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Sample records for rapidly growing mycobacteria

  1. [Infections due to rapidly growing mycobacteria].

    PubMed

    García-Martos, Pedro; García-Agudo, Lidia

    2012-04-01

    Rapidly growing mycobacteria (RGM) are ubiquitous in nature and widely distributed in water, soil and animals. During the past three decades we have observed a notable increment of infections caused by RGM, both localized and disseminated, as well as nosocomial outbreaks of contaminated medical equipment. The microbiological diagnosis of RGM infections includes direct microscopic observation and culture. The taxonomic identification is performed by phenotypic, biochemical, chromatographic and molecular biology techniques. The treatment differs from that of other mycobacteriosis like tuberculosis, owing to the variable in vitro susceptibility of the species of this group. The RGM are resistant to conventional antituberculous drugs, but can be susceptible to broad spectrum antimicrobial agents. In this study we comment on the significant aspects of human infections by RGM, including their biology, epidemiology, pathology, microbiological diagnosis, taxonomic identification, antimicrobial susceptibility and treatment.

  2. Evaluation of INNO-LiPA mycobacteria v2 assay for identification of rapidly growing mycobacteria.

    PubMed

    García-Agudo, Lidia; Jesús, Iría; Rodríguez-Iglesias, Manuel; García-Martos, Pedro

    2011-07-01

    A total of 54 rapidly growing mycobacteria (RGM) isolated from patients attended in the two hospitals of Cádiz Bay (Spain) were selected during a seven-year-period (2000-2006) in order to evaluate the INNO-LiPA Mycobacteria v2 assay for mycobacterial identification, based on the reverse hybridization principle. The strains were cultured in Löwenstein-Jensen and Middlebrook 7H9 media and identified to the species level by sequencing of the 16S rRNA, PCR-restriction enzyme analysis of the hsp65 gene, conventional tests and INNO-LiPA Mycobacteria v2 assay. By the molecular methods we identified a total of 12 different species: 23 Mycobacterium fortuitum, 11 M. chelonae, 10 M. abscessus, 2 M. senegalense, 1 M. alvei, 1 M. brumae, 1 M. mageritense, 1 M. mucogenicum, 1 M. neoaurum, 1 M. peregrinum, 1 M. septicum and 1 M. smegmatis. Fifty two strains (96.3%) were correctly identified by conventional techniques and 47 strains (87.0%) by INNO-LiPA Mycobacteria v2 assay. We find INNO-LiPA Mycobacteria v2 assay simple to perform but it provides few advantages in comparison with conventional methods and sometimes needs complementary tests to identify Mycobacterium fortuitum complex, M. chelonae complex and specific species due to the great heterogeneity in the RGM group.

  3. Evaluation of INNO-LiPA mycobacteria v2 assay for identification of rapidly growing mycobacteria

    PubMed Central

    García-Agudo, Lidia; Jesús, Iría; Rodríguez-Iglesias, Manuel; García-Martos, Pedro

    2011-01-01

    A total of 54 rapidly growing mycobacteria (RGM) isolated from patients attended in the two hospitals of Cádiz Bay (Spain) were selected during a seven-year-period (2000–2006) in order to evaluate the INNO-LiPA Mycobacteria v2 assay for mycobacterial identification, based on the reverse hybridization principle. The strains were cultured in Löwenstein-Jensen and Middlebrook 7H9 media and identified to the species level by sequencing of the 16S rRNA, PCR-restriction enzyme analysis of the hsp65 gene, conventional tests and INNO-LiPA Mycobacteria v2 assay. By the molecular methods we identified a total of 12 different species: 23 Mycobacterium fortuitum, 11 M. chelonae, 10 M. abscessus, 2 M. senegalense, 1 M. alvei, 1 M. brumae, 1 M. mageritense, 1 M. mucogenicum, 1 M. neoaurum, 1 M. peregrinum, 1 M. septicum and 1 M. smegmatis. Fifty two strains (96.3%) were correctly identified by conventional techniques and 47 strains (87.0%) by INNO-LiPA Mycobacteria v2 assay. We find INNO-LiPA Mycobacteria v2 assay simple to perform but it provides few advantages in comparison with conventional methods and sometimes needs complementary tests to identify Mycobacterium fortuitum complex, M. chelonae complex and specific species due to the great heterogeneity in the RGM group. PMID:24031745

  4. Evaluation of Various Culture Media for Detection of Rapidly Growing Mycobacteria from Patients with Cystic Fibrosis

    PubMed Central

    Preece, Clair L.; Wichelhaus, Thomas A.; Perry, Audrey; Jones, Amanda L.; Cummings, Stephen P.; Hogardt, Michael

    2016-01-01

    Isolation of nontuberculous mycobacteria (NTM) from the sputum of patients with cystic fibrosis (CF) is challenging due to overgrowth by rapidly growing species that colonize the lungs of patients with CF. Extended incubation on Burkholderia cepacia selective agar (BCSA) has been recommended as an expedient culture method for the isolation of rapidly growing NTM in this setting. The aim of this study was to assess five selective media designed for the isolation of Burkholderia cepacia complex, along with two media designed for the isolation of mycobacteria (rapidly growing mycobacteria [RGM] medium and Middlebrook 7H11 agar), for their abilities to isolate NTM. All seven media were challenged with 147 isolates of rapidly growing mycobacteria and 185 isolates belonging to other species. RGM medium was then compared with the most selective brand of BCSA for the isolation of NTM from 224 sputum samples from patients with CF. Different agars designed for the isolation of B. cepacia complex varied considerably in their inhibition of other bacteria and fungi. RGM medium supported the growth of all isolates of mycobacteria and was more selective than any other medium. NTM were recovered from 17 of 224 sputum samples using RGM medium, compared with only 7 samples using the most selective brand of BCSA (P = 0.023). RGM medium offers a superior option, compared to other selective agars, for the isolation of rapidly growing mycobacteria from the sputum of patients with CF. Furthermore, the convenience of using RGM medium enables routine screening for rapidly growing NTM in all submitted sputum samples from patients with CF. PMID:27098962

  5. Structural analysis of biofilm formation by rapidly and slowly growing nontuberculous mycobacteria

    EPA Science Inventory

    Mycobacterium avium complex (MAC) and rapidly growing mycobacteria (RGM) such as M. abscessus, M. mucogenicum, M. chelonae and M. fortuitum, implicated in healthcare-associated infections, are often isolated from potable water supplies as part of the microbial flora. To understa...

  6. Structural analysis of biofilm formation by rapidly and slowly growing nontuberculous mycobacteria

    EPA Science Inventory

    Mycobacterium avium complex (MAC) and rapidly growing mycobacteria (RGM) such as M. abscessus, M. mucogenicum, M. chelonae and M. fortuitum, implicated in healthcare-associated infections, are often isolated from potable water supplies as part of the microbial flora. To understa...

  7. Pacemaker pocket infection due to Mycobacterium goodii, a rapidly growing mycobacteria.

    PubMed

    Yoo, David K; Hosseini-Moghaddam, Seyed M

    2017-01-10

    A woman aged 74 years with an implanted dual-chamber pacemaker presented with pacemaker site infection after failing empiric antimicrobial therapy. The pathogen was later identified as Mycobacterium goodii, a rapidly growing mycobacteria species. The pacemaker was subsequently removed and the patient was treated with oral ciprofloxacin and doxycycline with clinical improvement. In this article, we describe a rare case of pacemaker site infection by M. goodii. 2017 BMJ Publishing Group Ltd.

  8. Management of Infections with Rapidly Growing Mycobacteria after Unexpected Complications of Skin and Subcutaneous Surgical Procedures

    PubMed Central

    Lim, Jong Min; Kim, Jong Hwan

    2012-01-01

    Background Infection caused by rapidly growing mycobacteria (RGM) is not uncommon, and the prevalence of RGM infection has been increasing. Clinical diagnosis is difficult because there are no characteristic clinical features. There is also no standard antibiotic regimen for treating RGM infection. A small series of patients with RGM infections was studied to examine their treatments and outcomes. Methods A total of 5 patients who had developed postoperative infections from January 2009 to December 2010 were retrospectively reviewed. Patients were initially screened using a mycobacteria rapid screening test (polymerase chain reaction [PCR]-reverse blot hybridization assay). To confirm mycobacterial infection, specimens were cultured for nontuberculous mycobacteria and analyzed by 16 S ribosomal RNA and rpoB gene PCR. Results The patients were treated with intravenous antibiotics during hospitalization, and oral antibiotics were administered after discharge. The mean duration of follow-up was 9 months, and all patients were completely cured of infection with a regimen of a combination of antibiotics plus surgical treatment. Although none of the patients developed recurrence, there were complications at the site of infection, including hypertrophic scarring, pigmentation, and disfigurement. Conclusions Combination antibiotic therapy plus drainage of surgical abscesses appeared to be effective for the RGM infections seen in our patients. Although neither the exact dosage nor a standardized regimen has been firmly established, we propose that our treatment can provide an option for the management of rapidly growing mycobacterial infection. PMID:22783486

  9. Mycobacteriocins produced by rapidly growing mycobacteria are Tween-hydrolyzing esterases.

    PubMed Central

    Saito, H; Tomioka, H; Watanabe, T; Yoneyama, T

    1983-01-01

    Smegmatocin, a protein produced by Mycobacterium smegmatis ATCC 14468, was found to have an esterase activity, hydrolyzing Tween 80, polyoxyethylene sorbitan monooleate, added to the assay medium for various "bacteriocins" from mycobacteria. Because M. diernhoferi ATCC 19340 (indicator strain for smegmatocin) is highly susceptible to oleic acid and smegmatocin requires Tween 80 for manifestation of its anti-M. diernhoferi activity, it is likely that smegmatocin-mediated antimicrobial action is caused by oleic acid generated by hydrolysis of Tween 80 by the inherent esterase action of smegmatocin. Other mycobacteriocins from rapidly growing mycobacteria also have inherent esterase activity against Tween 80 and require Tween 80 for expression of antimycobacterial action. Smegmatocin was found to hydrolyze various polyoxyethylene (sorbitan) fatty acyl esters but not sorbitan monooleate and glyceryl esters. Images PMID:6826523

  10. Development of a rapid ATP bioluminescence assay for biocidal susceptibility testing of rapidly growing mycobacteria.

    PubMed

    Kapoor, Renuka; Yadav, Jagjit S

    2010-10-01

    An ATP-based biocide susceptibility assay for mycobacteria was developed by optimizing the cell lysis and assay conditions. Compared to the conventional agar plating method, the assay was rapid (1.5 h) and showed high sensitivity and specificity as determined by receiver operating characteristic (ROC) analysis. The test species, Mycobacterium immunogenum, M. chelonae, and M. abscessus, showed various susceptibilities to the glutaraldehyde- and isothiazolone-based test biocides.

  11. In vitro activity of extracts and constituents of Pelagonium against rapidly growing mycobacteria.

    PubMed

    Seidel, Veronique; Taylor, Peter W

    2004-06-01

    Extracts of the roots of plants of the Geraniaceae family have been used for many years in South Africa as native herbal remedies and there is circumstantial evidence for efficacy in the treatment of pulmonary tuberculosis. We have examined dried roots of Pelargonium reniforme and P. sidoides for antibacterial activity against rapidly growing mycobacteria. Fractions with activity against Mycobacterium aurum and M. smegmatis were obtained from both plant species by bioassay-guided fractionation of n-hexane extracts and were found to contain mixtures of straight-chain fatty acids. Analysis by gas chromatography-mass spectrometry (GC-MS) of the corresponding fatty acid methyl esters revealed structures with chain lengths ranging from C12 to C26. Unsaturated compounds were analysed as the corresponding dimethyl disulfide adducts to determine double-bond positions. Active mixtures differed in the relative abundance of their components, but all contained 16:0 (palmitic), Delta9-18:1 (oleic) and Delta9,12-18:2 (linoleic acid) as the major components. When tested against M. aurum, M. smegmatis and other rapidly growing mycobacteria (M. fortuitum, M. abscessus and M. phlei), all saturated compounds except 12:0 were devoid of antimycobacterial activity, whereas unsaturated compounds showed antimycobacterial activity related to their degree of unsaturation, their chain length and the bacterial species tested. The most potent compound was linoleic acid, with MIC of 2 mg/l against M. aurum. Copyright 2004 Elsevier B.V.

  12. Variables Affecting Results of Sodium Chloride Tolerance Test for Identification of Rapidly Growing Mycobacteria

    PubMed Central

    Conville, Patricia S.; Witebsky, Frank G.

    1998-01-01

    The sodium chloride tolerance test is often used in the identification of rapidly growing mycobacteria, particularly for distinguishing between Mycobacterium abscessus and Mycobacterium chelonae. This test, however, is frequently unreliable for the identification of some species. In this study we examined the following variables: medium manufacturer, inoculum concentration, and atmosphere and temperature of incubation. Results show that reliability is improved if the test and control slants are inoculated with an organism suspension spectrophotometrically equal to a 1 McFarland standard. Slants should be incubated at 35°C in ambient air and checked weekly for 4 weeks. Growth on control slants should be critically evaluated to determine the adequacy of the inoculum; colonies should number greater than 50. Salt-containing media should be examined carefully to detect pinpoint or tiny colonies, and colonies should number greater than 50 for a positive reaction. Concurrent use of a citrate slant may be helpful for distinguishing between M. abscessus and M. chelonae. Molecular methodologies are probably the most reliable means for the identification of rapidly growing mycobacteria and should be used, if possible, when unequivocal species identification is of particular importance. PMID:9620376

  13. Antimicrobial susceptibility of rapidly growing mycobacteria using the rapid colorimetric method.

    PubMed

    Ramis, I B; Cnockaert, M; von Groll, A; Nogueira, C L; Leão, S C; Andre, E; Simon, A; Palomino, J C; da Silva, P E A; Vandamme, P; Martin, A

    2015-07-01

    Drug susceptibility testing (DST) of rapidly growing mycobacteria (RGM) are recommended for guiding the antimicrobial therapy. We have evaluated the use of resazurin in Mueller-Hinton medium (MHR) for MIC determination of RGM and compared the results with those obtained with the reference standard broth microdilution in Mueller-Hinton (MH) and with the resazurin microtiter assay (REMA) in 7H9 broth. The MIC of eight drugs: amikacin (AMI), cefoxitin (FOX), ciprofloxacin (CIP), clarithromycin (CLA), doxycycline (DOX), linezolid (LZD), moxifloxacin (MXF) and trimethoprim-sulfamethoxazole (TMP-SMX) were evaluated against 76 RGM (18 species) using three methods (MH, MHR, and REMA) in a 96-well plate format incubated at 37 °C over 3-5 days. Results obtained in the MH plates were interpreted by the appearance of turbidity at the bottom of the well before adding the resazurin. MHR and 7H9-REMA plates were read by visual observation for a change in color from blue to pink. The majority of results were obtained at day 5 for MH and 1 day after for MHR and 7H9-REMA. However, the preliminary experiment on time to positivity results using the reference strain showed that the resazurin can be added to the MH at day 2 to produce the results at day 3, but future studies with large sets of strains are required to confirm this suggestion. A high level of agreement (kappa 1.000-0.884) was obtained between the MH and the MHR. Comparison of results obtained with 7H9-REMA, on the other hand, revealed several discrepancies and a lower level of agreement (kappa 1.000-0.111). The majority of the strains were resistant to DOX and TMP-SMX, and the most active antimicrobials for RGM were AMI and FOX. In the present study, MHR represented an excellent alternative for MIC determination of RGM. The results could be read reliably, more easily, and more quickly than with the classical MH method.

  14. Mercuric reductase activity and evidence of broad-spectrum mercury resistance among clinical isolates of rapidly growing mycobacteria

    SciTech Connect

    Steingrube, V.A.; Wallace, R.J. Jr.; Steele, L.C.; Pang, Y.J. )

    1991-05-01

    Resistance to mercury was evaluated in 356 rapidly growing mycobacteria belonging to eight taxonomic groups. Resistance to inorganic Hg2+ ranged from 0% among the unnamed third biovariant complex of Mycobacterium fortuitum to 83% among M. chelonae-like organisms. With cell extracts and 203Hg(NO3)2 as the substrate, mercuric reductase (HgRe) activity was demonstrable in six of eight taxonomic groups. HgRe activity was inducible and required NADPH or NADH and a thiol donor for optimai activity. Species with HgRe activity were also resistant to organomercurial compounds, including phenylmercuric acetate. Attempts at intraspecies and intragenus transfer of HgRe activity by conjugation or transformation were unsuccessful. Mercury resistance is common in rapidly growing mycobacteria and appears to function via the same inducible enzyme systems already defined in other bacterial species. This system offers potential as a strain marker for epidemiologic investigations and for studying genetic systems in rapidly growing mycobacteria.

  15. Skin and Soft Tissue Infection due to Rapidly Growing Mycobacteria: Case Series and Literature Review

    PubMed Central

    Yu, Jung Re; Lee, Keun Hwa; Kim, Jinseok; Sung, Jae Kyung; Kim, Young Ree; Kim, Jae Wang

    2013-01-01

    Background Nontuberculous mycobacteria (NTM) are ubiquitous in soil and water. Most NTM cause disease in humans only rarely unless some aspect of host defense is impaired. Recently, rapidly growing mycobacteria (RGM) is not uncommon, and the prevalence of RGM infection has been increasing. RGM causes a wide spectrum of pulmonary and extrapulmonary diseases and has been shown as an important source for opportunistic infection. Materials and Methods We report 5 patients of skin and soft tissue infection due to RGM in tertiary medical center in Jeju Island and analyzed 21 patients of skin and soft tissue infection due to RGM in Republic of Korea. Clinical, microbiological and epidemiological data were collected from each patient. NTM isolates were identified using conventional and molecular methods including 16S rDNA gene sequencing. Results The mean age of the RGM patients (n=26) was 54.9 ± 15.9 years and 73% were women. Mycobacterium fortuitum complex was the most common (12/26). Antimicrobial resistance for clarithromycin and quinolone were 12% and 60%, respectively. Clarithromycin based therapy was done in 46%. The mean duration of treatment was 21.2 ± 8.7 weeks. Conclusions Many cases can be cured after therapy for 4-7 month with at least 2 or 3 antibiotics according to in vitro susceptibility. Recent increasing of NTM cases suggests that species and subspecies identification is epidemiologically important, especially related to medical procedure, and surgery. PMID:24265954

  16. Rapidly growing non-tuberculous mycobacteria infection of prosthetic knee joints: A report of two cases.

    PubMed

    Kim, Manyoung; Ha, Chul-Won; Jang, Jae Won; Park, Yong-Beom

    2017-08-01

    Non-tuberculous mycobacteria (NTM) cause prosthetic knee joint infections in rare cases. Infections with rapidly growing non-tuberculous mycobacteria (RGNTM) are difficult to treat due to their aggressive clinical behavior and resistance to antibiotics. Infections of a prosthetic knee joint by RGNTM have rarely been reported. A standard of treatment has not yet been established because of the rarity of the condition. In previous reports, diagnoses of RGNTM infections in prosthetic knee joints took a long time to reach because the condition was not suspected, due to its rarity. In addition, it is difficult to identify RGNTM in the lab because special identification tests are needed. In previous reports, after treatment for RGNTM prosthetic infections, knee prostheses could not be re-implanted in all cases but one, resulting in arthrodesis or resection arthroplasty; this was most likely due to the aggressiveness of these organisms. In the present report, two cases of prosthetic knee joint infection caused by RGNTM (Mycobacterium abscessus) are described that were successfully treated, and in which prosthetic joints were finally reimplanted in two-stage revision surgery. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. In vitro susceptibilities of rapidly growing mycobacteria to newer antimicrobial agents.

    PubMed Central

    Khardori, N; Nguyen, H; Rosenbaum, B; Rolston, K; Bodey, G P

    1994-01-01

    The in vitro antimicrobial susceptibilities of 42 isolates of rapidly growing mycobacteria (Mycobacterium fortuitum, M. chelonae, and Mycobacterium species [other than M. fortuitum and M. chelonae]) to nine quinolones, including newer agents, two new aminoglycosides, and an aminocyclitol (trospectomycin) were determined by a broth microdilution method. The new quinolones, PD 117596, PD 127391, and PD 117558, showed excellent in vitro activities against M. fortuitum (MICs for 90% of isolates [MIC90s], 0.06, 0.06, and 0.12 microgram/ml, respectively). The MIC90 of ciprofloxacin for M. fortuitum was 0.5 microgram/ml. Only 14 to 28% of isolates of M. chelonae were susceptible to various quinolones. Most isolates of all three species were susceptible to the new aminoglycosides SCH 21420 and SCH 22591. The MIC90s of trospectomycin were 8 micrograms/ml for M. chelonae, 32 micrograms/ml for Mycobacterium species, and > 64 micrograms/ml for M. fortuitum. PMID:8141567

  18. In-vitro evaluation of the adhesion to polypropylene sutures of non-pigmented, rapidly growing mycobacteria.

    PubMed

    Zamora, N; Esteban, J; Kinnari, T J; Celdrán, A; Granizo, J J; Zafra, C

    2007-09-01

    The ability of non-pigmented, rapidly growing mycobacteria (NPRGM) to attach to polypropylene sutures was evaluated using an in-vitro assay. Thirty clinical isolates and five culture collection strains of NPRGM, together with Staphylococcus epidermidis ATCC 35983, were tested. Mycobacterium fortuitum and Mycobacterium chelonae showed the highest attachment ability, which differed significantly from the results obtained with Mycobacterium peregrinum. According to these results, NPRGM are able to attach to polypropylene sutures, and the species implicated most frequently in human infection showed increased levels of attachment in comparison with the other mycobacteria studied.

  19. [Assessment of in vitro susceptibility to antimicrobials of rapidly growing mycobacteria by E-test].

    PubMed

    García-Agudo, Lidia; García-Martos, Pedro; Jesús, Iría; Rodríguez-Iglesias, Manuel

    2009-07-01

    Rapidly growing mycobacteria (RGM) are considered opportunistic pathogens. An increasing number of post traumatic or surgical infections are caused by these microorganisms. To determine the antimicrobial susceptibility of RGM using the E-test method. A total of 54 isolates of RGM was obtained from several clinical samples and selected for this study Strains were identified to the species level by phenotypic and biochemical characteristics, PCR-restriction enzyme analysis of the hsp65 gene (PRA) and sequencing of the 16S rRNA. Susceptibility was investigated by E-test to amikacin, cefoxitin, ciprofioxacin, clarithromycin, imipenem, quinupristin/dalfopristin, linezolid and tigecycline. Twelve different species of RGM were identified: Mycobacterium fortuitum (23 strains), M chelonae (11), M abscessus (10), Msenegalense (2), Malvei (1), Mbrumae (1), Mmageritense (1), mucogenicum (1), M neoaurum (1), Mperegrinum (1), M septicum (1) y M smegmatis (1). All the strains were inhibited by low concentrations of amikacin and tigecycline. Susceptibility to cefoxitin, fluoroquinolones, clarithromycin, imipenem and linezolid was variable. All but two strains were resistant to quinupristin/ dalfopristin. Due to the uneven antimicrobial susceptibility of different species of RGM, an antimicrobial susceptibility test is mandatory for these microorganisms. The E-test method is well suited to determine minimum inhibitory concentrations.

  20. Clinical and Taxonomic Status of Pathogenic Nonpigmented or Late-Pigmenting Rapidly Growing Mycobacteria

    PubMed Central

    Brown-Elliott, Barbara A.; Wallace, Richard J.

    2002-01-01

    The history, taxonomy, geographic distribution, clinical disease, and therapy of the pathogenic nonpigmented or late-pigmenting rapidly growing mycobacteria (RGM) are reviewed. Community-acquired disease and health care-associated disease are highlighted for each species. The latter grouping includes health care-associated outbreaks and pseudo-outbreaks as well as sporadic disease cases. Treatment recommendations for each species and type of disease are also described. Special emphasis is on the Mycobacterium fortuitum group, including M. fortuitum, M. peregrinum, and the unnamed third biovariant complex with its recent taxonomic changes and newly recognized species (including M. septicum, M. mageritense, and proposed species M. houstonense and M. bonickei). The clinical and taxonomic status of M. chelonae, M. abscessus, and M. mucogenicum is also detailed, along with that of the closely related new species, M. immunogenum. Additionally, newly recognized species, M. wolinskyi and M. goodii, as well as M. smegmatis sensu stricto, are included in a discussion of the M. smegmatis group. Laboratory diagnosis of RGM using phenotypic methods such as biochemical testing and high-performance liquid chromatography and molecular methods of diagnosis are also discussed. The latter includes PCR-restriction fragment length polymorphism analysis, hybridization, ribotyping, and sequence analysis. Susceptibility testing and antibiotic susceptibility patterns of the RGM are also annotated, along with the current recommendations from the National Committee for Clinical Laboratory Standards (NCCLS) for mycobacterial susceptibility testing. PMID:12364376

  1. Outbreak of Rapidly Growing Nontuberculous Mycobacteria Among Patients Undergoing Cosmetic Surgery in the Dominican Republic.

    PubMed

    Green, Daniel A; Whittier, Susan; Greendyke, William; Win, Cindy; Chen, Xiaowei; Hamele-Bena, Diane

    2017-01-01

    Rapidly growing nontuberculous mycobacteria (RG-NTM), which can contaminate inadequately sterilized medical instruments, have been known to cause serious postsurgical skin and soft tissue infections that often are characterized by a prolonged incubation period and a disfiguring clinical course. Historically, these infections have been associated with surgical procedures performed outside the United States. The Centers for Disease Control and Prevention recently reported an outbreak of RG-NTM infections among women who underwent cosmetic surgery in the Dominican Republic. Because of the large Dominican American community in upper Manhattan, we have recently observed a number of these cases at NewYork-Presbyterian Hospital/Columbia University Medical Center. We highlight the case of a 55-year-old woman who developed a postsurgical RG-NTM infection after bilateral breast reduction in the Dominican Republic; she experienced progressive deformity of her left breast until the causative pathogen was identified 20 months after her initial surgery. To assist in the timely diagnosis and treatment of these infections, we aim to promote greater awareness among physicians who are likely to encounter such patients. We present the pathologic findings of a review of 7 cases of RG-NTM infections seen at NewYork-Presbyterian Hospital/Columbia University Medical Center and discuss the diagnostic and therapeutic challenges associated with these infections, such as prolonged incubation periods, the need for acid-fast stains and mycobacterial cultures, and the combination of surgical therapy and lengthy antibiotic courses that are often required for treatment.

  2. Epidemiology of infections due to nonpigmented rapidly growing mycobacteria diagnosed in an urban area.

    PubMed

    Esteban, J; Martín-de-Hijas, N Z; Fernandez, A-I; Fernandez-Roblas, R; Gadea, I

    2008-10-01

    The objective was to determine the incidence, clinical significance, and epidemiology of the isolates of nonpigmented, rapidly growing mycobacteria (NPRGM) in Madrid, Spain. Patients with new isolates of NPRGM during 2005 were selected prospectively for review of clinical charts. Clinical significance was analyzed according internationally accepted criteria. Randomly amplified polymorphic DNA (RAPD) was used for the genotyping of the isolates. NPRGM were identified in 70 patients (1.51 cases/100,000 inhabitants). The species were M. abscessus (in 5 patients), M. chelonae (in 9), M. fortuitum (in 40), M. peregrinum (in 9), M. mageritense (in 5), M. mucogenicum (in 2), and M. alvei (in 1 patient). The isolates were clinically significant in 17 cases (24.3%, 0.39 cases/100,000 inhabitants): in 4 cases of M. abscessus, in 5 of M. chelonae, and in 9 of M. fortuitum. Only 10.7% of the respiratory isolates were significant, whereas 75% of the nonrespiratory ones were significant (p < 0.001). RAPD analysis showed no relationship among the 74 strains available for the study. No characteristic resistance pattern could be found, although 4 strains appeared to be resistant to amikacin. Significant isolates were mainly nonrespiratory ones. The most significant species was M. abscessus. No relationship between the various isolates was detected, ruling out interhuman transmission between these cases.

  3. Rapid-Growing Mycobacteria Infections in Medical Tourists: Our Experience and Literature Review.

    PubMed

    Singh, Mansher; Dugdale, Caitlin M; Solomon, Isaac H; Huang, Anne; Montgomery, Mary W; Pomahac, Bohdan; Yawetz, Sigal; Maguire, James H; Talbot, Simon G

    2016-09-01

    "Medical tourism" has gained popularity over the past few decades. This is particularly common with patients seeking elective cosmetic surgery in the developing world. However, the risk of severe and unusual infectious complications appears to be higher than for patients undergoing similar procedures in the United States. The authors describe their experience with atypical mycobacterial infections in cosmetic surgical patients returning to the United States postoperatively. A review of patient medical records presenting with infectious complications after cosmetic surgery between January 2010 and July 2015 was performed. Patients presenting with mycobacterial infections following cosmetic surgery were reviewed in detail. An extensive literature review was performed for rapid-growing mycobacteria (RGM) related to cosmetic procedures. Between January 2010 and July 2015, three patients presented to our institution with culture-proven Mycobacterium abscessus at the sites of recent cosmetic surgery. All had surgery performed in the developing world. The mean age of these patients was 36 years (range, 29-44 years). There was a delay of up to 16 weeks between the initial presentation and correct diagnosis. All patients were treated with surgical drainage and combination antibiotics with complete resolution. We present series of patients with mycobacterial infections after cosmetic surgery in the developing world. This may be related to the endemic nature of these bacteria and/or inadequate sterilization or sterile technique. Due to low domestic incidence of these infections, diagnosis may be difficult and/or delayed. Consulting physicians should have a low threshold to consider atypical etiologies in such scenarios. 5 Therapeutic. © 2016 The American Society for Aesthetic Plastic Surgery, Inc. Reprints and permission: journals.permissions@oup.com.

  4. Analysis of a panel of rapidly growing mycobacteria for resistance to aldehyde-based disinfectants.

    PubMed

    De Groote, Mary Ann; Gibbs, Sara; de Moura, Vinicius Calado Nogueira; Burgess, Winona; Richardson, Kris; Kasperbauer, Shannon; Madinger, Nancy; Jackson, Mary

    2014-08-01

    After several accounts across the globe of mycobacteria outbreaks associated with medical procedures and aldehyde disinfectants resistance, we undertook an analysis of mycobacteria isolated from patients seen in a hospital in the United States between 1994 and 2008 to determine prevalence of resistance to aldehyde-based disinfectants. Out of the 117 clinical isolates screened, 6 isolates belonging to the emerging Mycobacterium abscessus group were found to display significant levels of resistance to glutaraldehyde and ortho-phthalaldehyde. Copyright © 2014 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

  5. Prosthetic joint infections secondary to rapidly growing mycobacteria: Two case reports and a review of the literature.

    PubMed

    Henry, Michael W; Miller, Andy O; Kahn, Barbara; Windsor, Russel E; Brause, Barry D

    2016-01-01

    Rapidly growing mycobacteria (RGM) are a rare but treatable cause of prosthetic joint infections. This study reports on two patients comprising three prosthetic joint infections caused by RGM successfully treated at the institution. With removal of the infected prosthetic joint and judicious use of prolonged courses of antibiotics, patients with prosthetic joint infections secondary to RGM can both be cured and retain function of the affected joint. In addition, this study identified 40 additional cases reported during an extensive review of the literature and provide a summary of these cases. These infections can present within days of arthroplasty or can develop only decades after the index surgery. The clinical presentations often mimic those of more routine bacterial prosthetic joint infections.

  6. Analysis of a Panel of Rapidly Growing Mycobacteria for Resistance to Aldehyde-based Disinfectants

    PubMed Central

    De Groote, Mary Ann; Gibbs, Sara; de Moura, Vinicius Calado Nogueira; Burgess, Winona; Richardson, Kris; Kasperbauer, Shannon; Madinger, Nancy; Jackson, Mary

    2014-01-01

    Summary After several accounts across the globe of mycobacterial outbreaks associated with medical procedures and aldehyde disinfectants resistance, we undertook an analysis of mycobacteria isolated from patients seen in a hospital in the United States between 1994 and 2008 to determine prevalence of resistance to aldehyde-based disinfectants. Out of the 117 clinical isolates screened, six isolates belonging to the emerging Mycobacterium abscessus group were found to display significant levels of resistance to glutaraldehyde and ortho-phthalaldehyde. PMID:25087149

  7. Mycobacterial contamination of metalworking fluids: involvement of a possible new taxon of rapidly growing mycobacteria.

    PubMed

    Moore, J S; Christensen, M; Wilson, R W; Wallace, R J; Zhang, Y; Nash, D R; Shelton, B

    2000-01-01

    Contamination of air and metalworking fluid (MWF) systems with a rapidly growing mycobacterium (RGM) was detected in 1995 in a single manufacturing plant with recent cases of hypersensitivity pneumonitis (HP). Extensive environmental sampling was performed to determine the extent of the contamination and its variability over time. RGM were present in multiple indoor air samples, 100% of the central MWF storage tanks, and 75% of the freestanding cutting, drilling, and grinding machines. With one exception, contamination was limited to a recently introduced formulation (brand) of semisynthetic MWF used in 95% of the facility's machining operations. In general, the mycobacterial counts were stable over time, with the degree of contamination ranging from 10(2)-10(7) colony forming units (CFU)/mL. A few systems were culture positive for the mycobacterium (> 10(1) CFU/mL), changed to culture negative (< 10(1) CFU/mL), then changed back to culture positive without explanation. Samples obtained from diluted (5%) but unused MWF, a replenishment line with 2% unused MWF, an MWF pasteurizer, city water, and deionized water were culture negative for this species of mycobacterium. Inoculation and growth studies demonstrated that this mycobacterium does not grow in liquid samples of 5% unused MWF. By molecular techniques, the mycobacterial isolates consisted of a single strain and represented a previously undescribed taxon closely related to Mycobacterium chelonae/abscessus. The relationship of this mycobacterium to the cases of HP is unknown.

  8. Effect of Antibiotics and Antibiofilm Agents in the Ultrastructure and Development of Biofilms Developed by Nonpigmented Rapidly Growing Mycobacteria.

    PubMed

    Muñoz-Egea, María-Carmen; García-Pedrazuela, María; Mahillo-Fernandez, Ignacio; Esteban, Jaime

    2016-01-01

    We analyze the effect of amikacin, ciprofloxacin, and clarithromycin, alone and associated with N-acetylcysteine (NAC) and Tween 80, at different times and concentrations in nonpigmented rapidly growing mycobacteria (NPRGM) biofilms. For this purpose, confocal laser scanning microscopy and image analysis were used to study the development and behavior of intrinsic autofluorescence, covered area, thickness, and cell viability in NPRGM biofilms after adding antibiotics alone and associated with antibiofilm agents. In this study, ciprofloxacin is the most active antibiotic against this type of biofilm and thickness is the most affected parameter. NAC and Tween 80 combined with antibiotics exert a synergistic effect in increasing the percentage of dead bacteria and also reducing the percentage of covered surface and thickness of NPRGM biofilms. Tween 80 seems to be an antibiofilm agent more effective than NAC due to its higher reduction in the percentage of cover surface and thickness. In conclusion, the results obtained in this work show that phenotypic parameters (thickness, percentage of covered surface, autofluorescence, percentage of live/dead bacteria) are affected by combining antibiotics and antibiofilm agents, ciprofloxacin and Tween 80 being the most active agents against NPRGM biofilms.

  9. [Rapid identification of mycobacteria using laser fluorescence].

    PubMed

    Ivanova, M A; Makarova, M V; Vasil'ev, E; Aleksandrov, M T; Pashkov, E P

    2009-01-01

    To develop rapid method of identification of mycobacteria based on laser fluorescence. Characteristics of laser-induced fluorescence of 19 bacteria species, including 17 species of mycobacteria, were studied. Identification of microorganisms was performed using measurement of spectral-fluorescent characteristics. Library of spectral-fluorescent characteristics of mycobacteria in different concentrations ratios and associations was created, which formed the basis of database for identification of mycobacteria by laser-fluorescent method. Principles of diagnostic algorithm of indication and differentiation of mycobacteria using this method were developed. Effect of myramistin for increasing the intensity of mycobacteria fluorescence, account of the diffracting characteristics of medium for adjustment of spectral characteristics of mycobacteria and processing of data by factor analysis are needed. Efficacy of the method was 80 - 90%. Principles of rapid identification of mycobacteria and their associations developed on the basis of laser-fluorescent method are experimentally founded and tested on unknown cultures of mycobacteria and objectively prove the possibility to apply this method for express identification of mycobacteria belonging to M. tuberculosis complex as well as non-tuberculous mycobacteria.

  10. Alpha-1-antitrypsin (AAT) anomalies are associated with lung disease due to rapidly growing mycobacteria and AAT inhibits Mycobacterium abscessus infection of macrophages.

    PubMed

    Chan, Edward D; Kaminska, Aleksandra M; Gill, Wendy; Chmura, Kathryn; Feldman, Nicole E; Bai, Xiyuan; Floyd, Corinne M; Fulton, Kayte E; Huitt, Gwen A; Strand, Matthew J; Iseman, Michael D; Shapiro, Leland

    2007-01-01

    Rapidly growing mycobacteria (RGM) are ubiquitous in the environment but cause lung disease in only a fraction of exposed individuals. This variable susceptibility to disease implies vulnerability to RGM infection due to weakness in host defense. Since most persons who contract RGM lung disease have no known host defense defect, it is likely that uncharacterized host deficiencies exist that predispose to RGM infection. Alpha-1-antitrypsin (AAT) is a host factor that may protect individuals from respiratory infections. Therefore, we assessed AAT protein anomalies as a risk factor for RGM lung disease. In a cohort of 100 patients with RGM lung disease, Mycobacterium (M.) abscessus was the most prevalent organism, isolated in 64 (64%) subjects. Anomalous AAT proteins were present in 27% of the cohort, which is 1.6 times the estimated prevalence of anomalous AAT proteins in the United States population (p=0.008). In in vitro studies, both AAT and a synthetic inhibitor of serine proteases suppressed M. abscessus infection of monocyte-derived macrophages by up to 65% (p<0.01). AAT may be an anti-RGM host-defense factor, and anomalous AAT phenotypes or AAT deficiency may constitute risk factors for pulmonary disease due to RGM.

  11. Point-of-use membrane filtration and hyperchlorination to prevent patient exposure to rapidly growing mycobacteria in the potable water supply of a skilled nursing facility.

    PubMed

    Williams, Margaret M; Chen, Tai-Ho; Keane, Tim; Toney, Nadege; Toney, Sean; Armbruster, Catherine R; Butler, W Ray; Arduino, Matthew J

    2011-09-01

    Healthcare-associated outbreaks and pseudo-outbreaks of rapidly growing mycobacteria (RGM) are frequently associated with contaminated tap water. A pseudo-outbreak of Mycobacterium chelonae-M. abscessus in patients undergoing bronchoscopy was identified by 2 acute care hospitals. RGM was identified in bronchoscopy specimens of 28 patients, 25 of whom resided in the same skilled nursing facility (SNF). An investigation ruled out bronchoscopy procedures, specimen collection, and scope reprocessing at the hospitals as sources of transmission. To identify the reservoir for RGM within the SNF and evaluate 2 water system treatments, hyperchlorination and point-of-use (POU) membrane filters, to reduce RGM. A comparative in situ study of 2 water system treatments to prevent RGM transmission. An SNF specializing in care of patients requiring ventilator support. RGM and heterotrophic plate count (HPC) bacteria were examined in facility water before and after hyperchlorination and in a subsequent 24-week assessment of filtered water by colony enumeration on Middlebrook and R2A media. Mycobacterium chelonae was consistently isolated from the SNF water supply. Hyperchlorination reduced RGM by 1.5 log(10) initially, but the population returned to original levels within 90 days. Concentration of HPC bacteria also decreased temporarily. RGM were reduced below detection level in filtered water, a 3-log(10) reduction. HPC bacteria were not recovered from newly installed filters, although low quantities were found in water from 2-week-old filters. POU membrane filters may be a feasible prevention measure for healthcare facilities to limit exposure of sensitive individuals to RGM in potable water systems.

  12. Diversity, Community Composition, and Dynamics of Nonpigmented and Late-Pigmenting Rapidly Growing Mycobacteria in an Urban Tap Water Production and Distribution System

    PubMed Central

    Dubrou, S.; Konjek, J.; Macheras, E.; Welté, B.; Guidicelli, L.; Chignon, E.; Joyeux, M.; Gaillard, J. L.; Heym, B.; Tully, T.

    2013-01-01

    Nonpigmented and late-pigmenting rapidly growing mycobacteria (RGM) have been reported to commonly colonize water production and distribution systems. However, there is little information about the nature and distribution of RGM species within the different parts of such complex networks or about their clustering into specific RGM species communities. We conducted a large-scale survey between 2007 and 2009 in the Parisian urban tap water production and distribution system. We analyzed 1,418 water samples from 36 sites, covering all production units, water storage tanks, and distribution units; RGM isolates were identified by using rpoB gene sequencing. We detected 18 RGM species and putative new species, with most isolates being Mycobacterium chelonae and Mycobacterium llatzerense. Using hierarchical clustering and principal-component analysis, we found that RGM were organized into various communities correlating with water origin (groundwater or surface water) and location within the distribution network. Water treatment plants were more specifically associated with species of the Mycobacterium septicum group. On average, M. chelonae dominated network sites fed by surface water, and M. llatzerense dominated those fed by groundwater. Overall, the M. chelonae prevalence index increased along the distribution network and was associated with a correlative decrease in the prevalence index of M. llatzerense, suggesting competitive or niche exclusion between these two dominant species. Our data describe the great diversity and complexity of RGM species living in the interconnected environments that constitute the water production and distribution system of a large city and highlight the prevalence index of the potentially pathogenic species M. chelonae in the distribution network. PMID:23835173

  13. In vitro interaction of some drug combinations to inhibit rapidly growing mycobacteria isolates from cats and dogs and these isolates' susceptibility to cefovecin and clofazimine.

    PubMed

    Bennie, C J M; To, J L K; Martin, P A; Govendir, M

    2015-01-01

    To investigate whether selected drug combinations used to treat rapidly growing mycobacteria (RGM) have drug-drug interactions that affect efficacy and to investigate each isolate's susceptibility to cefovecin and clofazimine, individually. In vitro susceptibility testing of bacterial isolates. Initially, five feline isolates and one canine isolate from both Mycobacterium fortuitum and M. smegmatis clusters (n = 12) underwent microbroth susceptibility testing to individual drugs to establish minimum inhibitory concentrations (MIC) of cefovecin, ciprofloxacin, clarithromycin, clofazimine, doxycycline, enrofloxacin, trimethoprim and sulfanilamide (the latter two as a combination). Checkerboard assays were then performed for susceptible M. smegmatis isolates for the following combinations: clarithromycin (one isolate only) versus enrofloxacin, clarithromycin vs doxycycline, clarithromycin vs trimethoprim/sulfanilamide; enrofloxacin vs doxycycline (six isolates); enrofloxacin vs trimethoprim/sulfanilamide (six isolates). Susceptible M. fortuitum isolates were tested against enrofloxacin versus doxycycline (four isolates only). All six M. fortuitum isolates were susceptible to enrofloxacin, but only four of six were susceptible to doxycycline. All six M. smegmatis isolates were susceptible to doxycycline, enrofloxacin and trimethoprim/sulfanilamide. A single isolate from the 12, a M. smegmatis isolate, was susceptible to clarithromycin. The fractional inhibitory concentration of each drug ranged from 0.64 to 1.84, indicating that neither synergism nor antagonism was evident. All 12 isolates were resistant to cefovecin. The clofazimine MIC50 to inhibit isolate growth was approximately 3.3 μg/mL for both strains. Drugs commonly used for treatment of RGM, when tested as combinations, do not appear to antagonise one another in vitro. Cefovecin is not efficacious for treatment of RGM infections. © 2015 Australian Veterinary Association.

  14. Diversity, community composition, and dynamics of nonpigmented and late-pigmenting rapidly growing mycobacteria in an urban tap water production and distribution system.

    PubMed

    Dubrou, S; Konjek, J; Macheras, E; Welté, B; Guidicelli, L; Chignon, E; Joyeux, M; Gaillard, J L; Heym, B; Tully, T; Sapriel, G

    2013-09-01

    Nonpigmented and late-pigmenting rapidly growing mycobacteria (RGM) have been reported to commonly colonize water production and distribution systems. However, there is little information about the nature and distribution of RGM species within the different parts of such complex networks or about their clustering into specific RGM species communities. We conducted a large-scale survey between 2007 and 2009 in the Parisian urban tap water production and distribution system. We analyzed 1,418 water samples from 36 sites, covering all production units, water storage tanks, and distribution units; RGM isolates were identified by using rpoB gene sequencing. We detected 18 RGM species and putative new species, with most isolates being Mycobacterium chelonae and Mycobacterium llatzerense. Using hierarchical clustering and principal-component analysis, we found that RGM were organized into various communities correlating with water origin (groundwater or surface water) and location within the distribution network. Water treatment plants were more specifically associated with species of the Mycobacterium septicum group. On average, M. chelonae dominated network sites fed by surface water, and M. llatzerense dominated those fed by groundwater. Overall, the M. chelonae prevalence index increased along the distribution network and was associated with a correlative decrease in the prevalence index of M. llatzerense, suggesting competitive or niche exclusion between these two dominant species. Our data describe the great diversity and complexity of RGM species living in the interconnected environments that constitute the water production and distribution system of a large city and highlight the prevalence index of the potentially pathogenic species M. chelonae in the distribution network.

