A spontaneous genomic deletion in Listeria ivanovii identifies LIPI‐2, a species‐specific pathogenicity island encoding sphingomyelinase and numerous internalins

Summary Listeria ivanovii differs from the human pathogen Listeria monocytogenes in that it specifically affects ruminants, causing septicaemia and abortion but not meningo‐encephalitis. The genetic characterization of spontaneous L. ivanovii mutants lacking the virulence factor SmcL (sphingomyelinase) led us to identify LIPI‐2, the first species‐specific pathogenicity island from Listeria. Besides SmcL, this 22 kb chromosomal locus encodes 10 internalin (Inl) proteins: i‐InlB1 and ‐B2 are large/surface‐associated Inls similar to L. monocytogenes InlB; i‐InlE to –L are small/excreted (SE)‐Inls, i‐InlG being a tandem fusion of two SE‐Inls. Except i‐inlB1, all LIPI‐2 inl genes are controlled by the virulence regulator, PrfA. LIPI‐2 is inserted into a tRNA locus and is unstable – half of it deleting at ≈10−4 frequency with a portion of contiguous DNA. The spontaneous mutants were attenuated in vivo in mice and lambs and showed impaired intracellular growth and apoptosis induction in bovine MDBK cells. Targeted knock‐out mutations associated the virulence defect with LIPI‐2 genes. The region between the core genome loci ysnB‐tRNAarg and ydeI flanking LIPI‐2 contained different gene complements in the different Listeria spp. and even serovars of L. monocytogenes, including remnants of the PSA bacteriophage int gene in serovar 4b, indicating it is a hot spot for horizontal genome diversification. LIPI‐2 is conserved in L. ivanovii ssp. ivanovii and londoniensis, suggesting an early acquisition during the species’ evolution. LIPI‐2 is likely to play an important role in the pathogenic and host tropism of L. ivanovii.