinical recurring <i>C. jejuni</i> genotypes across several countries. A major concern in studying epidemiological dynamics is resolving the large genomic diversity of strains circulating in the environment and various reservoirs, challenging to achieve with isolation techniques. Here, we applied a passive-filtration method to obtain isolates and in parallel recovered genotypes from metagenomic sequencing data from associated filter sweeps. For genotyping mixed strains, a reference-based computational workflow to predict allelic profiles of nine extended-MLST loci was utilized. We validated the pipeline by sequencing artificial mixtures of <i>C. jejuni</i> strains and observed the highest prediction accuracy when including obtained isolates as references. By analyzing metagenomic samples, we were able to detect over 20% additional genetic diversity and observed an over 50% increase in the potential to connect genotypes across wild-bird samples. With an optimized filtration method and a computational approach for genotyping strain mixtures, we provide the foundation for future studies assessing <i>C. jejuni</i> diversity in environmental and clinical settings at improved throughput and resolution.