  15. Deep brain stimulator infection by a novel rapid growing mycobacterium.

    PubMed

    Moritz, Donna C; Harrington, Amanda T; Slavin, Konstantin; Gomez, Christy; Jarrett, Olamide D

    2017-09-20

    Devise-related infections after deep brain stimulator implantation are not uncommon. However, infections due to mycobacteria have not been reported in the medical literature. We describe the first reported case of DBS infection due to a novel rapidly growing mycobacteria, most closely resembling Mycobacterium goodii, by rpoB gene sequencing.

  16. Differential Macrophage Response to Slow- and Fast-Growing Pathogenic Mycobacteria

    PubMed Central

    Helguera-Repetto, A. Cecilia; Chacon-Salinas, Rommel; Cerna-Cortes, Jorge F.; Rivera-Gutierrez, Sandra; Ortiz-Navarrete, Vianney; Estrada-Garcia, Iris; Gonzalez-y-Merchand, Jorge A.

    2014-01-01

    Nontuberculous mycobacteria (NTM) have recently been recognized as important species that cause disease even in immunocompetent individuals. The mechanisms that these species use to infect and persist inside macrophages are not well characterised. To gain insight concerning this process we used THP-1 macrophages infected with M. abscessus, M. fortuitum, M. celatum, and M. tuberculosis. Our results showed that slow-growing mycobacteria gained entrance into these cells with more efficiency than fast-growing mycobacteria. We have also demonstrated that viable slow-growing M. celatum persisted inside macrophages without causing cell damage and without inducing reactive oxygen species (ROS), as M. tuberculosis caused. In contrast, fast-growing mycobacteria destroyed the cells and induced high levels of ROS. Additionally, the macrophage cytokine pattern induced by M. celatum was different from the one induced by either M. tuberculosis or fast-growing mycobacteria. Our results also suggest that, in some cases, the intracellular survival of mycobacteria and the immune response that they induce in macrophages could be related to their growth rate. In addition, the modulation of macrophage cytokine production, caused by M. celatum, might be a novel immune-evasion strategy used to survive inside macrophages that is different from the one reported for M. tuberculosis. PMID:24949482

  17. Role of GenoType(®) Mycobacterium Common Mycobacteria/Additional Species Assay for Rapid Differentiation Between Mycobacterium tuberculosis Complex and Different Species of Non-Tuberculous Mycobacteria.

    PubMed

    Singh, Amresh Kumar; Maurya, Anand Kumar; Umrao, Jyoti; Kant, Surya; Kushwaha, Ram Awadh Singh; Nag, Vijaya Laskshmi; Dhole, Tapan N

    2013-07-01

    Mycobacterium tuberculosis complex (MTBC) and non-tuberculous mycobacteria (NTM) may or may not have same clinical presentations, but the treatment regimens are always different. Laboratory differentiation between MTBC and NTM by routine methods are time consuming and cumbersome to perform. We have evaluated the role of GenoType(®) Mycobacterium common mycobacteria/additional species (CM/AS) assay for differentiation between MTBC and different species of NTM in clinical isolates from tuberculosis (TB) cases. A total of 1080 clinical specimens were collected from January 2010 to June 2012. Diagnosis was performed by Ziehl-Neelsen staining followed by culture in BacT/ALERT 3D system (bioMerieux, France). A total of 219 culture positive clinical isolates (BacT/ALERT(®) MP cultures) were selected for differentiation by p-nitrobenzoic acid (PNB) sensitivity test as and BIO-LINE SD Ag MPT64 TB test considering as the gold standard test. Final identification and differentiation between MTBC and different species of NTM were further confirmed by GenoType(®) Mycobacterium CM/AS assay (Hain Lifescience, Nehren, Germany). Out of 219 BacT/ALERT(®) MP culture positive isolates tested by PNB as 153 MTBC (69.9%) and by GenoType(®) Mycobacterium CM/AS assay as 159 (72.6%) MTBC and remaining 60 (27.4%) were considered as NTM species. The GenoType(®) Mycobacterium CM/AS assay was proved 99.3% sensitive and 98.3% specific for rapid differentiation of MTBC and NTM. The most common NTM species were; Mycobacterium fortuitum 20 (33.3%) among rapid growing mycobacteria and Mycobacterium intracellulare 11 (18.3%) among slow growing mycobacteria. The GenoType(®) Mycobacterium assay makes rapid and accurate identification of NTM species as compared with different phenotypic and molecular diagnostic tool and helps in management of infections caused by different mycobacteria.

  18. [Rapidly growing mycobacteria infection after videosurgical procedures - the glutaraldehyde hypothesis].

    PubMed

    Lorena, Nádia Suely de Oliveira; Duarte, Rafael Silva; Pitombo, Marcos Bettini

    2009-07-01

    Between August 2006 and February 2007, in the state of Rio de Janeiro, Brazil, a massive outbreak of RGM infections after video laparoscopy was mainly associated to the recently described Mycobacterium massiliense species. All confirmed and probable cases reports described the use of high-level disinfection of medical devices by using 2% glutaraldehyde (2% GA) for 30 min before the surgical procedures. We investigated the susceptibility of the M. massiliense isolates recovered during the outbreak to high-level disinfection after 30 min, 1h, 6h and 10h of exposure to the commercial disinfectants. Reference strains for official mycobactericidal tests such as Mycobacterium abscessus, Mycobacterium bovis, Mycobacterium chelonae, Mycobacterium neoaurum and Mycobacterium smegmatis were included as controls. Although all the reference strains were eliminated in 30 min of exposure to 2% GA, we observed the recovery of all M. massiliense clinical isolates even after 10h of exposure. This study suggests that failures in high-level disinfection and the high tolerance of these M. massiliense clinical strains to the 2% GA were strongly associated to the magnitude of the outbreak.

  19. A Novel Rapidly Growing Mycobacterium Species Causing an Abdominal Cerebrospinal Fluid Pseudocyst Infection

    PubMed Central

    Hussain, Cory K.; de Man, Tom J. B.; Toney, Nadege C.; Kamboj, Kamal; Balada-Llasat, Joan-Miquel; Wang, Shu-Hua

    2016-01-01

    Nontuberculous mycobacteria (NTM) are a rare cause of ventriculoperitoneal shunt infections. We describe the isolation and identification of a novel, rapidly growing, nonpigmented NTM from an abdominal cerebrospinal fluid pseudocyst. The patient presented with fevers, nausea, and abdominal pain and clinically improved after shunt removal. NTM identification was performed by amplicon and whole-genome sequencing. PMID:27704004

  20. Rapid differentiation of mycobacteria by simplex real-time PCR with melting temperature calling analysis.

    PubMed

    Lin, L; Yin, X; Wang, Q

    2015-09-01

    This study aimed to develop a rapid, simple and cost-effective method for the differentiation of Mycobacterium species. A total of 80 clinical mycobacterial isolates belonging to 12 different species and 16 reference strains of 16 different species were differentiated by the simplex real-time PCR coupled with melting temperature calling analysis. By comparing their melting profiles with those of the reference strains, all clinical mycobacterial isolates were differentiated as Mycobacterium tuberculosis complex or nontuberculous mycobacteria, and the latter were further divided into five groups. In comparison with 16S-23S internal transcribed spacer sequencing method as the gold standard method, both sensitivity and specificity of the assay were 100% when it was used for the differentiation between Myco. tuberculosis complex and nontuberculous mycobacteria. The simplex real-time PCR coupled with melting temperature calling analysis could be an alternative method for the differentiation between Myco. tuberculosis complex and nontuberculous mycobacteria. Rapid differentiation of mycobacteria could shorten the diagnostic time of mycobacterial diseases. It is also helpful for achieving optimal therapy and appropriate patient management. © 2015 The Society for Applied Microbiology.

  1. Use of Microelectronic Array Technology for Rapid Identification of Clinically Relevant Mycobacteria

    PubMed Central

    Sanguinetti, Maurizio; Novarese, Linda; Posteraro, Brunella; Ranno, Stefania; De Carolis, Elena; Pecorini, Giovanni; Lucignano, Barbara; Ardito, Fausta; Fadda, Giovanni

    2005-01-01

    We developed a new method based on the Nanochip microelectronic array technology for identification of various clinically relevant mycobacterial species. PCR-amplified rRNA genes obtained from 270 positive Mycobacteria Growth Indicator Tube cultures were successfully tested by hybridizing them with species-selective probes, and the results agreed with those of conventional identification methods. The system is rapid and accurate and opens new perspectives in clinical diagnostics. PMID:16333127

  2. Use of microelectronic array technology for rapid identification of clinically relevant mycobacteria.

    PubMed

    Sanguinetti, Maurizio; Novarese, Linda; Posteraro, Brunella; Ranno, Stefania; De Carolis, Elena; Pecorini, Giovanni; Lucignano, Barbara; Ardito, Fausta; Fadda, Giovanni

    2005-12-01

    We developed a new method based on the Nanochip microelectronic array technology for identification of various clinically relevant mycobacterial species. PCR-amplified rRNA genes obtained from 270 positive Mycobacteria Growth Indicator Tube cultures were successfully tested by hybridizing them with species-selective probes, and the results agreed with those of conventional identification methods. The system is rapid and accurate and opens new perspectives in clinical diagnostics.

  3. Rapid recovery of mycobacteria from clinical specimens using automated radiometric technic

    SciTech Connect

    Park, C.H.; Hixon, D.L.; Ferguson, C.B.; Hall, S.L.; Risheim, C.C.; Cook, C.B.

    1984-03-01

    Automated radiometric technic (BACTEC Johnston Laboratories, Towson, MD) was compared with conventional mycobacterial culture procedure (Lowenstein-Jensen plus Gruft modification of Lowenstein-Jensen) in this study of 1,000 clinical specimens. In addition, 8-azaguanine inhibition was tested by radiometric technic as a rapid procedure for the differentiation of Mycobacterium tuberculosis from other mycobacterial species. A total of 59 mycobacteria was recovered. Of 28 clinically significant isolates (M. tuberculosis, M. kansasii, M. avium, M. fortuitum), the BACTEC system detected 26 (93%). Conventional methods recovered 23 (82%). The BACTEC system required an average of seven days to recover M. tuberculosis from smear-positive specimens compared with 18 days required by Lowenstein-Jensen or Gruft slants. From smear-negative specimens, the BACTEC detected M. tuberculosis in an average of 20 days versus 28 days by conventional procedure. All 20 isolates of M. tuberculosis were inhibited by 8-azaguanine, whereas 39 isolates of mycobacteria other than M. tuberculosis were not inhibited. The BACTEC system accomplishes more rapid recovery of mycobacteria and provides a higher yield than conventional methods.

  4. Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis.

    PubMed Central

    Telenti, A; Marchesi, F; Balz, M; Bally, F; Böttger, E C; Bodmer, T

    1993-01-01

    A method for the rapid identification of mycobacteria to the species level was developed on the basis of evaluation by the polymerase chain reaction (PCR) of the gene encoding for the 65-kDa protein. The method involves restriction enzyme analysis of PCR products obtained with primers common to all mycobacteria. Using two restriction enzymes, BstEII and HaeIII, medically relevant and other frequent laboratory isolates were differentiated to the species or subspecies level by PCR-restriction enzyme pattern analysis. PCR-restriction enzyme pattern analysis was performed on isolates (n = 330) from solid and fluid culture media, including BACTEC, or from frozen and lyophilized stocks. The procedure does not involve hybridization steps or the use of radioactivity and can be completed within 1 working day. Images PMID:8381805

  5. Isolation and characterization of a unique group of slowly growing mycobacteria: description of Mycobacterium lentiflavum sp. nov.

    PubMed Central

    Springer, B; Wu, W K; Bodmer, T; Haase, G; Pfyffer, G E; Kroppenstedt, R M; Schröder, K H; Emler, S; Kilburn, J O; Kirschner, P; Telenti, A; Coyle, M B; Böttger, E C

    1996-01-01

    A distinct group of slowly growing mycobacteria was identified on the basis of growth characteristics, biochemical and lipid profiles, and nucleic acid analyses. The isolates showed growth at 22 to 37 degrees C, yellow pigmentation, and negative tests for Tween 80 hydrolysis, nicotinic acid, nitrate reductase, and urease; tests for arylsulfatase, pyrazinamidase, and heat-stable catalase were variable. Analysis of cellular fatty acids by gas-liquid chromatography and mycolic acids by thin-layer chromatography and high-performance liquid chromatography indicated a distinctive pattern which was unlike those of other species. Determination of the 16S rRNA gene sequence showed a unique sequence closely related to Mycobacterium simiae and M. genavense. On the basis of DNA homology studies, we suggest that these organisms are representatives of a novel species, for which the name M. lentiflavum sp. nov. is proposed. PMID:8727884

  6. Rapid identification of mycolic acid patterns of mycobacteria by high-performance liquid chromatography using pattern recognition software and a Mycobacterium library.

    PubMed Central

    Glickman, S E; Kilburn, J O; Butler, W R; Ramos, L S

    1994-01-01

    Current methods for identifying mycobacteria by high-performance liquid chromatography (HPLC) require a visual assessment of the generated chromatographic data, which often involves time-consuming hand calculations and the use of flow charts. Our laboratory has developed a personal computer-based file containing patterns of mycolic acids detected in 45 species of Mycobacterium, including both slowly and rapidly growing species, as well as Tsukamurella paurometabolum and members of the genera Corynebacterium, Nocardia, Rhodococcus, and Gordona. The library was designed to be used in conjunction with a commercially available pattern recognition software package, Pirouette (Infometrix, Seattle, Wash.). Pirouette uses the K-nearest neighbor algorithm, a similarity-based classification method, to categorize unknown samples on the basis of their multivariate proximities to samples of a preassigned category. Multivariate proximity is calculated from peak height data, while peak heights are named by retention time matching. The system was tested for accuracy by using 24 species of Mycobacterium. Of the 1,333 strains evaluated, > or = 97% were correctly identified. Identification of M. tuberculosis (n = 649) was 99.85% accurate, and identification of the M. avium complex (n = 211) was > or = 98% accurate; > or = 95% of strains of both double-cluster and single-cluster M. gordonae (n = 47) were correctly identified. This system provides a rapid, highly reliable assessment of HPLC-generated chromatographic data for the identification of mycobacteria. PMID:8195387

  7. Rapid identification of mycobacteria and rapid detection of drug resistance in Mycobacterium tuberculosis in cultured isolates and in respiratory specimens.

    PubMed

    Yam, Wing-Cheong; Siu, Kit-Hang Gilman

    2013-01-01

    Recent advances in molecular biology and better understanding of the genetic basis of drug resistance have allowed rapid identification of mycobacteria and rapid detection of drug resistance of Mycobacterium tuberculosis present in cultured isolates or in respiratory specimens. In this chapter, several simple nucleic acid amplification-based techniques are introduced as molecular approach for clinical diagnosis of tuberculosis. A one-tube nested IS6110-based polymerase chain reaction (PCR) is used for M. tuberculosis complex identification; the use of a multiplex allele-specific PCR is demonstrated to detect the isoniazid resistance; PCR-sequencing assays are applied for rifampicin and ofloxacin resistance detection and 16S rDNA sequencing is utilized for identification of mycobacterial species from cultures of acid fast bacilli (AFB). Despite the high specificity and sensitivity of the molecular techniques, mycobacterial culture remains the "Gold Standard" for tuberculosis diagnosis. Negative results of molecular tests never preclude the infection or the presence of drug resistance. These technological advancements are, therefore, not intended to replace the conventional tests, but rather have major complementary roles in tuberculosis diagnosis.

  8. Multistate US Outbreak of Rapidly Growing Mycobacterial Infections Associated with Medical Tourism to the Dominican Republic, 2013-2014(1).

    PubMed

    Schnabel, David; Esposito, Douglas H; Gaines, Joanna; Ridpath, Alison; Barry, M Anita; Feldman, Katherine A; Mullins, Jocelyn; Burns, Rachel; Ahmad, Nina; Nyangoma, Edith N; Nguyen, Duc B; Perz, Joseph F; Moulton-Meissner, Heather A; Jensen, Bette J; Lin, Ying; Posivak-Khouly, Leah; Jani, Nisha; Morgan, Oliver W; Brunette, Gary W; Pritchard, P Scott; Greenbaum, Adena H; Rhee, Susan M; Blythe, David; Sotir, Mark

    2016-08-01

    During 2013, the Maryland Department of Health and Mental Hygiene in Baltimore, MD, USA, received report of 2 Maryland residents whose surgical sites were infected with rapidly growing mycobacteria after cosmetic procedures at a clinic (clinic A) in the Dominican Republic. A multistate investigation was initiated; a probable case was defined as a surgical site infection unresponsive to therapy in a patient who had undergone cosmetic surgery in the Dominican Republic. We identified 21 case-patients in 6 states who had surgery in 1 of 5 Dominican Republic clinics; 13 (62%) had surgery at clinic A. Isolates from 12 (92%) of those patients were culture-positive for Mycobacterium abscessus complex. Of 9 clinic A case-patients with available data, all required therapeutic surgical intervention, 8 (92%) were hospitalized, and 7 (78%) required ≥3 months of antibacterial drug therapy. Healthcare providers should consider infection with rapidly growing mycobacteria in patients who have surgical site infections unresponsive to standard treatment.

  9. Synergistic activity of rifampicin and ethambutol against slow-growing nontuberculous mycobacteria is currently of questionable clinical significance.

    PubMed

    van Ingen, Jakko; Hoefsloot, Wouter; Mouton, Johan W; Boeree, Martin J; van Soolingen, Dick

    2013-07-01

    A key issue in the treatment of disease caused by slow-growing nontuberculous mycobacteria is the limited association between in vitro minimum inhibitory concentrations (MICs) of rifampicin and ethambutol alone and the in vivo outcome of treatment with these drugs. Combined susceptibility testing to rifampicin and ethambutol could provide a more realistic view of the efficacy of these drugs. In this study, Mycobacterium avium (n = 5), Mycobacterium chimaera (n = 6), Mycobacterium intracellulare (n = 4), Mycobacterium xenopi (n = 4), Mycobacterium malmoense (n = 3) and Mycobacterium simiae (n = 2) clinical isolates were selected and the MICs of rifampicin and ethambutol alone and in combination were measured using the Middlebrook 7H10 agar dilution method. Synergy was defined as a fractional inhibitory concentration index ≤ 0.5. Rifampicin and ethambutol showed synergistic activity against the majority of M. avium (4/5), M. chimaera (5/6) and M. intracellulare (3/4) isolates and 1 of 2 eligible M. malmoense isolates. No synergistic activity was measured against M. xenopi and M. simiae. Synergy was neither universal for all species nor for all isolates of one species; it thus needs to be tested for rather than assumed. Even if this synergy exists in vivo, it is questionable whether the MICs to the combined drugs can be overcome by the drug exposure attained by current regimens at the recommended dosages. New dosing strategies for rifampicin and ethambutol should be studied to increase the exposure to these drugs and thus maximise their impact.

  10. Lipoproteins of slow-growing Mycobacteria carry three fatty acids and are N-acylated by apolipoprotein N-acyltransferase BCG_2070c.

    PubMed

    Brülle, Juliane K; Tschumi, Andreas; Sander, Peter

    2013-10-05

    Lipoproteins are virulence factors of Mycobacterium tuberculosis. Bacterial lipoproteins are modified by the consecutive action of preprolipoprotein diacylglyceryl transferase (Lgt), prolipoprotein signal peptidase (LspA) and apolipoprotein N- acyltransferase (Lnt) leading to the formation of mature triacylated lipoproteins. Lnt homologues are found in Gram-negative and high GC-rich Gram-positive, but not in low GC-rich Gram-positive bacteria, although N-acylation is observed. In fast-growing Mycobacterium smegmatis, the molecular structure of the lipid modification of lipoproteins was resolved recently as a diacylglyceryl residue carrying ester-bound palmitic acid and ester-bound tuberculostearic acid and an additional amide-bound palmitic acid. We exploit the vaccine strain Mycobacterium bovis BCG as model organism to investigate lipoprotein modifications in slow-growing mycobacteria. Using Escherichia coli Lnt as a query in BLASTp search, we identified BCG_2070c and BCG_2279c as putative lnt genes in M. bovis BCG. Lipoproteins LprF, LpqH, LpqL and LppX were expressed in M. bovis BCG and BCG_2070c lnt knock-out mutant and lipid modifications were analyzed at molecular level by matrix-assisted laser desorption ionization time-of-flight/time-of-flight analysis. Lipoprotein N-acylation was observed in wildtype but not in BCG_2070c mutants. Lipoprotein N- acylation with palmitoyl and tuberculostearyl residues was observed. Lipoproteins are triacylated in slow-growing mycobacteria. BCG_2070c encodes a functional Lnt in M. bovis BCG. We identified mycobacteria-specific tuberculostearic acid as further substrate for N-acylation in slow-growing mycobacteria.

  11. Lipoproteins of slow-growing Mycobacteria carry three fatty acids and are N-acylated by Apolipoprotein N-Acyltransferase BCG_2070c

    PubMed Central

    2013-01-01

    Background Lipoproteins are virulence factors of Mycobacterium tuberculosis. Bacterial lipoproteins are modified by the consecutive action of preprolipoprotein diacylglyceryl transferase (Lgt), prolipoprotein signal peptidase (LspA) and apolipoprotein N- acyltransferase (Lnt) leading to the formation of mature triacylated lipoproteins. Lnt homologues are found in Gram-negative and high GC-rich Gram-positive, but not in low GC-rich Gram-positive bacteria, although N-acylation is observed. In fast-growing Mycobacterium smegmatis, the molecular structure of the lipid modification of lipoproteins was resolved recently as a diacylglyceryl residue carrying ester-bound palmitic acid and ester-bound tuberculostearic acid and an additional amide-bound palmitic acid. Results We exploit the vaccine strain Mycobacterium bovis BCG as model organism to investigate lipoprotein modifications in slow-growing mycobacteria. Using Escherichia coli Lnt as a query in BLASTp search, we identified BCG_2070c and BCG_2279c as putative lnt genes in M. bovis BCG. Lipoproteins LprF, LpqH, LpqL and LppX were expressed in M. bovis BCG and BCG_2070c lnt knock-out mutant and lipid modifications were analyzed at molecular level by matrix-assisted laser desorption ionization time-of-flight/time-of-flight analysis. Lipoprotein N-acylation was observed in wildtype but not in BCG_2070c mutants. Lipoprotein N- acylation with palmitoyl and tuberculostearyl residues was observed. Conclusions Lipoproteins are triacylated in slow-growing mycobacteria. BCG_2070c encodes a functional Lnt in M. bovis BCG. We identified mycobacteria-specific tuberculostearic acid as further substrate for N-acylation in slow-growing mycobacteria. PMID:24093492

  12. Mycobacteria Isolated from Angkor Monument Sandstones Grow Chemolithoautotrophically by Oxidizing Elemental Sulfur

    PubMed Central

    Kusumi, Asako; Li, Xian Shu; Katayama, Yoko

    2011-01-01

    To characterize sulfate-producing microorganisms from the deteriorated sandstones of Angkor monuments in Cambodia, strains of Mycobacterium spp. were isolated from most probable number-positive cultures. All five strains isolated were able to use both elemental sulfur (S0) for chemolithoautotrophic growth and organic substances for chemoorganoheterotrophic growth. Results of phylogenetic and phenotypic analyses indicated that all five isolates were rapid growers of the genus Mycobacterium and were most similar to Mycobacterium cosmeticum and Mycobacterium pallens. Chemolithoautotrophic growth was further examined in the representative strain THI503. When grown in mineral salts medium, strain THI503 oxidized S0 to thiosulfate and sulfate; oxidation was accompanied by a decrease in the pH of the medium from 4.7 to 3.6. The link between sulfur oxidation and energy metabolism was confirmed by an increase in ATP. Fluorescence microscopy of DAPI-stained cells revealed that strain THI503 adheres to and proliferates on the surface of sulfur particles. The flexible metabolic ability of facultative chemolithoautotrophs enables their survival in nutrient-limited sandstone environments. PMID:21747806

  13. Mycobacterium gilvum illustrates size-correlated relationships between mycobacteria and Acanthamoeba polyphaga.

    PubMed

    Lamrabet, Otmane; Drancourt, Michel

    2013-03-01

    Mycobacteria are isolated from soil and water environments, where free-living amoebae live. Free-living amoebae are bactericidal, yet some rapidly growing mycobacteria are amoeba-resistant organisms that survive in the amoebal trophozoites and cysts. Such a capacity has not been studied for the environmental rapidly growing organism Mycobacterium gilvum. We investigated the ability of M. gilvum to survive in the trophozoites of Acanthamoeba polyphaga strain Linc-AP1 by using optical and electron microscopy and culture-based microbial enumerations in the presence of negative controls. We observed that 29% of A. polyphaga cells were infected by M. gilvum mycobacteria by 6 h postinfection. Surviving M. gilvum mycobacteria did not multiply and did not kill the amoebal trophozoites during a 5-day coculture. Extensive electron microscopy observations indicated that M. gilvum measured 1.4 ± 0.5 μm and failed to find M. gilvum organisms in the amoebal cysts. Further experimental study of two other rapidly growing mycobacteria, Mycobacterium rhodesiae and Mycobacterium thermoresistibile, indicated that both measured <2 μm and exhibited the same amoeba-mycobacterium relationships as M. gilvum. In general, we observed that mycobacteria measuring <2 μm do not significantly grow within and do not kill amoebal trophozoites, in contrast to mycobacteria measuring >2 μm (P < 0.05). The mechanisms underlying such an observation remain to be determined.

  14. Mycobacterium gilvum Illustrates Size-Correlated Relationships between Mycobacteria and Acanthamoeba polyphaga

    PubMed Central

    Lamrabet, Otmane

    2013-01-01

    Mycobacteria are isolated from soil and water environments, where free-living amoebae live. Free-living amoebae are bactericidal, yet some rapidly growing mycobacteria are amoeba-resistant organisms that survive in the amoebal trophozoites and cysts. Such a capacity has not been studied for the environmental rapidly growing organism Mycobacterium gilvum. We investigated the ability of M. gilvum to survive in the trophozoites of Acanthamoeba polyphaga strain Linc-AP1 by using optical and electron microscopy and culture-based microbial enumerations in the presence of negative controls. We observed that 29% of A. polyphaga cells were infected by M. gilvum mycobacteria by 6 h postinfection. Surviving M. gilvum mycobacteria did not multiply and did not kill the amoebal trophozoites during a 5-day coculture. Extensive electron microscopy observations indicated that M. gilvum measured 1.4 ± 0.5 μm and failed to find M. gilvum organisms in the amoebal cysts. Further experimental study of two other rapidly growing mycobacteria, Mycobacterium rhodesiae and Mycobacterium thermoresistibile, indicated that both measured <2 μm and exhibited the same amoeba-mycobacterium relationships as M. gilvum. In general, we observed that mycobacteria measuring <2 μm do not significantly grow within and do not kill amoebal trophozoites, in contrast to mycobacteria measuring >2 μm (P < 0.05). The mechanisms underlying such an observation remain to be determined. PMID:23275502

  15. An Experimental Model for the Rapid Screening of Compounds with Potential Use Against Mycobacteria.

    PubMed

    Costa, Sofia Santos; Lopes, Elizeth; Azzali, Elisa; Machado, Diana; Coelho, Tatiane; da Silva, Pedro Eduardo Almeida; Viveiros, Miguel; Pieroni, Marco; Couto, Isabel

    2016-11-01

    Infections caused by Mycobacterium tuberculosis and other mycobacteria are major challenges for global public health. Particularly worrisome are infections caused by multidrug-resistant bacteria, which are increasingly difficult to treat because of the loss of efficacy of the current antibacterial agents, a problem that continues to escalate worldwide. There has been a limited interest and investment on the development of new antibacterial agents in the past decades. This has led to the current situation, in which there is an urgent demand for innovative therapeutic alternatives to fight infections caused by multidrug-resistant pathogens, such as multidrug-resistant tuberculosis. The identification of compounds that can act as adjuvants in antimycobacterial therapeutic regimens is an appealing strategy to restore the efficacy lost by some of the antibiotics currently used and shorten the duration of the therapeutic regimen. In this work, by setting Mycobacterium smegmatis as a model organism, we have developed a methodological strategy to identify, in a fast and simple approach, compounds with antimycobacterial activity or with potential adjuvant properties, by either inhibition of efflux or other unrelated mechanisms. Such an approach may increase the rate of identification of promising molecules, to be further explored in pathogenic models for their potential use either as antimicrobials or as adjuvants, in combination with available therapeutic regimens for the treatment of mycobacterial infections. This method allowed us to identify a new molecule that shows promising activity as an efflux inhibitor in M. smegmatis.

  16. Nontuberculous Mycobacteria in Respiratory Tract Infections, Eastern Asia

    PubMed Central

    van Ingen, Jakko; Hsueh, Po-Ren; Van Hung, Nguyen; Dekhuijzen, P.N. Richard; Boeree, Martin J.; van Soolingen, Dick

    2011-01-01

    To characterize the distribution of nontuberculous mycobacteria (NTM) species isolated from pulmonary samples from persons in Asia and their association with pulmonary infections, we reviewed the literature. Mycobacterium avium complex bacteria were most frequently isolated (13%–81%) and were the most common cause of pulmonary NTM disease (43%–81%). Also pathogenic were rapidly growing mycobacteria (M. chelonae, M. fortuitum, M. abscessus). Among all NTM isolated from pulmonary samples, 31% (582/1,744) were considered clinically relevant according to American Thoracic Society diagnostic criteria. Most patients were male (79%) and had a history of tuberculosis (37%). In Asia, high prevalence of rapidly growing mycobacteria and a history of tuberculosis are distinct characteristics of pulmonary NTM disease. This geographic variation is not well reflected in the American Thoracic Society criteria for NTM infections and could be incorporated in future guidelines. PMID:21392422

  17. Nontuberculous Mycobacteria in Saudi Arabia and Gulf Countries: A Review

    PubMed Central

    2017-01-01

    Nontuberculous Mycobacteria (NTM) are causing growing health problems worldwide. This is indicated by an increasing amount of scientific reports showing not only well-identified species reemerging but also emergence of new species. The emergence and reemergence of NTM are particularly worrying in developing countries due to scarce published data and improper identification. Here we aimed to examine the main epidemiological aspects and diagnostic challenges associated with NTM in countries of the Gulf Cooperation Council (GCC) and compare these findings to the international arena findings. Data revealed that countries of the GCC are largely dominated by rapidly growing mycobacteria species such as M. fortuitum (29%) and M. abscessus (17%) with high rate of definitive respiratory diseases. On the other hand, most of the developed countries are dominated by slowly growing mycobacteria such as MAC, M. kansasii, and M. gordonae. More efforts are needed, however, to gain insights into NTM issues in countries of the GCC. PMID:28348502

  18. Multistate US Outbreak of Rapidly Growing Mycobacterial Infections Associated with Medical Tourism to the Dominican Republic, 2013–20141

    PubMed Central

    Esposito, Douglas H.; Gaines, Joanna; Ridpath, Alison; Barry, M. Anita; Feldman, Katherine A.; Mullins, Jocelyn; Burns, Rachel; Ahmad, Nina; Nyangoma, Edith N.; Nguyen, Duc B.; Perz, Joseph F.; Moulton-Meissner, Heather A.; Jensen, Bette J.; Lin, Ying; Posivak-Khouly, Leah; Jani, Nisha; Morgan, Oliver W.; Brunette, Gary W.; Pritchard, P. Scott; Greenbaum, Adena H.; Rhee, Susan M.; Blythe, David; Sotir, Mark

    2016-01-01

    During 2013, the Maryland Department of Health and Mental Hygiene in Baltimore, MD, USA, received report of 2 Maryland residents whose surgical sites were infected with rapidly growing mycobacteria after cosmetic procedures at a clinic (clinic A) in the Dominican Republic. A multistate investigation was initiated; a probable case was defined as a surgical site infection unresponsive to therapy in a patient who had undergone cosmetic surgery in the Dominican Republic. We identified 21 case-patients in 6 states who had surgery in 1 of 5 Dominican Republic clinics; 13 (62%) had surgery at clinic A. Isolates from 12 (92%) of those patients were culture-positive for Mycobacterium abscessus complex. Of 9 clinic A case-patients with available data, all required therapeutic surgical intervention, 8 (92%) were hospitalized, and 7 (78%) required ≥3 months of antibacterial drug therapy. Healthcare providers should consider infection with rapidly growing mycobacteria in patients who have surgical site infections unresponsive to standard treatment. PMID:27434822

  19. Occurrence of Mycobacteria in Water Treatment Lines and in Water Distribution Systems

    PubMed Central

    Le Dantec, Corinne; Duguet, Jean-Pierre; Montiel, Antoine; Dumoutier, Nadine; Dubrou, Sylvie; Vincent, Véronique

    2002-01-01

    The frequency of recovery of atypical mycobacteria was estimated in two treatment plants providing drinking water to Paris, France, at some intermediate stages of treatment. The two plants use two different filtration processes, rapid and slow sand filtration. Our results suggest that slow sand filtration is more efficient for removing mycobacteria than rapid sand filtration. In addition, our results show that mycobacteria can colonize and grow on granular activated carbon and are able to enter distribution systems. We also investigated the frequency of recovery of mycobacteria in the water distribution system of Paris (outside buildings). The mycobacterial species isolated from the Paris drinking water distribution system are different from those isolated from the water leaving the treatment plants. Saprophytic mycobacteria (present in 41.3% of positive samples), potentially pathogenic mycobacteria (16.3%), and unidentifiable mycobacteria (54.8%) were isolated from 12 sites within the Paris water distribution system. Mycobacterium gordonae was preferentially recovered from treated surface water, whereas Mycobacterium nonchromogenicum was preferentially recovered from groundwater. No significant correlations were found among the presence of mycobacteria, the origin of water, and water temperature. PMID:12406720

  20. Occurrence of mycobacteria in water treatment lines and in water distribution systems.

    PubMed

    Le Dantec, Corinne; Duguet, Jean-Pierre; Montiel, Antoine; Dumoutier, Nadine; Dubrou, Sylvie; Vincent, Véronique

    2002-11-01

    The frequency of recovery of atypical mycobacteria was estimated in two treatment plants providing drinking water to Paris, France, at some intermediate stages of treatment. The two plants use two different filtration processes, rapid and slow sand filtration. Our results suggest that slow sand filtration is more efficient for removing mycobacteria than rapid sand filtration. In addition, our results show that mycobacteria can colonize and grow on granular activated carbon and are able to enter distribution systems. We also investigated the frequency of recovery of mycobacteria in the water distribution system of Paris (outside buildings). The mycobacterial species isolated from the Paris drinking water distribution system are different from those isolated from the water leaving the treatment plants. Saprophytic mycobacteria (present in 41.3% of positive samples), potentially pathogenic mycobacteria (16.3%), and unidentifiable mycobacteria (54.8%) were isolated from 12 sites within the Paris water distribution system. Mycobacterium gordonae was preferentially recovered from treated surface water, whereas Mycobacterium nonchromogenicum was preferentially recovered from groundwater. No significant correlations were found among the presence of mycobacteria, the origin of water, and water temperature.

  1. [Effect of heat-staining procedure on the gram staining properties of mycobacteria].

    PubMed

    Nakamura, M; Harano, Y; Koga, T

    1991-03-01

    Since the establishment of Gram stain by H.C.Y. Gram in 1884, it has been widely and routinely used as an aid for differentiation of bacteria. The bacteria are divided into three categories by the staining properties; Gram-positive, -negative, and -indefinite. All the text books in the world describe that mycobacteria such as M. tuberculosis are Gram-positive. By the merest chance, however, it was found that M. lepraemurium grown in tissues was not stained by the routinely used Gram staining method. Therefore, we tried to stain some of the mycobacteria by the Gram staining procedure which is widely used at present. The results obtained indicated that the mycobacteria tested were divided into three groups; the unstainable group such as M. leprae and M. lepraemurium, the Gram-positive and difficult-to-stain group which involves such slow growing mycobacteria as M. tuberculosis, M. avium, and M. intracellulare, and the Gram-indefinite group which contains such rapid growing mycobacteria as M. phlei, M. smegmatis, and M. chelonae. However, if Gram stain is carried out by the heating procedure at the first staining step, all the mycobacteria would become Gram-positive. Therefore, we emphasize that Gram staining of mycobacteria should be performed by the heating procedure.

  2. Application of a dual target PCR-high resolution melting (HRM) method for rapid nontuberculous mycobacteria identification.

    PubMed

    Chen, Jonathan Hk; Cheng, Vincent Cc; She, Kevin Kk; Yam, Wing-Cheong; Yuen, Kwok-Yung

    2017-01-01

    Species differentiation of nontuberculous mycobacteria (NTM) has long been a difficult task in clinical laboratories. This study demonstrated and evaluated a simple and cost-effective method using the real-time PCR with high-resolution melting (PCR-HRM) analysis technique, which could differentiate at least 14 different medically related NTM.

  3. DNA-based methodologies for rapid detection, quantification, and species- or strain-level identification of respiratory pathogens (Mycobacteria and Pseudomonads) in metalworking fluids.

    PubMed

    Yadav, Jagjit S; Khan, Izhar U H; Fakhari, Farnaz; Soellner, Mathew B

    2003-11-01

    Mycobacteria and pseudomonads occurring in modern metalworking fluids (MWF) have been implicated in occupational health hazards as causal agents for hypersensitivity pneumonitis (HP) and other respiratory illnesses in machine workers exposed to these fluids and their aerosols. Unlike the conventional cultural and biochemical methods, which are often slow and ambiguous and detect only culturable cells, DNA-based methods offer a time-saving alternative for reliable detection and identification of both culturable and nonculturable bacteria in MWF and for selective quantification of individual genera of pathogens of interest in these fluids. This is the first report on DNA-based direct detection of mycobacteria and pseudomonads in MWF without culturing. Genus-specific PCR approach was successfully applied for screening of field MWF samples originating from different industrial users for detection of mycobacteria or pseudomonads including both culturable and nonculturable cells. PCR in combination with amplicon DNA sequencing led to the identification of Mycobacterium chelonae, Pseudomonas nitroreducens, and an undefined Pseudomonas species from these fluids. Genome fingerprinting by pulsed-field gel electrophoresis (PFGE) on Mycobacterium isolates further showed that the isolates represented three strains of M. chelonae although the possibility of one of the strains being clonal with M. immunogenum cannot be excluded. In parallel efforts, a quantitative competitive PCR method developed based on the Pseudomonas-specific PCR was applied to quantify total P. fluorescens cells in contaminated metalworking fluid and MWF aerosol without culturing. The DNA-based protocols developed in this study will allow rapid screening of field MWF samples for the presence of both culturable and nonculturable cells and thus facilitate effective fluid management and timely exposure assessment.

  4. Epidemiology of infection by nontuberculous mycobacteria. VII. Absence of mycobacteria in southeastern groundwaters.

    PubMed

    Martin, E C; Parker, B C; Falkinham, J O

    1987-08-01

    Using the fluorochrome auramine-O, direct microscopic counts of log phase cells of Mycobacterium avium, M. intracellulare, and M. scrofulaceum (MAIS) group showed excellent correlations with viable spread-plate counts. Accordingly, an enumeration of total acid-fast and MAIS cells by stain and culture (respectively) in groundwaters from three United States regions that differ in their incidence of human infection by MAIS was undertaken. Of 30 state-monitored, undisinfected wells, 11 were in Georgia coastal plain (high incidence), 10 were in the Virginia coastal plain (intermediate incidence), and 9 were in Montgomery County, Virginia (low incidence). Total acid-fast cells ranged widely between 280 to 5,367 per ml among the groundwaters, and with one exception showed no correlations or trends between regions of different incidence of human infection, or to total bacterial cell counts or colony-forming units. The exception was that the proportions of acid-fast cells relative to total cells were higher in the Georgia groundwaters. However, despite the relatively high auramine-O counts, few mycobacteria were recovered by culture. Of 12 wells yielding mycobacteria, 9 yielded rapidly growing and 4 slowly growing mycobacteria. Only one well in Montgomery County, Virginia (region of low incidence) yielded a MAIS isolate, albeit at low density. This research supports the conclusion that clean groundwaters are unlikely sources of MAIS infection in humans in the southeastern United States.

  5. Rapidly Growing Thyroid Mass in an Immunocompromised Young Male Adult

    PubMed Central

    Santiago, Mónica; Martinez, José Hernán; Palermo, Coromoto; Figueroa, Carlos; Torres, Oberto; Trinidad, Rafael; Gonzalez, Eva; Miranda, Maria de Lourdes; Garcia, Miosotis; Villamarzo, Guillermo

    2013-01-01

    We describe a 20-year-old man diagnosed with a myelodysplastic syndrome (MDS), admitted to our hospital due to pancytopenia and fever of undetermined origin after myelosuppression with chemotherapy. Disseminated aspergillosis (DIA) was suspected when he developed skin and lung involvement. A rapidly growing mass was detected on the left neck area, during hospitalization. A thyroid ultrasound reported a 3.7 × 2.5 × 2.9 cm oval heterogeneous structure, suggestive of an abscess versus a hematoma. Fine needle aspiration of the thyroid revealed invasion of aspergillosis. Fungal thyroiditis is a rare occurrence. Thyroid fungal infection is difficult to diagnose; for this reason it is rarely diagnosed antemortem. To our knowledge, this is the 10th case reported in the literature in an adult where the diagnosis of fungal invasion to the thyroid was able to be corroborated antemortem by fine needle aspiration biopsy. PMID:23936688

  6. 16S-23S Internal Transcribed Spacer Region PCR and Sequencer-Based Capillary Gel Electrophoresis has Potential as an Alternative to High Performance Liquid Chromatography for Identification of Slowly Growing Nontuberculous Mycobacteria

    PubMed Central

    Subedi, Shradha; Kong, Fanrong; Jelfs, Peter; Gray, Timothy J.; Xiao, Meng; Sintchenko, Vitali; Chen, Sharon C-A

    2016-01-01

    Accurate identification of slowly growing nontuberculous mycobacteria (SG-NTM) of clinical significance remains problematic. This study evaluated a novel method of SG-NTM identification by amplification of the mycobacterial 16S-23S rRNA internal transcribed spacer (ITS) region followed by resolution of amplified fragments by sequencer-based capillary gel electrophoresis (SCGE). Fourteen American Type Culture Collection (ATCC) strains and 103 clinical/environmental isolates (total n = 24 species) of SG-NTM were included. Identification was compared with that achieved by high performance liquid chromatography (HPLC), in-house PCR and 16S/ITS sequencing. Isolates of all species yielded a SCGE profile comprising a single fragment length (or peak) except for M. scrofulaceum (two peaks). SCGE peaks of ATCC strains were distinct except for peak overlap between Mycobacterium kansasii and M. marinum. Of clinical/environmental strains, unique peaks were seen for 7/17 (41%) species (M. haemophilum, M. kubicae, M. lentiflavum, M. terrae, M. kansasii, M. asiaticum and M. triplex); 3/17 (18%) species were identified by HPLC. There were five SCGE fragment length types (I–V) each of M. avium, M. intracellulare and M. gordonae. Overlap of fragment lengths was seen between M. marinum and M. ulcerans; for M. gordonae SCGE type III and M. paragordonae; M. avium SCGE types III and IV, and M. intracellulare SCGE type I; M. chimaera, M. parascrofulaceum and M. intracellulare SCGE types III and IV; M. branderi and M. avium type V; and M. vulneris and M. intracellulare type V. The ITS-SCGE method was able to provide the first line rapid and reproducible species identification/screening of SG-NTM and was more discriminatory than HPLC. PMID:27749897

  7. 16S-23S Internal Transcribed Spacer Region PCR and Sequencer-Based Capillary Gel Electrophoresis has Potential as an Alternative to High Performance Liquid Chromatography for Identification of Slowly Growing Nontuberculous Mycobacteria.

