Attenuated strains of mycobacteria can be exploited to determine genes essential for their pathogenesis and persistence. To this goal, we sequenced the genome of H37Ra, an attenuated variant of Mycobacterium tuberculosis H37Rv strain. Comparison with H37Rv revealed three unique coding region polymorphisms. One polymorphism was located in the DNA-binding domain of the transcriptional regulator PhoP, causing the protein's diminished DNA-binding capacity. Temporal gene expression profiles showed that several genes with reduced expression in H37Ra were also repressed in an H37Rv phoP knockout strain. At later time points, genes of the dormancy regulon, typically expressed in a state of nonreplicating persistence, were upregulated in H37Ra. Complementation of H37Ra with H37Rv phoP partially restored its persistence in a murine macrophage infection model. Our approach demonstrates the feasibility of identifying minute but distinct differences between isogenic strains and illustrates the consequences of single point mutations on the survival stratagem of M. tuberculosis.

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To investigate the prevalence of types of meticillin-resistant Staphylococcus aureus (MRSA) in slaughter pigs in German abattoirs, nasal swabs were collected from a total of 1026 pigs in five abattoirs after stunning in the course of two studies, and examined for MRSA. Study 1 included four abattoirs; study 2 was carried out in one large abattoir. Isolates were tested for antimicrobial susceptibility and characterised using spa-typing, multilocus sequence typing (MLST) and typing of the staphylococcal cassette chromosome, SCCmec. Overall, MRSA was isolated from 70.8 per cent of 520 samples in study 1 and from 49.0 per cent of 506 samples in study 2. The proportion of positive samples varied substantially between the abattoirs in study 1. Most isolates belonged to spa-types t011 and t034 and SCCmec types III and V. MLST of selected isolates revealed that they were all MLST ST398. Besides beta-lactams, 100 per cent of the isolates were resistant to tetracycline, 80.5 per cent were resistant to erythromycin and 80.7 per cent were resistant to clindamycin. Less than 5 per cent of the isolates were resistant to other antimicrobials.