[Peculiar "chestnuts in burrs" formation in MGIT cultures of pulmonary Mycobacterium xenopi cases].

We report 3 patients whose sputum and bronchoalveolar lavage fluid (BALF) cultures for acid fast bacteria in MGIT liquid media grew colonies of Mycobacterium xenopi (M. xenopi) with a characteristic chestnut burr like appearance. Patients I, II, and III were a 74-year-old man, 47-year-old woman, and 62-year-old woman, respectively. Chest X ray showed a pulmonary cavity in each case. Patient I had a history of pulmonary and renal tuberculosis. The past medical history of patient II was unremarkable. Patient III had a history of lung cancer. Eight sputum samples and 4 BALF samples from patient I, 3 sputum samples and 1 BALF sample from patient II, and 4 sputum samples from patient III were positive for acid fast bacteria, and the organism was identified as M. xenopi in 9 samples. Smears of these MGIT-positive cultures were stained by the Ziehl Neelsen method, and examined under a microscope. Large and small, spherical shaped, 15-100 microm clusters of thin, elongated bacteria, with a chestnut burr-like or spherical moss like and partly budding appearance, were scattered throughout the smear preparation. Although only 34 cases of M. xenopi infection were reported in Japan between 1984 and 2005, the number of reported cases has been on the increase in recent years. Since no report from Japan, Europe, or the United States have noted the characteristic appearance of M. xenopi in cultures, we consider that the feature described in this communication is useful to presumptively identify M. xenopi.

Interactions between Mycobacterium xenopi, amoeba and human cells.

Outbreaks due to Mycobacterium xenopi have been linked with contaminated water. M. xenopi has been shown to interact with the biofilm formed in water distribution systems and to be hosted by free-living Acanthamoeba. The present study investigated the interaction between M. xenopi and A. polyphaga amoeba, and between M. xenopi and human fibroblast HEL cells. Examination using the light microscopy together with electronic and confocal microscopy demonstrated that M. xenopi was located within the amoeba and in HEL cells. The Light Cycler measurement of the M. xenopi:A. polyphaga DNA ratio and the M. xenopi:HEL cell DNA ratio demonstrated intra-amoebal and intracellular growth of M. xenopi with doubling-times of five-days and 10 days, respectively. Intra-amoebal M. xenopi survived protozoan encystment and germination. These data demonstrate that M. xenopi is a facultative intra-amoebal and intracellular pathogen. Testing intra-amoebal M. xenopi might be necessary to properly evaluate decontamination procedures for hospital water supply systems in order to achieve eradication.

Mycobacterium xenopi multiplies within human macrophages and enhances HIV replication in vitro.

Mycobacterium xenopi can cause opportunistic infections, particularly in persons infected with human immunodeficiency virus type 1 (HIV-1). The primary focus of this effort was to determine if M. xenopi isolates could survive and grow in human peripheral blood macrophage (MPhi), and if these isolates could promote the replication of HIV-1 in vitro. M. xenopi bacilli survived and replicated 10-fold within 48 h in human MPhi while avirulent Mycobacterium smegmatis, did not grow within the MPhi. M. xenopi bacilli when cultured with peripheral blood mononuclear cells enhanced HIV-1 replication 30- and 50-fold with the macrophage-tropic HIV-1(Ba-L) and 50- and 75-fold with T-cell-tropic strain HIV-1(LAI) by 6 days post-infection when compared to M. smegmatis. The enhanced HIV replication was associated with increased production of TNF-alpha. Partial inhibition of HIV-1 induction was observed using a neutralizing anti-TNF-alpha monoclonal antibody, pentoxifylline, and matrix metalloproteinase (MMP) inhibitor I. Similar mechanisms of pathogenesis among mycobacterial species may help elucidate better treatment approaches in HIV co-infected persons.

Mycobacterium xenopi and drinking water biofilms.

The ability of Mycobacterium xenopi to colonize an experimental drinking water distribution system (a Propella reactor) was investigated. M. xenopi was present in the biofilm within an hour following its introduction. After 9 weeks, it was always present in the outlet water (1 to 10 CFU 100 ml(-1)) and inside the biofilm (10(2) to 10(3) CFU cm(-2)). Biofilms may be considered reservoirs for the survival of M. xenopi.

[Negative effect of the components of the lysis-centrifugation system in the growth of mycobacteria in MGIT and Septi-Chek AFB liquid media]. / Efecto negativo de los componentes del sistema lisis-centrifugación en el crecimiento de micobacterias en los medios líquidos MGIT y Septi-Chek AFB.

BACKGROUND: The lysis-centrifugation system (Isolator system) is a technique with excellent results in the recovery of mycobacteria from blood specimens. This system consists mainly of saponin (SAP), polypropylenglycol (PPG), and sodium polianthol sulfonate (SPS). The objective of this work was to determine the effect of SAP, PPG, and SPS on the growth of Mycobacterium avium, M. kansasii, M. tuberculosis, and M. xenopi in fluid culture media MGIT and Septi-Chek AFB. METHODS: Two concentrations each of SAP, PPG, and SPS were prepared, and were added in 0.1 ml amounts (alone, in pairs and in combination) to fluid media MGIT and Septi-Chek AFB. Fluid culture media were then in individually inoculated with two different concentrations (10(3) and 10(5) CFU/ml) of each of the four mycobacterial strains used in this study. Culture media were incubated at 37 degrees C and were checked for growth daily. RESULTS: SAP, PPG, and SPS did not inhibit growth of mycobacteria but growth of these strains was indeed retarded (a lengthier time was required for detection of bacterial growth compared with the positive control). Final concentrations of SAP, PPG, and SPS which retarded mycobacterial growth varied, depending upon species, mycobacterial inoculum size, and fluid culture media used. CONCLUSIONS: Components included in the lysy-centrifugation system (SAP, PPG, and SPS), either alone or in combination retarded growth of M. avium, M. kansasii, M. tuberculosis, and M. xenopi in 10(3) and 10(5) CFU/ml concentrations in fluid culture media MGIT and Septi-Chek AFB. These results suggest that strategies should be adopted to decrease the concentrations of these three components, present in the sediment of the processed blood by the Isolator System, which eventually are going to be added to fluid media MGIT and Septi-Check AFB.