    PubMed

    Subedi, Shradha; Kong, Fanrong; Jelfs, Peter; Gray, Timothy J; Xiao, Meng; Sintchenko, Vitali; Chen, Sharon C-A

    2016-01-01

    Accurate identification of slowly growing nontuberculous mycobacteria (SG-NTM) of clinical significance remains problematic. This study evaluated a novel method of SG-NTM identification by amplification of the mycobacterial 16S-23S rRNA internal transcribed spacer (ITS) region followed by resolution of amplified fragments by sequencer-based capillary gel electrophoresis (SCGE). Fourteen American Type Culture Collection (ATCC) strains and 103 clinical/environmental isolates (total n = 24 species) of SG-NTM were included. Identification was compared with that achieved by high performance liquid chromatography (HPLC), in-house PCR and 16S/ITS sequencing. Isolates of all species yielded a SCGE profile comprising a single fragment length (or peak) except for M. scrofulaceum (two peaks). SCGE peaks of ATCC strains were distinct except for peak overlap between Mycobacterium kansasii and M. marinum. Of clinical/environmental strains, unique peaks were seen for 7/17 (41%) species (M. haemophilum, M. kubicae, M. lentiflavum, M. terrae, M. kansasii, M. asiaticum and M. triplex); 3/17 (18%) species were identified by HPLC. There were five SCGE fragment length types (I-V) each of M. avium, M. intracellulare and M. gordonae. Overlap of fragment lengths was seen between M. marinum and M. ulcerans; for M. gordonae SCGE type III and M. paragordonae; M. avium SCGE types III and IV, and M. intracellulare SCGE type I; M. chimaera, M. parascrofulaceum and M. intracellulare SCGE types III and IV; M. branderi and M. avium type V; and M. vulneris and M. intracellulare type V. The ITS-SCGE method was able to provide the first line rapid and reproducible species identification/screening of SG-NTM and was more discriminatory than HPLC.

  8. Rapid and accurate identification of Mycobacterium tuberculosis complex and common non-tuberculous mycobacteria by multiplex real-time PCR targeting different housekeeping genes.

    PubMed

    Nasr Esfahani, Bahram; Rezaei Yazdi, Hadi; Moghim, Sharareh; Ghasemian Safaei, Hajieh; Zarkesh Esfahani, Hamid

    2012-11-01

    Rapid and accurate identification of mycobacteria isolates from primary culture is important due to timely and appropriate antibiotic therapy. Conventional methods for identification of Mycobacterium species based on biochemical tests needs several weeks and may remain inconclusive. In this study, a novel multiplex real-time PCR was developed for rapid identification of Mycobacterium genus, Mycobacterium tuberculosis complex (MTC) and the most common non-tuberculosis mycobacteria species including M. abscessus, M. fortuitum, M. avium complex, M. kansasii, and the M. gordonae in three reaction tubes but under same PCR condition. Genetic targets for primer designing included the 16S rDNA gene, the dnaJ gene, the gyrB gene and internal transcribed spacer (ITS). Multiplex real-time PCR was setup with reference Mycobacterium strains and was subsequently tested with 66 clinical isolates. Results of multiplex real-time PCR were analyzed with melting curves and melting temperature (T (m)) of Mycobacterium genus, MTC, and each of non-tuberculosis Mycobacterium species were determined. Multiplex real-time PCR results were compared with amplification and sequencing of 16S-23S rDNA ITS for identification of Mycobacterium species. Sensitivity and specificity of designed primers were each 100 % for MTC, M. abscessus, M. fortuitum, M. avium complex, M. kansasii, and M. gordonae. Sensitivity and specificity of designed primer for genus Mycobacterium was 96 and 100 %, respectively. According to the obtained results, we conclude that this multiplex real-time PCR with melting curve analysis and these novel primers can be used for rapid and accurate identification of genus Mycobacterium, MTC, and the most common non-tuberculosis Mycobacterium species.

  9. On the killing of mycobacteria by macrophages.

    PubMed

    Jordao, Luisa; Bleck, Christopher K E; Mayorga, Luis; Griffiths, Gareth; Anes, Elsa

    2008-02-01

    Both pathogenic and non-pathogenic mycobacteria are internalized into macrophage phagosomes. Whereas the non-pathogenic types are invariably killed by all macrophages, the pathogens generally survive and grow. Here, we addressed the survival, production of nitrogen intermediates (RNI) and intracellular trafficking of the non-pathogenic Mycobacterium smegmatis, the pathogen-like, BCG and the pathogenic M. bovis in different mouse, human and bovine macrophages. The bacteriocidal effects of RNI were restricted for all bacterial species to the early stages of infection. EM analysis showed clearly that all the mycobacteria remained within phagosomes even at late times of infection. The fraction of BCG and M. bovis found in mature phagolysosomes rarely exceeded 10% of total, irrespective of whether bacteria were growing, latent or being killed, with little correlation between the extent of phagosome maturation and the degree of killing. Theoretical modelling of our data identified two different potential sets of explanations that are consistent with our results. The model we favour is one in which a small but significant fraction of BCG is killed in an early phagosome, then maturation of a small fraction of phagosomes with both live and killed bacteria, followed by extremely rapid killing and digestion of the bacteria in phago-lysosomes.

  10. Deoxyfluoro-D-trehalose (FDTre) analogues as potential PET probes for imaging mycobacterial infection: rapid synthesis and purification, conformational analysis, and uptake by mycobacteria

    PubMed Central

    Rundell, Sarah R.; Wagar, Zachary L.; Meints, Lisa M.; Olson, Claire D.; O’Neill, Mara K.; Piligian, Brent F.; Poston, Anne W.; Hood, Robin J.; Woodruff, Peter J.

    2016-01-01

    Mycobacterium tuberculosis, the etiological agent of human tuberculosis, requires the non-mammalian disaccharide trehalose for growth and virulence. Recently, detectable trehalose analogues have gained attention as probes for studying trehalose metabolism and as potential diagnostic imaging agents for mycobacterial infections. Of particular interest are deoxy-[18F]fluoro-D-trehalose (18F-FDTre) analogues, which have been suggested as possible positron emission tomography (PET) probes for in vivo imaging of M. tuberculosis infection. Here, we report progress toward this objective, including the synthesis and conformational analysis of four non-radioactive deoxy-[19F]fluoro-D-trehalose (19F-FDTre) analogues, as well as evaluation of their uptake by M. smegmatis. The rapid synthesis and purification of several 19F-FDTre analogues was accomplished in high yield using a one-step chemoenzymatic method. Conformational analysis of the 19F-FDTre analogues using NMR and molecular modeling methods showed that fluorine substitution had a negligible effect on the conformation of the native disaccharide, suggesting that fluorinated analogues may be successfully recognized and processed by trehalose metabolic machinery in mycobacteria. To test this hypothesis and to evaluate a possible route for delivery of FDTre probes specifically to mycobacteria, we showed that 19F-FDTre analogues are actively imported into M. smegmatis via the trehalose-specific transporter SugABC-LpqY. Finally, to demonstrate the applicability of these results to the efficient preparation and use of short-lived 18F-FDTre PET radiotracers, we carried out 19F-FDTre synthesis, purification, and administration to M. smegmatis in 1 hour. PMID:27560008

  11. Rapidly growing Mycobacterium infections after cosmetic surgery in medical tourists: the Bronx experience and a review of the literature.

    PubMed

    Cusumano, Lucas R; Tran, Vivy; Tlamsa, Aileen; Chung, Philip; Grossberg, Robert; Weston, Gregory; Sarwar, Uzma N

    2017-10-01

    Medical tourism is increasingly popular for elective cosmetic surgical procedures. However, medical tourism has been accompanied by reports of post-surgical infections due to rapidly growing mycobacteria (RGM). The authors' experience working with patients with RGM infections who have returned to the USA after traveling abroad for cosmetic surgical procedures is described here. Patients who developed RGM infections after undergoing cosmetic surgeries abroad and who presented at the Montefiore Medical Center (Bronx, New York, USA) between August 2015 and June 2016 were identified. A review of patient medical records was performed. Four patients who presented with culture-proven RGM infections at the sites of recent cosmetic procedures were identified. All patients were treated with a combination of antibiotics and aggressive surgical treatment. This case series of RGM infections following recent cosmetic surgeries abroad highlights the risks of medical tourism. Close monitoring of affected patients by surgical and infectious disease specialties is necessary, as aggressive surgical debridement combined with appropriate antibiotic regimens is needed to achieve cure. Given the increasing reports of post-surgical RGM infections, consultants should have a low threshold for suspecting RGM, as rapid diagnosis may accelerate the initiation of targeted treatment and minimize morbidity. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  12. Species of environmental mycobacteria differ in their abilities to grow in human, mouse, and carp macrophages and with regard to the presence of mycobacterial virulence genes, as observed by DNA microarray hybridization.

    PubMed

    Harriff, Melanie J; Wu, Martin; Kent, Michael L; Bermudez, Luiz E

    2008-01-01

    There are many species of environmental mycobacteria (EM) that infect animals that are important to the economy and research and that also have zoonotic potential. The genomes of very few of these bacterial species have been sequenced, and little is known about the molecular mechanisms by which most of these opportunistic pathogens cause disease. In this study, 18 isolates of EM isolated from fish and humans (including strains of Mycobacterium avium, Mycobacterium peregrinum, Mycobacterium chelonae, and Mycobacterium salmoniphilum) were examined for their abilities to grow in macrophage lines from humans, mice, and carp. Genomic DNA from 14 of these isolates was then hybridized against DNA from an M. avium reference strain, with a custom microarray containing virulence genes of mycobacteria and a selection of representative genes from metabolic pathways. The strains of EM had different abilities to grow within the three types of cell lines, which grouped largely according to the host from which they were isolated. Genes identified as being putatively absent in some of the strains included those with response regulatory functions, cell wall compositions, and fatty acid metabolisms as well as a recently identified pathogenicity island important to macrophage uptake. Further understanding of the role these genes play in host specificity and pathogenicity will be important to gain insight into the zoonotic potential of certain EM as well as their mechanisms of virulence.

  13. Routine Use of PCR–Reverse Cross-Blot Hybridization Assay for Rapid Identification of Mycobacterium Species Growing in Liquid Media

    PubMed Central

    Sanguinetti, M.; Posteraro, B.; Ardito, F.; Zanetti, S.; Cingolani, A.; Sechi, L.; De Luca, A.; Ortona, L.; Fadda, G.

    1998-01-01

    A PCR–reverse cross-blot hybridization assay procedure that is able to rapidly identify 13 species of clinically relevant mycobacteria was evaluated for routine use in the identification of acid-fast isolates growing in BACTEC 460 TB (12B and 13A) and BACTEC 9000 MB (Myco/F) liquid media. Eight of the probes used were already described by Kox et al. (L. F. F. Kox et al., J. Clin. Microbiol. 33:3225–3233, 1995). In addition, we used six other probes specific for M. chelonae, M. malmoense or M. szulgai, M. genavense, M. gordonae, M. terrae, and M. marinum/M. ulcerans that we designed ourselves. This procedure allowed us to identify 459 mycobacterial species directly from broth cultures of 5,466 clinical samples collected over 1 year and processed with the radiometric or nonradiometric BACTEC system. Our results were in agreement with those obtained by conventional identification methods and also with those obtained by mycolic acid analysis by high-performance liquid chromatography. This assay seems to be a reliable procedure for the routine identification of mycobacteria, providing an accurate identification of mycobacterial isolates more rapidly than conventional tests, with remarkable implications for an efficacious specific antimycobacterial therapy. PMID:9620371

  14. Advantages of Using Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry as a Rapid Diagnostic Tool for Identification of Yeasts and Mycobacteria in the Clinical Microbiological Laboratory

    PubMed Central

    Chen, Jonathan H. K.; Yam, Wing-Cheong; Ngan, Antonio H. Y.; Fung, Ami M. Y.; Woo, Wai-Lan; Yan, Mei-Kum; Choi, Garnet K. Y.; Ho, Pak-Leung; Cheng, Vincent C. C.

    2013-01-01

    Yeast and mycobacteria can cause infections in immunocompromised patients and normal hosts. The rapid identification of these organisms can significantly improve patient care. There has been an increasing number of studies on using matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) for rapid yeast and mycobacterial identifications. However, studies on direct comparisons between the Bruker Biotyper and bioMérieux Vitek MS systems for the identification of yeast and mycobacteria have been limited. This study compared the performance of the two systems in their identification of 98 yeast and 102 mycobacteria isolates. Among the 98 yeast isolates, both systems generated species-level identifications in >70% of the specimens, of which Candida albicans was the most commonly cultured species. At a genus-level identification, the Biotyper system identified more isolates than the Vitek MS system for Candida (75/78 [96.2%]versus 68/78 [87.2%], respectively; P = 0.0426) and non-Candida yeasts (18/20 [90.0%]versus 7/20 [35.0%], respectively; P = 0.0008). For mycobacterial identification, the Biotyper system generated reliable identifications for 89 (87.3%) and 64 (62.8%) clinical isolates at the genus and species levels, respectively, from solid culture media, whereas the Vitek MS system did not generate any reliable identification. The MS method differentiated 12/21 clinical species, despite the fact that no differentiation between Mycobacterium abscessus and Mycobacterium chelonae was found by using 16S rRNA gene sequencing. In summary, the MALDI-TOF MS method provides short turnaround times and a standardized working protocol for the identification of yeast and mycobacteria. Our study demonstrates that MALDI-TOF MS is suitable as a first-line test for the identification of yeast and mycobacteria in clinical laboratories. PMID:24048537

  15. “Mycobacterium massilipolynesiensis” sp. nov., a rapidly-growing mycobacterium of medical interest related to Mycobacterium phlei

    PubMed Central

    Phelippeau, M.; Asmar, S.; Osman, D. Aboubaker; Sassi, M.; Robert, C.; Michelle, C.; Musso, D.; Drancourt, M.

    2017-01-01

    In French Polynesia, respiratory tract clinical isolate M26, displayed unusual phenotype and contradictory phylogenetic affiliations, suggesting a hitherto unidentified rapidly-growing Mycobacterium species. The phenotype of strain M26 was further characterized and its genome sequenced. Strain M26 genome consists in a 5,732,017-bp circular chromosome with a G + C% of 67.54%, comprising 5,500 protein-coding genes and 52 RNA genes (including two copies of the 16 S rRNA gene). One region coding for a putative prophage was also predicted. An intriguing characteristic of strain M26’s genome is the large number of genes encoding polyketide synthases and nonribosomal peptide synthases. Phylogenomic analysis showed that strain M26’s genome is closest to the Mycobacterium phlei genome with a 76.6% average nucleotide identity. Comparative genomics of 33 Mycobacterium genomes yielded 361 genes unique to M26 strain which functional annotation revealed 84.21% of unknown function and 3.88% encoding lipid transport and metabolism; while 48.87% of genes absent in M26 strain have unknown function, 9.5% are implicated in transcription and 19% are implicated in transport and metabolism. Strain M26’s unique phenotypic and genomic characteristics indicate it is representative of a new species named “Mycobacterium massilipolynesiensis”. Looking for mycobacteria in remote areas allows for the discovery of new Mycobacterium species. PMID:28074866

  16. Primary amyloid goiter mimicking rapid growing thyroid malignancy.

    PubMed

    Joung, Kyong Hye; Park, Jae-Yong; Kim, Koon Soon; Koo, Bon Seok

    2014-02-01

    Amyloid accumulation in the thyroid gland leading to a clinically detectable mass, known as amyloid goiter, is a rare condition associated with primary amyloidosis. Moreover, a localized primary amyloid goiter involving only the thyroid gland is rarer still. Here, we report a patient with a localized primary amyloid goiter that had grown rapidly, causing dysphagia and dyspnea on exercise, and confused us with malignancy such as anaplastic carcinoma. After surgery, no further symptoms occurred. A diagnosis of amyloid goiter was established on microscopic examination. In patients with a rapidly enlarging thyroid gland presenting with dysphagia, dyspnea, or hoarseness, amyloid goiter and malignancy should both be suspected, even when systemic amyloidosis is not suspected.

  17. Glasses crystallize rapidly at free surfaces by growing crystals upward.

    PubMed

    Sun, Ye; Zhu, Lei; Kearns, Kenneth L; Ediger, Mark D; Yu, Lian

    2011-04-12

    The crystallization of glasses and amorphous solids is studied in many fields to understand the stability of amorphous materials, the fabrication of glass ceramics, and the mechanism of biomineralization. Recent studies have found that crystal growth in organic glasses can be orders of magnitude faster at the free surface than in the interior, a phenomenon potentially important for understanding glass crystallization in general. Current explanations differ for surface-enhanced crystal growth, including released tension and enhanced mobility at glass surfaces. We report here a feature of the phenomenon relevant for elucidating its mechanism: Despite their higher densities, surface crystals rise substantially above the glass surface as they grow laterally, without penetrating deep into the bulk. For indomethacin (IMC), an organic glass able to grow surface crystals in two polymorphs (α and γ), the growth front can be hundreds of nanometers above the glass surface. The process of surface crystal growth, meanwhile, is unperturbed by eliminating bulk material deeper than some threshold depth (ca. 300 nm for α IMC and less than 180 nm for γ IMC). As a growth strategy, the upward-lateral growth of surface crystals increases the system's surface energy, but can effectively take advantage of surface mobility and circumvent slow growth in the bulk.

  18. Glasses crystallize rapidly at free surfaces by growing crystals upward

    PubMed Central

    Sun, Ye; Zhu, Lei; Kearns, Kenneth L.; Ediger, Mark D.; Yu, Lian

    2011-01-01

    The crystallization of glasses and amorphous solids is studied in many fields to understand the stability of amorphous materials, the fabrication of glass ceramics, and the mechanism of biomineralization. Recent studies have found that crystal growth in organic glasses can be orders of magnitude faster at the free surface than in the interior, a phenomenon potentially important for understanding glass crystallization in general. Current explanations differ for surface-enhanced crystal growth, including released tension and enhanced mobility at glass surfaces. We report here a feature of the phenomenon relevant for elucidating its mechanism: Despite their higher densities, surface crystals rise substantially above the glass surface as they grow laterally, without penetrating deep into the bulk. For indomethacin (IMC), an organic glass able to grow surface crystals in two polymorphs (α and γ), the growth front can be hundreds of nanometers above the glass surface. The process of surface crystal growth, meanwhile, is unperturbed by eliminating bulk material deeper than some threshold depth (ca. 300 nm for α IMC and less than 180 nm for γ IMC). As a growth strategy, the upward-lateral growth of surface crystals increases the system’s surface energy, but can effectively take advantage of surface mobility and circumvent slow growth in the bulk. PMID:21444775

  19. Intraoral tumor with rapid growing. Report of a case.

    PubMed

    González-Martín-Moro, Javier; Cebrián-Carretero, Jose Luis; Gómez-García, Elena; del Castillo-Pardo de Vera, Jose Luis; del Val, Daniel

    2005-01-01

    The appearance of an intraoral mass is common in our specialty. Most are benign lesions, but some are primary malignancies. Metastases account for less than 1% of all oral malignancies. An 86 year old woman was referred to our department with a large, asymptomatic, intraoral, fast-growing mass. She had no previous cancer history or other relevant physical findings. The radiology studies showed underlying bone erosion. The histological study showed a metastatic adenocarcinoma with a suspected origin in the abdomen. We were unable to identify it by non invasive diagnostic procedures. Given the patient's general status and despite the ominous prognosis of such lesions, we decided not to perform any aggressive therapy beyond removing the oral mass, in order to maintain her quality of life. There have been no local recurrences until this time.

  20. E-cigarettes: a rapidly growing Internet phenomenon.

    PubMed

    Yamin, Cyrus K; Bitton, Asaf; Bates, David W

    2010-11-02

    Electronic cigarettes (e-cigarettes) aerosolize nicotine and produce a vapor that emulates that of cigarettes but purportedly has fewer traditional toxins than secondhand smoke. Although e-cigarettes are widely sold online and by retailers, new research suggests that they may contain unexpected toxins and may provide unreliable nicotine delivery. Many countries have already banned or strictly regulated e-cigarettes. Currently in the United States, e-cigarettes are exempt from regulation as drug-delivery devices. Meanwhile, the presence of e-cigarettes on the Internet, including in Web searches, virtual user communities, and online stores where people sell e-cigarettes on commission, is increasing rapidly. Physicians should be aware of the popularity, questionable efficacy claims, and safety concerns of e-cigarettes so that they may counsel patients against use and advocate for research to inform an evidence-based regulatory approach.

  1. [Phyllodes tumour: a rare, rapidly growing breast tumour].

    PubMed

    den Exter, Paul L; Hornstra, Bonne J; Vree, Robbert

    2009-01-01

    A 40-year-old woman presented at the breast outpatient clinic with a giant tumour of her left breast. The size, rapid growth and radiological characteristics of the lesion led us to suspect a phyllodes tumour. A histological examination of a needle biopsy confirmed this diagnosis. An additional CT scan revealed no signs of metastases. We performed a mastectomy during which a tumour measuring 48 x 33 x 25 cm was resected. Histological examination revealed a borderline phyllodes tumour. Phyllodes tumours are rare fibroepithelial neoplasms of the breast and pre-operatively these are often difficult to differentiate from fibroadenomas. Phyllodes tumours have a variable clinical course with the ability to metastasize and a propensity to recur locally. Complete excision with wide margins is essential to prevent local recurrence. In our case, the surgical margins were limited and our patient was therefore treated with postoperative radiation therapy.

  2. Rapid Identification of Mycobacteria and Drug-Resistant Mycobacterium tuberculosis by Use of a Single Multiplex PCR and DNA Sequencing

    PubMed Central

    Pérez-Osorio, Ailyn C.; Boyle, David S.; Ingham, Zachary K.; Ostash, Alla; Gautom, Romesh K.; Colombel, Craig; Houze, Yolanda

    2012-01-01

    Tuberculosis (TB) remains a significant global health problem for which rapid diagnosis is critical to both treatment and control. This report describes a multiplex PCR method, the Mycobacterial IDentification and Drug Resistance Screen (MID-DRS) assay, which allows identification of members of the Mycobacterium tuberculosis complex (MTBC) and the simultaneous amplification of targets for sequencing-based drug resistance screening of rifampin-resistant (rifampinr), isoniazidr, and pyrazinamider TB. Additionally, the same multiplex reaction amplifies a specific 16S rRNA gene target for rapid identification of M. avium complex (MAC) and a region of the heat shock protein 65 gene (hsp65) for further DNA sequencing-based confirmation or identification of other mycobacterial species. Comparison of preliminary results generated with MID-DRS versus culture-based methods for a total of 188 bacterial isolates demonstrated MID-DRS sensitivity and specificity as 100% and 96.8% for MTBC identification; 100% and 98.3% for MAC identification; 97.4% and 98.7% for rifampinr TB identification; 60.6% and 100% for isoniazidr TB identification; and 75.0% and 98.1% for pyrazinamider TB identification. The performance of the MID-DRS was also tested on acid-fast-bacterium (AFB)-positive clinical specimens, resulting in sensitivity and specificity of 100% and 78.6% for detection of MTBC and 100% and 97.8% for detection of MAC. In conclusion, use of the MID-DRS reduces the time necessary for initial identification and drug resistance screening of TB specimens to as little as 2 days. Since all targets needed for completing the assay are included in a single PCR amplification step, assay costs, preparation time, and risks due to user errors are also reduced. PMID:22162548

  3. Rapidly growing aortic arch aneurysm in Behcet's disease.

    PubMed

    Kojima, Nozomi; Sakano, Yasuhito; Ohki, Shin-Ichi; Misawa, Yoshio

    2011-03-01

    We present a patient with a nine-year history of Behçet's disease (BD), who developed a rapidly expanding aneurysm of the aortic arch. Three-dimensional computed tomography demonstrated a saccular aortic arch aneurysm with a maximal diameter of 5 cm. No bacteria were detected by serial blood cultures. The aneurysm, however, showed a multi-lobular cavity, mimicking an infectious aneurysm. Therefore, we prescribed antibacterial agents for one week. The patient still had a high-fever and an elevated C-reactive protein level thereafter. Aortic arch replacement was performed emergently. Because we were unable to determine whether the aneurysm was caused by infection or BD, the implanted prosthetic graft and the anastomotic sites were covered with a pedicle graft of the greater omentum, and we continued to administer antibacterial agents for four weeks postoperatively. The pathological examination showed neither bacteria nor cystic medial necrosis in the resected aortic wall. Inflammatory changes with eosinophilic infiltration were recognized mainly around the adventitia near the aneurysm. The patient had a favorable postoperative course without any complications.

  4. Mycobacterium saopaulense sp. nov., a rapidly growing mycobacterium closely related to members of the Mycobacterium chelonae–Mycobacterium abscessus group

    PubMed Central

    Nogueira, Christiane Lourenço; Whipps, Christopher M.; Matsumoto, Cristianne Kayoko; Chimara, Erica; Droz, Sara; Tortoli, Enrico; de Freitas, Denise; Cnockaert, Margo; Palomino, Juan Carlos; Martin, Anandi; Vandamme, Peter

    2015-01-01

    Five isolates of non-pigmented, rapidly growing mycobacteria were isolated from three patients and, in an earlier study, from zebrafish. Phenotypic and molecular tests confirmed that these isolates belong to the Mycobacterium chelonae–Mycobacterium abscessus group, but they could not be confidently assigned to any known species of this group. Phenotypic analysis and biochemical tests were not helpful for distinguishing these isolates from other members of the M. chelonae–M. abscessus group. The isolates presented higher drug resistance in comparison with other members of the group, showing susceptibility only to clarithromycin. The five isolates showed a unique PCR restriction analysis pattern of the hsp65 gene, 100 % similarity in 16S rRNA gene and hsp65 sequences and 1–2 nt differences in rpoB and internal transcribed spacer (ITS) sequences. Phylogenetic analysis of a concatenated dataset including 16S rRNA gene, hsp65, and rpoB sequences from type strains of more closely related species placed the five isolates together, as a distinct lineage from previously described species, suggesting a sister relationship to a group consisting of M. chelonae, Mycobacterium salmoniphilum, Mycobacterium franklinii and Mycobacterium immunogenum. DNA–DNA hybridization values >70 % confirmed that the five isolates belong to the same species, while values < 70 % between one of the isolates and the type strains of M. chelonae and M. abscessus confirmed that the isolates belong to a distinct species. The polyphasic characterization of these isolates, supported by DNA–DNA hybridization results, demonstrated that they share characteristics with M. chelonae–M. abscessus members, but constitute a different species, for which the name Mycobacterium saopaulense sp. nov. is proposed. The type strain is EPM 10906T ( = CCUG 66554T = LMG 28586T = INCQS 0733T). PMID:26358475

  5. Susceptibility Testing of Antibiotics That Degrade Faster than the Doubling Time of Slow-Growing Mycobacteria: Ertapenem Sterilizing Effect versus Mycobacterium tuberculosis

    PubMed Central

    Srivastava, Shashikant; van Rijn, Sander P.; Wessels, A. Mireille A.; Gumbo, Tawanda

    2016-01-01

    Drug susceptibility tests (DSTs) for Mycobacterium tuberculosis require at least 7 days of incubation. Drugs that are unstable at 37°C, such as ertapenem, are likely to be degraded before killing or inhibiting slow-growing bacteria. This would alter the MICs of these drugs, including ertapenem, leading to falsely high MICs. Here, we describe a new strategy we developed to perform DSTs and measure MICs for such unstable compounds. PMID:26926650

  6. In vitro activity of bedaquiline against nontuberculous mycobacteria in China.

    PubMed

    Pang, Yu; Zheng, Huiwen; Tan, Yaoju; Song, Yuanyuan; Zhao, Yanlin

    2017-02-27

    The main goal of our study was to evaluate in vitro drug susceptibility of bedaquiline against six prevalent pathogenic nontuberculous mycobacteria (NTM) diseases in China. In addition, we investigated the potential molecular mechanism contributing to the bedaquiline resistance in these different NTM species. For slowly growing mycobacteria (SGM), bedaquiline exhibited the highest activity against Mycobacterium avium, the MIC50 and MIC90 were 0.03 and 16 mg/L, respectively. Of rapidly growing mycobacteria (RGM), Mycobacterium abscessus subsp. abscessus (M. abscessus) and Mycobacterium abscessus subsp. massiliense (M. massiliense) seemed more susceptible to bedaquiline than Mycobacterium fortuitum, with the MIC50s and MIC90s of 0.13 and >16 mg/L for both species. On the basis of bimodal distributions of the bedaquiline MICs, we proposed the following epidemiological cut-off (ECOFF) values: 1.0 mg/L for SGM and 2.0 mg/L for RGM. There were 14 (29.8%), 41 (27.2%), 33 (39.3%), 44 (20.2%), 42 (25.8%) and 7 (31.8%) isolates resistant to bedaquiline for M. avium,Mycobacterium intracellulare, Mycobacterium kansasii, M. abscessus, M. massiliense, and M. fortuitum, respectively. No significant difference was observed in the proportion of bedaquiline resistance among these species (P>0.05). The genetic mutations were observed in 74 (10.8%) isolates, while all nucleotide substitutions belonged to synonymous mutations. In conclusion, our data demonstrate that bedaquiline shows moderate in vitro activity against NTM species. Using the proposed ECOFF values, we could distinguish between bedaquiline resistant and susceptible strains by broth dilution method. In addition, no nonsynonymous mutations are identified in the atpE gene conferring bedaquiline resistance in all six NTM species.

  7. Ghost mycobacteria on Gram stain.

    PubMed Central

    Trifiro, S; Bourgault, A M; Lebel, F; René, P

    1990-01-01

    The Gram stain is a key tool in diagnostic microbiology. Its usefulness with respect to mycobacteria is undefined. The neutrality of mycobacteria other than Mycobacterium tuberculosis on Gram staining of various clinical specimens is described. Images PMID:1688872

  8. Rapid-growing juvenile xanthogranuloma on the scalp in 18-month-old girl.

    PubMed

    Park, Young Woo; Koh, Eun Jeong; Choi, Ha Young

    2011-09-01

    Juvenile xanthogranuloma (JXG) is an uncommon histiocytic cutaneous lesion. An 18-month-old girl visited our clinic due to rapid growing orange-yellowish lesion on scalp. Enlarging time from 1 mm to 12 mm was just 8 weeks. We excised the tumor and adjacent normal tissue. Histopathological study showed numerous eosinophils and Touton giant cells within the lesion. Immunohistochemical study revealed positive immunoreactivity for CD68 in most areas. No recurrence was seen during 12 months after resection. We report a case with rapidly growing JXG on scalp with peculiar histopathologic findings.

  9. The genomics of mycobacteria.

    PubMed

    Viale, M N; Zumárraga, M J; Araújo, F R; Zarraga, A M; Cataldi, A A; Romano, M I; Bigi, F

    2016-04-01

    The species Mycobacterium bovis and Mycobacterium avium subspecies paratuberculosis are the causal agents, respectively, of tuberculosis and paratuberculosis in animals. Both mycobacteria, especially M. bovis, are also important to public health because they can infect humans. In recent years, this and the impact of tuberculosis and paratuberculosis on animal production have led to significant advances in knowledge about both pathogens and their host interactions. This article describes the contribution of genomics and functional genomics to studies of the evolution, virulence, epidemiology and diagnosis of both these pathogenic mycobacteria.

  10. Public Relations in Rapidly Growing Suburban Districts: Insights from Texas Superintendents Bonny Cain and Doug Otto

    ERIC Educational Resources Information Center

    Decman, John M.

    2005-01-01

    Throughout Texas, enrollments in suburban school districts have been rising, and in some instances, the increases have been substantial. Causes of this demographic trend are multifaceted, but in most instances they are an intricate mix of state population growth and urban flight. Rapidly growing suburban districts, like all Texas districts,…

  11. Vibrio natriegens: A Rapidly Growing Micro-Organism Ideally Suited for Class Experiments

    ERIC Educational Resources Information Center

    Mullenger, L.; Gill, Nijole R.

    1973-01-01

    Describes five microbiological experiments using the marine organism Vibrio natriegens. This organism is highly suitable for laboratory work because it is non-pathogenic and grows extremely rapidly, having the distinction of the lowest mean generation time yet recorded (9.8 minutes). (JR)

  12. Public Relations in Rapidly Growing Suburban Districts: Insights from Texas Superintendents Bonny Cain and Doug Otto

    ERIC Educational Resources Information Center

    Decman, John M.

    2005-01-01

    Throughout Texas, enrollments in suburban school districts have been rising, and in some instances, the increases have been substantial. Causes of this demographic trend are multifaceted, but in most instances they are an intricate mix of state population growth and urban flight. Rapidly growing suburban districts, like all Texas districts,…

  13. Vibrio natriegens: A Rapidly Growing Micro-Organism Ideally Suited for Class Experiments

    ERIC Educational Resources Information Center

    Mullenger, L.; Gill, Nijole R.

    1973-01-01

    Describes five microbiological experiments using the marine organism Vibrio natriegens. This organism is highly suitable for laboratory work because it is non-pathogenic and grows extremely rapidly, having the distinction of the lowest mean generation time yet recorded (9.8 minutes). (JR)

  14. Quality Requirements in Rapidly Growing Higher Education Systems: The Israeli Example.

    ERIC Educational Resources Information Center

    Mehrez, Abraham; Mizrahi, Shlomo

    2000-01-01

    Offers a quality-policy planning model for academic institutions, noting that rapidly growing higher education systems are often characterized by market failure in terms of program quality, because controlling bodies have incomplete information about program quality and low sensitivity to quality variations. Applies this framework to Israel,…

  15. Growth physiology of mycobacteria in modified Dubos liquid medium.

    PubMed

    Tsukamura, M

    1990-01-01

    Although mycobacteria grow in Dubos liquid medium showing an arithmetic linear growth, the initial few days of growth were found to correspond to an 'induction' period. In this period, rapid increase of the amount of growth occurred, whereas increase of the number of colony-forming units remained at a low level. This finding shows that the rapid increase of the amount of growth is accompanied by rapid death of multiplied bacteria. In a successive period, which was considered to correspond to the logarithmic growth phase, a 1:1 correspondence existed between the amount of growth and the number of colony-forming units. The induction period is not considered to be a lag phase, in which the bacteria grow slowly, but a period of unbalanced relationship between the growth and the viability. Even when we inoculated different sizes of bacteria, the amounts of growth became similar in both inoculations after several days of incubation. However, the number of colony-forming units remained always smaller in the use of small inocula than in the use of large inocula. In the use of small inocula, much more rapid increase of the amount of growth occurred. However, this rapid increase gave rise to rapid death of bacteria.

  16. An Inhibitor of Exported Mycobacterium tuberculosis Glutamine Synthetase Selectively Blocks the Growth of Pathogenic Mycobacteria in Axenic Culture and in Human Monocytes: Extracellular Proteins as Potential Novel Drug Targets

    PubMed Central

    Harth, Günter; Horwitz, Marcus A.

    1999-01-01

    Mycobacterium tuberculosis and other pathogenic mycobacteria export abundant quantities of proteins into their extracellular milieu when growing either axenically or within phagosomes of host cells. One major extracellular protein, the enzyme glutamine synthetase, is of particular interest because of its link to pathogenicity. Pathogenic mycobacteria, but not nonpathogenic mycobacteria, export large amounts of this protein. Interestingly, export of the enzyme is associated with the presence of a poly-l-glutamate/glutamine structure in the mycobacterial cell wall. In this study, we investigated the influence of glutamine synthetase inhibitors on the growth of pathogenic and nonpathogenic mycobacteria and on the poly-l-glutamate/glutamine cell wall structure. The inhibitor l-methionine-S-sulfoximine rapidly inactivated purified M. tuberculosis glutamine synthetase, which was 100-fold more sensitive to this inhibitor than a representative mammalian glutamine synthetase. Added to cultures of pathogenic mycobacteria, l-methionine- S-sulfoximine rapidly inhibited extracellular glutamine synthetase in a concentration-dependent manner but had only a minimal effect on cellular glutamine synthetase, a finding consistent with failure of the drug to cross the mycobacterial cell wall. Remarkably, the inhibitor selectively blocked the growth of pathogenic mycobacteria, all of which release glutamine synthetase extracellularly, but had no effect on nonpathogenic mycobacteria or nonmycobacterial microorganisms, none of which release glutamine synthetase extracellularly. The inhibitor was also bacteriostatic for M. tuberculosis in human mononuclear phagocytes (THP-1 cells), the pathogen's primary host cells. Paralleling and perhaps underlying its bacteriostatic effect, the inhibitor markedly reduced the amount of poly-l-glutamate/glutamine cell wall structure in M. tuberculosis. Although it is possible that glutamine synthetase inhibitors interact with additional extracellular

  17. Insertional mutagenesis and illegitimate recombination in mycobacteria.

    PubMed Central

    Kalpana, G V; Bloom, B R; Jacobs, W R

    1991-01-01

    Mycobacteria, particularly Mycobacterium tuberculosis, Mycobacterium leprae, and Mycobacterium avium, are major pathogens of man. Although insertional mutagenesis has been an invaluable genetic tool for analyzing the mechanisms of microbial pathogenesis, it has not yet been possible to apply it to the mycobacteria. To overcome intrinsic difficulties in directly manipulating the genetics of slow-growing mycobacteria, including M. tuberculosis and bacille Calmette-Guérin (BCG) vaccine strains, we developed a system for random shuttle mutagenesis. A genomic library of Mycobacterium smegmatis was subjected to transposon mutagenesis with Tn5 seq1, a derivative of Tn5, in Escherichia coli and these transposon-containing recombinant plasmids were reintroduced into mycobacterial chromosomes by homologous recombination. This system has allowed us to isolate several random auxotrophic mutants of M. smegmatis. To extend this strategy to M. tuberculosis and BCG, targeted mutagenesis was performed using a cloned BCG methionine gene that was subjected to Tn5 seq1 mutagenesis in E. coli and reintroduced into the mycobacteria. Surprisingly for prokaryotes, both BCG and M. tuberculosis were found to incorporate linear DNA fragments into illegitimate sites throughout the mycobacterial genomes at a frequency of 10(-5) to 10(-4) relative to the number of transformants obtained with autonomously replicating vectors. Thus the efficient illegitimate recombination of linear DNA fragments provides the basis for an insertional mutagenesis system for M. tuberculosis and BCG. Images PMID:2052623

  18. Nontuberculous Mycobacteria Isolation from Clinical and Environmental Samples in Iran: Twenty Years of Surveillance.

    PubMed

    Velayati, Ali Akbar; Farnia, Parissa; Mozafari, Mohadese; Mirsaeidi, Mehdi

    2015-01-01

    Nontuberculous mycobacteria (NTM) are opportunistic pathogens that are widely distributed in the environment. There is a lack of data on species distribution of these organisms from Iran. This study consists of a review of NTM articles published in Iran between the years 1992 and 2014. In this review, 20 articles and 14 case reports were identified. Among the 20 articles, 13 (65%) studies focused on NTM isolates from clinical specimens, 6 (30%) studies examined NTM isolates from environmental samples, and one (5%) article included both clinical and environmental isolates. M. fortuitum (229/997; 23%) was recorded as the most prevalent and rapid growing mycobacteria (RGM) species in both clinical (28%) and environmental (19%) isolated samples (P < 0.05). Among slow growing mycobacteria (SGM), M. simiae (103/494; 21%) demonstrated a higher frequency in clinical samples whereas in environmental samples it was M. flavescens (44/503; 9%). These data represent information from 14 provinces out of 31 provinces of Iran. No information is available in current published data on clinical or environmental NTM from the remaining 17 provinces in Iran. These results emphasize the potential importance of NTM as well as the underestimation of NTM frequency in Iran. NTM is an important clinical problem associated with significant morbidity and mortality in Iran. Continued research is needed from both clinical and environmental sources to help clinicians and researchers better understand and address NTM treatment and prevention.

  19. Rapid phenotypic assay of antimycobacterial susceptibility pattern by direct mycobacteria growth indicator tube and phage amplified biological assay compared to BACTEC 460 TB.

    PubMed

    El-Sayed Zaki, Maysaa; Goda, Tarek

    2007-03-01

    The performance of antimycobacterial susceptibility testing for the first line drugs (isoniazid, streptomycine, rifampicin and ethambutol) with mycobacteria growth indicator tube (MGIT) and by bacteriophage amplified biological assay by FAST-plaque TB-MDR were compared to automated radiometric BACTEC 460 TB system. This study was carried on 84 sputum samples of positive Zhiel-Neelsen (ZN) smears. Sputum samples were subjected to culture and antimycobacterial susceptibility testing by BACTEC 460 TB. Samples were also tested by direct susceptibility tests for isoniazid (INH), ethambutol, rifampicin (RIF) and streptomycine by MGIT. Sensitive and resistant isolates for RIF were further studied by FAST-plaque TB-MDR for RIF resistance. The commonest resistance pattern by BACTEC 460 TB was for INH (32%) followed by RIF (24%) either alone or in combination with other drugs. Multiple drugs resistance was 20%. The agreement between BACTEC 460 TB and direct MGIT for resistant strains was 100% for INH and ethambutol, 91.7% for rifampicin, 80% for streptomycine and was 90% for MDR. FAST-plaque TB-MDR detected correctly all RIF resistant strains and 97.2% of the sensitive strains. For majority of strains direct susceptibility tests were available within 6.34-6.404 days (95% confidence interval) with direct mycobacteria growth tube, while results for FAST-plaque TB-MDR appear within 10.5-11.5 days from the time that the sputum was received in the laboratory (95% confidence interval). From this study, we could conclude that direct MGIT AST is the quickest method for screening antimycobacterial susceptibility pattern for the drugs commonly used (INH, RIF, etambutol, streptomycin) as results were available within 6.34-6.404 days. Also FAST-plaque TB-MDR method is accurate for detection of rifampicin resistance after primary culture which can be used as a surrogate marker for presence of MDR strains and the results were available within 10.5-11.5 days.

  20. Factors Affecting Phage D29 Infection: A Tool to Investigate Different Growth States of Mycobacteria

    PubMed Central

    Swift, Benjamin M. C.; Gerrard, Zara E.; Huxley, Jonathan N.; Rees, Catherine E. D.

    2014-01-01

    Bacteriophages D29 and TM4 are able to infect a wide range of mycobacteria, including pathogenic and non-pathogenic species. Successful phage infection of both fast- and slow-growing mycobacteria can be rapidly detected using the phage amplification assay. Using this method, the effect of oxygen limitation during culture of mycobacteria on the success of phage infection was studied. Both D29 and TM4 were able to infect cultures of M. smegmatis and Mycobacterium avium subspecies paratuberculosis (MAP) grown in liquid with aeration. However when cultures were grown under oxygen limiting conditions, only TM4 could productively infect the cells. Cell attachment assays showed that D29 could bind to the cells surface but did not complete the lytic cycle. The ability of D29 to productively infect the cells was rapidly recovered (within 1 day) when the cultures were returned to an aerobic environment and this recovery required de novo RNA synthesis. These results indicated that under oxygen limiting conditions the cells are entering a growth state which inhibits phage D29 replication, and this change in host cell biology which can be detected by using both phage D29 and TM4 in the phage amplification assay. PMID:25184428

  1. Factors affecting phage D29 infection: a tool to investigate different growth states of mycobacteria.

    PubMed

    Swift, Benjamin M C; Gerrard, Zara E; Huxley, Jonathan N; Rees, Catherine E D

    2014-01-01

    Bacteriophages D29 and TM4 are able to infect a wide range of mycobacteria, including pathogenic and non-pathogenic species. Successful phage infection of both fast- and slow-growing mycobacteria can be rapidly detected using the phage amplification assay. Using this method, the effect of oxygen limitation during culture of mycobacteria on the success of phage infection was studied. Both D29 and TM4 were able to infect cultures of M. smegmatis and Mycobacterium avium subspecies paratuberculosis (MAP) grown in liquid with aeration. However when cultures were grown under oxygen limiting conditions, only TM4 could productively infect the cells. Cell attachment assays showed that D29 could bind to the cells surface but did not complete the lytic cycle. The ability of D29 to productively infect the cells was rapidly recovered (within 1 day) when the cultures were returned to an aerobic environment and this recovery required de novo RNA synthesis. These results indicated that under oxygen limiting conditions the cells are entering a growth state which inhibits phage D29 replication, and this change in host cell biology which can be detected by using both phage D29 and TM4 in the phage amplification assay.

  2. Rapidly growing pigmented tumor on a scalp nevus sebaceous of a pediatric patient: Observation or excision.

    PubMed

    Gaitan-Gaona, Francisco; Said, Mirra C; Galvan-Linares, Aldo; Palafox-Vigil, Gloria; Valdes-Rodriguez, Rodrigo

    2014-07-15

    A 14-year-old girl presented with a new, rapidly growing, pigmented tumor on a previously existing yellowish, verrucous plaque on the scalp. The patient received complete surgical excision. Routine histology ruled out basal cell carcinoma (BCC) and the histological diagnosis was pigmented trichoblastoma arising in nevus sebaceous (NS). It is important to define management for new lesions developing in pediatric patients with existing nevus sebaceus.

