Mycobacterium ulcerans Experimental Dormancy.

Whether Mycobacterium ulcerans, the etiological agent of Buruli ulcer in numerous tropical countries, would exist in a dormant state as reported for closely related Mycobacterium species, has not been established. Six M. ulcerans strains were exposed to a progressive depletion in oxygen for 2 months, using the Wayne model of dormancy previously described for M. tuberculosis, and further examined by microscopy after staining of dynamic, dormant, and dead mycobacteria (DDD staining), microcalorimetry and subculture in the presence of dead and replicative M. ulcerans as controls. Mycobacterium ulcerans CU001 strain died during the progressive oxygen depletion and four of five remaining strains exhibited Nile red-stained intracellular lipid droplets and a 14- to 20-day regrowth when exposed to ambient air, consistent with dormancy. A fifth M. ulcerans 19423 strain stained negative in DDD staining and slowly regrew in 27 days. Three tested M. ulcerans strains yielded microcalorimetric pattern similar to that of the negative (dead) homologous controls, differing from that of the homologous positive (replicative) controls. The relevance of these experimental observations, suggesting a previously unreported dormancy state of M. ulcerans, warrants further investigations in the natural ecological niches where M. ulcerans thrive as well as in Buruli ulcer lesions.

Optimized MALDI TOF Mass Spectrometry Identification of Francisella tularensis Subsp. holarctica.

Francisella tularensis is a tier 1 agent causing the zoonosis tularemia. This highly infectious Gram-negative bacterium is occasionally isolated from human samples (especially blood samples) in routine clinical microbiology laboratories. A rapid and accurate method for identifying this pathogen is needed in order to optimize the infected patient's healthcare management and prevent contamination of the laboratory personnel. MALDI TOF mass spectrometry has become the gold standard for the rapid identification of most human pathogens. However, F. tularensis identification using such technology and commercially available databases is currently considered unreliable. Real-time PCR-based methods for rapid detection and accurate identification of F. tularensis are not available in many laboratories. As a national reference center for tularemia, we developed a MALDI TOF database allowing accurate identification of the species F. tularensis and its differentiation from the closely related neighbor species F. tularensis subsp. novicida and F. philomiragia. The sensitivity and specificity of this database were validated by testing 71 F. tularensis strains and 165 strains from 63 species not belonging to the Francisella genus. We obtained accurate identification at the species level and differentiation of all the tested bacterial strains. In particular, F. tularensis could be accurately differentiated from other small Gram-negative bacilli occasionally isolated from human samples, including species of the HACEK group and Brucella melitensis.

Disseminated Mycobacterium ulcerans Infection in Wild Grasscutters (Thryonomys swinderianus), Côte d'Ivoire.

Buruli ulcer is an infectious disease provoking chronic, disabling skin ulcers in mammals and humans. Buruli ulcer is caused by Mycobacterium ulcerans, an environmental mycobacterium synthesizing a toxin called mycolactone responsible for the pathogenicity. The reservoirs and the modes of transmission of M. ulcerans remain elusive, limiting the prophylaxis capabilities in rural areas in endemic countries. In Australia, several studies have demonstrated the probable role of possums as reservoirs. In Côte d'Ivoire, some studies have speculated on the potential role of grasscutters in the transmission cycle of M. ulcerans. In this study, we detected M. ulcerans-specific sequences in rectal contents and spleens collected in wild grasscutters hunted in Buruli ulcer-endemic area in Côte d'Ivoire, but not in farmed negative control animals and in domesticated animals, namely, pigs, goats, cattle, and dogs, living in close contact with the local population. Some grasscutters exhibited the same sequence pattern in the feces and spleen. These observations confirm the asymptomatic gut carriage of M. ulcerans in this mammal species. Moreover, these observations suggest the dissemination of M. ulcerans from the gut to the spleen in grasscutters. These observations suggest that, in some mammals, M. ulcerans is not only an inoculated pathogen but also a translocating invasive pathogen.