Comparison of Methods for Processing Drinking Water Samples for the Isolation of
            <i>Mycobacterium avium</i>
            and
            <i>Mycobacterium intracellulare</i>

Thomson, Rachel; Carter, Robyn; Gilpin, Chris; Coulter, Chris; Hargreaves, Megan

doi:10.1128/aem.02009-07

Cited by 47 publications

(40 citation statements)

References 30 publications

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“…This makes the failure to recover M. intracellulare from household water samples in the United States not likely to be a function of either geography or inadequate sampling. There is no evidence that the decontamination procedure or the growth medium inhibits the growth of M. intracellulare (18). ITS sequence analysis detects only the most abundant microbial species.…”

Section: Discussionmentioning

confidence: 99%

Absence of Mycobacterium intracellulare and Presence of Mycobacterium chimaera in Household Water and Biofilm Samples of Patients in the United States with Mycobacterium avium Complex Respiratory Disease

et al. 2013

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f Recent studies have shown that respiratory isolates from pulmonary disease patients and household water/biofilm isolates of Mycobacterium avium could be matched by DNA fingerprinting. To determine if this is true for Mycobacterium intracellulare, household water sources for 36 patients with Mycobacterium avium complex (MAC) lung disease were evaluated. MAC household water isolates from three published studies that included 37 additional MAC respiratory disease patients were also evaluated. Species identification was done initially using nonsequencing methods with confirmation by internal transcribed spacer (ITS) and/or partial 16S rRNA gene sequencing. M. intracellulare was identified by nonsequencing methods in 54 respiratory cultures and 41 household water/biofilm samples. By ITS sequencing, 49 (90.7%) respiratory isolates were M. intracellulare and 4 (7.4%) were Mycobacterium chimaera. In contrast, 30 (73%) household water samples were M. chimaera, 8 (20%) were other MAC X species (i.e., isolates positive with a MAC probe but negative with species-specific M. avium and M. intracellulare probes), and 3 (7%) were M. avium; none were M. intracellulare. In comparison, M. avium was recovered from 141 water/biofilm samples. These results indicate that M. intracellulare lung disease in the United States is acquired from environmental sources other than household water. Nonsequencing methods for identification of nontuberculous mycobacteria (including those of the MAC) might fail to distinguish closely related species (such as M. intracellulare and M. chimaera). This is the first report of M. chimaera recovery from household water. The study underscores the importance of taxonomy and distinguishing the many species and subspecies of the MAC. P revious studies have suggested household water (especially from bathroom showers) as a source of the Mycobacterium avium complex (MAC), which causes chronic lung disease (1, 2, 3, 4, 5). Both M. avium and M. intracellulare have been recovered from sputum, sinus, and household or potable water samples in multiple countries, including the United States and Japan (1, 4-8). In the majority of these studies, either hybridization probe methods (AccuProbe; Hologic Gen-Probe, Inc., San Diego, CA) or a multiplex 16S rRNA gene PCR was used for identification (1, 5, 9, 10). Isolates positive with the MAC probe but negative with the species-specific M. avium and M. intracellulare probes are collectively referred to as MAC X species.Currently We recently completed a study of variable-number tandemrepeat (VNTR) typing of clinical isolates of M. intracellulare using ITS sequencing to confirm the species designation (7). We then began household water biofilm studies for some of our patients and performed species identification and VNTR genotyping for MAC isolates as needed.Preliminary studies suggested that respiratory isolates of M. intracellulare indeed belonged to that species based on ITS sequencing (7) but that biofilm isolates thought to be M. intracellulare were actually M. chimaera ...

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Section: Discussionmentioning

confidence: 99%

Absence of Mycobacterium intracellulare and Presence of Mycobacterium chimaera in Household Water and Biofilm Samples of Patients in the United States with Mycobacterium avium Complex Respiratory Disease

et al. 2013

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“…Water samples were processed as previously described (9,19). After analysis of water sampling revealed an increased yield for rapidly growing mycobacteria using liquid medium (MGIT tubes) (9), these were included for the last 6 houses that were sampled.…”

Section: Methodsmentioning

confidence: 99%

Isolation of Nontuberculous Mycobacteria (NTM) from Household Water and Shower Aerosols in Patients with Pulmonary Disease Caused by NTM

et al. 2013

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“…This microbial diversity makes it likely that nontarget species will overgrow NTM in nutrient-rich medium. Several studies have been conducted to determine the optimum decontamination method for inhibiting the growth of nontarget bacteria in NTM assays, although most of the methods were developed for clinical samples (2,8,20,42,56). Moreover, no clear consensus for treatment of environmental samples has emerged from these studies.…”

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Comparison of Culture Methods for Isolation of Nontuberculous Mycobacteria from Surface Waters

Radomski

Cambau

Moulin

et al. 2010

Appl Environ Microbiol

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The environment is the likely source of most nontuberculous mycobacteria (NTM) involved in human infections, especially pulmonary, skin, and soft tissue infections. In order to measure the prevalence of NTM in different aquatic ecosystems, we tried to standardize the culture methods used for surface water testing since many procedures have been described previously. Cultivation of mycobacteria requires long-term incubation in rich media and inactivation of rapidly growing microorganisms whose growth impedes observation of mycobacterial colonies. Consequently, the two criteria used for evaluation of the methods examined were (i) the rate of inhibition of nontarget microorganisms and (ii) the efficiency of recovery of mycobacteria. We compared the competitive growth of Mycobacterium chelonae and M. avium with nontarget microorganisms on rich Middlebrook 7H11-mycobactin medium after treatment by several chemical decontamination methods that included acids, bases, detergent, or cetylpyridinium chloride (CPC) with and without an antibiotic cocktail, either PANTA (40 U/ml polymyxin, 4 g/ml amphotericin B, 16 g/ml nalidixic acid, 4 g/ml trimethoprim, and 4 g/ml azlocillin) or PANTAV (PANTA plus 10 g/ml vancomycin). Our results showed that treatment for 30 min with CPC (final concentration, 0.05%) of water concentrated by centrifugation, followed by culture on a rich medium supplemented with PANTA, significantly decreased the growth of nontarget microorganisms (the concentrations were 6.2 ؎ 0.4 log 10 CFU/liter on Middlebrook 7H11j medium and 4.2 ؎ 0.2 log 10 CFU/liter on Middlebrook 7H11j medium containing PANTA [P < 0.001]), while the effect of this procedure on NTM was not as great (the concentrations of M. chelonae on the two media were 7.0 ؎ 0.0 log 10 CFU/liter and 6.9 ؎ 0.0 log 10 CFU/liter, respectively, and the concentrations of M. avium were 9.1 ؎ 0.0 log 10 CFU/liter and 8.9 ؎ 0.0 log 10 CFU/liter, respectively). We propose that this standardized culture procedure could be used for detection of NTM in aquatic samples.

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Comparison of Methods for Processing Drinking Water Samples for the Isolation of Mycobacterium avium and Mycobacterium intracellulare

Cited by 47 publications

References 30 publications

Absence of Mycobacterium intracellulare and Presence of Mycobacterium chimaera in Household Water and Biofilm Samples of Patients in the United States with Mycobacterium avium Complex Respiratory Disease

Absence of Mycobacterium intracellulare and Presence of Mycobacterium chimaera in Household Water and Biofilm Samples of Patients in the United States with Mycobacterium avium Complex Respiratory Disease

Isolation of Nontuberculous Mycobacteria (NTM) from Household Water and Shower Aerosols in Patients with Pulmonary Disease Caused by NTM

Comparison of Culture Methods for Isolation of Nontuberculous Mycobacteria from Surface Waters

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