  3. Rapidly growing glandular papilloma associated with mucus production: a case report.

    PubMed

    Suzuki, Shigeki; Goto, Taichiro; Emoto, Katsura; Hayashi, Yuichiro

    2014-05-22

    Pulmonary glandular papillomas are rare neoplasms, and their very slow or absent growth over time generally facilitates establishing the diagnosis. In an 84-year-old woman who underwent surgery for sigmoid colon cancer, a growing solitary pulmonary nodule was identified on postoperative follow-up computed tomography. A computer tomography-guided needle biopsy was performed under suspicion that the nodule was malignant. The histopathological findings suggested a glandular papilloma. Right basilar segmentectomy was carried out, and the lesion was completely resected. Postoperative histopathological examination revealed a benign glandular papilloma accompanied by mucus retention in the surrounding alveolar region. A malignant neoplasm is usually suspected when a pulmonary tumor shows rapid growth. However, glandular papillomas associated with mucus retention also tend to grow in some cases, and should be included in the differential diagnosis.

  4. Rapidly growing glandular papilloma associated with mucus production: a case report

    PubMed Central

    2014-01-01

    Background Pulmonary glandular papillomas are rare neoplasms, and their very slow or absent growth over time generally facilitates establishing the diagnosis. Case presentation In an 84-year-old woman who underwent surgery for sigmoid colon cancer, a growing solitary pulmonary nodule was identified on postoperative follow-up computed tomography. A computer tomography-guided needle biopsy was performed under suspicion that the nodule was malignant. The histopathological findings suggested a glandular papilloma. Right basilar segmentectomy was carried out, and the lesion was completely resected. Postoperative histopathological examination revealed a benign glandular papilloma accompanied by mucus retention in the surrounding alveolar region. Conclusions A malignant neoplasm is usually suspected when a pulmonary tumor shows rapid growth. However, glandular papillomas associated with mucus retention also tend to grow in some cases, and should be included in the differential diagnosis. PMID:24886616

  5. Disseminated Cryptococcosis with Rapidly Growing Lung Nodules in an End-stage Renal Disease Patient

    PubMed Central

    Inaba, Atsushi; Okada, Akira; Yoshida, Taiko; Itoyama, Satoshi; Nakai, Tatsuro; Hisada, Tetsuya; Takano, Hideki

    2017-01-01

    A 73-year-old man with type 2 diabetes mellitus and end-stage renal disease was diagnosed with acute myocardial infarction. He required continuous dialysis after percutaneous coronary intervention. Subsequently, multiple nodules were discovered in both lungs for the first time, and Cryptococcus neoformans was isolated from the patient's sputum, blood, bilateral pleural fluid, and cerebrospinal fluid cultures, resulting in a diagnosis of disseminated cryptococcosis. This case represents an invaluable example of disseminated cryptococcosis with rapidly growing lung nodules in a dialysis patient, and illustrates that dialysis causes innate immune disorder and the reactivation of cryptococcosis. PMID:28154287

  6. Rapidly Growing Right Ventricular Outflow Tract Mass in Patient with Sarcomatoid Renal Cell Carcinoma

    PubMed Central

    Hwang, Jongmin; Choi, Kyung Un; Kim, Jeong Su; Hwang, Ki Won; Lee, Sang Hyun; Chon, Min Ku; Lee, Soo Yong; Lee, Dae Sung

    2016-01-01

    Cardiac metastasis from renal cell carcinoma (RCC) without inferior vena cava (IVC) involvements is extremely rare with few reported cases. Sarcomatoid RCC with rhabdoid feature is a rare pathologic type of RCC having aggressive behavior due to great metastatic potential. Here, we report a case of rapidly growing cardiac metastasis of RCC which brought on right ventricular outflow tract (RVOT) obstruction without IVC and right atrial involvement in a 61-year-old woman. Cardiac arrest occurred during radical nephrectomy and echocardiography revealed mass nearly obstructing the RVOT which was not recognized by preoperative echocardiography 1 month ago. Postoperative immunohistochemical evaluation of renal mass revealed sarcomatoid RCC with rhabdoid feature. PMID:28090262

  7. Disseminated Cryptococcosis with Rapidly Growing Lung Nodules in an End-stage Renal Disease Patient.

    PubMed

    Inaba, Atsushi; Okada, Akira; Yoshida, Taiko; Itoyama, Satoshi; Nakai, Tatsuro; Hisada, Tetsuya; Takano, Hideki

    2017-01-01

    A 73-year-old man with type 2 diabetes mellitus and end-stage renal disease was diagnosed with acute myocardial infarction. He required continuous dialysis after percutaneous coronary intervention. Subsequently, multiple nodules were discovered in both lungs for the first time, and Cryptococcus neoformans was isolated from the patient's sputum, blood, bilateral pleural fluid, and cerebrospinal fluid cultures, resulting in a diagnosis of disseminated cryptococcosis. This case represents an invaluable example of disseminated cryptococcosis with rapidly growing lung nodules in a dialysis patient, and illustrates that dialysis causes innate immune disorder and the reactivation of cryptococcosis.

  8. Overweight, obesity, and inactivity and urban design in rapidly growing Chinese cities.

    PubMed

    Day, Kristen; Alfonzo, Mariela; Chen, Yufei; Guo, Zhan; Lee, Karen K

    2013-05-01

    China faces rising rates of overweight, obesity, and physical inactivity among its citizens. Risk is highest in China's rapidly growing cities and urban populations. Current urban development practices and policies in China heighten this risk. These include policies that support decentralization in land use planning; practices of neighborhood gating; and policies and practices tied to motor vehicle travel, transit planning, and bicycle and pedestrian infrastructure. In this paper, we review cultural, political, and economic issues that influence overweight, obesity, and inactivity in China. We examine key urban planning features and policies that shape urban environments that may compromise physical activity as part of everyday life, including walking and bicycling. We review the empirical research to identify planning and design strategies that support physical activity in other high-density cities in developing and developed countries. Finally, we identify successful strategies to increase physical activity in another growing, high-density city - New York City - to suggest strategies that may have relevance for rapidly urbanizing Chinese cities.

  9. Non-tuberculosis mycobacteria in Africa

    PubMed Central

    Zykov, M. P.; Roulet, H.

    1967-01-01

    The third study in a series on the prevalence of non-tuberculosis mycobacteria in Africa is devoted to the investigation of the formamidase activity of 288 cultures of mycobacteria, already typed by a battery of standard tests as pathogenic or atypical (184 strains) and saprophytic (104 strains). Of the latter, 96 (92.3%) were formamidase-positive, as compared with only 6 (3.3%) of the former. A close correlation was observed between the speed of growth on Löwenstein-Jensen medium and formamidase activity, 98 (96.1%) of the positive strains showing visible growth within 1-3 days. The relation between formamidase activity and growth on nutrient media was less clear-cut, however, and it was concluded that for the routine differentiation of saprophytic from other mycobacteria the formamidase test should be combined with simple tests such as speed of growth on L-J medium and ability to grow on nutrient media. Russel's method and Nessler's reagent for the detection of ammonia in the formamidase test were compared; the authors consider the former to be preferable, since the reaction is easier to read. PMID:4173140

  10. Nontuberculous Mycobacteria, Zambia

    PubMed Central

    van der Sande, Marianne A.B.; de Graaff, Cas S.; Parkinson, Shelagh; Verbrugh, Henri A.; Petit, Pieter L.C.; van Soolingen, Dick

    2009-01-01

    Clinical relevance of nontuberculous mycobacteria (NTM) isolated from 180 chronically ill patients and 385 healthy controls in Zambia was evaluated to examine the contribution of these isolates to tuberculosis (TB)–like disease. The proportion of NTM-positive sputum samples was significantly higher in the patient group than in controls; 11% and 6%, respectively (p<0.05). NTM-associated lung disease was diagnosed for 1 patient, and a probable diagnosis was made for 3 patients. NTM-positive patients and controls were more likely to report vomiting and diarrhea and were more frequently underweight than the NTM-negative patients and controls. Chest radiographs of NTM-positive patients showed deviations consistent with TB more frequently than those of controls. The most frequently isolated NTM was Mycobacterium avium complex. Multiple, not previously identified mycobacteria (55 of 171 NTM) were isolated from both groups. NTM probably play an important role in the etiology of TB-like diseases in Zambia. PMID:19193268

  11. Energetics of Respiration and Oxidative Phosphorylation in Mycobacteria

    PubMed Central

    Hards, Kiel; Vilchèze, Catherine; Hartman, Travis; Berney, Michael

    2014-01-01

    Mycobacteria inhabit a wide range of intracellular and extracellular environments. Many of these environments are highly dynamic and therefore mycobacteria are faced with the constant challenge of redirecting their metabolic activity to be commensurate with either replicative growth or a non-replicative quiescence. A fundamental feature in this adaptation is the ability of mycobacteria to respire, regenerate reducing equivalents and generate ATP via oxidative phosphorylation. Mycobacteria harbor multiple primary dehydrogenases to fuel the electron transport chain and two terminal respiratory oxidases, an aa3-type cytochrome c oxidase and cytochrome bd-type menaquinol oxidase, are present for dioxygen reduction coupled to the generation of a protonmotive force. Hypoxia leads to the downregulation of key respiratory complexes, but the molecular mechanisms regulating this expression are unknown. Despite being obligate aerobes, mycobacteria have the ability to metabolize in the absence of oxygen and a number of reductases are present to facilitate the turnover of reducing equivalents under these conditions (e.g. nitrate reductase, succinate dehydrogenase/fumarate reductase). Hydrogenases and ferredoxins are also present in the genomes of mycobacteria suggesting the ability of these bacteria to adapt to an anaerobic-type of metabolism in the absence of oxygen. ATP synthesis by the membrane-bound F1FO-ATP synthase is essential for growing and non-growing mycobacteria and the enzyme is able to function over a wide range of protonmotive force values (aerobic to hypoxic). The discovery of lead compounds that target respiration and oxidative phosphorylation in Mycobacterium tuberculosis highlights the importance of this area for the generation of new front line drugs to combat tuberculosis. PMID:25346874

  12. A novel protein extraction method for identification of mycobacteria using MALDI-ToF MS.

    PubMed

    Adams, La'Tonzia L; Salee, Parichat; Dionne, Kim; Carroll, Karen; Parrish, Nicole

    2015-12-01

    Commercial extraction methods for identification of mycobacteria using MALDI-ToF MS are laborious and time consuming. We have developed a novel extraction method which utilizes a bead beater and zirconia/silica beads to significantly shorten the existing protocol. This novel method provides a more rapid extraction of mycobacteria versus the commercial standard.

  13. Patient Report and Review of Rapidly Growing Mycobacterial Infection after Cardiac Device Implantation.

    PubMed

    Phadke, Varun K; Hirsh, David S; Goswami, Neela D

    2016-03-01

    Mycobacterial infections resulting from cardiac implantable electronic devices are rare, but as more devices are implanted, these organisms are increasingly emerging as causes of early-onset infections. We report a patient with an implantable cardioverter-defibrillator pocket and associated bloodstream infection caused by an organism of the Mycobacterium fortuitum group, and we review the literature regarding mycobacterial infections resulting from cardiac device implantations. Thirty-two such infections have been previously described; most (70%) were caused by rapidly growing species, of which M. fortuitum group species were predominant.When managing such infections, clinicians should consider the potential need for extended incubation of routine cultures or dedicated mycobacterial cultures for accurate diagnosis; combination antimicrobial drug therapy, even for isolates that appear to be macrolide susceptible, because of the potential for inducible resistance to this drug class; and the arrhythmogenicity of the antimicrobial drugs traditionally recommended for infections caused by these organisms.

  14. Rapid DNA, RNA and protein extraction protocols optimized for slow continuously growing yeast cultures.

    PubMed

    Sasidharan, Kalesh; Amariei, Cornelia; Tomita, Masaru; Murray, Douglas B

    2012-08-01

    Conventional extraction protocols for yeast have been developed for relatively rapid-growing low cell density cultures of laboratory strains and often do not have the integrity for frequent sampling of cultures. Therefore, these protocols are usually inefficient for cultures under slow growth conditions or of non-laboratory strains. We have developed a combined mechanical and chemical disruption procedure using vigorous bead-beating that can consistently disrupt yeast cells (> 95%), irrespective of cell cycle and metabolic state. Using this disruption technique coupled with quenching, we have developed DNA, RNA and protein extraction protocols that are optimized for a large number of samples from slow-growing high-density industrial yeast cultures. Additionally, sample volume, the use of expensive reagents/enzymes, handling times and incubations were minimized. We have tested the reproducibility of our methods using triplicate/time-series extractions and compared these with commonly used protocols or commercially available kits. Moreover, we utilized a simple flow-cytometric approach to estimate the mitochondrial DNA copy number. Based on the results, our methods have shown higher reproducibility, yield and quality. Copyright © 2012 John Wiley & Sons, Ltd.

  15. Rapid urbanization and the growing threat of violence and conflict: a 21st century crisis.

    PubMed

    Patel, Ronak B; Burkle, Frederick M

    2012-04-01

    As the global population is concentrated into complex environments, rapid urbanization increases the threat of conflict and insecurity. Many fast-growing cities create conditions of significant disparities in standards of living, which set up a natural environment for conflict over resources. As urban slums become a haven for criminal elements, youth gangs, and the arms trade, they also create insecurity for much of the population. Specific populations, such as women, migrants, and refugees, bear the brunt of this lack of security, with significant impacts on their livelihoods, health, and access to basic services. This lack of security and violence also has great costs to the general population, both economic and social. Cities have increasingly become the battlefield of recent conflicts as they serve as the seats of power and gateways to resources. International agencies, non-governmental organizations, and policy-makers must act to stem this tide of growing urban insecurity. Protecting urban populations and preventing future conflict will require better urban planning, investment in livelihood programs for youth, cooperation with local communities, enhanced policing, and strengthening the capacity of judicial systems.

  16. Intra-urban air pollution in a rapidly growing Sahelian city.

    PubMed

    Lindén, J; Boman, J; Holmer, B; Thorsson, S; Eliasson, I

    2012-04-01

    In this paper we analyze spatial and temporal variations of air pollution (PM(1), PM(2.5), PM(10), CO, NO(x), O(3), Toluene and Benzene) and climate in areas of different development typology in Ouagadougou, Burkina Faso. Analyses are based on measurements from fixed sites and car traverse measurements during field studies in 2007 and 2010. Large spatial and temporal variations were found, showing a generally poor air quality situation, with extreme levels of PM(10), commonly exceeding air quality guidelines of WHO. Pollution levels increase considerably with increased atmospheric stability. Important sources were transported dust and re-suspension of dust from unpaved roads, but also traffic emissions and biomass burning. The spatial variations are examined with focus on effects for variations in potential exposure depending on for example area of residence and daily activity pattern, showing that great differences are likely to exist. Ouagadougou, like most developing countries worldwide, currently experiences an extremely rapid population growth in combination with limited financial means. This is likely to create increasingly harmful air pollution situations for the rapidly growing populations of these areas, and shows an urgent need for increased understanding of the pollution situation as well as development of mitigation strategies.

  17. Animal Models of Mycobacteria Infection

    PubMed Central

    Ordway, Diane J.; Orme, Ian M.

    2011-01-01

    This unit describes the infection of mice and guinea pigs with mycobacteria via various routes, as well as necropsy methods for the determination of mycobacterial loads within target organs. Additionally, methods for cultivating mycobacteria and preparing stocks are described. The protocols outlined are primarily used for M. tuberculosis, but can also be used for the study of other non-tuberculosis mycobacterial species. PMID:18432756

  18. Water as a source of potentially pathogenic mycobacteria.

    PubMed

    Goslee, S; Wolinsky, E

    1976-03-01

    The mycobacterial flora of 321 water samples was explored to evaluate the role of this part of the environment as a possible source of human mycobacterial disease. The samples included natural waters, waters treated to make them suitable for drinking, and waters in contact with animals. Water from the city aquarium contained the greatest abundance of mycobacteria, with an average of 3.5 strains per sample. The highest yield of positive cultures came from samples in contact with zoo animals and with fish. The majority of the isolated strains were slowly growing mycobacteria; 80 were Mycobacterium gordonae, and 34 of thse belong to a new serotype. Forty-seven cultures were members of the M. avian-intracellulare-scrofulaceum complex, of which 11 were typable by agglutination. From this study and from the work of others, it is concluded that water may be contaminated with potentially pathogenic mycobacteria and thus may serve as a source of human disease.

  19. Rapidly growing tropical trees mobilize remarkable amounts of nitrogen, in ways that differ surprisingly among species

    PubMed Central

    Russell, Ann E.; Raich, James W.

    2012-01-01

    Fast-growing forests such as tropical secondary forests can accumulate large amounts of carbon (C), and thereby play an important role in the atmospheric CO2 balance. Because nitrogen (N) cycling is inextricably linked with C cycling, the question becomes: Where does the N come from to match high rates of C accumulation? In unique experimental 16-y-old plantations established in abandoned pasture in lowland Costa Rica, we used a mass-balance approach to quantify N accumulation in vegetation, identify sources of N, and evaluate differences among tree species in N cycling. The replicated design contained four broad-leaved evergreen tree species growing under similar environmental conditions. Nitrogen uptake was rapid, reaching 409 (±30) kg⋅ha−1⋅y−1, double the rate reported from a Puerto Rican forest and greater than four times that observed at Hubbard Brook Forest (New Hampshire, USA). Nitrogen amassed in vegetation was 874 (±176) kg⋅ha−1, whereas net losses of soil N (0–100 cm) varied from 217 (±146) to 3,354 (±915) kg⋅ha−1 (P = 0.018) over 16 y. Soil C:N, δ13C values, and N budgets indicated that soil was the main source of biomass N. In Vochysia guatemalensis, however, N fixation contributed >60 kg⋅ha−1⋅y−1. All species apparently promoted soil N turnover, such that the soil N mean residence time was 32–54 y, an order of magnitude lower than the global mean. High rates of N uptake were associated with substantial N losses in three of the species, in which an average of 1.6 g N was lost for every gram of N accumulated in biomass. PMID:22689942

  20. Rapidly growing tropical trees mobilize remarkable amounts of nitrogen, in ways that differ surprisingly among species.

    PubMed

    Russell, Ann E; Raich, James W

    2012-06-26

    Fast-growing forests such as tropical secondary forests can accumulate large amounts of carbon (C), and thereby play an important role in the atmospheric CO(2) balance. Because nitrogen (N) cycling is inextricably linked with C cycling, the question becomes: Where does the N come from to match high rates of C accumulation? In unique experimental 16-y-old plantations established in abandoned pasture in lowland Costa Rica, we used a mass-balance approach to quantify N accumulation in vegetation, identify sources of N, and evaluate differences among tree species in N cycling. The replicated design contained four broad-leaved evergreen tree species growing under similar environmental conditions. Nitrogen uptake was rapid, reaching 409 (± 30) kg · ha(-1) · y(-1), double the rate reported from a Puerto Rican forest and greater than four times that observed at Hubbard Brook Forest (New Hampshire, USA). Nitrogen amassed in vegetation was 874 (± 176) kg · ha(-1), whereas net losses of soil N (0-100 cm) varied from 217 (±146) to 3,354 (± 915) kg · ha(-1) (P = 0.018) over 16 y. Soil C:N, δ(13)C values, and N budgets indicated that soil was the main source of biomass N. In Vochysia guatemalensis, however, N fixation contributed >60 kg · ha(-1) · y(-1). All species apparently promoted soil N turnover, such that the soil N mean residence time was 32-54 y, an order of magnitude lower than the global mean. High rates of N uptake were associated with substantial N losses in three of the species, in which an average of 1.6 g N was lost for every gram of N accumulated in biomass.

  1. First Detection of Mycobacteria in African Rodents and Insectivores, Using Stratified Pool Screening▿ †

    PubMed Central

    Durnez, Lies; Eddyani, Miriam; Mgode, Georgies F.; Katakweba, Abdul; Katholi, Charles R.; Machang'u, Robert R.; Kazwala, Rudovik R.; Portaels, Françoise; Leirs, Herwig

    2008-01-01

    With the rising number of patients with human immunodeficiency virus (HIV)/AIDS in developing countries, the control of mycobacteria is of growing importance. Previous studies have shown that rodents and insectivores are carriers of mycobacteria. However, it is not clear how widespread mycobacteria are in these animals and what their role is in spreading them. Therefore, the prevalence of mycobacteria in rodents and insectivores was studied in and around Morogoro, Tanzania. Live rodents were trapped, with three types of live traps, in three habitats. Pieces of organs were pooled per habitat, species, and organ type (stratified pooling); these sample pools were examined for the presence of mycobacteria by PCR, microscopy, and culture methods. The mycobacterial isolates were identified using phenotypic techniques and sequencing. In total, 708 small mammals were collected, 31 of which were shrews. By pool prevalence estimation, 2.65% of the animals were carriers of mycobacteria, with a higher prevalence in the urban areas and in Cricetomys gambianus and the insectivore Crocidura hirta. Nontuberculous mycobacteria (Mycobacterium chimaera, M. intracellulare, M. arupense, M. parascrofulaceum, and Mycobacterium spp.) were isolated from C. gambianus, Mastomys natalensis, and C. hirta. This study is the first to report findings of mycobacteria in African rodents and insectivores and the first in mycobacterial ecology to estimate the prevalence of mycobacteria after stratified pool screening. The fact that small mammals in urban areas carry more mycobacteria than those in the fields and that potentially pathogenic mycobacteria were isolated identifies a risk for other animals and humans, especially HIV/AIDS patients, that have a weakened immune system. PMID:18065608

  2. First detection of mycobacteria in African rodents and insectivores, using stratified pool screening.

    PubMed

    Durnez, Lies; Eddyani, Miriam; Mgode, Georgies F; Katakweba, Abdul; Katholi, Charles R; Machang'u, Robert R; Kazwala, Rudovik R; Portaels, Françoise; Leirs, Herwig

    2008-02-01

    With the rising number of patients with human immunodeficiency virus (HIV)/AIDS in developing countries, the control of mycobacteria is of growing importance. Previous studies have shown that rodents and insectivores are carriers of mycobacteria. However, it is not clear how widespread mycobacteria are in these animals and what their role is in spreading them. Therefore, the prevalence of mycobacteria in rodents and insectivores was studied in and around Morogoro, Tanzania. Live rodents were trapped, with three types of live traps, in three habitats. Pieces of organs were pooled per habitat, species, and organ type (stratified pooling); these sample pools were examined for the presence of mycobacteria by PCR, microscopy, and culture methods. The mycobacterial isolates were identified using phenotypic techniques and sequencing. In total, 708 small mammals were collected, 31 of which were shrews. By pool prevalence estimation, 2.65% of the animals were carriers of mycobacteria, with a higher prevalence in the urban areas and in Cricetomys gambianus and the insectivore Crocidura hirta. Nontuberculous mycobacteria (Mycobacterium chimaera, M. intracellulare, M. arupense, M. parascrofulaceum, and Mycobacterium spp.) were isolated from C. gambianus, Mastomys natalensis, and C. hirta. This study is the first to report findings of mycobacteria in African rodents and insectivores and the first in mycobacterial ecology to estimate the prevalence of mycobacteria after stratified pool screening. The fact that small mammals in urban areas carry more mycobacteria than those in the fields and that potentially pathogenic mycobacteria were isolated identifies a risk for other animals and humans, especially HIV/AIDS patients, that have a weakened immune system.

  3. Rapidly growing giant cell tumor of bone in a skeletally immature girl.

    PubMed

    Akaike, Gensuke; Ueno, Teruko; Matsumoto, Seiichi; Motoi, Noriko; Matsueda, Kiyoshi

    2016-04-01

    Giant cell tumor of bone (GCTB) in skeletally immature patients is rare, and little is known regarding how fast GCTB can grow. We report a case of a 10-year-old skeletally immature girl with pathologically proven GCTB with obvious growth plate invasion that showed surprisingly rapid growth over only 14 days. A radiograph of the left knee revealed well-circumscribed, geographic bone destruction at the distal metaphysis of the femur with a focal cortical defect, suggesting a pathologic fracture. No abnormal mineralization or periosteal reaction was seen. A CT without contrast and an MRI demonstrated a homogeneous lesion with cortical disruption posteriorly and laterally with a slight soft tissue extension. Biopsy showed numerous multinucleated giant cells and spindle-shaped mononuclear cells without any sign of malignancy, suggesting GCTB. However, rapid lesion enlargement and destruction of the surrounding cortex were noted 14 days after biopsy. Considering the amount of bone destruction, traditional treatment of curettage and bone cement would not suffice to sustain structural strength. In addition, considering the patient's age, the tumor location, and the aggressive course, a malignant tumor, especially a giant cell-rich osteosarcoma, could not be excluded. Therefore, en bloc resection, including the growth plate and prosthetic replacement, were performed. Confirmation of GCTB was made from a pathologic evaluation, and a breach to the growth plate was identified. Since very little inflammatory reaction, degenerative change, or aneurysmal, bone, cyst-like change was found, the growth plate invasion was confirmed as due to GCTB extension, not due to the preoperative biopsy.

  4. The spatial biology of transcription and translation in rapidly growing Escherichia coli.

    PubMed

    Bakshi, Somenath; Choi, Heejun; Weisshaar, James C

    2015-01-01

    Single-molecule fluorescence provides high resolution spatial distributions of ribosomes and RNA polymerase (RNAP) in live, rapidly growing Escherichia coli. Ribosomes are more strongly segregated from the nucleoids (chromosomal DNA) than previous widefield fluorescence studies suggested. While most transcription may be co-translational, the evidence indicates that most translation occurs on free mRNA copies that have diffused from the nucleoids to a ribosome-rich region. Analysis of time-resolved images of the nucleoid spatial distribution after treatment with the transcription-halting drug rifampicin and the translation-halting drug chloramphenicol shows that both drugs cause nucleoid contraction on the 0-3 min timescale. This is consistent with the transertion hypothesis. We suggest that the longer-term (20-30 min) nucleoid expansion after Rif treatment arises from conversion of 70S-polysomes to 30S and 50S subunits, which readily penetrate the nucleoids. Monte Carlo simulations of a polymer bead model built to mimic the chromosomal DNA and ribosomes (either 70S-polysomes or 30S and 50S subunits) explain spatial segregation or mixing of ribosomes and nucleoids in terms of excluded volume and entropic effects alone. A comprehensive model of the transcription-translation-transertion system incorporates this new information about the spatial organization of the E. coli cytoplasm. We propose that transertion, which radially expands the nucleoids, is essential for recycling of 30S and 50S subunits from ribosome-rich regions back into the nucleoids. There they initiate co-transcriptional translation, which is an important mechanism for maintaining RNAP forward progress and protecting the nascent mRNA chain. Segregation of 70S-polysomes from the nucleoid may facilitate rapid growth by shortening the search time for ribosomes to find free mRNA concentrated outside the nucleoid and the search time for RNAP concentrated within the nucleoid to find transcription initiation

  5. Phagocytosis of mycobacteria by U937 cells: a rapid method for monitoring uptake and separating phagocytosed and free bacteria by magnetic beads.

    PubMed

    Whyte, J; Roberts, A D; Morley, K A; Sharp, R J; Marsh, P D

    2000-01-01

    A human-derived monocytic cell line (U937) was induced to phagocytose Mycobacterium phlei by the addition of phorbol myristate acetate (PMA) to the culture medium for 50-60 h. Cells not treated with PMA were unable to phagocytose M. phlei. Magnetic beads enabled a rapid and highly efficient separation of phagocytosed and free bacteria to be achieved, an approach which is particularly useful if colony plating is used to enumerate bacterial survival within phagocytic cells. Fluorescence-activated cell sorting (FACS) analysis showed that 98% of U937 cells contained viable bacteria after 3 h.

  6. An Automated System for Rapid Non-Destructive Enumeration of Growing Microbes

    PubMed Central

    London, Roanna; Schwedock, Julie; Sage, Andrew; Valley, Heather; Meadows, Jamie; Waddington, Michael; Straus, Don

    2010-01-01

    Background The power and simplicity of visual colony counting have made it the mainstay of microbiological analysis for more than 130 years. A disadvantage of the method is the long time required to generate visible colonies from cells in a sample. New rapid testing technologies generally have failed to maintain one or more of the major advantages of culture-based methods. Principal Findings We present a new technology and platform that uses digital imaging of cellular autofluorescence to detect and enumerate growing microcolonies many generations before they become visible to the eye. The data presented demonstrate that the method preserves the viability of the microcolonies it detects, thus enabling generation of pure cultures for microbial identification. While visual colony counting detects Escherichia coli colonies containing about 5×106 cells, the new imaging method detects E. coli microcolonies when they contain about 120 cells and microcolonies of the yeast Candida albicans when they contain only about 12 cells. We demonstrate that digital imaging of microcolony autofluorescence detects a broad spectrum of prokaryotic and eukaryotic microbes and present a model for predicting the time to detection for individual strains. Results from the analysis of environmental samples from pharmaceutical manufacturing plants containing a mixture of unidentified microbes demonstrate the method's improved test turnaround times. Conclusion This work demonstrates a new technology and automated platform that substantially shortens test times while maintaining key advantages of the current methods. PMID:20062794

  7. An automated system for rapid non-destructive enumeration of growing microbes.

    PubMed

    London, Roanna; Schwedock, Julie; Sage, Andrew; Valley, Heather; Meadows, Jamie; Waddington, Michael; Straus, Don

    2010-01-07

    The power and simplicity of visual colony counting have made it the mainstay of microbiological analysis for more than 130 years. A disadvantage of the method is the long time required to generate visible colonies from cells in a sample. New rapid testing technologies generally have failed to maintain one or more of the major advantages of culture-based methods. We present a new technology and platform that uses digital imaging of cellular autofluorescence to detect and enumerate growing microcolonies many generations before they become visible to the eye. The data presented demonstrate that the method preserves the viability of the microcolonies it detects, thus enabling generation of pure cultures for microbial identification. While visual colony counting detects Escherichia coli colonies containing about 5x10(6) cells, the new imaging method detects E. coli microcolonies when they contain about 120 cells and microcolonies of the yeast Candida albicans when they contain only about 12 cells. We demonstrate that digital imaging of microcolony autofluorescence detects a broad spectrum of prokaryotic and eukaryotic microbes and present a model for predicting the time to detection for individual strains. Results from the analysis of environmental samples from pharmaceutical manufacturing plants containing a mixture of unidentified microbes demonstrate the method's improved test turnaround times. This work demonstrates a new technology and automated platform that substantially shortens test times while maintaining key advantages of the current methods.

  8. A Rapidly Growing Instability in the Wake of Airfoils with Multiple Trailing Vortices

    NASA Astrophysics Data System (ADS)

    Ortega, Jason; Savas, Omer

    2000-11-01

    Dye flow visualization and two-dimensional PIV measurements are performed in the wakes of airfoils with multiple trailing vortices. The circulation-based Reynolds number of the vortices is of order 100,000. From the flow visualization data, an instability is observed to develop in the wake within 25 spans downstream of the airfoil. At approximately 50 spans, the instability becomes highly non-linear and three-dimensional, which, for some run parameters, results in an exchange of vorticity from opposite sides of the wake. By 200 spans, the dye is completely dispersed from the vortices and no coherent structures are visible in the wake. The PIV data indicate that the wake's "two-dimensional" kinetic energy decreases by a half at the onset of the non-linear behavior. By 200 spans, the "two-dimensional" kinetic energy is a fraction of its initial value. Further quantitative analyses demonstrate that at 200 spans, the resulting vortex core radius is on average 20of the individual vortices. The results of this study indicate that this rapidly growing instability has the potential for being a solution to the wake hazard problem

  9. Novel Flavohemoglobins of Mycobacteria

    PubMed Central

    Gupta, Sanjay; Pawaria, Sudesh; Lu, Changyuan; Yeh, Syun-Ru; Dikshit, Kanak L.

    2015-01-01

    Summary Flavohemoglobins (flavoHbs) constitute a distinct class of chimeric hemoglobins in which a globin domain is coupled with a ferredoxin reductase such as FAD- and NADH-binding modules. Structural features and active site of heme and reductase domains are highly conserved in various flavoHbs. A new class of flavoHbs, displaying crucial differences in functionally conserved regions of heme and reductase domains, have been identified in mycobacteria. Mining of microbial genome data indicated that the occurrence of such flavoHbs might be restricted to a small group of microbes unlike conventional flavoHbs that are widespread among prokaryotes and lower eukaryotes. One of the representative flavoHbs of this class, encoded by Rv0385 gene (MtbFHb) of Mycobacterium tuberculosis, has been cloned, expressed, and characterized. The ferric and deoxy spectra of MtbFHb displayed a hexacoordinate state indicating that its distal site may be occupied by an intrinsic amino acid or an external ligand and it may not be involved in nitric oxide detoxification. Phylogenetic analysis revealed that mycobacterial flavoHbs constitute a separate cluster distinct from conventional flavoHbs and may have novel function(s). PMID:21491561

  10. Comparison of two in-house real-time PCR assays with MTB Q-PCR Alert and GenoType MTBDRplus for the rapid detection of mycobacteria in clinical specimens.

    PubMed

    Seagar, Amie-Louise; Neish, Barry; Laurenson, Ian F

    2012-10-01

    An in-house IS6110 real-time PCR (IH IS6110), MTB Q-PCR Alert (Q-PCR) and GenoType MTBDRplus (MTBDR; Hain Lifescience) were compared for the direct detection of Mycobacterium tuberculosis complex (MTBC) in 87 specimens following automated NucliSENS easyMAG DNA extraction. This included 82 first smear-positive specimens and three smear-negative specimens. Another in-house real-time PCR with a Mycobacterium genus-specific probe for the internal transcribed spacer (ITS) region (IH ITS) was used to allow a full comparison with culture results. The sensitivities of IH IS6110, Q-PCR, MTBDR and IH ITS for MTBC detection were 100, 92, 87 and 87 %, respectively, compared with culture. Both IS6110-based real-time PCRs (in-house and Q-PCR) were similar in performance, with 91.2 % concordant results for MTBC detection. Inhibition rates were low, with zero to three specimens producing uninterpretable results. However, the Q-PCR failed to detect MTBC in five samples that were smear negative or had few acid-fast bacilli (one to 10 bacilli in 10 microscopic fields) detected by IH IS6110. IH ITS was the least sensitive assay but may be useful when used in conjunction with IS6110 PCR results to determine the presence of non-tuberculous mycobacteria in smear-negative specimens. None of the real-time PCR assays tested provided drug-resistance data. It was concluded that an IH IS6110 assay could easily be incorporated into the workflow of a diagnostic laboratory for rapid and accurate identification of MTBC from clinical specimens. The inclusion of an internal control and amplification of an ITS target enhance the diagnostic utility of the test.

  11. Mycobacteriophage cell binding proteins for the capture of mycobacteria

    PubMed Central

    Arutyunov, Denis; Singh, Upasana; El-Hawiet, Amr; Seckler, Henrique dos Santos; Nikjah, Sanaz; Joe, Maju; Bai, Yu; Lowary, Todd L; Klassen, John S; Evoy, Stephane; Szymanski, Christine M

    2014-01-01

    Slow growing Mycobacterium avium subsp. paratuberculosis (MAP) causes a deadly condition in cattle known as Johne's disease where asymptomatic carriers are the major source of disease transmission. MAP was also shown to be associated with chronic Crohn's disease in humans. Mycobacterium smegmatis is a model mycobacterium that can cause opportunistic infections in a number of human tissues and, rarely, a respiratory disease. Currently, there are no rapid, culture-independent, reliable and inexpensive tests for the diagnostics of MAP or M. smegmatis infections. Bacteriophages are viruses producing a number of proteins that effectively and specifically recognize the cell envelopes of their bacterial hosts. We demonstrate that the mycobacterial phage L5 minor tail protein Gp6 and lysin Gp10 are useful tools for the rapid capture of mycobacteria. Immobilized Gp10 was able to bind both MAP and M. smegmatis cells whereas Gp6 was M. smegmatis specific. Neither of the 2 proteins was able to capture E. coli, salmonella, campylobacter or Mycobacterium marinum cells. Gp6 was detected previously as a component of the phage particle and shows no homology to proteins with known function. Therefore, electrospray ionization mass spectrometry was used to determine whether recombinant Gp6 could bind to a number of chemically synthesized fragments of mycobacterial surface glycans. These findings demonstrate that mycobacteriophage proteins could be used as a pathogen capturing platform that can potentially improve the effectiveness of existing diagnostic methods. PMID:26713219

  12. Prevalence of nontuberculous mycobacteria isolated from environmental samples in Iran: A meta-analysis

    PubMed Central

    Khaledi, Azad; Bahador, Abbas; Esmaeili, Davood; Tafazoli, Alireza; Ghazvini, Kiarash; Mansury, Davood

    2016-01-01

    Background: While the most nontuberculous mycobacteria (NTMs) species are considered as opportunistic pathogens, some of them are related to several human infections. It is believed that environment is the main source for these infections. Distribution and scattering pattern of NTMs has not been well studied in Iran and a few studies about this subject have been done, so the aim of this study was to determine prevalence of NTMs in environmental samples from Iran. Materials and Methods: Data about prevalence of NTMs in environmental samples from Iran were obtained by searching databases. The studies presenting cross-sectional or cohort and the papers with sample size ≥30 were included. Then, the meta-analysis was performed using Comprehensive Meta-Analysis software and Cochran's Q and I2 tests. The strategy search was based PRISMA protocol is available online (PRISMA, http://www.prisma-statement.org). Results: The results of this meta-analysis showed that overall combined prevalence of NTMs in environmental samples from Iran was 38.3%. The frequency of NTM was higher in the north of Iran (73.2%). The most prevalent rapid-growing mycobacterium was Mycobacterium fortuitum (19.8%), and the most dominant slow-growing mycobacterium was Mycobacterium flavescens (16.8%). Conclusion: In regard to increasing incidence of disease in immunocompromised patients and existence of different types of mycobacteria species in environmental samples, efforts should be focused on measures that will specifically remove NTMs from habitats where susceptible individuals are exposed. PMID:27904603

  13. Prevalence of Non-Tuberculous Mycobacteria in Hospital Waters of Major Cities of Khuzestan Province, Iran

    PubMed Central

    Khosravi, Azar Dokht; Hashemi Shahraki, Abdolrazagh; Hashemzadeh, Mohammad; Sheini Mehrabzadeh, Rasa; Teimoori, Ali

    2016-01-01

    Non-tuberculous mycobacteria (NTM) are among the emerging pathogens in immunocompromised individuals including hospitalized patients. So, it is important to consider hospitals water supplies as a source for infection. The aim of this study was to determine the prevalence of NTM in the hospital aquatic systems of Khuzestan, South west of Iran. In total, 258 hospital water samples were collected and examined. After initial sample processing, sediment of each sample were inoculated into two Lowenstein-Jensen medium. The positive cultures were studied with phenotypic tests including growth rate, colony morphology, and pigmentation, with subsequent PCR- restriction enzyme analysis (PRA) and rpoB gene sequence analysis. Mycobacterial strains were isolated from 77 samples (29.8%), comprising 52 (70.1%) rapid growing, and 25 (32.4%) slow growing mycobacteria. Based on the overall results, M. fortuitum (44.1%) was the most common mycobacterial species in hospital water samples, followed by M. gordonae (n = 13, 16.8%) and M. senegalense (n = 5, 7.7%). In conclusion, current study demonstrated the NTM strains as one of the major parts of hospital water supplies with probable potential source for nosocomial infections. This finding also help to shed light on to the dynamics of the distribution and diversity of NTM in the water system of hospitals in the region of study. PMID:27148491

  14. Identification of different subtypes of rapid growing Atypical Mycobacterium from water and soil sources: Using PCR-RFLP using hsp65 and rRNA 16s-23s genes.

    PubMed

    Varahram, Mohammad; Farnia, Parissa; Saif, Shima; Marashian, Mehran; Farnia, Poopak; Ghanavi, Jaladein; Velayati, Ali Akbar

    2016-12-01

    Nontuberculosis mycobacteria (NTM) are a diverse group of microorganisms that cause a variety of diseases in humans including skin, respiratory, and gastrointestinal tract infection. Generally, NTM are classified into two categories: rapid (<7days) and slow growing (>7days). In this study, we aimed to investigate NTM frequency and prevalence in environmental samples. Additionally, we tried to identify various subtypes of isolated rapid growing mycobacteria (RGM). Through a prospective descriptive cross-sectional study, water and soil samples were gathered from four neighboring towns around Tehran, the capital of Iran, at different geographic directions. Every 100m(2) of the studied areas gave one sample containing 6g of soil in 3-5cm depth deposited in 50mL sterile water as sampling media. After digestion and decontamination, DNA from culture-positive specimens (RGM) were extracted using phenol-chloroform methods. Then the molecular identification of species and subspecies were performed using 16s-23s rRNA and hsp65 gene. In total, 341 RGM were found, out of which 322 (94.4%) were identified and 20 (5.8%) could not be identified. The most frequent RGM was, Mycobacterium fortuitum (72; 22%), Mycobacterium senegalense (58; 17.7%), Mycobacterium parafortuitum (44; 13.4%) and Mycobacterium conceptionense type 1 (24; 7.2%), and Mycobacterium cheloni type 1 (20; 6.0%). As shown in Table 1, M. fortuitum had more subtypes (8), and the frequency of subtypes 1 (27.7%), 4 (16.6%), and 5 (13.8%) were higher. Among subtypes of M. senegalense, subtype 1 had a higher frequency (70.4%) in comparison to subtype 2 (29.5%). M. cheloni had just one subtype. Our results showed M. fortuitum as the most prominent strain isolated from environmental samples. The frequency was similar in different places, irrespective of climatic variations. Availability of various subtypes of M. fortuitum might indicate a large circulation of this RGM in soil and water of Iranian territory. This high

  15. An aircraft study of rapid precipitation development and electrification in a growing convective cloud

    NASA Astrophysics Data System (ADS)

    Willis, P. T.; Hallett, J.; Black, R. A.; Hendricks, W.

    The rapid initial precipitation growth and initial electrification of a convective cloud, growing as a new cell on the upshear side of a cloud system in Florida, is traced from radar data and aircraft penetrations at the -7°C to -10°C level. This study combines radar, microphysical and electrical measurements so that an examination of the interactions between the cloud dynamics, microphysics and electrification is possible. The first pass (-7°C) was characterized by a strong 23 m/s updraft, all liquid cloud water, no precipitation, and no significant electrification. In the 300 s between the two penetrations, precipitation developed very rapidly from < 15 dBZ to < 45 dBZ, and the vertical component of the electric field increased from below the measurement threshold to - 25 kv/m. The second penetration, which started at - 7°C and ended at - 10°C, was still exclusively updraft, but with lesser peak velocities and a more complex structure; i.e., no downdraft, but with relative minima in the updraft. The microphysics of the second pass displayed a segment of exclusively cloud liquid water (no precipitation size hydrometeors), a small segment of all liquid precipitation size hydrometeors, a small region of mixed hydrometeors and an extensive region of graupel hydrometeors, ranging in size from 100 μm to several mm. High cloud liquid water coexisted with the liquid and graupel hydrometeors in the strong updrafts. The electrification was observed to occur exclusively in the segments of the cloud pass where graupel were observed. Within this graupel region, where the graupel often coexisted with supercooled cloud liquid water, significant electric field occurred only at relative minima in the updraft. These relative velocity minima were also minima in the cloud liquid water content. The observed updraft velocities in these relative minima were close to balance velocities for the observed larger graupel hydrometeors. The strongest updrafts, where the formation and

  16. Impact of human activities on the ecology of nontuberculous mycobacteria.

    PubMed

    Falkinham, Joseph O

    2010-06-01

    Nontuberculous mycobacteria (NTM) are environmental opportunistic pathogens of humans and animals. They are found in a wide variety of habitats to which humans are exposed, including drinking water distribution systems and household water and plumbing. In that regard, they are distinct from their obligate pathogenic relatives, the members of the Mycobacterium tuberculosis complex. Owing to the presence of NTM in the human environment, human activities have had direct impacts on their ecology and thereby their epidemiology. NTM are oligotrophic, able to grow at low organic matter concentrations and over a wide range of temperatures, and even at low oxygen concentrations. Thus, NTM are normal inhabitants of natural waters and drinking waters. Discovery of the presence of NTM-polluted soils is not surprising in light of the ability of NTM to degrade a variety of hydrocarbon pollutants. A major human activity selecting for the growth and predominance of mycobacteria in habitats is disinfection. In comparison to other bacteria, NTM are disinfectant, heavy metal and antibiotic resistant. Therefore, the use of any antimicrobial agent selects for mycobacteria. Use of disinfectant in drinking water treatment selects for mycobacteria that can grow and come to proliferate in drinking water distribution systems in the absence of disinfectant-sensitive competing microorganisms. NTM selection may also occur as a consequence of antibiotics in drinking water sources.

