Genetic features of strains of Mycobacterium avium subsp. paratuberculosis circulating in the West of France deciphered by Whole-Genome Sequencing
Résumé
Introduction: Paratuberculosis is a chronic infection of the intestine, mainly the ileum, caused by Mycobacterium avium subsp. paratuberculosis (Map) in cattle and other ruminants. This enzootic disease is present worldwide and has a strong impact on the dairy cattle industry. For this species, the typing tools do not make it possible to investigate the genetic diversity of the strains. These limitations can be overcome by the application of Whole Genome Sequencing (WGS), particularly for clonal populations such as Map. WGS analyses can provide comprehensive genetic information, including information on genome evolution, discrimination of closely related strains and virulence determinants.
The purpose of the present study was to undertake a whole-genome analysis of Map strains to identify accurate phylogenetic relationships between isolates and establish correlations between genomic traits and epidemiological
data within a population of well documented-strains.
Materials and methods: A set of 200 animal field strains were selected based on a previous large-scale longitudinal study representative of the French Map population circulating in the West of France. Strains were isolated from bovine of breed Prim’Holstein or Normande naturally infected by Map. For each strain isolated, all information about the animal is available including: breed, herd prevalence, locations, serological status and excretion level. The genomic sequences of Map strains were obtained by Illumina MiSeq sequencing. Genomic sequences were assessed for potential contamination using Kraken2. Cleaned reads were aligned to the reference genome K-10 and SNP were called using FreeBayes. IQ-Tree was used to infer a SNP-based phylogeny.
Results, discussion and conclusion: This study provided 200 new genomes of French strains isolates from naturally infected animals. Pangenome analysis of this panel confirmed the degree of Map clonality. SNP analyzes provided accurate phylogeny able to distinguish each strain divided into 4 clusters independently of the breed of the cattle. Interestingly, the phylogeny clearly distinct the two major MLVA profiles. A phylogeny was inferred with French Map isolates and with other Map isolates found across the world. Relations between genetic traits and epidemiological data will be investigated to better understand the transmission dynamics of the disease