  17. Survey of solid waste generation and composition in a rapidly growing urban area in Central Nigeria.

    PubMed

    Sha'Ato, R; Aboho, S Y; Oketunde, F O; Eneji, I S; Unazi, G; Agwa, S

    2007-01-01

    The objective of this study was to carry out a field survey of the solid waste generation profile in parts of Makurdi, a rapidly growing urban city in north central Nigeria. The areas surveyed covered low, medium and high-density residential quarters, representing high/medium/low income groups in the area. Results of the survey show that the bulk ( approximately 82%) of the solid waste generated in the area originates from households, rather than from commercial, institutional or industrial premises. Of the waste from households, a substantial proportion consists of various putrescible materials (36-57%), with ash, dust and sand (combined) forming another significant proportion (21-41%). From the non-household sources, putrescible matter is also significant (23-45%), as is the combined ash/dust/sand fraction (32-36%). The quantity of plastics/cellophane materials from household and non-household sources was, however, comparable (6-10%). There was more paper from commercial and institutional premises (9-12%) than from household or small/medium scale industrial premises (2-4%). Glass (0.1-6.9%), metals (mostly cans and bottle corks, 0.7-3.4%) and textiles (0.3-6%) form only a minor proportion of the waste across generators. Waste generation rates were for households, 0.54kg/cap/day; for commercial, 0.018kg/m(2)/day; institutional, 0.015kg/m(2)/day while for small and medium scale industries, the rate was 0.47kg/m(2)/day. As there is no previous study of this kind in the Makurdi urban area, what is reported here may be taken as baseline for the entire area. The implications of the findings for solid waste management planning are discussed.

  18. A new biosensor for rapid BOD estimation by using immobilized growing cell beads.

    PubMed

    Su, Y C; Huang, J H; Liu, M L

    1986-04-01

    A closed, reactor-type sensor system for rapid estimation of BOD by the use of immobilized growing whole cells of a facultative bacterium, Bacillus polymyxa D-21, in kappa-carrageenan and an oxygen electrode is described. This system consists of a transformer, a recorder, a thermostated water bath (30 +/- 1 degree C), a magnetic stirrer (200 rpm), an oxygen electrode, and a flask containing 10 g (wet weight) immobilized cell beads of 2-3 mm in diameter. The total time required for an assay is less than 15 min. The calibration curves obtained show a linear relationship between the rate of oxygen consumption and the concentration of standard wastewater solution which contains equal amounts of glucose and L-glutamic acid each at a concentration below 10 mg/l. The measurement is independent of the pH of the wastewater solution under investigation. The rate of oxygen consumption was reproducible with an average relative error of 5.89% when a standard wastewater solution containing 12 mg/l of glucose and 12 mg/l of L-glutamic acid was assayed 54 times. When the immobilized cell beads were suspended in a 0.1 M phosphate buffer (pH 6.0) and stored at 25 degrees C, the response of the sensor system remained unchanged for about 30 days. However, based on the observed linear relationship between the rate of oxygen consumption and the concentration of standard wastewater solution, the sensor system should remain usable for at least 2 months. There is a good correlation between BOD5 determined by a conventional method and BOD5 determined by the proposed method when wastewater samples from a local fermentation factory were assayed.

  19. Evaluation of two line probe assays for rapid detection of Mycobacterium tuberculosis, tuberculosis (TB) drug resistance, and non-TB Mycobacteria in HIV-infected individuals with suspected TB.

    PubMed

    Luetkemeyer, Anne F; Kendall, Michelle A; Wu, Xingye; Lourenço, Maria Cristina; Jentsch, Ute; Swindells, Susan; Qasba, Sarojini S; Sanchez, Jorge; Havlir, Diane V; Grinsztejn, Beatriz; Sanne, Ian M; Firnhaber, Cynthia

    2014-04-01

    Limited performance data from line probe assays (LPAs), nucleic acid tests used for the rapid diagnosis of tuberculosis (TB), nontuberculosis mycobacteria (NTM), and Mycobacterium tuberculosis drug resistance are available for HIV-infected individuals, in whom paucibacillary TB is common. In this study, the strategy of testing sputum with GenoType MTBDRplus (MTBDR-Plus) and GenoType Direct LPA (Direct LPA) was compared to a gold standard of one mycobacterial growth indicator tube (MGIT) liquid culture. HIV-positive (HIV(+)) individuals with suspected TB from southern Africa and South America with <7 days of TB treatment had 1 sputum specimen tested with Direct LPA, MTBDR-Plus LPA, smear microscopy, MGIT, biochemical identification of mycobacterial species, and culture-based drug-susceptibility testing (DST). Of 639 participants, 59.3% were MGIT M. tuberculosis culture positive, of which 276 (72.8%) were acid-fast bacillus (AFB) smear positive. MTBDR-Plus had a sensitivity of 81.0% and a specificity of 100%, with sensitivities of 44.1% in AFB smear-negative versus 94.6% in AFB smear-positive specimens. For specimens that were positive for M. tuberculosis by MTBDR-Plus, the sensitivity and specificity for rifampin resistance were 91.7% and 96.6%, respectively, and for isoniazid (INH) they were 70.6% and 99.1%. The Direct LPA had a sensitivity of 88.4% and a specificity of 94.6% for M. tuberculosis detection, with a sensitivity of 72.5% in smear-negative specimens. Ten of 639 MGIT cultures grew Mycobacterium avium complex or Mycobacterium kansasii, half of which were detected by Direct LPA. Both LPA assays performed well in specimens from HIV-infected individuals, including in AFB smear-negative specimens, with 72.5% sensitivity for M. tuberculosis identification with the Direct LPA and 44.1% sensitivity with MTBDR-Plus. LPAs have a continued role for use in settings where rapid identification of INH resistance and clinically relevant NTM are priorities.

  20. Management of a rapidly growing peritoneal dialysis population at the First Affiliated Hospital of Sun Yat-sen University.

    PubMed

    Yu, Xueqing; Yang, Xiao; Huang, Naya

    2014-06-01

    Managing a rapidly growing peritoneal dialysis program with more than 1000 patients involves multiple challenges, labor constraints, logistics, and excessive geographic distance. This paper describes how Sun Yat-sen University, Guangzhou, China, manages those issues, while simultaneously improving quality of the care and, subsequently, clinical outcomes.

  1. Rapidly Growing Brtl/+ Mouse Model of Osteogenesis Imperfecta Improves Bone Mass and Strength with Sclerostin Antibody Treatment

    PubMed Central

    Sinder, Benjamin P.; Salemi, Joseph D.; Ominsky, Michael S.; Caird, Michelle S.; Marini, Joan C.; Kozloff, Kenneth M.

    2014-01-01

    Osteogenesis imperfecta (OI) is a heritable collagen-related bone dysplasia, characterized by brittle bones with increased fracture risk that presents most severely in children. Anti-resorptive bisphosphonates are frequently used to treat pediatric OI and controlled clinical trials have shown bisphosphonate therapy improves vertebral outcomes but has little benefit on long bone fracture rate. New treatments which increase bone mass throughout the pediatric OI skeleton would be beneficial. Sclerostin antibody (Scl-Ab) is a potential candidate anabolic therapy for pediatric OI and functions by stimulating osteoblastic bone formation via the canonical wnt signaling pathway. To explore the effect of Scl-Ab on the rapidly growing OI skeleton, we treated rapidly growing 3 week old Brtl/+ mice, harboring a typical heterozygous OI-causing Gly->Cys substitution on col1a1, for 5 weeks with Scl-Ab. Scl-Ab had anabolic effects in Brtl/+ and led to new cortical bone formation and increased cortical bone mass. This anabolic action resulted in improved mechanical strength to WT Veh levels without altering the underlying brittle nature of the material. While Scl-Ab was anabolic in trabecular bone of the distal femur in both genotypes, the effect was less strong in these rapidly growing Brtl/+ mice compared to WT. In conclusion, Scl-Ab was able to stimulate bone formation in a rapidly growing Brtl/+ murine model of OI, and represents a potential new therapy to improve bone mass and reduce fracture risk in pediatric OI. PMID:25445450

  2. Physiology of Mycobacteria

    PubMed Central

    Cook, Gregory M.; Berney, Michael; Gebhard, Susanne; Heinemann, Matthias; Cox, Robert A.; Danilchanka, Olga; Niederweis, Michael

    2013-01-01

    mycobacteria of course stems from the diseases they cause and, lest it be imagined that tuberculosis is a disease which has now been largely conquered and that leprosy is of relatively rare occurrence, current estimates for the number of case of tuberculosis and leprosy in the world today are 20,000,000 and 11,000,000, respectively (Bechelli and Dominguez, 1972). The annual estimated mortality rate is equally dramatic, namely 3,000,000 (World Health Organization, 1974). Also causing unease is the continuing isolation from tubercular patients of strains already resistant to one or more chemotherapeutic agent”. C. Ratledge (1976). PMID:19573696

  3. Rapid Mineralization of Benzo[a]pyrene by a Microbial Consortium Growing on Diesel Fuel

    PubMed Central

    Kanaly, Robert A.; Bartha, Richard; Watanabe, Kazuya; Harayama, Shigeaki

    2000-01-01

    A microbial consortium which rapidly mineralized the environmentally persistent pollutant benzo[a]pyrene was recovered from soil. The consortium cometabolically converted [7-14C]benzo[a]pyrene to 14CO2 when it was grown on diesel fuel, and the extent of benzo[a]pyrene mineralization was dependent on both diesel fuel and benzo[a]pyrene concentrations. Addition of diesel fuel at concentrations ranging from 0.007 to 0.2% (wt/vol) stimulated the mineralization of 10 mg of benzo[a]pyrene per liter 33 to 65% during a 2-week incubation period. When the benzo[a]pyrene concentration was 10 to 100 mg liter−1 and the diesel fuel concentration was 0.1% (wt/vol), an inoculum containing 1 mg of cell protein per liter (small inoculum) resulted in mineralization of up to 17.2 mg of benzo[a]pyrene per liter in 16 days. This corresponded to 35% of the added radiolabel when the concentration of benzo[a]pyrene was 50 mg liter−1. A radiocarbon mass balance analysis recovered 25% of the added benzo[a]pyrene solubilized in the culture suspension prior to mineralization. Populations growing on diesel fuel most likely promoted emulsification of benzo[a]pyrene through the production of surface-active compounds. The consortium was also analyzed by PCR-denaturing gradient gel electrophoresis of 16S rRNA gene fragments, and 12 dominant bands, representing different sequence types, were detected during a 19-day incubation period. The onset of benzo[a]pyrene mineralization was compared to changes in the consortium community structure and was found to correlate with the emergence of at least four sequence types. DNA from 10 sequence types were successfully purified and sequenced, and that data revealed that eight of the consortium members were related to the class Proteobacteria but that the consortium also included members which were related to the genera Mycobacterium and Sphingobacterium. PMID:11010861

  4. Effect of diet composition on pregnancy outcome in overnourished rapidly growing adolescent sheep.

    PubMed

    Wallace, Jacqueline M; Milne, John S; Redmer, Dale A; Aitken, Raymond P

    2006-12-01

    of exposure to high protein intakes. The data imply that it is high-energy intakes that are the primary cause of impaired placental development and adverse pregnancy outcome in rapidly growing adolescent sheep.

  5. Verification of Frequency in Species of Nontuberculous Mycobacteria in Kermanshah Drinking Water Supplies Using the PCR-Sequencing Method.

    PubMed

    Mohajeri, Parviz; Yazdani, Laya; Shahraki, Abdolrazagh Hashemi; Alvandi, Amirhoshang; Atashi, Sara; Farahani, Abbas; Almasi, Ali; Rezaei, Mansour

    2017-04-01

    Nontuberculous mycobacteria are habitants of environment, especially in aquatic systems. Some of them cause problems in immunodeficient patients. Over the last decade, 16S rRNA gene sequencing was established in 45 novel species of nontuberculous mycobacteria. Experiences revealed that this method underestimates the diversity, but does not distinguish between some of mycobacterium subsp. To recognize emerging rapidly growing mycobacteria and identify their subsp, rpoB gene sequencing has been developed. To better understand the transmission of nontuberculous mycobacterial species from drinking water and preventing the spread of illness with these bacteria, the aim of this study was to detect the presence of bacteria by PCR-sequencing techniques. Drinking water samples were collected from different areas of Kermanshah city in west of IRAN. After decontamination with cetylpyridinium chloride, samples were filtered with 0.45-micron filters, the filter transferred directly on growth medium waiting to appear in colonies, then DNA extraction and PCR were performed, and products were sent to sequencing. We found 35/110 (32%) nontuberculous mycobacterial species in drinking water samples, isolates included Mycobacterium goodii, Mycobacterium aurum, and Mycobacterium gastri with the most abundance (11.5%), followed by Mycobacterium smegmatis, Mycobacterium porcinum, Mycobacterium peregrinum, Mycobacterium mucogenicum, and Mycobacterium chelonae (8%). In this study, we recognized the evidence of contamination by nontuberculous mycobacteria in corroded water pipes. As a result of the high prevalence of these bacteria in drinking water in Kermanshah, this is important evidence of transmission through drinking water. This finding can also help public health policy makers control these isolates in drinking water supplies in Kermanshah.

  6. Pacemaker pocket infection due to environmental mycobacteria: Successful management of an outbreak and steps for prevention in future.

    PubMed

    Bharat, Vijaya; Hittinahalli, Vivek; Mishra, Meenakshi; Pradhan, Sridhar

    2016-01-01

    An outbreak of surgical site infection (SSI) due to environmental mycobacteria (EMB) occurred in a hospital in Eastern India. A quality improvement project (QIP) was undertaken to analyze the causes and prevent further outbreak. Step (1) Proof of the need: Four patients who had undergone pacemaker implantation consecutively during a 10-day period developed SSI. Step (2) Diagnostic journey: Since all patients developed SSI within 2 months of implantation, a common source of infection was likely. Atypical mycobacteria (AMB) were grown from surgical sites as well as from the surface of operation table, image intensifier, and lead aprons. It was a rapid growing variety that lacked pigment, a characteristic of EMB with pathogenic potential. The EMB was finally traced to its source, the overhead water tank. Step (3) Remedial journey: By thorough cleaning of the water tank and enriching its chlorine content, the EMB was eliminated from its source. Step (4) Holding the gains: Protocol for cleaning the water tank once in 3 months was made. A checklist was prepared to ensure compliance to asepsis protocol in the operation theater. In the ensuing 5 years, the infection did not recur. The bacteria that caused SSI were identified as EMB that grew in the water tank and contaminated the operation room. It could be eliminated by appropriate measures. Water is a potential reservoir for EMB. Use of the term 'environmental mycobacteria' instead of 'atypical mycobacteria' will generate awareness about contamination as the cause of SSI. Copyright © 2015 Cardiological Society of India. Published by Elsevier B.V. All rights reserved.

  7. Relation of Stem Diameter, Branch Basal Area, and Leaf Biomass in Rapidly Growing Loblolly Pine

    Treesearch

    Thomas M. Williams; Charles A. Gresham

    2004-01-01

    Twenty loblolly pines, growing in International Paper’s maximum growth experiment at Bainbridge GA, were destructively sampled at the end of the sixth growing season. Ten trees in the control and 10 in the maximum treatment were sampled. All trees were planted at a 2.4- by 3.6-m spacing and grown with complete competition control. The maximum trees also received...

  8. Malachite green interferes with postantibiotic recovery of mycobacteria.

    PubMed

    Gelman, Ekaterina; McKinney, John D; Dhar, Neeraj

    2012-07-01

    The genus Mycobacterium comprises slow-growing species with generation times ranging from hours to weeks. The protracted incubation time before colonies appear on solid culture medium can result in overgrowth by faster-growing microorganisms. To prevent contamination, the solid media used in laboratories and clinics for cultivation of mycobacteria contain the arylmethane compound malachite green, which has broad-spectrum antimicrobial activity. Malachite green has no impact on the plating efficiency of mycobacteria when cells are grown under normal conditions. However, we found that malachite green interfered with colony formation when bacteria were preexposed to antibiotics targeting cell wall biogenesis (isoniazid, ethionamide, ethambutol). This inhibitory effect of malachite green was not observed when bacteria were preexposed to antibiotics targeting cellular processes other than cell wall biogenesis (rifampin, moxifloxacin, streptomycin). Sputum specimens from tuberculosis patients are routinely evaluated on solid culture medium containing high concentrations of malachite green. This practice could lead to underestimation of bacterial loads and overestimation of chemotherapeutic efficacy.

  9. Nontuberculous Mycobacteria in Noncystic Fibrosis Bronchiectasis.

    PubMed

    Bonaiti, Giulia; Pesci, Alberto; Marruchella, Almerico; Lapadula, Giuseppe; Gori, Andrea; Aliberti, Stefano

    2015-01-01

    During the past decades, a growing interest has been raised in evaluating nontuberculous mycobacteria (NTM) in patients with noncystic fibrosis bronchiectasis (NCFBE). This paper reviews several aspects of the correlations between NTM and NCFBE, including pathogenesis, radiological features, diagnosis, and management. Bronchiectasis and NTM lung disease are connected, but which one comes first is still an unresolved question. The rate of NTM lung disease in NCFBE varies through the studies, from 5% to 30%. The most frequent species isolated is MAC. NCFBE patients affected by NTM infection frequently present coinfections, including both other different NTM species and microorganisms, such as P. aeruginosa. Once a diagnosis of NTM disease has been reached, the initiation of therapy is not always mandatory. NTM species isolated, patients' conditions, and disease severity and its evolution should be considered. Risk factors for disease progression in NCFBE patients with NTM are low body mass index, cavitary disease, consolidations, and macrolide resistance at presentation.

  10. Nontuberculous Mycobacteria in Noncystic Fibrosis Bronchiectasis

    PubMed Central

    Bonaiti, Giulia; Pesci, Alberto; Marruchella, Almerico; Lapadula, Giuseppe; Gori, Andrea

    2015-01-01

    During the past decades, a growing interest has been raised in evaluating nontuberculous mycobacteria (NTM) in patients with noncystic fibrosis bronchiectasis (NCFBE). This paper reviews several aspects of the correlations between NTM and NCFBE, including pathogenesis, radiological features, diagnosis, and management. Bronchiectasis and NTM lung disease are connected, but which one comes first is still an unresolved question. The rate of NTM lung disease in NCFBE varies through the studies, from 5% to 30%. The most frequent species isolated is MAC. NCFBE patients affected by NTM infection frequently present coinfections, including both other different NTM species and microorganisms, such as P. aeruginosa. Once a diagnosis of NTM disease has been reached, the initiation of therapy is not always mandatory. NTM species isolated, patients' conditions, and disease severity and its evolution should be considered. Risk factors for disease progression in NCFBE patients with NTM are low body mass index, cavitary disease, consolidations, and macrolide resistance at presentation. PMID:26106603

  11. Raman spectroscopic monitoring of the growth of pigmented and non-pigmented mycobacteria.

    PubMed

    Stöckel, Stephan; Stanca, Andrei Sebastian; Helbig, Jonathan; Rösch, Petra; Popp, Jürgen

    2015-11-01

    Raman microspectroscopy has increased in popularity in the field of microbiology because it allows a spectral fingerprinting of bacterial pathogens at an unrivaled speed, which is important for the early treatment of infectious diseases such as tuberculosis. An indispensable prerequisite for the success of this method is a profound knowledge, how the spectral profiles depend on the age of the bacteria. We therefore followed the growth of two rapidly growing Mycobacterium tuberculosis relatives, the pigmented Mycobacterium aurum, and the non-pigmented Mycobacterium smegmatis, by means of Raman microspectroscopy. Both species showed remarkable temporal changes in the single-bacteria Raman spectra: In the signatures of M. aurum, pigment-associated Raman signals could be detected not until 72 h of growth and also remained highly variable thereafter. The Raman spectra of M. smegmatis exhibited lipid signals presumably arising from mycolic acids, which are a hallmark feature of mycobacteria, but only after the bacteria reached the late stationary growth phase (>48 h). A principal component analysis thus classified the Raman spectra according to the cultivation age. In summary, these findings have to be reckoned with in future studies dealing with the identification of mycobacteria via Raman microspectroscopy. Graphical abstract Changes in the chemical composition of bacterial cells over growth time may influence the results of Raman spectroscopic studies of bacteria.

  12. The geographic diversity of nontuberculous mycobacteria isolated from pulmonary samples: an NTM-NET collaborative study.

    PubMed

    Hoefsloot, Wouter; van Ingen, Jakko; Andrejak, Claire; Angeby, Kristian; Bauriaud, Rosine; Bemer, Pascale; Beylis, Natalie; Boeree, Martin J; Cacho, Juana; Chihota, Violet; Chimara, Erica; Churchyard, Gavin; Cias, Raquel; Daza, Rosa; Daley, Charles L; Dekhuijzen, P N Richard; Domingo, Diego; Drobniewski, Francis; Esteban, Jaime; Fauville-Dufaux, Maryse; Folkvardsen, Dorte Bek; Gibbons, Noel; Gómez-Mampaso, Enrique; Gonzalez, Rosa; Hoffmann, Harald; Hsueh, Po-Ren; Indra, Alexander; Jagielski, Tomasz; Jamieson, Frances; Jankovic, Mateja; Jong, Eefje; Keane, Joseph; Koh, Wo-Jung; Lange, Berit; Leao, Sylvia; Macedo, Rita; Mannsåker, Turid; Marras, Theodore K; Maugein, Jeannette; Milburn, Heather J; Mlinkó, Tamas; Morcillo, Nora; Morimoto, Kozo; Papaventsis, Dimitrios; Palenque, Elia; Paez-Peña, Mar; Piersimoni, Claudio; Polanová, Monika; Rastogi, Nalin; Richter, Elvira; Ruiz-Serrano, Maria Jesus; Silva, Anabela; da Silva, M Pedro; Simsek, Hulya; van Soolingen, Dick; Szabó, Nora; Thomson, Rachel; Tórtola Fernandez, Teresa; Tortoli, Enrico; Totten, Sarah E; Tyrrell, Greg; Vasankari, Tuula; Villar, Miguel; Walkiewicz, Renata; Winthrop, Kevin L; Wagner, Dirk

    2013-12-01

    A significant knowledge gap exists concerning the geographical distribution of nontuberculous mycobacteria (NTM) isolation worldwide. To provide a snapshot of NTM species distribution, global partners in the NTM-Network European Trials Group (NET) framework (www.ntm-net.org), a branch of the Tuberculosis Network European Trials Group (TB-NET), provided identification results of the total number of patients in 2008 in whom NTM were isolated from pulmonary samples. From these data, we visualised the relative distribution of the different NTM found per continent and per country. We received species identification data for 20 182 patients, from 62 laboratories in 30 countries across six continents. 91 different NTM species were isolated. Mycobacterium avium complex (MAC) bacteria predominated in most countries, followed by M. gordonae and M. xenopi. Important differences in geographical distribution of MAC species as well as M. xenopi, M. kansasii and rapid-growing mycobacteria were observed. This snapshot demonstrates that the species distribution among NTM isolates from pulmonary specimens in the year 2008 differed by continent and differed by country within these continents. These differences in species distribution may partly determine the frequency and manifestations of pulmonary NTM disease in each geographical location.

  13. Gene probes versus classical methods in the identification of mycobacteria.

    PubMed

    Coelho, Andréa Gobetti Vieira; Zamarioli, Liliana Aparecida; Reis, Clemira Martins Pereira Vidal; Nascimento, Ana Carolina Chiou; Rodrigues, Juliana Dos Santos

    2008-11-01

    The emergence of tuberculosis/HIV co-infection and the increase in the number of cases of infection with nontuberculous mycobacteria (NTM) require rapid laboratory test results in the isolation and identification of mycobacteria. The objective of this study was to evaluate the identification of mycobacteria by means of gene probes in comparison with that obtained using classical biochemical methods. Between 2002 and 2004, 178 mycobacterial cultures, all testing positive for acid-fast bacilli, were analyzed. Samples were obtained from clinical specimens of patients with respiratory symptoms or with clinical suspicion of pulmonary tuberculosis/mycobacteriosis who were treated in the greater metropolitan area of Santos. The gene probe identified 137 samples (77%) as Mycobacterium tuberculosis complex and 41 (23%) as NTM. Discordant results between the methods (3%) were obtained only in the year of implementation (2002). When comparing the methods, the sensitivity, specificity, positive predictive value and negative predictive value of the gene probe method were 98%, 93%, 98% and 93%, respectively. Despite the cost, the identification of mycobacteria using the molecular technique is faster: maximum 3 h vs. 28-30 days for classical methods. The use of gene probes is a validated molecular technique. It is fast, easy to use and readily available on the market. It has high specificity and sensitivity, which justifies its implementation and routine use in referral laboratories, since it facilitates the diagnosis providing agile clinical interventions.

  14. Rapidly growing Brtl/+ mouse model of osteogenesis imperfecta improves bone mass and strength with sclerostin antibody treatment.

    PubMed

    Sinder, Benjamin P; Salemi, Joseph D; Ominsky, Michael S; Caird, Michelle S; Marini, Joan C; Kozloff, Kenneth M

    2015-02-01

    Osteogenesis imperfecta (OI) is a heritable collagen-related bone dysplasia, characterized by brittle bones with increased fracture risk that presents most severely in children. Anti-resorptive bisphosphonates are frequently used to treat pediatric OI and controlled clinical trials have shown that bisphosphonate therapy improves vertebral outcomes but has little benefit on long bone fracture rate. New treatments which increase bone mass throughout the pediatric OI skeleton would be beneficial. Sclerostin antibody (Scl-Ab) is a potential candidate anabolic therapy for pediatric OI and functions by stimulating osteoblastic bone formation via the canonical Wnt signaling pathway. To explore the effect of Scl-Ab on the rapidly growing OI skeleton, we treated rapidly growing 3week old Brtl/+ mice, harboring a typical heterozygous OI-causing Gly→Cys substitution on col1a1, for 5weeks with Scl-Ab. Scl-Ab had anabolic effects in Brtl/+ and led to new cortical bone formation and increased cortical bone mass. This anabolic action resulted in improved mechanical strength to WT Veh levels without altering the underlying brittle nature of the material. While Scl-Ab was anabolic in trabecular bone of the distal femur in both genotypes, the effect was less strong in these rapidly growing Brtl/+ mice compared to WT. In conclusion, Scl-Ab was able to stimulate bone formation in a rapidly growing Brtl/+ murine model of OI, and represents a potential new therapy to improve bone mass and reduce fracture risk in pediatric OI. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Isolation of Nontuberculous Mycobacteria from the Environment of Ghanian Communities Where Buruli Ulcer Is Endemic

    PubMed Central

    Aboagye, Samuel Yaw; Danso, Emelia; Ampah, Kobina Assan; Nakobu, Zuliehatu; Asare, Prince; Otchere, Isaac Darko; Röltgen, Katharina; Yirenya-Tawiah, Dzidzo

    2016-01-01

    ABSTRACT This study aimed to isolate nontuberculous mycobacterial species from environmental samples obtained from some selected communities in Ghana. To optimize decontamination, spiked environmental samples were used to evaluate four decontamination solutions and supplemented media, after which the best decontamination solution and media were used for the actual analysis. The isolates obtained were identified on the basis of specific genetic sequences, including heat shock protein 65, IS2404, IS2606, rpoB, and the ketoreductase gene, as needed. Among the methods evaluated, decontamination with 1 M NaOH followed by 5% oxalic acid gave the highest rate of recovery of mycobacteria (50.0%) and the lowest rate of contamination (15.6%). The cultivation medium that supported the highest rate of recovery of mycobacteria was polymyxin B-amphotericin B-nalidixic acid-trimethoprim-azlocillin–supplemented medium (34.4%), followed by isoniazid-supplemented medium (28.1%). Among the 139 samples cultivated in the main analysis, 58 (41.7%) yielded mycobacterial growth, 70 (50.4%) had no growth, and 11 (7.9%) had all inoculated tubes contaminated. A total of 25 different mycobacterial species were identified. Fifteen species (60%) were slowly growing (e.g., Mycobacterium ulcerans, Mycobacterium avium, Mycobacterium mantenii, and Mycobacterium malmoense), and 10 (40%) were rapidly growing (e.g., Mycobacterium chelonae, Mycobacterium fortuitum, and Mycobacterium abscessus). The occurrence of mycobacterial species in the various environmental samples analyzed was as follows: soil, 16 species (43.2%); vegetation, 14 species (38.0%); water, 3 species (8.0%); moss, 2 species (5.4%); snail, 1 species (2.7%); fungi, 1 species (2.7%). This study is the first to report on the isolation of M. ulcerans and other medically relevant nontuberculous mycobacteria from different environmental sources in Ghana. IMPORTANCE Diseases caused by mycobacterial species other than those that cause

  16. Evidence for Mycobacteria in Sarcoidosis

    PubMed Central

    Brownell, Isaac; Ramírez-Valle, Francisco; Sanchez, Miguel

    2011-01-01

    Despite its recognition as a distinct granulomatous disease for over a century, the etiology of sarcoidosis remains to be defined. Since the early 1900s, infectious agents have been suspected in causing sarcoidosis. For much of this time, mycobacteria were considered a likely culprit, yet until recently, the supporting evidence has been tenuous at best. In this review, we evaluate the reported association between mycobacteria and sarcoidosis. Historically, mycobacterial infection has been investigated using histologic stains, cultures of lesional tissue or blood, and identification of bacterial nucleic acids or bacterial antigens. More recently, advances in biochemical, molecular, and immunological methods have produced a more rigorous analysis of the antigenic drivers of sarcoidosis. The result of these efforts indicates that mycobacterial products likely play a role in at least a subset of sarcoidosis cases. This information, coupled with a better understanding of genetic susceptibility to this complex disease, has therapeutic implications. PMID:21659662

  17. Phylogenetic comparison of two polycyclic aromatic hydrocarbon-degrading mycobacteria.

    PubMed Central

    Govindaswami, M; Feldhake, D J; Kinkle, B K; Mindell, D P; Loper, J C

    1995-01-01

    Two mycobacterial strains previously isolated from fossil-fuel-contaminated environments and shown to degrade four- and/or five-ring polycyclic aromatic hydrocarbons were further characterized. The two strains, PYR-I and RJGII-135, had similar growth characteristics, colony morphologies, and scotochromogenic pigmentations. DNA amplification fingerprints obtained with total genomic DNA indicated some strain similarities but with several distinctly different bands. Moreover, phylogenetic analysis based upon essentially full-length 16S rRNA gene sequences separates the two strains as distinct species within the fast-growing group of mycobacteria. Although both strains are thermosensitive, strain PYR-I has the bulged U between positions 184 and 193 characteristic of thermotolerant mycobacteria. Both strains are of potential use for reintroduction into and bioremediation of polycyclic aromatic hydrocarbon-contaminated soils. PMID:7574631

  18. Phylogenetic comparison of two polycyclic aromatic hydrogen-degrading mycobacteria

    SciTech Connect

    Govindaswami, M.; Loper, J.C.; Feldhake, D.J.

    1995-09-01

    Two mycobacterial strains previously isolated from fossil-fuel-contaminated environments and shown to degrade four- and/or five-ring polycyclic aromatic hydrocarbons were further characterized. The two strains, PYR-I and RJGII-135, had similar growth characteristics, colony morphologies, and scotochromogenic pigmentations. DNA amplification fringerprints obtained with total genomic DNA indicated some strain similarities but with several distinctly different bands. Moreover, phylogenetic analysis based upon essentially full-length 16S rRNA gene sequences separates the two strains as distinct species within the fast-growing group of mycobacteria. Although both strains are thermosensitive, strain PYR-I has the bulged U between positions 184 and 193 characteristic of thermotolerant mycobacteria. Both strains are of potential use for reintroduction into and bioremediation of polycyclic aromatic hydrocarbon-contaminated soils. 26 refs., 4 figs., 1 tab.

  19. Rapidly growing primary gastric B-cell lymphoma after eradication of Helicobacter pylori.

    PubMed

    Furusyo, N; Kanamoto, K; Nakamura, S; Yao, T; Suekane, H; Yano, Y; Ariyama, I; Hayashi, J; Kashiwagi, S

    1999-10-01

    Helicobacter pylori (H. pylori) infection plays a decisive role in primary gastric B-cell lymphoma especially of mucosa-associated lymphoid tissue (MALT)-type. We treated a 47-year-old male patient with primary gastric B-cell lymphoma associated with H. pylori infection. Although antibiotic therapy for eradication of H. pylori caused great improvement in the low-grade MALT lymphoma-like lesion, the small areas of high-grade lesion rapidly formed a new bulky mass in only 8 weeks. This suggests that eradication of H. pylori is not effective for high-grade lymphoma.

  20. Antimicrobial susceptibility and MIC distribution of 41 drugs against clinical isolates from China and reference strains of nontuberculous mycobacteria.

    PubMed

    Li, Guilian; Pang, Hui; Guo, Qian; Huang, Mingxiang; Tan, Yanhong; Li, Chao; Wei, Jianhao; Xia, Yuanzhi; Jiang, Yi; Zhao, Xiuqin; Liu, Haican; Zhao, Li-Li; Liu, Zhiguang; Xu, Donglei; Wan, Kanglin

    2017-03-01

    To treat nontuberculous mycobacteria (NTM) infections more optimally, further research pertaining to mycobacterial susceptibility to antimicrobial agents is required. A total of 82 species of NTM reference strains and 23 species of NTM clinical isolates were included. Minimum inhibitory concentrations (MICs) for 41 drugs were determined using the microdilution method in cation-adjusted Mueller-Hinton broth. The results showed that most of the NTM were susceptible to aminoglycosides, quinolones, three macrolides (clarithromycin, azithromycin and roxithromycin), cefmetazole, linezolid and capreomycin. Rapidly growing mycobacterium strains were additionally susceptible to cefoxitin, clofazimine, rifapentine, doxycycline, minocycline, tigecycline, meropenem and sulfamethoxazole, whereas slowly growing mycobacterium strains were additionally susceptible to rifabutin. This study on the susceptibility of NTM includes the largest sample size of Chinese clinical isolates and reference strains. NTM species-specific drug susceptibility patterns suggested that it is urgent to identify the species of NTM, to normalise the treatment of NTM infectious disease and to clarify the resistance mechanisms of NTM.

  1. Cellular Midpalatal Suture Changes after Rapid Maxillary Expansion in Growing Subjects: A Case Report

    PubMed Central

    Caprioglio, Alberto; Fastuca, Rosamaria; Zecca, Piero Antonio; Beretta, Matteo; Mangano, Carlo; Piattelli, Adriano; Macchi, Aldo; Iezzi, Giovanna

    2017-01-01

    The present case report aimed to investigate immediate histologic changes in midpalatal suture in humans following rapid maxillary expansion compared to control. Three patients (mean age 8.3 ± 0.9 years) were enrolled in the case report and underwent midpalatal suture biopsy. Two patients underwent treatment before biopsy. The third patient did not show transversal maxillary deficiency and was enrolled as a control. Biopsy samples of midpalatal suture at 7 (subject 1) and 30 days (subject 2) after maxillary expansion as well as of one control (subject 3) were collected and processed for histology. In the control (subject 3) inter-digitations at the palatal suture gap were observed. At 7 days (subject 1) mature bone with small marrow spaces and trabecular bone with the peculiar storiform appearance inside the soft tissue and collagen fibers running parallel only in the central part were present. At 30 days (subject 2), a greater number of newly-formed bone trabeculae with a perpendicular orientation to the long axis of the suture could be seen. At 30 days the fibrous component of bone tissue was less represented compared to the sample at 7 days. Data from the preliminary histological results showed that bone formation was observed in the gap after rapid maxillary expansion, although the healing process was still ongoing. PMID:28287481

  2. Rapidly growing complex fibroadenoma with surrounding ductal hyperplasia mimics breast malignancy on serial F-18 FDG PET/CT imaging.

    PubMed

    Makis, William; Ciarallo, Anthony; Hickeson, Marc; Derbekyan, Vilma

    2011-07-01

    A 30-year-old woman was referred for an F-18 fluorodeoxyglucose (FDG) PET/CT to rule out lymphoma, and was found to have an incidental FDG-avid right breast nodule that grew significantly in size and FDG uptake on a subsequent scan, raising suspicion of a growing breast malignancy. Histologic evaluation showed a complex fibroadenoma with adenosis and surrounding ductal hyperplasia. Although variable F-18 FDG uptake in fibroadenomas has been described, a distinction between simple and complex fibroadenomas has not been made in the PET literature, even though complex fibroadenomas have a higher propensity to develop into malignancies. This case shows that a rapidly growing complex fibroadenoma can mimic a breast malignancy on serial F-18 FDG PET/CT scans, showing significant increase in both size and FDG-avidity on follow-up studies.

  3. Relative in vitro growth rates of duckweeds (Lemnaceae) - the most rapidly growing higher plants.

    PubMed

    Ziegler, P; Adelmann, K; Zimmer, S; Schmidt, C; Appenroth, K-J

    2015-01-01

    Relative growth rates (RGR), doubling times (DT) and relative weekly yields (RY) of 39 clones (ecotypes) from 13 species representing all five genera of duckweeds were determined under standardised cultivation conditions. RGR ranged overall from 0.153 to 0.519 day(-1) , DT from 1.34 to 4.54 days and RY from 2.9 to 37.8 week(-1) . The RGR and RY data can be compared directly to other published findings to only a limited extent on account of missing clonal designations for and limited accessibility to previously investigated clones, as well as the use of different data denominators. However, they are consistent with the published results of other comparative duckweed studies of similar scope in showing that RGR does not vary primarily at the level of the genus or species, but rather reflects the adaptation of individual clones to specific local conditions. The RGR data support the widely held assumption that duckweeds can grow faster than other higher plants and that they can thus surpass land-based agricultural crops in productivity. Duckweeds are highly promising for the production of biomass for nutrition and energy, but extensive clonal comparison will be required to identify the most suitable isolates for this purpose.

  4. Rapid growing clay coatings to reduce the fire threat of furniture.

    PubMed

    Kim, Yeon Seok; Li, Yu-Chin; Pitts, William M; Werrel, Martin; Davis, Rick D

    2014-02-12

    Layer-by-layer (LbL) assembly coatings reduce the flammability of textiles and polyurethane foam but require extensive repetitive processing steps to produce the desired coating thickness and nanoparticle fire retardant content that translates into a fire retardant coating. Reported here is a new hybrid bi-layer (BL) approach to fabricate fire retardant coatings on polyurethane foam. Utilizing hydrogen bonding and electrostatic attraction along with the pH adjustment, a fast growing coating with significant fire retardant clay content was achieved. This hybrid BL coating exhibits significant fire performance improvement in both bench scale and real scale tests. Cone calorimetry bench scale tests show a 42% and 71% reduction in peak and average heat release rates, respectively. Real scale furniture mockups constructed using the hybrid LbL coating reduced the peak and average heat release rates by 53% and 63%, respectively. This is the first time that the fire safety in a real scale test has been reported for any LbL technology. This hybrid LbL coating is the fastest approach to develop an effective fire retardant coating for polyurethane foam.

  5. Rapidly Growing Chest Wall Mass in a Case of Sporadic Metastatic Paraganglioma: Imaging With 4 Different PET Radiopharmaceuticals.

    PubMed

    Janssen, Ingo; Xekouki, Paraskevi; Nambuba, Joan; Chen, Clara C; Herscovitch, Peter; Millo, Corina M; Schrump, David S; Pacak, Karel

    2016-05-01

    Pheochromocytomas/paragangliomas (PGLs) are rare tumors and mostly benign. We report on a 32-year-old woman with metastatic PGL who was first diagnosed with an abdominal PGL at the age of 12 years. She soon developed metastatic disease and received several treatments including external beam radiation and chemotherapy. When she was referred to our institution in 2014, her major complaint was a rapidly growing chest wall mass on the left side. The patient was imaged at our institution with 4 different PET radiopharmaceuticals.

  6. GROWTH CHARACTERISTICS OF TUBERCLE BACILLI AND CERTAIN OTHER MYCOBACTERIA IN HELA CELLS

    PubMed Central

    Shepard, Charles C.

    1957-01-01

    By making use of the increased phagocytosis which follows the exposure of HeLa cells to tissue culture media containing selected horse sera, it was possible to introduce all of the mycobacterial species studied into the cells, where many of them proceeded to grow. Fully virulent strains of tubercle bacilli filled much of the cytoplasm in a few days and formed characteristic cords not seen with other strains. The strains said to be less virulent, R1Rv, BCG, H37Ra, and R1Ra, grew less rapidly and in characteristic patterns. Their rates of multiplication in HeLa cells were in the order named and correlated well with their reported pathogenicity for mice and guinea pigs. Six INH-resistant strains grew at rates characteristic of fully virulent strains. Among the "rapidly growing" species, M. phlei and M. smegmatis did not show evidence of growth in the cells, although M. fortuitum did. Some strains with optimal temperatures on bacteriological media below 37°C, M. balnei, M. marinum, and M. platypoecilus, grew rapidly in HeLa cells, especially at temperatures of 31 to 35°C. The growth patterns of the bacilli in HeLa cells appear sufficiently specific to be useful in differentiation among the mycobacteria. PMID:13385405

  7. Confinement-Induced Drug-Tolerance in Mycobacteria Mediated by an Efflux Mechanism.

    PubMed

    Luthuli, Brilliant B; Purdy, Georgiana E; Balagaddé, Frederick K

    2015-01-01

    Tuberculosis (TB) is the world's deadliest curable disease, responsible for an estimated 1.5 million deaths annually. A considerable challenge in controlling this disease is the prolonged multidrug chemotherapy (6 to 9 months) required to overcome drug-tolerant mycobacteria that persist in human tissues, although the same drugs can sterilize genetically identical mycobacteria growing in axenic culture within days. An essential component of TB infection involves intracellular Mycobacterium tuberculosis bacteria that multiply within macrophages and are significantly more tolerant to antibiotics compared to extracellular mycobacteria. To investigate this aspect of human TB, we created a physical cell culture system that mimics confinement of replicating mycobacteria, such as in a macrophage during infection. Using this system, we uncovered an epigenetic drug-tolerance phenotype that appears when mycobacteria are cultured in space-confined bioreactors and disappears in larger volume growth contexts. Efflux mechanisms that are induced in space-confined growth environments contribute to this drug-tolerance phenotype. Therefore, macrophage-induced drug tolerance by mycobacteria may be an effect of confined growth among other macrophage-specific mechanisms.

  8. Confinement-Induced Drug-Tolerance in Mycobacteria Mediated by an Efflux Mechanism

    PubMed Central

    Luthuli, Brilliant B.; Purdy, Georgiana E.; Balagaddé, Frederick K.

    2015-01-01

    Tuberculosis (TB) is the world’s deadliest curable disease, responsible for an estimated 1.5 million deaths annually. A considerable challenge in controlling this disease is the prolonged multidrug chemotherapy (6 to 9 months) required to overcome drug-tolerant mycobacteria that persist in human tissues, although the same drugs can sterilize genetically identical mycobacteria growing in axenic culture within days. An essential component of TB infection involves intracellular Mycobacterium tuberculosis bacteria that multiply within macrophages and are significantly more tolerant to antibiotics compared to extracellular mycobacteria. To investigate this aspect of human TB, we created a physical cell culture system that mimics confinement of replicating mycobacteria, such as in a macrophage during infection. Using this system, we uncovered an epigenetic drug-tolerance phenotype that appears when mycobacteria are cultured in space-confined bioreactors and disappears in larger volume growth contexts. Efflux mechanisms that are induced in space-confined growth environments contribute to this drug-tolerance phenotype. Therefore, macrophage-induced drug tolerance by mycobacteria may be an effect of confined growth among other macrophage-specific mechanisms. PMID:26295942

  9. Mycobacteria in nail salon whirlpool footbaths, California.

    PubMed

    Vugia, Duc J; Jang, Yvonne; Zizek, Candi; Ely, Janet; Winthrop, Kevin L; Desmond, Edward

    2005-04-01

    In 2000, an outbreak of Mycobacterium fortuitum furunculosis affected customers using whirlpool footbaths at a nail salon. We swabbed 30 footbaths in 18 nail salons from 5 California counties and found mycobacteria in 29 (97%); M. fortuitum was the most common. Mycobacteria may pose an infectious risk for pedicure customers.

  10. Mycobacteria in Nail Salon Whirlpool Footbaths, California

    PubMed Central

    Jang, Yvonne; Zizek, Candi; Ely, Janet; Winthrop, Kevin L.; Desmond, Edward

    2005-01-01

    In 2000, an outbreak of Mycobacterium fortuitum furunculosis affected customers using whirlpool footbaths at a nail salon. We swabbed 30 footbaths in 18 nail salons from 5 California counties and found mycobacteria in 29 (97%); M. fortuitum was the most common. Mycobacteria may pose an infectious risk for pedicure customers. PMID:15829204

  11. Methods for determining the antimicrobial susceptibility of mycobacteria.

    PubMed

    Alcaide, Fernando; Esteban, Jaime; González-Martin, Julià; Palacios, Juan-José

    2017-10-01

    Mycobacteria are a large group of microorganisms, multiple species of which are major causes of morbidity and mortality, such as tuberculosis and leprosy. At present, the emergence and spread of multidrug-resistant strains of Mycobacterium tuberculosis complex are one of the most serious health problems worldwide. Furthermore, in contrast to M. tuberculosis and Mycobacterium leprae, non-tuberculous mycobacteria (NTM) are more frequently isolated and, in many cases, treatment is based on drug susceptibility testing. This article is a review of the different methods to determine the in vitro drug susceptibility of M. tuberculosis complex and the most relevant NTM isolates. The molecular techniques currently used for rapid detection of resistance of clinical specimens are also analysed. Copyright © 2016 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  12. A rapidly growing moraine-dammed glacial lake on Ngozumpa Glacier, Nepal

    NASA Astrophysics Data System (ADS)

    Thompson, Sarah S.; Benn, Douglas I.; Dennis, Kathryn; Luckman, Adrian

    2012-04-01

    Moraine-dammed glacial lakes are becoming increasingly common in the Himalaya as a result of glacier mass loss, causing concern about glacier lake outburst flood risk. In addition to extant lakes, the potential exists for many more to form, as more glaciers ablate down to the level of potential moraine dams. In this paper, we document the recent rapid growth of, a moraine-dammed lake on Ngozumpa Glacier, Nepal. Using a combination of ground-based mapping and sonar surveys, aerial photographs (< 1 m resolution), and ASTER imagery (15 m resolution), processes and rates of lake expansion have been determined. The lake first formed between 1984 and 1992 when collapse of an englacial conduit allowed water to accumulate at the level of a gap in the lateral moraine, ~km from the glacier terminus. Lake growth was initially slow, but since 2001 it has undergone exponential growth at an average rate of 10% y-1. In 2009, the lake area was 300,000 m2, and its volume was at least 2.2 million m3. Calving, subaqueous melting, and melting of subaerial ice faces all contribute to the expansion of the lake; but large-scale, full-height slab calving is now the dominant contributor to growth. Comparison with other lakes in the region indicate that lake growth will likely continue unchecked whilst the spillway remains at its current level and may attain a volume of hundreds of millions of cubic metres within the next few decades.

  13. Method and apparatus for rapidly growing films on substrates using pulsed supersonic jets

    DOEpatents

    Eres, Diula; Lowndes, Douglas H.

    1992-01-01

    A method and apparatus for the rapid and economical deposition of uniform and high quality films upon a substrate for subsequent use in producing electronic devices, for example. The resultant films are either epitaxial (crystalline) or amorphous depending upon the incidence rate and the temperature and structure of the substrate. The deposition is carried out in a chamber maintained at about 10.sup.-6 Torr. A gaseous source of the material for forming the deposit is injected into the deposition chamber in the form of a pulsed supersonic jet so as to obtain a high incidence rate. The supersonic jet is produced by a pulsed valve between a relatively high presure reservoir, containing the source gaseous molecules, and the deposition chamber; the valve has a small nozzle orifice (e.g., 0.1-1.0 mm diameter). The type of deposit (crystalline amorphous) is then dependent upon the temperature and structure of the substrate. Very high deposition rates are achieved, and the deposit is very smooth and of uniform thickness. Typically the deposition rate is about 100 times that of much more expensive conventional molecular beam methods for deposition, and comparable to certain expensive plasma-assisted CVD methods of the art. The high growth rate of this method results in a reduced contamination of the deposit from other elements in the environment. The method is illustrated by the deposition of epitaxial and amorphour germanium films upon GaAs substrates.

  14. Protective effect of breastfeeding on diarrhea among children in a rapidly growing newly developed society.

    PubMed

    Ehlayel, Mohammad S; Bener, Abdulbari; Abdulrahman, Hatim M

    2009-01-01

    of infantile diarrhea. This observation is particularly important given the growing concern that, as an unwanted effect of 'modernization', breastfeeding is on the decline in Qatar and comparable populations elsewhere.

  15. Neonatal sepsis in a rapidly growing, tertiary neonatal intensive care unit: Trends over 18 years.

    PubMed

    Heo, Ju Sun; Shin, Seung Han; Jung, Young Hwa; Kim, Ee-Kyung; Choi, Eun Hwa; Kim, Han-Suk; Lee, Hoan Jong; Choi, Jung-Hwan

    2015-10-01

    We investigated changes in the admission patterns of neonatal intensive care units and the epidemiology of neonatal sepsis following the rapid expansion and improvements in neonatal intensive care. Data on the admission of neonates with culture-proven sepsis between 1996 and 2013 (period I, 1996-2005; period II, 2006-2013) were collected retrospectively. The admission of extremely low-birthweight (ELBW) infants increased between periods I and II (11.1 vs 28.7 infants per 1000 live births, P < 0.001). The survival rate of the ELBW infants improved (57.5 vs 80.1%, P < 0.001), and duration of hospital stay increased (median, 64 vs 80 days, P = 0.001). The incidence of sepsis among all infants and ELBW infants increased (all infants, 5.9 vs 12.7 cases per 1000 live births; ELBW infants, 189.5 vs 290.1 cases per 1000 live births). In ELBW infants, the incidence of sepsis caused by coagulase-negative Staphylococcus (CONS), significantly increased during period II (8.8 vs 25.4%, P = 0.039). On multivariate analysis, central vascular catheters and prolonged hospitalization were independently associated with increased sepsis rate, particularly CONS in ELBW infants. The inborn admission rate for ELBW infants has increased significantly and is accompanied by improved survival and longer hospital stay. The incidence of neonatal sepsis, particularly in ELBW infants, has also increased, and CONS has emerged as a major pathogen. Central vascular catheters and prolonged hospitalization could be independent risk factors for the increased sepsis rate, particularly sepsis due to CONS. © 2015 Japan Pediatric Society.

  16. Rapidly changing climatic conditions for wine grape growing in the Okanagan Valley region of British Columbia, Canada.

    PubMed

    Rayne, Sierra; Forest, Kaya

    2016-06-15

    A statistical analysis was conducted on long-term climate records for sites bordering Okanagan Lake in the Okanagan Valley viticultural region of British Columbia, Canada. Average wine grape growing season temperatures are increasing rapidly in the area over the post-1980 period at rates upwards of 7.0±1.3°C/century. Similar increases in the average dormant season temperature are evident. These temperature changes are likely some of the most extreme observed among the world's wine producing areas during the past few decades. Growing degree day base 10°C (GDD10) has increased by nearly 50% at some locations since the 1970s, resulting in major impacts on the corresponding climate classification for viticulture. If current climate trends continue, the southern and central portions of the region will likely enter Winkler region II within the next few decades, placing them in the same category as well-established warmer wine regions from France, Spain, Italy, and Australia. The large dormant season temperature increases over the last several decades have resulted in the area no longer being a cold season outlier when compared to most other cool-climate viticultural areas. Based on average growing season temperatures, the southern end of Okanagan Lake has moved out of the cool-climate viticultural classification and into the intermediate zone, while the central and northern regions are now at the cool/intermediate viticulture interface, similar to the historical positions of the Rhine Valley in Germany, northern Oregon in the United States, and the Loire Valley, Burgundy-Cote, Burgundy-Beaujolais, and Champagne appelations of France. The corresponding suitable grape species for the area have evolved into warmer region varietals during this time frame, having substantial economic impacts on producers. Increased temperatures are also expected to bring greater threats from agricultural pests, notably Pierce's disease from the bacterium Xylella fastidiosa.

  17. Selective killing of nonreplicating mycobacteria.

    PubMed

    Bryk, Ruslana; Gold, Benjamin; Venugopal, Aditya; Singh, Jasbir; Samy, Raghu; Pupek, Krzysztof; Cao, Hua; Popescu, Carmen; Gurney, Mark; Hotha, Srinivas; Cherian, Joseph; Rhee, Kyu; Ly, Lan; Converse, Paul J; Ehrt, Sabine; Vandal, Omar; Jiang, Xiuju; Schneider, Jean; Lin, Gang; Nathan, Carl

    2008-03-13

    Antibiotics are typically more effective against replicating rather than nonreplicating bacteria. However, a major need in global health is to eradicate persistent or nonreplicating subpopulations of bacteria such as Mycobacterium tuberculosis (Mtb). Hence, identifying chemical inhibitors that selectively kill bacteria that are not replicating is of practical importance. To address this, we screened for inhibitors of dihydrolipoamide acyltransferase (DlaT), an enzyme required by Mtb to cause tuberculosis in guinea pigs and used by the bacterium to resist nitric oxide-derived reactive nitrogen intermediates, a stress encountered in the host. Chemical screening for inhibitors of Mtb DlaT identified select rhodanines as compounds that almost exclusively kill nonreplicating mycobacteria in synergy with products of host immunity, such as nitric oxide and hypoxia, and are effective on bacteria within macrophages, a cellular reservoir for latent Mtb. Compounds that kill nonreplicating pathogens in cooperation with host immunity could complement the conventional chemotherapy of infectious disease.

  18. Natural Disasters and Nontuberculous Mycobacteria

    PubMed Central

    Bernhard, Jon N.; Chan, Edward D.

    2015-01-01

    Infectious diseases acquired by survivors of large-scale natural disasters complicate the recovery process. During events such as tsunamis, hurricanes, earthquakes, and tornados and well into the recovery period, victims often are exposed to water-soil mixtures that have relocated with indigenous microbes. Because nontuberculous mycobacteria (NTM) are ubiquitous in water and soil, there is potential for increased exposure to these organisms during natural disasters. In this hypothesis-driven commentary, we discuss the rise in NTM lung disease and natural disasters and examine the geographic overlap of NTM infections and disaster frequencies in the United States. Moreover, we show an increased number of positive NTM cultures from Louisiana residents in the years following three of the relatively recent epic hurricanes and posit that such natural disasters may help to drive the increased number of NTM infections. Finally, we advocate for increased environmental studies and surveillance of NTM infections before and after natural disasters. PMID:25644904

  19. Genomic characterization of Nontuberculous Mycobacteria

    PubMed Central

    Fedrizzi, Tarcisio; Meehan, Conor J.; Grottola, Antonella; Giacobazzi, Elisabetta; Fregni Serpini, Giulia; Tagliazucchi, Sara; Fabio, Anna; Bettua, Clotilde; Bertorelli, Roberto; De Sanctis, Veronica; Rumpianesi, Fabio; Pecorari, Monica; Jousson, Olivier; Tortoli, Enrico; Segata, Nicola

    2017-01-01

    Mycobacterium tuberculosis and Mycobacterium leprae have remained, for many years, the primary species of the genus Mycobacterium of clinical and microbiological interest. The other members of the genus, referred to as nontuberculous mycobacteria (NTM), have long been underinvestigated. In the last decades, however, the number of reports linking various NTM species with human diseases has steadily increased and treatment difficulties have emerged. Despite the availability of whole genome sequencing technologies, limited effort has been devoted to the genetic characterization of NTM species. As a consequence, the taxonomic and phylogenetic structure of the genus remains unsettled and genomic information is lacking to support the identification of these organisms in a clinical setting. In this work, we widen the knowledge of NTMs by reconstructing and analyzing the genomes of 41 previously uncharacterized NTM species. We provide the first comprehensive characterization of the genomic diversity of NTMs and open new venues for the clinical identification of opportunistic pathogens from this genus. PMID:28345639

  20. General Overview on Nontuberculous Mycobacteria, Biofilms, and Human Infection

    PubMed Central

    Faria, Sonia; Joao, Ines; Jordao, Luisa

    2015-01-01

    Nontuberculous mycobacteria (NTM) are emergent pathogens whose importance in human health has been growing. After being regarded mainly as etiological agents of opportunist infections in HIV patients, they have also been recognized as etiological agents of several infections on immune-competent individuals and healthcare-associated infections. The environmental nature of NTM and their ability to assemble biofilms on different surfaces play a key role in their pathogenesis. Here, we review the clinical manifestations attributed to NTM giving particular importance to the role played by biofilm assembly. PMID:26618006

  1. Pacemaker pocket infection due to environmental mycobacteria: Successful management of an outbreak and steps for prevention in future

    PubMed Central

    Bharat, Vijaya; Hittinahalli, Vivek; Mishra, Meenakshi; Pradhan, Sridhar

    2016-01-01

    Background An outbreak of surgical site infection (SSI) due to environmental mycobacteria (EMB) occurred in a hospital in Eastern India. Method A quality improvement project (QIP) was undertaken to analyze the causes and prevent further outbreak. Step (1) Proof of the need: Four patients who had undergone pacemaker implantation consecutively during a 10-day period developed SSI. Step (2) Diagnostic journey: Since all patients developed SSI within 2 months of implantation, a common source of infection was likely. Atypical mycobacteria (AMB) were grown from surgical sites as well as from the surface of operation table, image intensifier, and lead aprons. It was a rapid growing variety that lacked pigment, a characteristic of EMB with pathogenic potential. The EMB was finally traced to its source, the overhead water tank. Step (3) Remedial journey: By thorough cleaning of the water tank and enriching its chlorine content, the EMB was eliminated from its source. Step (4) Holding the gains: Protocol for cleaning the water tank once in 3 months was made. A checklist was prepared to ensure compliance to asepsis protocol in the operation theater. In the ensuing 5 years, the infection did not recur. Result The bacteria that caused SSI were identified as EMB that grew in the water tank and contaminated the operation room. It could be eliminated by appropriate measures. Interpretation Water is a potential reservoir for EMB. Use of the term ‘environmental mycobacteria’ instead of ‘atypical mycobacteria’ will generate awareness about contamination as the cause of SSI. PMID:26896269

  2. Utility of rpoB Gene Sequencing for Identification of Nontuberculous Mycobacteria in the Netherlands

    PubMed Central

    de Zwaan, Rina; van Ingen, Jakko

    2014-01-01

    In the Netherlands, clinical isolation of nontuberculous mycobacteria (NTM) has increased over the past decade. Proper identification of isolates is important, as NTM species differ strongly in clinical relevance. Most of the currently applied identification methods cannot distinguish between all different Mycobacterium species and complexes within species. rpoB gene sequencing exhibits a promising level of discrimination among rapidly and slowly growing mycobacteria, including the Mycobacterium avium complex. In this study, we prospectively compared rpoB gene sequencing with our routine algorithm of reverse line blot identification combined with partial 16S rRNA gene sequencing of 455 NTM isolates. rpoB gene sequencing identified 403 isolates to species level as 45 different known species and identified 44 isolates to complex level, and eight isolates remained unidentifiable to species level. In contrast, our reference reverse line blot assay with adjunctive 16S rRNA gene sequencing identified 390 isolates to species level (30 distinct species) and identified 56 isolates to complex level, and nine isolates remained unidentified. The higher discriminatory power of rpoB gene sequencing results largely from the distinction of separate species within complexes and subspecies. Also, Mycobacterium gordonae, Mycobacterium kansasii, and Mycobacterium interjectum were separated into multiple groupings with relatively low sequence similarity (98 to 94%), suggesting that these are complexes of closely related species. We conclude that rpoB gene sequencing is a more discriminative identification technique than the combination of reverse line blot and 16S rRNA gene sequencing and could introduce a major improvement in clinical care of NTM disease and the research on the epidemiology and clinical relevance of NTM. PMID:24808238

  3. Identification of mycobacteria by high-performance liquid chromatography.

    PubMed Central

    Butler, W R; Jost, K C; Kilburn, J O

    1991-01-01

    Mycolic acids extracted from saponified mycobacterial cells were examined as p-bromophenacyl esters by high-performance liquid chromatography (HPLC). Standard HPLC patterns were developed for species of Mycobacterium by examination of strains from culture collections and other well-characterized isolates. Relative retention times of peaks and peak height comparisons were used to develop a differentiation scheme that was 98% accurate for the species examined. A rapid, cost-effective HPLC method which offers an alternative approach to the identification of mycobacteria is described. PMID:1774251

  4. Molecular characterization of mycobacteria isolated from seals.

    PubMed

    Zumárraga, M J; Bernardelli, A; Bastida, R; Quse, V; Loureiro, J; Cataldi, A; Bigi, F; Alito, A; Castro Ramos, M; Samper, S; Otal, I; Martin, C; Romano, M I

    1999-09-01

    Tuberculosis (TB) was diagnosed in 10 seals from three species (Arctocephalus australis, Arctocephalus tropicalis and Otaria flavescens) found in South America. The mycobacteria isolated from these cases belonged to the Mycobacterium tuberculosis complex, as determined by RFLP using an IS6110 probe, spoligotyping, analysis of the 16S rRNA gene sequence and by PCR-restriction analysis of hsp65. Polymorphisms in gyrA, katG, oxyR and pncA were investigated in some of the isolates, as well as the presence of the MPB70 antigen. The insertion sequence IS6110 was present in three to seven copies in the genome of the mycobacteria isolated from seals. Using the IS6110 probe, six patterns (designated A, B, C, D, E and F) were identified from 10 different isolates. Patterns A and B were found for the mycobacteria isolated from two and four seals, respectively, indicating an epidemiological relationship between isolates grouped according to their IS6110 RFLP. The mycobacteria isolated from seals shared the majority of their IS6110 DNA-containing restriction fragments, and nine isolates had an identical spoligotype; only one isolate showed a minor difference in its spoligotype. In addition, none of these spoligotypes were found in other M. tuberculosis complex strains. These results suggest that the isolates from seals constitute a unique group of closely related strains. The mycobacteria isolated from seals showed polymorphisms at gyrA codon 95 and katG codon 463, as do group 1 M. tuberculosis, and M. bovis. Group 1 mycobacteria are associated with cluster cases. The spoligotypes found in the mycobacteria isolated from seals lack spacers 39-43, as does M. bovis, but the MPB70 antigen, which is highly expressed in M. bovis and minimally expressed in M. tuberculosis, was not detected in these mycobacteria. The mycobacteria isolated from seals also showed oxyR and pncA polymorphisms specific to M. tuberculosis. In conclusion, the mycobacteria that cause TB in seals in the South

  5. Examination of specimens for mycobacteria in clinical laboratories in 21 countries: a 10-year review of the UK National Quality Assessment Scheme for Mycobacteria Culture.

    PubMed

    Walton, C; Hawkey, P M; James, V L A

    2005-12-01

    Results from clinical diagnostic microbiology laboratories taking part in the UK National Quality Assessment Service (UK NEQAS) scheme for Mycobacteria Culture between 1993 and 2003 were evaluated and assessed to determine whether the perceived increase in the use of rapid methods is improving time-to-positive reporting of results. Four simulated sputum specimens containing mycobacteria in mixed cultures with normal commensal organisms were distributed three times a year. Participating laboratories were required to report on the presence of 'mycobacteria' and on the time required to obtain a positive result. The overall level of performance with the mycobacteria culture external quality assessment specimens remained consistently high, with an average success rate of 94% over 10 years. The mean time-to-positive decreased from 24 to 17 days during the previous 8 years. A survey questionnaire, circulated in 2002, addressed the use of continuous automated mycobacterial liquid culture (CAMLiC) and molecular methods. The increase in the use of rapid culture methods for the detection of Mycobacterium tuberculosis has resulted in an overall reduction in time-to-positive data reported by participants, and has provided an indication of participants' ability to meet the 21-day target recommended by the CDC for the detection and identification of M. tuberculosis.

  6. Miniaturized extinction culturing is the preferred strategy for rapid isolation of fast-growing methane-oxidizing bacteria.

    PubMed

    Hoefman, Sven; van der Ha, David; De Vos, Paul; Boon, Nico; Heylen, Kim

    2012-05-01

    Methane-oxidizing bacteria (MOB) have a large potential as a microbial sink for the greenhouse gas methane as well as for biotechnological purposes. However, their application in biotechnology has so far been hampered, in part due to the relative slow growth rate of the available strains. To enable the availability of novel strains, this study compares the isolation of MOB by conventional dilution plating with miniaturized extinction culturing, both performed after an initial enrichment step. The extinction approach rendered 22 MOB isolates from four environmental samples, while no MOB could be isolated by plating. In most cases, extinction culturing immediately yielded MOB monocultures making laborious purification redundant. Both type I (Methylomonas spp.) and type II (Methylosinus sp.) MOB were isolated. The isolated methanotrophic diversity represented at least 11 different strains and several novel species based on 16S rRNA gene sequence dissimilarity. These strains possessed the particulate (100%) and soluble (64%) methane monooxygenase gene. Also, 73% of the strains could be linked to a highly active fast-growing mixed MOB community. In conclusion, miniaturized extinction culturing was more efficient in rapidly isolating numerous MOB requiring little effort and fewer materials, compared with the more widely applied plating procedure. This miniaturized approach allowed straightforward isolation and could be very useful for subsequent screening of desired characteristics, in view of their future biotechnological potential.

  7. Miniaturized extinction culturing is the preferred strategy for rapid isolation of fast‐growing methane‐oxidizing bacteria

    PubMed Central

    Hoefman, Sven; van der Ha, David; De Vos, Paul; Boon, Nico; Heylen, Kim

    2012-01-01

    Summary Methane‐oxidizing bacteria (MOB) have a large potential as a microbial sink for the greenhouse gas methane as well as for biotechnological purposes. However, their application in biotechnology has so far been hampered, in part due to the relative slow growth rate of the available strains. To enable the availability of novel strains, this study compares the isolation of MOB by conventional dilution plating with miniaturized extinction culturing, both performed after an initial enrichment step. The extinction approach rendered 22 MOB isolates from four environmental samples, while no MOB could be isolated by plating. In most cases, extinction culturing immediately yielded MOB monocultures making laborious purification redundant. Both type I (Methylomonas spp.) and type II (Methylosinus sp.) MOB were isolated. The isolated methanotrophic diversity represented at least 11 different strains and several novel species based on 16S rRNA gene sequence dissimilarity. These strains possessed the particulate (100%) and soluble (64%) methane monooxygenase gene. Also, 73% of the strains could be linked to a highly active fast‐growing mixed MOB community. In conclusion, miniaturized extinction culturing was more efficient in rapidly isolating numerous MOB requiring little effort and fewer materials, compared with the more widely applied plating procedure. This miniaturized approach allowed straightforward isolation and could be very useful for subsequent screening of desired characteristics, in view of their future biotechnological potential. PMID:22070783

  8. Nontuberculous Mycobacteria Immune Reconstitution Syndrome

    PubMed Central

    Mogambery, J. C.; Motala, A.; Padayachee, K.; Jozi, C.; Dawood, H.

    2014-01-01

    The prevalence of nontuberculous mycobacteria infection (NTM) in Sub-Saharan Africa is estimated to be less than 1%. NTM is often underdiagnosed or misdiagnosed as tuberculosis in patients who present with immune reconstitution syndrome (IRS) following initiation of antiretroviral treatment (ART). Immune reconstitution syndrome is common in patients who start ART with low CD4 counts and high HIV viral load. Furthermore, Mycobacterium avium complex (MAC) commonly infects those with CD4 counts less than 50 cells/mm3. Three patients, with low baseline CD4 counts, presenting with NTM following the initiation of antiretroviral treatment are described in this case series. The first patient presented with disseminated NTM two weeks after commencing antiretroviral treatment. Acid fast bacilli were found in the liver, duodenum, and bone marrow and were suggestive of MAC microscopically. The second developed cervical lymphadenitis following the initiation of ART. Lymph node aspirate culture grew NTM. The last patient developed pancytopenia after 3 months of ART. AFB was seen on bone marrow biopsy. Culture of the bone marrow aspirate was suggestive of NTM. All three patients improved on ethambutol, clarithromycin, and rifampicin. NTM may be underdiagnosed in areas with a high TB prevalence and should be actively excluded by culture. PMID:25435881

  9. Nontuberculous mycobacteria immune reconstitution syndrome.

    PubMed

    Mogambery, J C; Motala, A; Padayachee, K; Jozi, C; Dawood, H

    2014-01-01

    The prevalence of nontuberculous mycobacteria infection (NTM) in Sub-Saharan Africa is estimated to be less than 1%. NTM is often underdiagnosed or misdiagnosed as tuberculosis in patients who present with immune reconstitution syndrome (IRS) following initiation of antiretroviral treatment (ART). Immune reconstitution syndrome is common in patients who start ART with low CD4 counts and high HIV viral load. Furthermore, Mycobacterium avium complex (MAC) commonly infects those with CD4 counts less than 50 cells/mm(3). Three patients, with low baseline CD4 counts, presenting with NTM following the initiation of antiretroviral treatment are described in this case series. The first patient presented with disseminated NTM two weeks after commencing antiretroviral treatment. Acid fast bacilli were found in the liver, duodenum, and bone marrow and were suggestive of MAC microscopically. The second developed cervical lymphadenitis following the initiation of ART. Lymph node aspirate culture grew NTM. The last patient developed pancytopenia after 3 months of ART. AFB was seen on bone marrow biopsy. Culture of the bone marrow aspirate was suggestive of NTM. All three patients improved on ethambutol, clarithromycin, and rifampicin. NTM may be underdiagnosed in areas with a high TB prevalence and should be actively excluded by culture.

  10. Comparison of culture and qPCR methods in detection of mycobacteria from drinking waters.

    PubMed

    Räsänen, Noora H J; Rintala, Helena; Miettinen, Ilkka T; Torvinen, Eila

    2013-04-01

    Environmental mycobacteria are common bacteria in man-made water systems and may cause infections and hypersensitivity pneumonitis via exposure to water. We compared a generally used cultivation method and a quantitative polymerase chain reaction (qPCR) method to detect mycobacteria in 3 types of drinking waters: surface water, ozone-treated surface water, and groundwater. There was a correlation between the numbers of mycobacteria obtained by cultivation and qPCR methods, but the ratio of the counts obtained by the 2 methods varied among the types of water. The qPCR counts in the drinking waters produced from surface or groundwater were 5 to 34 times higher than culturable counts. In ozone-treated surface waters, both methods gave similar counts. The ozone-treated drinking waters had the highest concentration of assimilable organic carbon, which may explain the good culturability. In warm tap waters, qPCR gave 43 times higher counts than cultivation, but both qPCR counts and culturable counts were lower than those in the drinking waters collected from the same sites. The TaqMan qPCR method is a rapid and sensitive tool for total quantitation of mycobacteria in different types of clean waters. The raw water source and treatments affect both culturability and total numbers of mycobacteria in drinking waters.

  11. Natural and acquired macrolide resistance in mycobacteria.

    PubMed

    Doucet-Populaire, F; Buriánková, K; Weiser, J; Pernodet, J-L

    2002-12-01

    The genus Mycobacterium contains two of the most important human pathogens, Mycobacterium tuberculosis and Mycobacterium leprae, the etiologic agents of tuberculosis and leprosy, respectively. Other mycobacteria are mostly saprophytic organisms, living in soil and water, but some of them can cause opportunistic infections. The increasing incidence of tuberculosis as well as infections with non-tuberculous mycobacteria (NTM) in AIDS patients has renewed interest in molecular mechanisms of drug resistance in these pathogens. Mycobacteria show a high degree of intrinsic resistance to most common antibiotics. For instance, species from the M. tuberculosis complex (MTC) are intrinsically resistant to macrolides. Nevertheless, some semi-synthetic macrolides as the erythromycin derivatives clarithromycin, azithromycin and most recently the ketolides, are active against NTM, particularly Mycobacterium avium, and some of them are widely used for infection treatment. However, shortly after the introduction of these new drugs, resistant strains appeared due to mutations in the macrolide target, the ribosome. The mycobacterial cell wall with its specific composition and structure is considered to be a major factor in promoting the natural resistance of mycobacteria to various antibiotics. However, to explain the difference in macrolide sensitivity between the MTC and NTM, the synergistic contribution of a specific resistance mechanism might be required, in addition to possible differences in cell wall permeability. This mini-review summarizes the current knowledge on the natural and acquired macrolide resistance in mycobacteria, gives an overview of potential mechanisms implicated in the intrinsic resistance and brings recent data concerning a macrolide resistance determinant in the MTC.

  12. BIOSYNTHESIS OF ERGOTHIONEINE AND HERCYNINE BY MYCOBACTERIA

    PubMed Central

    Genghof, Dorothy S.; Damme, Olga Van

    1964-01-01

    Genghof, Dorothy S. (Yeshiva University, New York, N.Y.), and Olga Van Damme. Biosynthesis of ergothioneine and hercynine by mycobacteria. J. Bacteriol. 87:852–862. 1964.—Ergothioneine and hercynine were found to be synthesized by a wide variety of mycobacteria grown in chemically defined media free from these compounds. The cultures examined included 53 recently isolated and laboratory strains of Mycobacterium tuberculosis, 26 “unclassified” mycobacteria (Runyon groups I to IV), and representatives of most other species in the genus. Purification and separation of the betaines was achieved by means of chromatography on two successive alumina columns. Photometric measurement of the diazotized effluents from the second column permitted amounts of each compound to be determined. Measurement of hercynine by this method was made possible for the first time by the development of a standard curve. The pathway of ergothioneine biosynthesis in mycobacteria, as judged by the use S35-sulfate and l-histidine-2-C14 as tracers, appears similar to that found in Neurospora crassa and Claviceps purpurea, that is, from histidine to ergothioneine via hercynine. None of a small group of bacteria other than mycobacteria was found to produce ergothioneine. Two strains of group A streptococci and one of Escherichia coli produced hercyninelike material, as yet unidentified. PMID:14137624

  13. Rapidly growing black holes and host galaxies in the distant Universe from the Herschel Radio Galaxy Evolution Project

    NASA Astrophysics Data System (ADS)

    Drouart, G.; De Breuck, C.; Vernet, J.; Seymour, N.; Lehnert, M.; Barthel, P.; Bauer, F. E.; Ibar, E.; Galametz, A.; Haas, M.; Hatch, N.; Mullaney, J. R.; Nesvadba, N.; Rocca-Volmerange, B.; Röttgering, H. J. A.; Stern, D.; Wylezalek, D.

    2014-06-01

    We present results from a comprehensive survey of 70 radio galaxies at redshifts 1 growing extremely rapidly, having SFR ≈ 100-5000 M⊙ yr-1 and ṀBH ≈ 1-100 M⊙ yr-1. The mean specific SFRs (sSFR) of radio galaxies at z> 2.5 are higher than the sSFR of typical star forming galaxies over the same redshift range, but are similar or perhaps lower than the galaxy population for radio galaxies at z< 2.5. By comparing the sSFR and the specific ṀBH (sṀBH), we conclude that black holes in radio loud AGN are already, or soon will be, overly massive compared to their host galaxies in terms of expectations from the local MBH-MGal relation. In order to catch up with the black hole, the galaxies require about an order of magnitude more time to grow in mass at the observed SFRs compared to the time the black hole is actively accreting

  14. Mycobacterium sediminis sp. nov. and Mycobacterium arabiense sp. nov., two rapidly growing members of the genus Mycobacterium.

    PubMed

    Zhang, Dao-Feng; Chen, Xiu; Zhang, Xiao-Mei; Zhi, Xiao-Yang; Yao, Ji-Cheng; Jiang, Yi; Xiong, Zhi; Li, Wen-jun

    2013-11-01

    Two novel isolates of rapidly growing, Gram-stain-positive, non-chromogenic species of the genus Mycobacterium, strain YIM M13028(T) from a sediment sample collected from the South China Sea (19° 30.261' N 111° 0.247' E) at a depth of 42 m and strain YIM 121001(T) from a coastal zone sand sample collected in Dubai, United Arab Emirates, were obtained in our laboratory. Their taxonomic positions were determined by a polyphasic approach. Good growth of the two strains was observed at 28 °C and pH 7.0 with 0-2 % NaCl on tryptic soy agar medium. Both strains formed round orange-red colonies, strain YIM M13028(T) had a rough surface, while YIM 121001(T) was smooth. Cellular fatty acids, whole-cell protein profiles and TLC analysis of their mycolic acids show significant differences from reference stains. Phenotypic characteristics and multilocus sequence analysis (MLSA) of 16S rRNA gene, hsp65, rpoB and 16S-23S internal transcribed spacer (ITS) sequences indicated that both strains YIM M13028(T) and YIM 121001(T) belong to the genus Mycobacterium. DNA-DNA hybridization values revealed a low relatedness (<70 %) of the two isolates with the type strains Mycobacterium neoaurum DSM 44074(T) and Mycobacterium hodleri DSM 44183(T). The low DNA-DNA hybridization values (40.4±3.5 %) between strains YIM M13028(T) and YIM 121001(T) and phenotypic distinctiveness indicated that the two strains were representatives of different novel species of the genus Mycobacterium. The names proposed for these novel species are Mycobacterium sediminis sp. nov. and Mycobacterium arabiense sp. nov., and the type strains are YIM M13028(T) ( = DSM 45643(T) = KCTC 19999(T)) and YIM 121001(T) ( = DSM 45768(T) = JCM 18538(T)), respectively.

  15. Effects of landscape change on fish assemblage structure in a rapidly growing metropolitan area in North Carolina, USA

    USGS Publications Warehouse

    Kennen, J.G.; Chang, M.; Tracy, B.H.

    2005-01-01

    We evaluated a comprehensive set of natural and land-use attributes that represent the major facets of urban development at fish monitoring sites in the rapidly growing Raleigh-Durham, North Carolina metropolitan area. We used principal component and correlation analysis to obtain a nonredundant subset of variables that extracted most variation in the complete set. With this subset of variables, we assessed the effect of urban growth on fish assemblage structure. We evaluated variation in fish assemblage structure with nonmetric multidimensional scaling (NMDS). We used correlation analysis to identify the most important environmental and landscape variables associated with significant NMDS axes. The second NMDS axis is related to many indices of land-use/land-cover change and habitat. Significant correlations with proportion of largest forest patch to total patch size (r = -0.460, P < 0.01), diversity of patch types (r = 0.554, P < 0.001), and population density (r = 0.385, P < 0.05) helped identify NMDS axis 2 as a disturbance gradient. Positive and negative correlations between the abundance of redbreast sunfish Lepomis auritus and bluehead chub Nocomis leptocephalus, respectively, and NMDS axis 2 also were evident. The North Carolina index of biotic integrity and many of its component metrics were highly correlated with urbanization. These results indicate that aquatic ecosystem integrity would be optimized by a comprehensive integrated management strategy that includes the preservation of landscape function by maximizing the conservation of contiguous tracts of forested lands and vegetative cover in watersheds. ?? 2005 by the American Fisheries Society.

  16. Detection of Mycobacteria by Culture and DNA-Based Methods in Animal-Derived Food Products Purchased at Spanish Supermarkets

    PubMed Central

    Sevilla, Iker A.; Molina, Elena; Tello, Maitane; Elguezabal, Natalia; Juste, Ramón A.; Garrido, Joseba M.

    2017-01-01

    Mycobacteria include obligate and opportunistic pathogens that cause significant human and animal disease. The burden of tuberculosis has been largely reduced in developed territories but remains a huge problem worldwide. The significance of nontuberculous mycobacteria is growing considerably, especially in developed regions with higher life expectancy and more therapy-related immunosuppressed individuals. Due to their robustness mycobacteria can contaminate animal products by direct transmission from infected individuals or by environmental contamination during processing. The situation at market level is poorly known. Most studies analyzing commercially available foods are limited to a small or local scale and mainly focused on a particular mycobacterial species. There is a need to investigate if animal products that have passed the established controls to be for sale at main supermarkets could represent a route of contact with any mycobacteria. Thus, our goal was to study the prevalence of mycobacteria in these foods to assess if this could represent a source of human exposure. Five stores from the main supermarket chains in Spain were selected. 138 dairy and 119 meat products were purchased. All were processed using culture and multiplex real-time PCR methods. Additional molecular methods were used to specifically identify any positive result. Mycobacterium avium subsp. hominissuis (2), M. avium subsp. avium (1), and M. fortuitum (1) were isolated from powdered infant formula and ground beef, chicken sausage, and mortadella cold cut, respectively. Mycobacterial DNA (M. avium, M. tuberculosis complex and other nontuberculous mycobacteria) was detected in 15% of dairy products and 2% of meat products. These results show that the prevalence of viable mycobacteria in foods of animal origin obtained at the supermarket was not substantial although a considerable proportion of them contained mycobacterial DNA. Contact with mycobacteria through this route could be

  17. Detection of Mycobacteria by Culture and DNA-Based Methods in Animal-Derived Food Products Purchased at Spanish Supermarkets.

    PubMed

    Sevilla, Iker A; Molina, Elena; Tello, Maitane; Elguezabal, Natalia; Juste, Ramón A; Garrido, Joseba M

    2017-01-01

    Mycobacteria include obligate and opportunistic pathogens that cause significant human and animal disease. The burden of tuberculosis has been largely reduced in developed territories but remains a huge problem worldwide. The significance of nontuberculous mycobacteria is growing considerably, especially in developed regions with higher life expectancy and more therapy-related immunosuppressed individuals. Due to their robustness mycobacteria can contaminate animal products by direct transmission from infected individuals or by environmental contamination during processing. The situation at market level is poorly known. Most studies analyzing commercially available foods are limited to a small or local scale and mainly focused on a particular mycobacterial species. There is a need to investigate if animal products that have passed the established controls to be for sale at main supermarkets could represent a route of contact with any mycobacteria. Thus, our goal was to study the prevalence of mycobacteria in these foods to assess if this could represent a source of human exposure. Five stores from the main supermarket chains in Spain were selected. 138 dairy and 119 meat products were purchased. All were processed using culture and multiplex real-time PCR methods. Additional molecular methods were used to specifically identify any positive result. Mycobacterium avium subsp. hominissuis (2), M. avium subsp. avium (1), and M. fortuitum (1) were isolated from powdered infant formula and ground beef, chicken sausage, and mortadella cold cut, respectively. Mycobacterial DNA (M. avium, M. tuberculosis complex and other nontuberculous mycobacteria) was detected in 15% of dairy products and 2% of meat products. These results show that the prevalence of viable mycobacteria in foods of animal origin obtained at the supermarket was not substantial although a considerable proportion of them contained mycobacterial DNA. Contact with mycobacteria through this route could be

  18. Small RNAs in mycobacteria: an unfolding story

    PubMed Central

    Haning, Katie; Cho, Seung Hee; Contreras, Lydia M.

    2014-01-01

    Mycobacteria represent a class of powerful pathogens, including those causing tuberculosis and leprosy, which continue to be worldwide health challenges. In the last 20 years, an abundance of non-coding, small RNAs (sRNAs) have been discovered in model bacteria and gained significant attention as regulators of cellular responses, including pathogenesis. Naturally, a search in mycobacteria followed, revealing over 200 sRNAs thus far. Characterization of these sRNAs is only beginning, but differential expression under environmental stresses suggests relevance to mycobacterial pathogenesis. This review provides a comprehensive overview of the current knowledge of sRNAs in mycobacteria, including historical perspective and techniques used for identification and characterization. PMID:25105095

  19. Nontuberculous Mycobacteria Lymphadenitis: A Case Report

    PubMed Central

    Varnam, Meera; Fernandez, Cristina

    2016-01-01

    Atypical mycobacteria, also known as nontuberculous mycobacteria (NTM) includes acid-fast bacteria other than Mycobacterium tuberculosis. NTM can be isolated from a variety of environmental sources including water, food products, domestic animals, and soil; human exposure is typically from soil to the oral cavity and respiratory tract. Diagnosis of NTM is suspected in children less than five years old with subacute, unilateral, non-tender cervicofacial lymphadenitis in combination with a history of water exposure, penetrating injection, as well as negative routine cultures or response to antistaphylococcal and antistreptococcal antibiotics. The course of the disease is variable and can involve eruption of the lymph node and tract formation with drainage. Management of nontuberculous mycobacteria can include surgical and antimycobacterial therapy. We present a case of a two-year-old African American girl who presented to the clinic with anterior ear lobe and submandibular lymphadenitis due to suspected NTM. PMID:27909634

  20. Distribution of Nontuberculous Mycobacteria strains

    PubMed Central

    2013-01-01

    Aim Mycobacteria other than tuberculosis (MOTT) cause increasingly serious infections especially in immunosuppressive patients by direct transmission from the environment or after colonization. However, identification of these species is difficult because of the cost and difficulties in defining to species level. Identification and distribution of these species can help clinician in the choice of treatment. Materials and methods A total of 90 MOTT strains obtained from four different centers were included in the study. These strains were identified by sequence analysis of 16S rRNA and Hsp65 genetic regions. Results Accordingly, within the 90 MOTT strains, 17 different species were identified. In order of frequency, these species were M. gordonea (n = 21), M. abscessus (n = 13), M. lentiflavum (n = 9), M. fortuitum (n = 8), M. intracellulare (n = 6), M. kumamotonense (n = 6), M. neoaurum (n = 5), M. chimaera (n = 5), M. alvei (n = 5), M. peregrinum (n = 3), M. canariasense (n = 3), M. flavescens (n = 1), M. mucogenicum (n = 1), M. chelona (n = 1), M. elephantis (n = 1), M. terrae (n = 1) and M. xenopi (n = 1). Most frequently identified MOTT species according to the geographical origin were as follows: M. abscessus was the most common species either in Istanbul or Malatya regions (n = 6, n = 6, consequently). While M. kumamotonense was the most frequent species isolated from Ankara region (n = 6), M. gordonea was the most common for Samsun region (n = 14). Conclusion Our study revealed that frequency of MOTT varies depending on the number of clinical samples and that frequency of these species were affected by the newly identified species as a result of the use of novel molecular methods. In conclusion, when establishing diagnosis and treatment methods, it is important to know that infections caused by unidentified MOTT species may vary according to the regions in Turkey. The results

  1. Infections Caused by Non-Tuberculous Mycobacteria in Recipients of Hematopoietic Stem Cell Transplantation

    PubMed Central

    Al-Anazi, Khalid Ahmed; Al-Jasser, Asma M.; Al-Anazi, Waleed Khalid

    2014-01-01

    Non-tuberculous mycobacteria (NTM) are acid-fast bacteria that are ubiquitous in the environment and can colonize soil, dust particles, water sources, and food supplies. They are divided into rapidly growing mycobacteria such as Mycobacterium fortuitum, Mycobacterium chelonae, and Mycobacterium abscessus as well as slowly growing species such as Mycobacterium avium, Mycobacterium kansasii, and Mycobacterium marinum. About 160 different species, which can cause community acquired and health care-associated infections, have been identified. NTM are becoming increasingly recognized in recipients of hematopoietic stem cell transplantation (HSCT) with incidence rates ranging between 0.4 and 10%. These infections are 50–600 times commoner in transplant recipients than in the general population and the time of onset ranges from day 31 to day 1055 post-transplant. They have been reported following various forms of HSCT. Several risk factors predispose to NTM infections in recipients of stem cell transplantation and these are related to the underlying medical condition and its treatment, the pre-transplant conditioning therapies as well as the transplant procedure and its complications. Clinically, NTM may present with: unexplained fever, lymphadenopathy, osteomyelitis, soft tissue and skin infections, central venous catheter infections, bacteremia, lung, and gastrointestinal tract involvement. However, disseminated infections are commonly encountered in severely immunocompromised individuals and bloodstream infections are almost always associated with catheter-related infections. It is usually difficult to differentiate colonization from true infection, thus, the threshold for starting therapy remains undetermined. Respiratory specimens such as sputum, pleural fluid, and bronchoalveolar lavage in addition to cultures of blood, bone, skin, and soft tissues are essential diagnostically. Susceptibility testing of mycobacterial isolates is a basic component of optimal care

  2. OCCURRENCE OF NONTUBERCULOUS MYCOBACTERIA IN ENVIRONMENTAL SAMPLES

    EPA Science Inventory

    Nontuberculous mycobacteria (NTM) are a major cause of opportunistic infection in immunocompromised hosts. Because there is no evidence of person-to-person transmission and NTM have been found in drinking water, the environment is considered a likely source of infection. In this ...

  3. EXAMINATION OF BOTTLED WATER FOR NONTUBERCULOUS MYCOBACTERIA

    EPA Science Inventory

    The objective of this study was to examine bottled water for the presence of nontuberculous mycobacteria as a potential source of infection in AIDS patients. Twenty brands of bottled water commonly used in the Los Angeles area were tested for the presence of nontuberculous mycoba...

  4. EXAMINATION OF BOTTLED WATER FOR NONTUBERCULOUS MYCOBACTERIA

    EPA Science Inventory

    The objective of this study was to examine bottled water for the presence of nontuberculous mycobacteria as a potential source of infection in AIDS patients. Twenty brands of bottled water commonly used in the Los Angeles area were tested for the presence of nontuberculous mycoba...

  5. OCCURRENCE OF NONTUBERCULOUS MYCOBACTERIA IN ENVIRONMENTAL SAMPLES

    EPA Science Inventory

    Nontuberculous mycobacteria (NTM) are a major cause of opportunistic infection in immunocompromised hosts. Because there is no evidence of person-to-person transmission and NTM have been found in drinking water, the environment is considered a likely source of infection. In this ...

  6. Clinical Relevance of Nontuberculous Mycobacteria, Oman

    PubMed Central

    Al-Mahruqi, Sara H.; Al-Busaidy, Suleiman; Boeree, Martin J.; Al-Zadjali, Samiya; Patel, Arti; Dekhuijzen, P.N. Richard; van Soolingen, Dick

    2009-01-01

    Little is known about the clinical relevance of nontuberculous mycobacteria (NTM) in the Arabian Peninsula. We assessed the prevalence and studied a random sample of isolates at a reference laboratory in Muscat, Oman. NTM cause disease in this region, and their prevalence has increased. PMID:19193276

  7. Factors Influencing Numbers of Mycobacterium avium, Mycobacterium intracellulare, and Other Mycobacteria in Drinking Water Distribution Systems

    PubMed Central

    Falkinham, Joseph O.; Norton, Cheryl D.; LeChevallier, Mark W.

    2001-01-01

    Eight water distribution systems were sampled over an 18-month period (528 water and 55 biofilm samples) to measure the frequency of recovery and number of mycobacteria, particularly Mycobacterium avium and Mycobacterium intracellulare, in raw source waters before and after treatment and within the distribution system. The systems were chosen to assess the influence of source water, treatment, and assimilable organic carbon levels on mycobacterial numbers. Overall, mycobacterial recovery from the systems was low (15% of samples). Numbers of mycobacteria ranged from 10 to 700,000 CFU liter−1. The number of M. avium in raw waters was correlated with turbidity. Water treatment substantially reduced the number of mycobacteria in raw waters by 2 to 4 log units. Mycobacterial numbers were substantially higher in the distribution system samples (average, 25,000-fold) than in those collected immediately downstream from the treatment facilities, indicating that mycobacteria grow in the distribution system. The increase in mycobacterial numbers was correlated with assimilable organic carbon and biodegradable organic carbon levels (r2 = 0.65, P = 0.03). Although M. intracellulare was seldom recovered from water samples, it was frequently recovered (six of eight systems) in high numbers from biofilms (average, 600 CFU/cm2). Evidently, the ecological niches of M. avium and M. intracellulare are distinct. PMID:11229914

  8. Temporal and intrinsic factors of rifampicin tolerance in mycobacteria

    PubMed Central

    Richardson, Kirill; Bennion, Owen T.; Tan, Shumin; Hoang, Anh N.; Cokol, Murat; Aldridge, Bree B.

    2016-01-01

    Mycobacteria grow and divide asymmetrically, creating variability in growth pole age, growth properties, and antibiotic susceptibilities. Here, we investigate the importance of growth pole age and other growth properties in determining the spectrum of responses of Mycobacterium smegmatis to challenge with rifampicin. We used a combination of live-cell microscopy and modeling to prospectively identify subpopulations with altered rifampicin susceptibility. We found two subpopulations that had increased susceptibility. At the initiation of treatment, susceptible cells were either small and at early stages of the cell cycle, or large and in later stages of their cell cycle. In contrast to this temporal window of susceptibility, tolerance was associated with factors inherited at division: long birth length and mature growth poles. Thus, rifampicin response is complex and due to a combination of differences established from both asymmetric division and the timing of treatment relative to cell birth. PMID:27357669

  9. Detection of Mycobacteria, Mycobacterium avium Subspecies, and Mycobacterium tuberculosis Complex by a Novel Tetraplex Real-Time PCR Assay

    PubMed Central

    Molina, Elena; Elguezabal, Natalia; Pérez, Valentín; Garrido, Joseba M.

    2015-01-01

    Mycobacterium tuberculosis complex, Mycobacterium avium, and many other nontuberculous mycobacteria are worldwide distributed microorganisms of major medical and veterinary importance. Considering the growing epidemiologic significance of wildlife-livestock-human interrelation, developing rapid detection tools of high specificity and sensitivity is vital to assess their presence and accelerate the process of diagnosing mycobacteriosis. Here we describe the development and evaluation of a novel tetraplex real-time PCR for simultaneous detection of Mycobacterium genus, M. avium subspecies, and M. tuberculosis complex in an internally monitored single assay. The method was evaluated using DNA from mycobacterial (n = 38) and nonmycobacterial (n = 28) strains, tissues spiked with different CFU amounts of three mycobacterial species (n = 57), archival clinical samples (n = 233), and strains isolated from various hosts (n = 147). The minimum detectable DNA amount per reaction was 50 fg for M. bovis BCG and M. kansasii and 5 fg for M. avium subsp. hominissuis. When spiked samples were analyzed, the method consistently detected as few as 100 to 1,000 mycobacterial CFU per gram. The sensitivity and specificity values for the panel of clinical samples were 97.5 and 100% using a verified culture-based method as the reference method. The assays performed on clinical isolates confirmed these results. This PCR was able to identify M. avium and M. tuberculosis complex in the same sample in one reaction. In conclusion, the tetraplex real-time PCR we designed represents a highly specific and sensitive tool for the detection and identification of mycobacteria in routine laboratory diagnosis with potential additional uses. PMID:25588660

  10. Detection of mycobacteria, Mycobacterium avium subspecies, and Mycobacterium tuberculosis complex by a novel tetraplex real-time PCR assay.

    PubMed

    Sevilla, Iker A; Molina, Elena; Elguezabal, Natalia; Pérez, Valentín; Garrido, Joseba M; Juste, Ramón A

    2015-03-01

    Mycobacterium tuberculosis complex, Mycobacterium avium, and many other nontuberculous mycobacteria are worldwide distributed microorganisms of major medical and veterinary importance. Considering the growing epidemiologic significance of wildlife-livestock-human interrelation, developing rapid detection tools of high specificity and sensitivity is vital to assess their presence and accelerate the process of diagnosing mycobacteriosis. Here we describe the development and evaluation of a novel tetraplex real-time PCR for simultaneous detection of Mycobacterium genus, M. avium subspecies, and M. tuberculosis complex in an internally monitored single assay. The method was evaluated using DNA from mycobacterial (n = 38) and nonmycobacterial (n = 28) strains, tissues spiked with different CFU amounts of three mycobacterial species (n = 57), archival clinical samples (n = 233), and strains isolated from various hosts (n = 147). The minimum detectable DNA amount per reaction was 50 fg for M. bovis BCG and M. kansasii and 5 fg for M. avium subsp. hominissuis. When spiked samples were analyzed, the method consistently detected as few as 100 to 1,000 mycobacterial CFU per gram. The sensitivity and specificity values for the panel of clinical samples were 97.5 and 100% using a verified culture-based method as the reference method. The assays performed on clinical isolates confirmed these results. This PCR was able to identify M. avium and M. tuberculosis complex in the same sample in one reaction. In conclusion, the tetraplex real-time PCR we designed represents a highly specific and sensitive tool for the detection and identification of mycobacteria in routine laboratory diagnosis with potential additional uses.

  11. Porins in the Cell Wall of Mycobacteria

    NASA Astrophysics Data System (ADS)

    Trias, Joaquim; Jarlier, Vincent; Benz, Roland

    1992-11-01

    The cell wall of mycobacteria is an efficient permeability barrier that makes mycobacteria naturally resistant to most antibiotics. Liposome swelling assays and planar bilayer experiments were used to investigate the diffusion process of hydrophilic molecules through the cell wall of Mycobacterium chelonae and identify the main hydrophilic pathway. A 59-kilodalton cell wall protein formed a water-filled channel with a diameter of 2.2 nanometers and an average single-channel conductance equal to 2.7 nanosiemens in 1 M potassium chloride. These results suggest that porins can be found in the cell wall of a Gram-positive bacterium. A better knowledge of the hydrophilic pathways should help in the design of more effective antimycobacterial agents.

  12. Mycobacteria in pond waters as a source of non-specific reactions to bovine tuberculin in New Zealand.

    PubMed

    Kazda, J; Cook, B R

    1988-12-01

    When 71 samples were collected from ponds throughout New Zealand, 35 (49.3%) were found to contain mycobacteria. The majority of these strains (62.9%) belonged to a homogeneous group (tentative designation H-Group, which differed from any known mycobacterial species. Mycobacteria of this H-group had also been found in sphagnum vegetation growing in the immediate vicinity of many of the ponds. H-Group mycobacteria induce sensitization in guinea pigs against bovine tuberculin. The PPD sensitin prepared from these mycobacteria gave rise to larger reactions in guinea pigs than did bovine tuberculin when used in the same concentrations (500 and 50 TU). The possible sensitization of cattle to bovine tuberculin via drinking water containing H-Group mycobacteria, is discussed. The larger size of the delayed hypersensitivity reactions in guinea pigs using the same concentrations of bovine and homologous tuberculin, suggests that comparative intradermal testing might enable this non-specific reaction to be distinguished from the specific reaction developed during bovine tuberculosis infection in cattle.

  13. Growing Pains from Rapid Growth: A Historical Case Study of George Fox University from 1983 to 2003

    ERIC Educational Resources Information Center

    Railsback, Gary L.

    2007-01-01

    This article is a historical case study of George Fox University (GFU) in Newberg, Oregon. Using organizational lifecycle as a theoretical framework, George Fox University had a long and delayed childhood in that it remained a small and struggling institution for most of the 20th century, and then experienced rapid growth in the late 1980s. This…

  14. Growing Pains from Rapid Growth: A Historical Case Study of George Fox University from 1983 to 2003

    ERIC Educational Resources Information Center

    Railsback, Gary L.

    2007-01-01

    This article is a historical case study of George Fox University (GFU) in Newberg, Oregon. Using organizational lifecycle as a theoretical framework, George Fox University had a long and delayed childhood in that it remained a small and struggling institution for most of the 20th century, and then experienced rapid growth in the late 1980s. This…

  15. BIOLOGICAL AND CHEMICAL STUDIES ON MYCOBACTERIA

    PubMed Central

    Fregnan, G. B.; Smith, D. W.; Randall, H. M.

    1961-01-01

    Fregnan, G. B. (University of Wisconsin, Madison), D. W. Smith, and H. M. Randall. Biological and chemical studies on mycobacteria. Relationship of colony morphology to mycoside content for Mycobacterium kansasii and Mycobacterium fortuitum. J. Bacteriol. 82:517–527. 1961.—Using a suitable technique and an adequate medium it was possible to show a unique and uniform type of colony characteristic for Mycobacterium kansasii (photochromogen) and for Mycobacterium fortuitum strains freshly isolated either from patients, or from the soil, or kept in our stock culture collection for several years. New symbols have been proposed to represent these colony types. It was demonstrated that colony morphology is closely related to the specific mycoside present in a given strain; for example, M. kansasii strains showed in each instance colony type K and mycoside A, and M. fortuitum strains showed colony type F and mycoside F. Attention is called to the importance of the technique and the medium used. No change in colony morphology resulted from incubation in the presence of air containing 5 to 10% CO2, although this improved growth. Better growth of mycobacteria occurred in the presence of glycerol, or lipids of a human strain of mycobacteria, or sodium bicarbonate, but the specificity of colony form was lost. Images PMID:13894938

  16. [Application of mass spectrometry in mycobacteria].

    PubMed

    Alcaide, Fernando; Palop-Borrás, Begoña; Domingo, Diego; Tudó, Griselda

    2016-06-01

    To date, more than 170 species of mycobacteria have been described, of which more than one third may be pathogenic to humans, representing a significant workload for microbiology laboratories. These species must be identified in clinical practice, which has long been a major problem due to the shortcomings of conventional (phenotypic) methods and the limitations and complexity of modern methods largely based on molecular biology techniques. The aim of this review was to briefly describe different aspects related to the use of MALDI-TOF (matrix-assisted laser desorption ionization time-of-flight) mass spectrometry (MS) for the identification of mycobacteria. Several difficulties are encountered with the use of this methodology in these microorganisms mainly due to the high pathogenicity of some mycobacteria and the peculiar structure of their cell wall, requiring inactivation and special protein extraction protocols. We also analysed other relevant aspects such as culture media, the reference methods employed (gold standard) in the final identification of the different species, the cut-off used to accept data as valid, and the databases of the different mass spectrometry systems available. MS has revolutionized diagnosis in modern microbiology; however, specific improvements are needed to consolidate the use of this technology in mycobacteriology. Copyright © 2016 Elsevier España, S.L.U. All rights reserved.

  17. Mycobacteria entry and trafficking into endothelial cells.

    PubMed

    Baltierra-Uribe, Shantal Lizbeth; García-Vásquez, Manuel de Jesús; Castrejón-Jiménez, Nayeli Shantal; Estrella-Piñón, Mayra Patricia; Luna-Herrera, Julieta; García-Pérez, Blanca Estela

    2014-09-01

    Endothelial cells are susceptible to infection by mycobacteria, but the endocytic mechanisms that mycobacteria exploit to enter host cells and their mechanisms of intracellular transport are completely unknown. Using pharmacological inhibitors, we determined that the internalization of Mycobacterium tuberculosis (MTB), Mycobacterium smegmatis (MSM), and Mycobacterium abscessus (MAB) is dependent on the cytoskeleton and is differentially inhibited by cytochalasin D, nocodazole, cycloheximide, wortmannin, and amiloride. Using confocal microscopy, we investigated their endosomal trafficking by analyzing Rab5, Rab7, LAMP-1, and cathepsin D. Our results suggest that MSM exploits macropinocytosis to enter endothelial cells and that the vacuoles containing these bacteria fuse with lysosomes. Conversely, the entry of MTB seems to depend on more than one endocytic route, and the observation that only a subset of the intracellular bacilli was associated with phagolysosomes suggests that these bacteria are able to inhibit endosomal maturation to persist intracellularly. The route of entry for MAB depends mainly on microtubules, which suggests that MAB uses a different trafficking pathway. However, MAB is also able to inhibit endosomal maturation and can replicate intracellularly. Together, these findings provide the first evidence that mycobacteria modulate proteins of host endothelial cells to enter and persist within these cells.

  18. Evaluation of MALDI Biotyper Mycobacteria Library v3.0 for Identification of Nontuberculous Mycobacteria

    PubMed Central

    Ruiz-Serrano, M. Jesús; Ruiz, Adrián; Timke, Markus; Kostrzewa, Markus; Bouza, Emilio

    2016-01-01

    Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has demonstrated its ability to promptly identify nontuberculous mycobacteria using the Mycobacteria Library v2.0. However, some species are particularly difficult to identify reliably using this database, providing a low log(score). In this study, the identification power of an updated Mycobacteria Library (v3.0) has been evaluated. Overall, 109 NTM isolates were analyzed with both databases. The v3.0 database allowed a high-level confidence in the identification [log(score) value, ≥1.8] of 91.7% of the isolates versus 83.5% with the v2.0 version (P < 0.01). PMID:26842704

  19. Detection and identification of aquatic mycobacteria in formalin-fixed, paraffin-embedded fish tissues.

    PubMed

    Pourahmad, F; Thompson, K D; Adams, A; Richards, R H

    2009-05-01

    The isolation of mycobacteria from field samples is problematic, and isolation of the bacterium is sometimes not even attempted. The detection of mycobacteria through traditional histology using formalin-fixed, paraffin-embedded (FFPE) tissues is neither sensitive nor specific. However, detection of mycobacterial DNA from FFPE specimens, suspected of being infected with mammalian mycobacteriosis, is a routine clinical procedure. In the present study, a polymerase chain reaction (PCR)-based method was used to detect and identify mycobacteria in FFPE specimens sampled from fish suspected of being infected with fish mycobacteriosis. A total of 45 fish tissue samples, comprising of 12 tissue samples obtained from experimentally infected fish and the remainder from fish naturally infected with mycobacteria, were analysed using a PCR protocol which amplifies a fragment of the mycobacterial 65 kDa heat-shock protein (hsp65) gene. PCR-restriction enzyme analysis and/or sequencing were employed to further analyse the PCR amplicons. The PCR results were compared with those obtained by histology and culture. Mycobacterial DNA was detected in 34 of the 45 samples examined, of which 16 samples (47%) showed granulomatous reactions on histological examination. Using histology as the gold standard, no false-negative PCR results were obtained. Also, considering the presence or absence of granulomas as a diagnostic criterion, the sensitivity and specificity of PCR in 42 of the FFPE tissues were 16/16 (100%) and 8/26 (approximately 30.8%), respectively. Corresponding microbiological cultures were available for 15 cases, of which 13 were pure Mycobacterium cultures. Of these, 13 were PCR positive (100% sensitivity and 50% specificity). The PCR-based methods used here proved sensitive, specific and rapid for the detection of mycobacteria in routinely processed paraffin wax-embedded and formalin-fixed histological samples, and the results of the study suggest that this method has

  20. Looking in amoebae as a source of mycobacteria.

    PubMed

    Drancourt, M

    2014-12-01

    Mycobacteria exhibit various relationships with amoebae, ranging from the killing of one partner by the other one, to amoebae hosting mycobacteria in trophozoites and cysts. This observation indicates that poorly described biological factors affect the relationships, including mycobacterial cell-wall glycolipids and the size of the mycobacteria. Experimental observations indicate that a majority of environmental, opportunistic mycobacteria but also obligate pathogens including Mycobacterium tuberculosis, Mycobacterium leprae and Mycobacterium ulcerans are inter-amoebal organisms. Amoebae may give opportunities for genetic exchanges between mycobacteria, sympatric intra-amoebal organisms and the amoebae themselves. Amoebae clearly protect opportunistic mycobacterial pathogens during their environmental life but their role for obligate mycobacterial infection remains to be established. Accordingly, water was the source for emerging, community-acquired and health care-associated infection with amoeba-resisting mycobacteria of the Mycobacterium avium, Mycobacterium abscessus and Mycobacterium fortuitum groups, among others. Amoebae are organisms where mycobacteria can be found and, accordingly, amoeba co-culture can be used for the isolation of mycobacteria from environmental and clinical specimens. Looking in amoebae may help recovering new species of mycobacteria.

  1. The positive effect of soybean protein hydrolysates-calcium complexes on bone mass of rapidly growing rats.

    PubMed

    Lv, Ying; Liu, He; Ren, Jianhua; Li, Xin; Guo, Shuntang

    2013-08-01

    It was previously found that soybean protein hydrolysates (SPHs) can bind with calcium to form soluble complexes and promote calcium uptake by Caco-2 cells. However, the role of SPHs-calcium complexes on bone mass still needs to be explored. Fast growing male and female rats (n = 72) were assigned to eight groups: Control, lactic acid calcium (LCa), SPHs-calcium complexes (SPHCa) and casein phosphopeptides calcium (CPPCa). After four weeks treatment, oral administration of SPHCa significantly increased femur BMD of rats compared with Control and LCa groups (P < 0.05), while there are no obvious difference on the BMD of femur and lumbar vertebrae between SPHCa and CPPCa groups. Also, SPHCa showed a tendency to improve the mechanical properties of vertebra lumber for female rats. These findings suggest that the SPHs-calcium complexes might have positive effects on bone accretion of fast growing animals. This study brings new insight for better understanding the role of soybean protein itself on bone mass.

  2. METHYL KETONE METABOLISM IN HYDROCARBON-UTILIZING MYCOBACTERIA

    PubMed Central

    Lukins, H. B.; Foster, J. W.

    1963-01-01

    Lukins, H. B. (University of Texas, Austin) and J. W. Foster. Methyl ketone metabolism in hydrocarbon-utilizing mycobacteria. J. Bacteriol. 85: 1074–1087. 1963.—Species of Mycobacterium especially M. smegmatis 422, produced the homologous methyl ketones during the oxidation of propane, n-butane, n-pentane, or n-hexane. A carrier-trapping experiment demonstrated the formation of 2-undecanone, as well as 1,11-undecanedioic acid, during the oxidation of undecane-1-C14. Aliphatic alkane-utilizing mycobacteria were able to grow at the expense of several aliphatic methyl ketones as sole sources of carbon. Other ketones which did not support growth were oxidized by resting bacterial suspensions. M. smegmatis 422 cells grown on propane or acetone were simultaneously adapted to oxidize both substrates, as well as n-propanol. n-Propanol cells were unadapted to propane or acetone. Acetone produced from propane in a medium enriched in D2O contained a negligible quantity of D, presumably eliminating propylene as an intermediate in the oxidation. Cells grown at the expense of alkanes or methyl ketones in the presence of O218 had a higher content of O18 than did cells grown on terminally oxidized compounds, e.g., primary alcohols or fatty acids. An oxygenase reaction is postulated for the attack on methyl ketones. Acetol was isolated and characterized as an oxidation product of acetone by M. smegmatis 422. Acetol-grown cells had a higher O18 content than did n-propanol cells, and its utilization appears to involve at least one oxygenase reaction. Acetol produced from acetone in the presence of O218 was not enriched in the isotope, indicating the occurrence of exchange reactions or of oxygenation reactions at a later stage in the assimilation of acetone and acetol. PMID:14043998

  3. Gallium Disrupts Iron Metabolism of Mycobacteria Residing within Human Macrophages

    PubMed Central

    Olakanmi, Oyebode; Britigan, Bradley E.; Schlesinger, Larry S.

    2000-01-01

    Mycobacterium tuberculosis and M. avium complex (MAC) enter and multiply within monocytes and macrophages in phagosomes. In vitro growth studies using standard culture media indicate that siderophore-mediated iron (Fe) acquisition plays a critical role in the growth and metabolism of both M. tuberculosis and MAC. However, the applicability of such studies to conditions within the macrophage phagosome is unclear, due in part to the absence of experimental means to inhibit such a process. Based on the ability of gallium (Ga3+) to concentrate within mononuclear phagocytes and on evidence that Ga disrupts cellular Fe-dependent metabolic pathways by substituting for Fe3+ and failing to undergo redox cycling, we hypothesized that Ga could disrupt Fe acquisition and Fe-dependent metabolic pathways of mycobacteria. We find that Ga(NO3)3 and Ga-transferrin produce an Fe-reversible concentration-dependent growth inhibition of M. tuberculosis strains and MAC grown extracellularly and within human macrophages. Ga is bactericidal for M. tuberculosis growing extracellularly and within macrophages. Finally, we provide evidence that exogenously added Fe is acquired by intraphagosomal M. tuberculosis and that Ga inhibits this Fe acquisition. Thus, Ga(NO3)3 disruption of mycobacterial Fe metabolism may serve as an experimental means to study the mechanism of Fe acquisition by intracellular mycobacteria and the role of Fe in intracellular survival. Furthermore, given the inability of biological systems to discriminate between Ga and Fe, this approach could have broad applicability to the study of Fe metabolism of other intracellular pathogens. PMID:10992462

  4. Detection of fastidious mycobacteria in human intestines by the polymerase chain reaction.

    PubMed

    Dumonceau, J M; Van Gossum, A; Adler, M; Van Vooren, J P; Fonteyne, P A; De Beenhouwer, H; Portaels, F

    1997-05-01

    The aim of this study was to determine whether difficult-to-grow mycobacteria are present in human intestines. Intestinal tissue samples were subjected to both mycobacterial culture and a polymerase chain reaction (PCR) assay. After detection by PCR, species identity was determined by hybridizing the amplified 16S rRNA gene fragments with species-specific oligonucleotides. Intestinal biopsies from 63 patients with noninflammatory bowel diseases (n = 22), Crohn's disease (n = 31), or ulcerative colitis (n = 10) were analyzed. Culture and PCR revealed mycobacteria in four (6%) and 25 (40%) samples, respectively. Samples positive by PCR were negative with all probes specific to nine common cultivable species but were positive with Mycobacterium genavense-specific probe in 68% of cases. Mycobacterial isolates were identified as Mycobacterium gordonae and Mycobacterium chelonae. Findings were similar in Crohn's disease samples compared to non-Chron's disease samples. This study shows that difficult-to-grow mycobacteria can be detected by PCR in large and similar proportions of inflamed intestinal tissue from patients with inflammatory bowel disease and intestinal tissue that appears normal from patients with noninflammatory bowel disease.

  5. Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Identification of Mycobacteria in Routine Clinical Practice

    PubMed Central

    El Khéchine, Amel; Couderc, Carine; Flaudrops, Christophe; Raoult, Didier; Drancourt, Michel

    2011-01-01

    Background Non-tuberculous mycobacteria recovered from respiratory tract specimens are emerging confounder organisms for the laboratory diagnosis of tuberculosis worldwide. There is an urgent need for new techniques to rapidly identify mycobacteria isolated in clinical practice. Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS) has previously been proven to effectively identify mycobacteria grown in high-concentration inocula from collections. However, a thorough evaluation of its use in routine laboratory practice has not been performed. Methodology We set up an original protocol for the MALDI-TOF MS identification of heat-inactivated mycobacteria after dissociation in Tween-20, mechanical breaking of the cell wall and protein extraction with formic acid and acetonitrile. By applying this protocol to as few as 105 colony-forming units of reference isolates of Mycobacterium tuberculosis, Mycobacterium avium, and 20 other Mycobacterium species, we obtained species-specific mass spectra for the creation of a local database. Using this database, our protocol enabled the identification by MALDI-TOF MS of 87 M. tuberculosis, 25 M. avium and 12 non-tuberculosis clinical isolates with identification scores ≥2 within 2.5 hours. Conclusions Our data indicate that MALDI-TOF MS can be used as a first-line method for the routine identification of heat-inactivated mycobacteria. MALDI-TOF MS is an attractive method for implementation in clinical microbiology laboratories in both developed and developing countries. PMID:21935444

  6. [Evaluation of the distribution of non-tuberculous mycobacteria strains isolated in National Tuberculosis Reference Laboratory in 2009-2010, Turkey].

    PubMed

    Albayrak, Nurhan; Simşek, Hülya; Sezen, Figen; Arslantürk, Ahmet; Tarhan, Gülnur; Ceyhan, Ismail

    2012-10-01

    Non-tuberculous mycobacteria (NTM) are commonly encountered environmental bacteria, and most of them are associated with lung diseases. Diagnosis of infections caused by NTM is based on clinical, radiological and microbiological findings. The aim of this study was to investigate the distribution of non-tuberculous mycobacterial species isolated from clinical specimens as etiologic agents. The NTM strains isolated from clinical specimens in National Tuberculosis Reference Laboratory (NTRL), together with the strains that were sent to NTRL for the advanced identification of non-tuberculous mycobacterial species that have clinical or microbiological significance, were analysed retrospectively. The strains belonged to January 2009 - December 2010 period. If the same NTM type was isolated more than once in the clinical specimens of a patient, then it was defined microbiologically as a causative agent. Identification of mycobacteria species was performed by using a commercial line-probe assay (GenoType Mycobacterium CM/AS; Hain Lifescience, Germany). In our study, pulmonary and non-pulmonary samples obtained from 206 patients yielded mycobacterial growth in their cultures, and of them 24 (11.7%) were identified as NTM. On the other hand, 51 of the 101 samples sent to NTRL for identification were confirmed as NTM. Of the patients who were found to be infected with NTM (n= 75), 59 (78.7%) were male and the mean age was 50.9 ± 18.8 years. The most frequently identified NTM species was M.fortuitum (33.3%, n= 25), followed by M.abscessus (18.7%, n= 14), M.gordonae (10.7%, n= 8) and M.avium (%8; n= 6). The other types of NTM species identified in our laboratory were M.chelonae (n= 3), M.intracellulare (n= 3), M.kansasii (n= 3), M.peregrinum (n= 2), M.scrofulaceum (n= 2), M.szulgai (n= 2), M.celatum (n= 1), M.haemophilum (n= 1), M.smegmatis (n= 1) and M.xenopi (n= 1). Rapidly growing NTM species (M.fortuitum and M.abscessus) were the most frequent (52%) species isolated in our

  7. Novel Diagnostic Algorithm for Identification of Mycobacteria Using Genus-Specific Amplification of the 16S-23S rRNA Gene Spacer and Restriction Endonucleases

    PubMed Central

    Roth, Andreas; Reischl, Udo; Streubel, Anna; Naumann, Ludmila; Kroppenstedt, Reiner M.; Habicht, Marion; Fischer, Marga; Mauch, Harald

    2000-01-01

    A novel genus-specific PCR for mycobacteria with simple identification to the species level by restriction fragment length polymorphism (RFLP) was established using the 16S-23S ribosomal RNA gene (rDNA) spacer as a target. Panspecificity of primers was demonstrated on the genus level by testing 811 bacterial strains (122 species in 37 genera from 286 reference strains and 525 clinical isolates). All mycobacterial isolates (678 strains among 48 defined species and 5 indeterminate taxons) were amplified by the new primers. Among nonmycobacterial isolates, only Gordonia terrae was amplified. The RFLP scheme devised involves estimation of variable PCR product sizes together with HaeIII and CfoI restriction analysis. It yielded 58 HaeIII patterns, of which 49 (84%) were unique on the species level. Hence, HaeIII digestion together with CfoI results was sufficient for correct identification of 39 of 54 mycobacterial taxons and one of three or four of seven RFLP genotypes found in Mycobacterium intracellulare and Mycobacterium kansasii, respectively. Following a clearly laid out diagnostic algorithm, the remaining unidentified organisms fell into five clusters of closely related species (i.e., the Mycobacterium avium complex or Mycobacterium chelonae-Mycobacterium abscessus) that were successfully separated using additional enzymes (TaqI, MspI, DdeI, or AvaII). Thus, next to slowly growing mycobacteria, all rapidly growing species studied, including M. abscessus, M. chelonae, Mycobacterium farcinogenes, Mycobacterium fortuitum, Mycobacterium peregrinum, and Mycobacterium senegalense (with a very high 16S rDNA sequence similarity) were correctly identified. A high intraspecies sequence stability and the good discriminative power of patterns indicate that this method is very suitable for rapid and cost-effective identification of a wide variety of mycobacterial species without the need for sequencing. Phylogenetically, spacer sequence data stand in good agreement with 16S r

  8. Notes from the field: rapidly growing nontuberculous Mycobacterium wound infections among medical tourists undergoing cosmetic surgeries in the Dominican Republic--multiple states, March 2013-February 2014.

    PubMed

    Schnabel, David; Gaines, Joanna; Nguyen, Duc B; Esposito, Douglas H; Ridpath, Alison; Yacisin, Kari; Poy, Joe A; Mullins, Jocelyn; Burns, Rachel; Lijewski, Virginia; McElroy, Nora P; Ahmad, Nina; Harrison, Cassandra; Parinelli, Ellen J; Beaudoin, Amanda L; Posivak-Khouly, Leah; Pritchard, Scott; Jensen, Bette J; Toney, Nadege C; Moulton-Meissner, Heather A; Nyangoma, Edith N; Barry, Anita M; Feldman, Katherine A; Blythe, David; Perz, Joseph F; Morgan, Oliver W; Kozarsky, Phyllis; Brunette, Gary W; Sotir, Mark

    2014-03-07

    In August 2013, the Maryland Department of Health and Mental Hygiene (MDHMH) was notified of two persons with rapidly growing nontuberculous mycobacterial (RG-NTM) surgical-site infections. Both patients had undergone surgical procedures as medical tourists at the same private surgical clinic (clinic A) in the Dominican Republic the previous month. Within 7 days of returning to the United States, both sought care for symptoms that included surgical wound abscesses, clear fluid drainage, pain, and fever. Initial antibiotic therapy was ineffective. Material collected from both patients' wounds grew Mycobacterium abscessus exhibiting a high degree of antibiotic resistance characteristic of this organism.

  9. An aerosol climatology for a rapidly growing arid region (southern Arizona): Major aerosol species and remotely sensed aerosol properties

    PubMed Central

    Sorooshian, Armin; Wonaschütz, Anna; Jarjour, Elias G.; Hashimoto, Bryce I.; Schichtel, Bret A.; Betterton, Eric A.

    2014-01-01

    This study reports a comprehensive characterization of atmospheric aerosol particle properties in relation to meteorological and back trajectory data in the southern Arizona region, which includes two of the fastest growing metropolitan areas in the United States (Phoenix and Tucson). Multiple data sets (MODIS, AERONET, OMI/TOMS, MISR, GOCART, ground-based aerosol measurements) are used to examine monthly trends in aerosol composition, aerosol optical depth (AOD), and aerosol size. Fine soil, sulfate, and organics dominate PM2.5 mass in the region. Dust strongly influences the region between March and July owing to the dry and hot meteorological conditions and back trajectory patterns. Because monsoon precipitation begins typically in July, dust levels decrease, while AOD, sulfate, and organic aerosol reach their maximum levels because of summertime photochemistry and monsoon moisture. Evidence points to biogenic volatile organic compounds being a significant source of secondary organic aerosol in this region. Biomass burning also is shown to be a major contributor to the carbonaceous aerosol budget in the region, leading to enhanced organic and elemental carbon levels aloft at a sky-island site north of Tucson (Mt. Lemmon). Phoenix exhibits different monthly trends for aerosol components in comparison with the other sites owing to the strong influence of fossil carbon and anthropogenic dust. Trend analyses between 1988 and 2009 indicate that the strongest statistically significant trends are reductions in sulfate, elemental carbon, and organic carbon, and increases in fine soil during the spring (March–May) at select sites. These results can be explained by population growth, land-use changes, and improved source controls. PMID:24707452

  10. Radiometric selective inhibition tests for differentiation of Mycobacterium tuberculosis, Mycobacterium bovis, and other mycobacteria.

    PubMed Central

    Gross, W M; Hawkins, J E

    1985-01-01

    In the context of a busy reference laboratory, radiometric selective inhibition tests were evaluated for rapid differentiation of Mycobacterium tuberculosis and Mycobacterium bovis and of the M. tuberculosis complex from other mycobacteria. p-Nitro-alpha-acetylamino-beta-hydroxypropiophenone at 5 micrograms and hydroxylamine hydrochloride at 62.5 and 125 micrograms per ml of 7H12 medium were used to separate the M. tuberculosis complex from other mycobacteria (MOTT bacilli). Since it is important epidemiologically to distinguish M. tuberculosis from M. bovis, susceptibility to 1 microgram of thiophene-2-carboxylic acid per ml was also determined radiometrically. By using these three agents as selective inhibitors, M. tuberculosis, M. bovis, and MOTT bacilli were differentiated with a high degree of specificity by a BACTEC radiometric procedure. Results of tests performed on clinical isolates submitted on solid medium to our reference laboratory were available within 5 days. PMID:3921561

  11. Mycobacterium bovis BCG mycobacteria--new application.

    PubMed

    Kowalewicz-Kulbat, Magdalena; Pestel, Joël; Biet, Franck; Locht, Camille; Tonnel, André-Bernard; Druszczyńska, Magdalena; Rudnicka, Wiesława

    2006-01-01

    The polarized response of T helper-2 (Th2) lymphocytes to an allergen is considered to be the main cause of the pathogenesis of asthma. In this study, we asked a question whether M. bovis BCG mycobacteria which are known for the preferential stimulation of T helper-1 (Th1) immunity, diminish the effector functions of Th2 cells from allergic patients upon stimulation with a common house dust mite Der p-1 allergen. Our results allow a positive answer to this question. We demonstrate that BCG modulates the dendritic cell-dependent allergen presentation process and switches naive T lymphocytes towards an anti-allergic Th1 profile.

  12. A Multi-Level Approach to Modeling Rapidly Growing Mega-Regions as a Coupled Human-Natural System

    NASA Astrophysics Data System (ADS)

    Koch, J. A.; Tang, W.; Meentemeyer, R. K.

    2013-12-01

    concept of our modeling approach and describe its strengths and weaknesses. We furthermore use empirical data for the states of North and South Carolina to demonstrate how the modeling framework can be applied to a large, heterogeneous study system with diverse decision-making agents. Grimm et al. (2005) Pattern-Oriented Modeling of Agent-Based Complex Systems: Lessons from Ecology. Science 310, 987-991. Liu et al. (2013) Framing Sustainability in a Telecoupled World. Ecology and Society 18(2), 26. Meentemeyer et al. (2013) FUTURES: Multilevel Simulations of Merging Urban-Rural Landscape Structure Using a Stochastic Patch-Growing Algorithm. Annals of the Association of American Geographers 103(4), 785-807.

  13. Ownership or leasing of CT scanners by nonradiologist physicians: a rapidly growing trend that raises concern about self-referral.

    PubMed

    Levin, David C; Rao, Vijay M; Parker, Laurence; Frangos, Andrea J; Sunshine, Jonathan H

    2008-12-01

    The aim of this study was to examine recent nationwide trends in the ownership or leasing of computed tomographic (CT) scanners in private offices by nonradiologist physicians. The Medicare Part B fee-for-service data sets for 2001 though 2006 were used to identify all CT scans performed in nonhospital, private-office settings. Ownership or leasing of CT scans was determined by tabulating all global and technical-component-only claims. Professional-component claims were excluded. The specialty of the owner or lessee was determined using Medicare's physician specialty codes. Procedure volume trends and growth rates among all nonradiologist physicians as a group were compared with those among radiologists. Individual specialty volume trends and growth rates were also studied. From 2001 to 2006, Medicare private-office CT scan volume in facilities owned by radiologists increased by 85%. CT scan volume in facilities owned or leased by nonradiologist physicians as a group increased by 263%. The nonradiologic specialties with the largest volumes in 2006 were primary care (192,255 scans), internal medicine subspecialties other than cardiology and medical oncology (184,991 scans), urology (125,850 scans), cardiology (104,739 scans), and medical oncology (61,976 scans). Excluding CT scans performed in independent diagnostic testing facilities (for which physician ownership cannot be determined), nonradiologists' private-office CT market share rose from 16% in 2001 to 28% in 2006. The majority of Medicare private-office CT scans are done in facilities owned by radiologists. However, nonradiologist physicians are acquiring or leasing CT scanners in increasing numbers, and the growth trend is much more rapid among them than it is among radiologists (85% among radiologists from 2001 to 2006, compared with 263% among nonradiologists). As a result, nonradiologists' market share has increased considerably. At a time when both cost containment and reduction in radiation exposure

  14. A Framework Predicting Water Availability in a Rapidly Growing, Semi-Arid Region under Future Climate Change

    NASA Astrophysics Data System (ADS)

    Han, B.; Benner, S. G.; Glenn, N. F.; Lindquist, E.; Dahal, K. R.; Bolte, J.; Vache, K. B.; Flores, A. N.

    2014-12-01

    Climate change can lead to dramatic variations in hydrologic regime, affecting both surface water and groundwater supply. This effect is most significant in populated semi-arid regions where water availability are highly sensitive to climate-induced outcomes. However, predicting water availability at regional scales, while resolving some of the key internal variability and structure in semi-arid regions is difficult due to the highly non-linearity relationship between rainfall and runoff. In this study, we describe the development of a modeling framework to evaluate future water availability that captures elements of the coupled response of the biophysical system to climate change and human systems. The framework is built under the Envision multi-agent simulation tool, characterizing the spatial patterns of water demand in the semi-arid Treasure Valley area of Southwest Idaho - a rapidly developing socio-ecological system where urban growth is displacing agricultural production. The semi-conceptual HBV model, a population growth and allocation model (Target), a vegetation state and transition model (SSTM), and a statistically based fire disturbance model (SpatialAllocator) are integrated to simulate hydrology, population and land use. Six alternative scenarios are composed by combining two climate change scenarios (RCP4.5 and RCP8.5) with three population growth and allocation scenarios (Status Quo, Managed Growth, and Unconstrained Growth). Five-year calibration and validation performances are assessed with Nash-Sutcliffe efficiency. Irrigation activities are simulated using local water rights. Results show that in all scenarios, annual mean stream flow decreases as the projected rainfall increases because the projected warmer climate also enhances water losses to evapotranspiration. Seasonal maximum stream flow tends to occur earlier than in current conditions due to the earlier peak of snow melting. The aridity index and water deficit generally increase in the

  15. Rapid Detection of Mycobacteria in Patients with HIV Infection.

    DTIC Science & Technology

    1992-06-30

    Mycobacterium paratuberculosis strains by restriction endonuclease analysis and ENA hybridization. J. Clin. Microbiol. 28:1591-1596. 5. Chiodini, R.J...1990. Characterization of Mvcobacterium paratuberculosis and organisms of the Mycobacterium avium complex by restriction polymorphism of the rim gene

  16. AAA+ Machines of Protein Destruction in Mycobacteria.

    PubMed

    Alhuwaider, Adnan Ali H; Dougan, David A

    2017-01-01

    The bacterial cytosol is a complex mixture of macromolecules (proteins, DNA, and RNA), which collectively are responsible for an enormous array of cellular tasks. Proteins are central to most, if not all, of these tasks and as such their maintenance (commonly referred to as protein homeostasis or proteostasis) is vital for cell survival during normal and stressful conditions. The two key aspects of protein homeostasis are, (i) the correct folding and assembly of proteins (coupled with their delivery to the correct cellular location) and (ii) the timely removal of unwanted or damaged proteins from the cell, which are performed by molecular chaperones and proteases, respectively. A major class of proteins that contribute to both of these tasks are the AAA+ (ATPases associated with a variety of cellular activities) protein superfamily. Although much is known about the structure of these machines and how they function in the model Gram-negative bacterium Escherichia coli, we are only just beginning to discover the molecular details of these machines and how they function in mycobacteria. Here we review the different AAA+ machines, that contribute to proteostasis in mycobacteria. Primarily we will focus on the recent advances in the structure and function of AAA+ proteases, the substrates they recognize and the cellular pathways they control. Finally, we will discuss the recent developments related to these machines as novel drug targets.

  17. Mycobacteria isolated from Chesapeake Bay fish.

    PubMed

    Stine, C B; Kane, A S; Baya, A M

    2010-01-01

    Mycobacteriosis in fish can result in ulcers, emaciation, and in some cases death. Mycobacteria have been previously isolated from a variety of Chesapeake Bay fish species, and the current study was designed to identify potential host specificity and location fidelity of mycobacterial isolates. Mycobacteria were isolated from wild fish of the Chesapeake Bay collected from the Upper Bay, the Choptank River, Herring Bay, the Chicamacomico River, the Pocomoke River and the Potomac River in 2003-2006. Mycobacterial isolates were recovered from striped bass, Morone saxatilis, Atlantic menhaden, Brevoortia tyrannus, white perch, Morone americana, summer flounder, Paralichthys dentatus, spot, Leiostomus xanthurus, largemouth bass, Micropterus salmoides, channel catfish, Ictalurus punctatus, common carp, Cyprinus carpio carpio, spotted seatrout, Cynoscion nebulosus, killifish, Fundulus sp., blueback herring, Alosa aestivalis, American gizzard shad, Dorosoma cepedianum and American silver perch, Bairdiella chrysoura. Twenty-nine well-defined mycobacterial groups resulted from gas chromatography dendrogram clustering of isolates. The majority of groups included more than one host species and more than one site of collection. However, four groups contained only striped bass isolates, three of which were similar to M. shottsii. Therefore, multiple Chesapeake Bay fish species are colonized with multiple mycobacterial isolates, of which few appear to be host or location specific.

  18. MYCOBACTOCIDIN, A NEW ANTIBIOTIC ACTIVE AGAINST MYCOBACTERIA

    PubMed Central

    Fregnan, G. B.; Smith, D. W.

    1962-01-01

    Fregnan, G. B. (University of Wisconsin, Madison) and D. W. Smith. Mycobactocidin, a new antiboitic active against mycobacteria. J. Bacteriol. 83:1069–1076. 1962.—A new antibiotic, produced by a strain of Staphylococcus epidermidis, is described. The name mycobactocidin is proposed because of the specificity of the antibiotic against mycobacteria. Methods for extraction and purification of the antibiotic are described. The water-soluble fraction is shown to be active in vitro. No toxicity could be detected in mice after intraperitoneal or subcutaneous injection. Enzymatic hydrolysis suggests a glyco-protein nature for the antibiotic. Mycobactocidin is soluble in distilled water at neutrality but is generally insoluble in organic solvents. The antibiotic is precipitated from aqueous solution by heavy metals. It is stable in the pH range of 2 to 8, but is precipitated at pH 3 to 4. An aqueous solution of the antibiotic is stable for several days at 5 C, and for longer periods if lyophilized and stored at −20 C. It does not pass through a dialysis membrane and its activity is retained after 15 to 20 hr of dialysis at 5 C. The activity was shown to be unrelated to the formation of hydrogen peroxide. Images PMID:16561941

  19. Non-Tuberculous Mycobacteria. An Emerging Threat?

    PubMed

    Martínez González, Susana; Cano Cortés, Arantxa; Sota Yoldi, Luis Alfonso; García García, José María; Alba Álvarez, Luz María; Palacios Gutiérrez, Juan José

    2017-10-01

    Non-tuberculous mycobacteria (NTM) isolates are becoming more common. The main objective of our study was to establish the number and diversity of NTM species in our region and their distribution according to the source sample, age and gender of the patients, and to analyse clinically significant isolates. Prospective study of all NTM isolated in Asturias from 2005 to 2012. Samples were processed following internationally accepted guidelines. Statistical analysis was based on Fisher's exact test for 2×2 contingency tables. A total of 3,284 mycobacteria were isolated: 1,499 Mycobacterium tuberculosis complex (MTB) and 1,785 NTM.During the study, NTM isolation rates increased while MTB isolation decreased. NTM were more frequent in men (P<.001). M.gordonae was the most frequently isolated species but did not cause disease in any case. NTM isolates from 212 patients were associated with clinically significant disease (17.1%). M.kansasii and M.avium were most commonly associated with disease. The number of M.kansasii isolates from men was statistically significant (P<.01). In our study, NTM isolates increased by 35%, compared with a 21% decline in cases of MTB. Both isolation of NTM and clinically significant cases were more common in men. Only 17.1% of NTM isolates were associated with disease, most commonly M.avium complex and M.kansasii. Copyright © 2017 SEPAR. Publicado por Elsevier España, S.L.U. All rights reserved.

  20. Pulmonary Disease Due to Nontuberculous Mycobacteria

    PubMed Central

    Glassroth, Jeffrey

    2015-01-01

    Since pulmonary nontuberculous mycobacteria (PNTM) lung disease was last reviewed in CHEST in 2008, new information has emerged spanning multiple domains, including epidemiology, transmission and pathogenesis, clinical presentation, diagnosis, and treatment. The overall prevalence of PNTM is increasing, and in the United States, areas of highest prevalence are clustered in distinct geographic locations with common environmental and socioeconomic factors. Although the accepted paradigm for transmission continues to be inhalation from the environment, provocative reports suggest that person-to-person transmission may occur. A panoply of host factors have been investigated in an effort to elucidate why infection from this bacteria develops in ostensibly immunocompetent patients, and there has been clarification that immunocompetent patients exhibit different histopathology from immunocompromised patients with nontuberculous mycobacteria infection. It is now evident that Mycobacterium abscessus, an increasingly prevalent cause of PNTM lung disease, can be classified into three separate subspecies with differing genetic susceptibility or resistance to macrolides. Recent publications also raise the possibility of improved control of PNTM through enhanced adherence to current treatment guidelines as well as new approaches to treatment and even prevention. These and other recent developments and insights that may inform our approach to PNTM lung disease are reviewed and discussed. PMID:26225805

  1. Mycobacteria in Terrestrial Small Mammals on Cattle Farms in Tanzania

    PubMed Central

    Durnez, Lies; Katakweba, Abdul; Sadiki, Harrison; Katholi, Charles R.; Kazwala, Rudovick R.; Machang'u, Robert R.; Portaels, Françoise; Leirs, Herwig

    2011-01-01

    The control of bovine tuberculosis and atypical mycobacterioses in cattle in developing countries is important but difficult because of the existence of wildlife reservoirs. In cattle farms in Tanzania, mycobacteria were detected in 7.3% of 645 small mammals and in cow's milk. The cattle farms were divided into “reacting” and “nonreacting” farms, based on tuberculin tests, and more mycobacteria were present in insectivores collected in reacting farms as compared to nonreacting farms. More mycobacteria were also present in insectivores as compared to rodents. All mycobacteria detected by culture and PCR in the small mammals were atypical mycobacteria. Analysis of the presence of mycobacteria in relation to the reactor status of the cattle farms does not exclude transmission between small mammals and cattle but indicates that transmission to cattle from another source of infection is more likely. However, because of the high prevalence of mycobacteria in some small mammal species, these infected animals can pose a risk to humans, especially in areas with a high HIV-prevalence as is the case in Tanzania. PMID:21785686

  2. Formation of intracytoplasmic membrane system of mycobacteria related to cell division.

    PubMed

    IMAEDA, T; OGURA, M

    1963-01-01

    Imaeda, Tamotsu (Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela) and Mitua Ogura. Formation of intracytoplasmic membrane system of mycobacteria related to cell division. J. Bacteriol. 85:150-163. 1963.-Mycobacterium leprae, M. lepraemurium, and a Mycobacterium sp. were observed with an electron microscope. In these bacilli, the three-dimensional structure of the intracytoplasmic membrane system consists of tubular infoldings of the invaginated plasma membrane. The moderately dense substance, presumably representing the cell-wall precursor, is found in the membranous system, especially in the rapid growth phase of mycobacteria. This system always shows an intimate relationship with cell division. A low-density zone, probably corresponding to the low-density substance which coats the cell wall, appears in the connecting regions of the system and in the longitudinal portion of the cell wall. These zones extend centripetally, and the separation of the cell wall occurs after the two zones meet. Based on these results, we hypothesize that the intracytoplasmic membrane system may produce cell-wall material during cell division of mycobacteria.

  3. Increased case finding of tuberculosis from sputum and sputum deposits after magnetic bead concentration of mycobacteria.

    PubMed

    Liu, Jun; Sun, Zhan-Qiang; Pei, Hao; Zhang, Shi-Liang; Zhang, Shu-Lin; Wilson, Stuart; De Smet, Koen; Cerullakandiyil, Noushad; Thayyullathil, Thanveer; Al-Suwaidi, Zubaida; Song, Yan-Zheng

    2013-05-01

    Concentration of mycobacteria from sputum by centrifugation prior to acid-fast microscopy increases case finding compared to direct microscopy of the sputum (direct smear). However, centrifugation has to be performed outside the safety cabinet and many laboratories do not have access to a centrifuge. Magnetic bead extraction of the mycobacteria is an alternative method that can be performed in a cabinet with just a magnet. Magnetic TB-Bead (Microsens Medtech Ltd) extraction of mycobacteria from sputum prior to microscopy was compared to direct smear on 78 sputum samples. Microscopy of the TB-Bead extracts identified all of 26 of the direct smear positive samples either with the same microscopy score or, in 19/27 of samples, with an increased microscopy score which aided microscopy detection. In addition, microscopy of the TB-Bead extracts identified 10 additional positive samples compared to direct smear; which represents a statistically significant increase in case finding of 38% (p = 0.002) compared to direct smear. In a separate study, TB-Beads enabled further 4 positive samples to be detected from 30 centrifuged pellets that were originally smear negative; two of these were subsequently found to be positive when the original deposits were reinvestigated by smear microscopy. By concentrating mycobacteria from sputum and sputum deposits, TB-Beads have been demonstrated to increase the number of positive sputum samples which could increase case-finding. The TB-Bead method is simple and rapid and compatible with use within a safety cabinet. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. PCR based detection of mycobacteria in paraffin wax embedded material routinely processed for morphological examination.

    PubMed Central

    Frevel, T; Schäfer, K L; Tötsch, M; Böcker, W; Dockhorn-Dworniczak, B

    1999-01-01

    BACKGROUND: The incidence of mycobacterial infections has increased during the past five years. A prompt diagnosis is indispensable for initiating appropriate treatment. Because culturing of mycobacteria takes three to six weeks and sensitivity of microscopic detection of acid fast bacilli is low, amplification methods provide promising possibilities. Recently, the polymerase chain reaction (PCR) has been shown to be useful for confirming a mycobacterial infection, especially in cases with unexpected histological findings or lack of suitable material for culturing. AIMS: To evaluate the impact of PCR based techniques in the detection of mycobacterial infections in uncultured routine histological specimens as an alternative to surgical pathology. METHODS: Two hundred and twenty nine formalin fixed and paraffin wax embedded samples from 141 patients with clinical or histological suspicion of a mycobacterial infection were investigated using three different PCR assays and Southern blotting. PCR results were compared with histology and culture and the patients' clinical findings. RESULTS: When using culture as the reference method, the sensitivity for the detection of mycobacteria of the tuberculosis complex was 90%, specificity was 92%, the positive predictive value was 81%, and the negative predictive value was 96%. The sensitivity for the detection of nontuberculous mycobacteria was 100% and specificity was 78%, the positive predictive value was 26%, and the negative predictive value was 100%. The patients' clinical findings supported the PCR positive results, indicating a mycobacterial infection in 11 of 18 initially culture negative cases and in 21 of 35 PCR positive cases without culture results. CONCLUSIONS: These results indicate that PCR based techniques are sensitive, specific, and rapid methods for the detection of mycobacteria in routinely processed paraffin wax embedded and formalin fixed histological samples. PMID:10748878

  5. Catecholates and mixed catecholate hydroxamates as artificial siderophores for mycobacteria.

    PubMed

    Wittmann, Steffen; Heinisch, Lothar; Scherlitz-Hofmann, Ina; Stoiber, Thomas; Ankel-Fuchs, Dorothe; Möllmann, Ute

    2004-02-01

    Different mono-, bis- or triscatecholates and mixed mono- or biscatecholate hydroxamates were synthesized as potential siderophores for mycobacteria. SiderOphore activity was tested by growth promotion assays using wild type strains and iron transport mutants of mycobacteria as well as Gram-negative bacteria. Some triscatecholates and biscatecholate hydroxamates were active in mutants of Mycobacterium smegmatis deficient in mycobactin and exochelin biosynthesis or exochelin permease, respectively, indicating an uptake route independent of the exochelin/mycobactin pathway. Structure activity relationships were studied. Ampicillin conjugates of some of these compounds were inactive against mycobacteria but active against Gram-negative bacteria.

  6. Complete plasmid sequence carrying type IV-like and type VII secretion systems from an atypical mycobacteria strain

    PubMed Central

    Morgado, Sergio Mascarenhas; Marín, Michel Abanto; Freitas, Fernanda S; Fonseca, Erica Lourenço; Vicente, Ana Carolina Paulo

    2017-01-01

    The genus Mycobacterium is highly diverse and ubiquitous in nature, comprehending fast- and slow-growing species with distinct impact in public health. The plasmid-mediated horizontal gene transfer represents one of the major events in bacteria evolution. Here, we report the complete sequence of a 160,489 bp circular plasmid (pCBMA213_2) from an atypical and fast-growing environmental mycobacteria. This is a unique plasmid, in comparison with the characterised mycobacteria plasmids, harboring a type IV-like and ESX-P2 type VII secretion systems. pCBMA213_2 can be further explored for evolutionary and conjugation studies as well as a tool to manipulate DNA within this bacteria genus. PMID:28591314

  7. Complete plasmid sequence carrying type IV-like and type VII secretion systems from an atypical mycobacteria strain.

    PubMed

    Morgado, Sergio Mascarenhas; Marín, Michel Abanto; Freitas, Fernanda S; Fonseca, Erica Lourenço; Vicente, Ana Carolina Paulo

    2017-07-01

    The genus Mycobacterium is highly diverse and ubiquitous in nature, comprehending fast- and slow-growing species with distinct impact in public health. The plasmid-mediated horizontal gene transfer represents one of the major events in bacteria evolution. Here, we report the complete sequence of a 160,489 bp circular plasmid (pCBMA213_2) from an atypical and fast-growing environmental mycobacteria. This is a unique plasmid, in comparison with the characterised mycobacteria plasmids, harboring a type IV-like and ESX-P2 type VII secretion systems. pCBMA213_2 can be further explored for evolutionary and conjugation studies as well as a tool to manipulate DNA within this bacteria genus.

  8. Mycobacteria Clumping Increase Their Capacity to Damage Macrophages

    PubMed Central

    Brambilla, Cecilia; Llorens-Fons, Marta; Julián, Esther; Noguera-Ortega, Estela; Tomàs-Martínez, Cristina; Pérez-Trujillo, Miriam; Byrd, Thomas F.; Alcaide, Fernando; Luquin, Marina

    2016-01-01

    The rough morphotypes of non-tuberculous mycobacteria have been associated with the most severe illnesses in humans. This idea is consistent with the fact that Mycobacterium tuberculosis presents a stable rough morphotype. Unlike smooth morphotypes, the bacilli of rough morphotypes grow close together, leaving no spaces among them and forming large aggregates (clumps). Currently, the initial interaction of macrophages with clumps remains unclear. Thus, we infected J774 macrophages with bacterial suspensions of rough morphotypes of M. abscessus containing clumps and suspensions of smooth morphotypes, primarily containing isolated bacilli. Using confocal laser scanning microscopy and electron microscopy, we observed clumps of at least five rough-morphotype bacilli inside the phagocytic vesicles of macrophages at 3 h post-infection. These clumps grew within the phagocytic vesicles, killing 100% of the macrophages at 72 h post-infection, whereas the proliferation of macrophages infected with smooth morphotypes remained unaltered at 96 h post-infection. Thus, macrophages phagocytose large clumps, exceeding the bactericidal capacities of these cells. Furthermore, proinflammatory cytokines and granuloma-like structures were only produced by macrophages infected with rough morphotypes. Thus, the present study provides a foundation for further studies that consider mycobacterial clumps as virulence factors. PMID:27757105

  9. Characterizing Non-Tuberculous Mycobacteria Infection in Bronchiectasis

    PubMed Central

    Faverio, Paola; Stainer, Anna; Bonaiti, Giulia; Zucchetti, Stefano C.; Simonetta, Edoardo; Lapadula, Giuseppe; Marruchella, Almerico; Gori, Andrea; Blasi, Francesco; Codecasa, Luigi; Pesci, Alberto; Chalmers, James D.; Loebinger, Michael R.; Aliberti, Stefano

    2016-01-01

    Chronic airway infection is a key aspect of the pathogenesis of bronchiectasis. A growing interest has been raised on non-tuberculous mycobacteria (NTM) infection. We aimed at describing the clinical characteristics, diagnostic process, therapeutic options and outcomes of bronchiectasis patients with pulmonary NTM (pNTM) disease. This was a prospective, observational study enrolling 261 adult bronchiectasis patients during the stable state at the San Gerardo Hospital, Monza, Italy, from 2012 to 2015. Three groups were identified: pNTM disease; chronic P. aeruginosa infection; chronic infection due to bacteria other than P. aeruginosa. NTM were isolated in 32 (12%) patients, and among them, a diagnosis of pNTM disease was reached in 23 cases. When compared to chronic P. aeruginosa infection, patients with pNTM were more likely to have cylindrical bronchiectasis and a “tree-in-bud” pattern, a history of weight loss, a lower disease severity and a lower number of pulmonary exacerbations. Among pNTM patients who started treatment, 68% showed a radiological improvement, and 37% achieved culture conversion without recurrence, while 21% showed NTM isolation recurrence. NTM isolation seems to be a frequent event in bronchiectasis patients, and few parameters might help to suspect NTM infection. Treatment indications and monitoring still remain an important area for future research. PMID:27854334

  10. Disinfectant Susceptibility Profiling of Glutaraldehyde-Resistant Nontuberculous Mycobacteria.

    PubMed

    Burgess, Winona; Margolis, Alyssa; Gibbs, Sara; Duarte, Rafael Silva; Jackson, Mary

    2017-07-01

    OBJECTIVE Activated alkaline glutaraldehyde (GTA) remains one of the most widely used high-level disinfectants worldwide. However, several reports have highlighted the potential for nontuberculous mycobacteria to develop high-level resistance to this product. Because aldehyde resistance may lead to cross-resistance to other biocides, we investigated the susceptibility profile of GTA-resistant Mycobacterium chelonae and M. abscessus isolates to various disinfectant chemistries. METHODS High-level disinfectants commonly used in the reprocessing of endoscopes and other heat-sensitive, semicritical medical equipment, including different formulations of aldehyde-based products and oxidizing agents, were tested against 10 slow- and fast-growing, GTA-susceptible and GTA-resistant, Mycobacterium isolates in suspension tests and carrier tests at different temperatures. RESULTS While peracetic acid- and hydrogen peroxide-based disinfectants (S40, Resert XL, Reliance DG) efficiently killed all of the Mycobacterium isolates, GTA- and ortho-phthalaldehyde-based products (ie, Cidex, Aldahol, Cidex OPA) showed variable efficacy against GTA-resistant strains despite the ability of some formulations (Aldahol) to overcome the resistance of some of these isolates, especially when the temperature was increased from 20°C to 25°C. CONCLUSIONS Application permitting, oxidizing chemistries may provide a safe alternative to aldehyde-based products, particularly in GTA-resistant mycobacterial outbreaks. Infect Control Hosp Epidemiol 2017;38:784-791.

  11. Characterizing Non-Tuberculous Mycobacteria Infection in Bronchiectasis.

    PubMed

    Faverio, Paola; Stainer, Anna; Bonaiti, Giulia; Zucchetti, Stefano C; Simonetta, Edoardo; Lapadula, Giuseppe; Marruchella, Almerico; Gori, Andrea; Blasi, Francesco; Codecasa, Luigi; Pesci, Alberto; Chalmers, James D; Loebinger, Michael R; Aliberti, Stefano

    2016-11-16

    Chronic airway infection is a key aspect of the pathogenesis of bronchiectasis. A growing interest has been raised on non-tuberculous mycobacteria (NTM) infection. We aimed at describing the clinical characteristics, diagnostic process, therapeutic options and outcomes of bronchiectasis patients with pulmonary NTM (pNTM) disease. This was a prospective, observational study enrolling 261 adult bronchiectasis patients during the stable state at the San Gerardo Hospital, Monza, Italy, from 2012 to 2015. Three groups were identified: pNTM disease; chronic P. aeruginosa infection; chronic infection due to bacteria other than P. aeruginosa. NTM were isolated in 32 (12%) patients, and among them, a diagnosis of pNTM disease was reached in 23 cases. When compared to chronic P. aeruginosa infection, patients with pNTM were more likely to have cylindrical bronchiectasis and a "tree-in-bud" pattern, a history of weight loss, a lower disease severity and a lower number of pulmonary exacerbations. Among pNTM patients who started treatment, 68% showed a radiological improvement, and 37% achieved culture conversion without recurrence, while 21% showed NTM isolation recurrence. NTM isolation seems to be a frequent event in bronchiectasis patients, and few parameters might help to suspect NTM infection. Treatment indications and monitoring still remain an important area for future research.

  12. Synthesis and biological activity of alkynoic acids derivatives against mycobacteria

    PubMed Central

    Vilchèze, Catherine; Leung, Lawrence W.; Bittman, Robert; Jacobs, William R.

    2015-01-01

    2-alkynoic acids have bactericidal activity against Mycobacterium smegmatis but their activity fall sharply as the length of the carbon chain increased. In this study, derivatives of 2- alkynoic acids were synthesized and tested against fast- and slow-growing mycobacteria. Their activity was first evaluated in M. smegmatis against their parental 2-alkynoic acids, as well as isoniazid, a first-line antituberculosis drug. The introduction of additional unsaturation or heteroatoms into the carbon chain enhanced the antimycobacterial activity of longer chain alkynoic acids (more than 19 carbons long). In contrast, although the modification of the carboxylic group did not improve the antimycobacterial activity, it significantly reduced the toxicity of the compounds against eukaryotic cells. Importantly, 4-(alkylthio)but-2-ynoic acids, had better bactericidal activity than the parental 2-alkynoic acids and on a par with isoniazid against the slow-grower Mycobacterium bovis BCG. These compounds had also low toxicity against eukaryotic cells, suggesting that they could be potential therapeutic agents against other types of topical mycobacterial infections causing skin diseases including Mycobacterium abscessus, Mycobacterium ulcerans, and Mycobacterium leprae. Moreover, they provide a possible scaffold for future drug development. PMID:26256431

  13. Thiol specific oxidative stress response in Mycobacteria.

    PubMed

    Dosanjh, Nirpjit S; Rawat, Mamta; Chung, Ji-Hae; Av-Gay, Yossef

    2005-08-01

    The cellular response of mycobacteria to thiol specific oxidative stress was studied in Mycobacterium bovis BCG cultures. Two-dimensional gel electrophoresis revealed that upon diamide treatment at least 60 proteins were upregulated. Fourteen of these proteins were identified by MALDI-MS; four proteins, AhpC, Tpx, GroEL2, and GroEL1 are functionally related to oxidative stress response; eight proteins, LeuC, LeuD, Rv0224c, Rv3029c, AsnB, Rv2971, PheA and HisH are classified as part of the bacterial intermediary metabolism and respiration pathways; protein EchA14 belong to lipid metabolism, and NrdE, belongs to the mycobacterial information pathway category. Reverse transcription followed by quantitative real time PCR in response to diamide stress demonstrated that protein expression is directly proportional to the corresponding gene transcription.

  14. Tetracycline-inducible gene regulation in mycobacteria

    PubMed Central

    Blokpoel, Marian C. J.; Murphy, Helen N.; O'Toole, Ronan; Wiles, Siouxsie; Runn, Ellen S. C.; Stewart, Graham R.; Young, Douglas B.; Robertson, Brian D.

    2005-01-01

    A system for the tetracycline-inducible regulation of gene expression in mycobacteria has been developed. We have sub-cloned the tetRO region from the Corynebacterium glutamicum TetZ locus into a mycobacterial shuttle plasmid, making expression of genes cloned downstream of tetRO responsive to tetracycline. Using the luxAB-encoded luciferase from Vibrio harveyi as a reporter (pMind-Lx), we observed a 40-fold increase in light output from Mycobacterium smegmatis cultures 2 h after adding 20 ng ml−1 of tetracycline. Similarly, exposure to the drug resulted in up to 20-fold increase in relative light units from M.bovis BCG carrying the reporter construct, and a 10-fold increase for M.tuberculosis. Tetracycline induction was demonstrated in log and stationary phase cultures. To evaluate whether this system is amenable to use in vivo, J774 macrophages were infected with M.bovis BCG[pMind-Lx], treated with amikacin to kill extracellular bacteria, and then incubated with tetracycline. A 10-fold increase in light output was measured after 24 h, indicating that intracellular bacteria are accessible and responsive to exogenously added tetracycline. To test the use of the tetracycline-inducible system for conditional gene silencing, mycobacteria were transformed with a pMind construct with tetRO driving expression of antisense RNA for the ftsZ gene. Bacterial cells containing the antisense construct formed filaments after 24 h exposure to tetracycline. These results demonstrate the potential of this tetracycline-regulated system for the manipulation of mycobacterial gene expression inside and outside cells. PMID:15687380

  15. Arylamine N-Acetyltransferases in Mycobacteria

    PubMed Central

    Sim, Edith; Sandy, James; Evangelopoulos, Dimitrios; Fullam, Elizabeth; Bhakta, Sanjib; Westwood, Isaac; Krylova, Anna; Lack, Nathan; Noble, Martin

    2008-01-01

    Polymorphic Human arylamine N-acetyltransferase (NAT2) inactivates the anti-tubercular drug isoniazid by acetyltransfer from acetylCoA. There are active NAT proteins encoded by homologous genes in mycobacteria including M. tuberculosis, M. bovis BCG, M. smegmatis and M. marinum. Crystallographic structures of NATs from M. smegmatis and M. marinum, as native enzymes and with isoniazid bound share a similar fold with the first NAT structure, Salmonella typhimurium NAT. There are three approximately equal domains and an active site essential catalytic triad of cysteine, histidine and aspartate in the first two domains. An acetyl group from acetylCoA is transferred to cysteine and then to the acetyl acceptor e.g. isoniazid. M. marinum NAT binds CoA in a more open mode compared with CoA binding to human NAT2. The structure of mycobacterial NAT may promote its role in synthesis of cell wall lipids, identified through gene deletion studies. NAT protein is essential for survival of M. bovis BCG in macrophage as are the proteins encoded by other genes in the same gene cluster (hsaA-D). HsaA-D degrade cholesterol, essential for mycobacterial survival inside macrophage. Nat expression remains to be fully understood but is co-ordinated with hsaA-D and other stress response genes in mycobacteria. Amide synthase genes in the streptomyces are also nat homologues. The amide synthases are predicted to catalyse intramolecular amide bond formation and creation of cyclic molecules, e.g. geldanamycin. Lack of conservation of the CoA binding cleft residues of M. marinum NAT suggests the amide synthase reaction mechanism does not involve a soluble CoA intermediate during amide formation and ring closure. PMID:18680471

  16. Germicidal Effect of Ultraviolet Irradiation on Paper Contaminated with Mycobacteria

    PubMed Central

    Huber, T. W.; Reddick, R. Anne; Kubica, G. P.

    1970-01-01

    An ultraviolet chamber for decontaminating single sheets of paper seeded with mycobacteria was investigated. The ultraviolet killing effect ranged from 50% for Mycobacterium kansasii to more than 99% for M. tuberculosis. PMID:4985435

  17. Epidemiology of nontuberculous mycobacteria, an emerging environmental pathogen

    EPA Science Inventory

    Nontuberculous mycobacteria (NTM) is an environmentally transmitted pathogen primarily associated with water and soil exposure. It is increasingly recognized in the developed world and may manifest as infection or colonization of multiple anatomic sites. Nontuberculous mycobacter...

  18. Epidemiology of nontuberculous mycobacteria, an emerging environmental pathogen

    EPA Science Inventory

    Nontuberculous mycobacteria (NTM) is an environmentally transmitted pathogen primarily associated with water and soil exposure. It is increasingly recognized in the developed world and may manifest as infection or colonization of multiple anatomic sites. Nontuberculous mycobacter...

  19. Comprehensive identification of conditionally essential genes in mycobacteria.

    PubMed

    Sassetti, C M; Boyd, D H; Rubin, E J

    2001-10-23

    An increasing number of microbial genomes have been completely sequenced, and the identified genes are categorized based on their homology to genes of known function. However, the function of a large number of genes cannot be determined on this basis alone. Here, we describe a technique, transposon site hybridization (TraSH), which allows rapid functional characterization by identifying the complete set of genes required for growth under different conditions. TraSH combines high-density insertional mutagenesis with microarray mapping of pools of mutants. We have made large pools of independent transposon mutants in mycobacteria by using a mariner-based transposon and efficient phage transduction. By using TraSH, we have defined the set of genes required for growth of Mycobacterium bovis bacillus Calmette-Guérin on minimal but not rich medium. Genes of both known and unknown functions were identified. Of the genes with known functions, nearly all were involved in amino acid biosynthesis. TraSH is a powerful method for categorizing gene function that should be applicable to a variety of microorganisms.

  20. Comparative genome analyses of mycobacteria give better insights into their evolution

    PubMed Central

    Wee, Wei Yee; Dutta, Avirup; Choo, Siew Woh

    2017-01-01

    Mycobacteria a genus of Actinobacteria are widespread in nature ranging from soil-dwelling saprophytes to human and animal pathogens. The rate of growth has been a classifying factor for the Mycobacterium spp., dividing them into the rapid growers and the slow growers. Here we have performed a comparative genome study of mycobacterial species in order to get better understanding of their evolution, particularly to understand the distinction between the rapid and slow growers. Our study shows that the slow growers had generally gained and lost more genes compared to the rapid growers. The slow growers might haved eventually lost genes (LivFGMH operon, shaACDEFG genes and MspA porin) that could contribute to the slow growth rate of the slow growers. The genes gained and lost in mycobacteria had eventually helped these bacteria to adapt to different environments and have led to the evolution of the present day rapid and slow growers. Our results also show high number of Mycobacterium abscessus specific genes (811 genes) and some of them are associated with the known bacterial quorum sensing genes that might be important for Mycobacterium abscessus to adapt and survive in variety of unfavorable environments. Mycobacterium abscessus also does not contains genes involved in the bacterial defense system and together with the quorum sensing genes may have contributed to the high gene gain rate of Mycobacterium abscessus. PMID:28291784

  1. DNA polymorphism in Mycobacterium paratuberculosis, "wood pigeon mycobacteria," and related mycobacteria analyzed by field inversion gel electrophoresis.

    PubMed Central

    Lévy-Frébault, V V; Thorel, M F; Varnerot, A; Gicquel, B

    1989-01-01

    Mycobacterium paratuberculosis strains, mycobacteria from patients suffering from Crohn's disease, "wood pigeon mycobacteria," and representatives of Mycobacterium avium-Mycobacterium intracellulare were compared by restriction endonuclease DraI digestion and field inversion gel electrophoresis. Characteristic profiles were seen for M. paratuberculosis, including isolates from patients suffering from Crohn's disease, for wood pigeon mycobacteria, and for M. avium-M. intracellulare serotypes 2, 16, 18, and 19. Two M. paratuberculosis strains used for vaccine production (St 18 and 316 F) presented patterns different from those of the other M. paratuberculosis strains. Strains St 18 yielded a pattern identical to that of the M. avium type strain serotype 2, whereas 316 F gave a unique pattern. The method developed in this study represents a useful taxonomic tool for the identification and classification of mycobacteria. Images PMID:2574186

  2. Genotypic identification of mycobacteria by nucleic acid sequence determination: report of a 2-year experience in a clinical laboratory.

    PubMed Central

    Kirschner, P; Springer, B; Vogel, U; Meier, A; Wrede, A; Kiekenbeck, M; Bange, F C; Böttger, E C

    1993-01-01

    Clinical isolates of Mycobacterium spp. were identified by direct sequence determination of 16S rRNA gene fragments amplified by polymerase chain reaction. Identification was based on a hypervariable region within the 16S rRNA gene in which mycobacterial species are characterized by species-specific nucleotide sequences. A manually aligned data base including the signature sequences of 52 species of mycobacteria easily allowed rapid and correct identification. The results of this study demonstrate that polymerase chain reaction-mediated direct sequence determination can be used as a rapid and reliable method for the identification of mycobacteria in the clinical laboratory. In addition, the prompt recognition of previously undescribed species is now feasible. PMID:7505291

  3. Characterization of rifampin-resistance in pathogenic mycobacteria.

    PubMed Central

    Williams, D L; Waguespack, C; Eisenach, K; Crawford, J T; Portaels, F; Salfinger, M; Nolan, C M; Abe, C; Sticht-Groh, V; Gillis, T P

    1994-01-01

    The emergence of rifampin-resistant strains of pathogenic mycobacteria has threatened the usefulness of this drug in treating mycobacterial diseases. Critical to the treatment of individuals infected with resistant strains is the rapid identification of these strains directly from clinical specimens. It has been shown that resistance to rifampin in Mycobacterium tuberculosis and Mycobacterium leprae apparently involves mutations in the rpoB gene encoding the beta-subunit of the RNA polymerases of these species. DNA sequences were obtained from a 305-bp fragment of the rpoB gene from 110 rifampin-resistant and 10 rifampin-susceptible strains of M. tuberculosis from diverse geographical regions throughout the world. In 102 of 110 rifampin-resistant strains 16 mutations affecting 13 amino acids were observed. No mutations were observed in rifampin-susceptible strains. No association was found between particular mutations in the rpoB gene and drug susceptibility patterns of multidrug-resistant M. tuberculosis strains. Drug-resistant M. tuberculosis strains from the same outbreak and exhibiting the same IS6110 DNA fingerprint and drug susceptibility pattern contained the same mutation in the rpoB gene. However, mutations are not correlated with IS6110 profiling outside of epidemics. The evolution of rifampin resistance as a consequence of mutations in the rpoB gene was documented in a patient who developed rifampin resistance during the course of treatment. Rifampin-resistant strains of M. leprae, Mycobacterium avium, and Mycobacterium africanum contained mutations in the rpoB gene similar to that documented for M. tuberculosis. This information served as the basis for developing a rapid DNA diagnostic assay (PCR-heteroduplex formation) for the detection of rifampin susceptibility of M. tuberculosis. Images PMID:7840574

  4. Single-cell dynamics of the chromosome replication and cell division cycles in mycobacteria.

    PubMed

    Santi, Isabella; Dhar, Neeraj; Bousbaine, Djenet; Wakamoto, Yuichi; McKinney, John D

    2013-01-01

    During the bacterial cell cycle, chromosome replication and cell division must be coordinated with overall cell growth in order to maintain the correct ploidy and cell size. The spatial and temporal coordination of these processes in mycobacteria is not understood. Here we use microfluidics and time-lapse fluorescence microscopy to measure the dynamics of cell growth, division and chromosome replication in single cells of Mycobacterium smegmatis. We find that single-cell growth is size-dependent (large cells grow faster than small cells), which implicates a size-control mechanism in cell-size homoeostasis. Asymmetric division of mother cells gives rise to unequally sized sibling cells that grow at different velocities but show no differential sensitivity to antibiotics. Individual cells are restricted to one round of chromosome replication per cell division cycle, although replication usually initiates in the mother cell before cytokinesis and terminates in the daughter cells after cytokinesis. These studies reveal important differences between cell cycle organization in mycobacteria compared with better-studied model organisms.

  5. A rapidly growing epidermoid cyst in an intrapancreatic accessory spleen treated by laparoscopic spleen-preserving distal pancreatectomy: Report of a case.

    PubMed

    Kumamoto, Yusuke; Kaizu, Takashi; Tajima, Hiroshi; Kubo, Hidefumi; Nishiyama, Ryo; Watanabe, Masahiko

    2015-06-02

    Epidermoid cysts arising in an intrapancreatic accessory spleen are exceedingly rare, furthermore the natural course of them is hardly known. We report a case correctly diagnosed with epidermoid cyst in an intrapancreatic accessory spleen, followed by 1 year observation, finally underwent surgical treatment. The patient presented with diarrhea. Contrast-enhanced computed tomography (CT) revealed a pancreatic cyst 20 mm in diameter, surrounded by a solid component showing the same enhancement as the spleen, suggesting the presence of an epidermoid cyst in an intrapancreatic accessory spleen. One year later, back discomfort developed, and a CT scan revealed that the cyst had grown to 38 mm in diameter. To obtain a definitive diagnosis, we performed a laparoscopic spleen-preserving distal pancreatectomy. The histopathological diagnosis was compatible with an epidermoid cyst in an intrapancreatic accessory spleen, which is benign. The postoperative course was uneventful. This case demonstrates that an epidermoid cyst arising in an intrapancreatic accessory spleen can rapidly grow, even if it is benign. Laparoscopic spleen-preserving distal pancreatectomy can be a useful procedure, with the advantages of low invasiveness and organ preservation, for the treatment of benign or low-grade malignant tumors located in the pancreatic body or tail.

  6. Survival of Environmental Mycobacteria in Acanthamoeba polyphaga

    PubMed Central

    Adékambi, Toïdi; Ben Salah, Skandar; Khlif, Mohamed; Raoult, Didier; Drancourt, Michel

    2006-01-01

    Free-living amoebae in water are hosts to many bacterial species living in such an environment. Such an association enables bacteria to select virulence factors and survive in adverse conditions. Waterborne mycobacteria (WBM) are important sources of community- and hospital-acquired outbreaks of nontuberculosis mycobacterial infections. However, the interactions between WBM and free-living amoebae in water have been demonstrated for only few Mycobacterium spp. We investigated the ability of a number (n = 26) of Mycobacterium spp. to survive in the trophozoites and cysts of Acanthamoeba polyphaga. All the species tested entered the trophozoites of A. polyphaga and survived at this location over a period of 5 days. Moreover, all Mycobacterium spp. survived inside cysts for a period of 15 days. Intracellular Mycobacterium spp. within amoeba cysts survived when exposed to free chlorine (15 mg/liter) for 24 h. These data document the interactions between free-living amoebae and the majority of waterborne Mycobacterium spp. Further studies are required to examine the effects of various germicidal agents on the survival of WBM in an aquatic environment. PMID:16957218

  7. Epidemiology of Nontuberculous Mycobacteria in French Polynesia

    PubMed Central

    Phelippeau, Michael; Aboubaker Osman, Djaltou; Musso, Didier

    2015-01-01

    As few data are available in the Pacific countries and territories of the Oceania region regarding nontuberculous mycobacteria, we retrospectively identified 87 such isolates from French Polynesia from 2008 to 2013 by hybridization using DNA-strip, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and partial rpoB gene sequencing. Partial rpoB gene sequencing classified 42/87 (48.3%) isolates in the Mycobacterium fortuitum complex, 28 (32.2%) in the Mycobacterium abscessus complex, 8 (9.2%) in the Mycobacterium mucogenicum complex, and 5 (5.7%) in the Mycobacterium avium complex. Two isolates were identified as Mycobacterium acapulcensis and Mycobacterium cosmeticum by partial 16S rRNA gene sequencing. One isolate, unidentified by MALDI-TOF MS and yielding less than 92% and 96% sequence similarity with rpoB and hsp65 reference sequences, respectively, was regarded as a potentially new species. Samples from three patients exhibiting ≥2 Mycobacterium porcinum isolates and from one patient with emphysema and a lung abscess exhibiting 2 Mycobacterium senegalense isolates fulfilled the American Thoracic Society microbiological criteria for nontuberculous mycobacterial lung infection. Remote geographic areas, such as French Polynesia, are potential sources for the discovery of new mycobacterial species. PMID:26400787

  8. Respiratory infections due to nontuberculous mycobacterias.

    PubMed

    Máiz Carro, Luis; Barbero Herranz, Esther; Nieto Royo, Rosa

    2017-09-15

    The most common infections caused by nontuberculous mycobacteria (NTM) are lung infections. The microorganisms causing these infections most frequently are Mycobacterium avium complex, Mycobacterium kansasii and Mycobacterium abscessus complex. Their incidence has increased in the last three decades. After identifying an NTM in the respiratory tract, clinical and radiological aspects must be considered to determine if isolations are clinically relevant. Predisposing conditions that could contribute to infection must also be investigated. Pulmonary disease due to NTM is presented in three clinical forms: a) pneumonitis due to hypersensitivity; b) fibrocavitary form; and c) nodular-bronchiectasic. The diagnosis of respiratory disease due to NTM does not make it obligatory to immediately initiate treatment. Before initiating the latter, other factors must be considered, such as age, comorbidities, life expectancy, due to the prolonged nature of treatments, with potential side effects and, in many cases, only a slight response to the treatment. Copyright © 2017 Elsevier España, S.L.U. All rights reserved.

  9. Environmental Nontuberculous Mycobacteria in the Hawaiian Islands

    PubMed Central

    Epperson, L. Elaine; Reynolds, Paul R.; Smith, Terry; Iakhiaeva, Elena; Bankowski, Matthew J.; Wallace, Richard J.; Chan, Edward D.; Falkinham, Joseph O.; Strong, Michael

    2016-01-01

    Lung disease caused by nontuberculous mycobacteria (NTM) is an emerging infectious disease of global significance. Epidemiologic studies have shown the Hawaiian Islands have the highest prevalence of NTM lung infections in the United States. However, potential environmental reservoirs and species diversity have not been characterized. In this cross-sectional study, we describe molecular and phylogenetic comparisons of NTM isolated from 172 household plumbing biofilms and soil samples from 62 non-patient households and 15 respiratory specimens. Although non-uniform geographic sampling and availability of patient information were limitations, Mycobacterium chimaera was found to be the dominant species in both environmental and respiratory specimens. In contrast to previous studies from the continental U.S., no Mycobacterium avium was identified. Mycobacterium intracellulare was found only in respiratory specimens and a soil sample. We conclude that Hawai’i’s household water sources contain a unique composition of Mycobacterium avium complex (MAC), increasing our appreciation of NTM organisms of pulmonary importance in tropical environments. PMID:27780201

  10. Environmental Nontuberculous Mycobacteria in the Hawaiian Islands.

    PubMed

    Honda, Jennifer R; Hasan, Nabeeh A; Davidson, Rebecca M; Williams, Myra D; Epperson, L Elaine; Reynolds, Paul R; Smith, Terry; Iakhiaeva, Elena; Bankowski, Matthew J; Wallace, Richard J; Chan, Edward D; Falkinham, Joseph O; Strong, Michael

    2016-10-01

    Lung disease caused by nontuberculous mycobacteria (NTM) is an emerging infectious disease of global significance. Epidemiologic studies have shown the Hawaiian Islands have the highest prevalence of NTM lung infections in the United States. However, potential environmental reservoirs and species diversity have not been characterized. In this cross-sectional study, we describe molecular and phylogenetic comparisons of NTM isolated from 172 household plumbing biofilms and soil samples from 62 non-patient households and 15 respiratory specimens. Although non-uniform geographic sampling and availability of patient information were limitations, Mycobacterium chimaera was found to be the dominant species in both environmental and respiratory specimens. In contrast to previous studies from the continental U.S., no Mycobacterium avium was identified. Mycobacterium intracellulare was found only in respiratory specimens and a soil sample. We conclude that Hawai'i's household water sources contain a unique composition of Mycobacterium avium complex (MAC), increasing our appreciation of NTM organisms of pulmonary importance in tropical environments.

  11. Characterization of non-tuberculosis mycobacteria by neutron radiography.

    PubMed

    da Silva, Jaqueline M; Crispim, Verginia Reis; da Silva, Marlei Gomes; Furtado, Vanessa Rodrigues; Duarte, Rafael Da Silva

    2013-07-01

    The genus Mycobacterium shares many characteristics with Corynebacterium and Actinomyces genera, among which the genomic guanine plus cytosine content and the production of long branched-chain fatty acids, known as mycolic acids are enhanced. Growth rate and optimal temperature of mycobacteria are variable. The genus comprises more than 140 known species; however Mycobacterium fortuitum, a fast growing nontuberculous mycobacterium, is clinically significant, because it has been associated to several lesions following surgery procedures such as liposuction, silicone breast and pacemaker implants, exposure to prosthetic materials besides sporadic lesions in the skin, soft tissues and rarely lungs. The objective of the present study is to reduce the time necessary for M. fortuitum characterization based on its morphology and the use of the neutron radiography technique substituting the classical biochemical assays. We also aim to confirm the utility of dendrimers as boron carriers. The samples were sterilized through conventional protocols using 10% formaldehyde. In the incubation process, two solutions with different molar ratios (10:1 and 20:1) of sodium borate and PAMAM G4 dendrimer and also pure sodium borate were used. After doping and sterilization procedures, the samples were deposited on CR-39 sheets, irradiated with a 4.6×10(5) n/cm(2)s thermal neutron flux for 30 min, from the J-9 irradiation channel of the Argonauta IEN/CNEN reactor. The images registered in the CR-39 were visualized in a Nikon E400 optical transmission microscope and captured by a Nikon Coolpix 995 digital camera. Developing the nuclear tracks registered in the CR-39 allowed a 1000× enlargement of mycobacterium images, facilitating their characterization, the use of more sophisticated equipment not being necessary. The use of neutron radiography technique reduced the time necessary for characterization. Doping with PAMAM dendrimer improved the visualization of NTM in neutron radiography

  12. Use of near-infrared reflectance spectroscopy for the rapid determination of the digestible energy and metabolizable energy content of corn fed to growing pigs.

    PubMed

    Li, Juntao; Li, Quanfeng; Li, Defa; Chen, Yiqiang; Wang, Xiaoxiao; Yang, Wunjun; Zhang, Liying

    2016-01-01

    The ability of near-infrared reflectance spectroscopy (NIRS) to determine the digestible energy (DE) and metabolizable energy (ME) content of corn fed to growing pigs was tested. One hundred and seventeen corn samples, comprising different planting regions and varieties were collected from all over China in a three-year period. The samples were randomly split into a calibration set (n = 88) and a validation set (n = 29). The actual and calculated DE and ME content of the corn samples was determined by digestion-metabolism experiments and the prediction equations of Noblet and Perez (J Anim Sci. 71:3389-98,1993). The samples were then subjected to NIRS scanning and calibrations were performed by the modified partial least square (MPLS) regression method based on 77 different spectral pre-treatments. The NIRS equations based on the actually determined and calculated DE and ME were built separately and then validated using validation samples. The NIRS equations obtained from actually determined DE, the coefficient of determination for calibration (RSQcal), cross-validation (R(2) CV), and validation (RSQv) were 0.89, 0.87 and 0.86, and these values for determined ME were 0.87, 0.86 and 0.86. For the NIRS equations built from calculated DE, the RSQcal, R(2) CV, and RSQv values were 0.88, 0.85 and 0.84, and these values for calculated ME were 0.86, 0.84 and 0.82. Except for the equation based on calculated ME (RPDv = 2.38, < 2.50), the other three equations built from actually determined energy and calculated DE produced good prediction performance (RPDv ranging from 2.53 to 2.69, > 2.50) when applied to validation samples. These results indicate that NIRS can be used as a quantitative method for the rapid determination of the available energy in corn fed to growing pigs, and the NIRS equations based on the actually determined energy produced better predictive performance than those built from calculated energy values.

  13. Delineating bacteriostatic and bactericidal targets in mycobacteria using IPTG inducible antisense expression.

    PubMed

    Kaur, Parvinder; Agarwal, Saurabh; Datta, Santanu

    2009-06-15

    In order to identify novel high value antibacterial targets it is desirable to delineate whether the inactivation of the target enzyme will lead to bacterial death or stasis. This knowledge is particularly important in slow growing organisms, like mycobacteria, where most of the viable anti-tubercular agents are bactericidal. A bactericidal target can be identified through the conditional deletion or inactivation of the target gene at a relatively high cell number and subsequently following the time course of survival for the bacteria. A simple protocol to execute conditional inactivation of a gene is by antisense expression. We have developed a mycobacteria specific IPTG inducible vector system and monitored the effect of antisense inhibition of several known essential genes in mycobacteria by following their survival kinetics. By this method, we could differentiate between genes whose down regulation lead to bacteriostatic or bactericidal effect. Targets for standard anti-tubercular drugs like inhA for isoniazid, rpoB and C for rifampicin, and gyr A/B for flouroquinolones were shown to be bactericidal. In contrast targets like FtsZ behaved in a bacteriostatic manner. Induction of antisense expression in embB and ribosomal RNA genes, viz., rplJ and rpsL showed only a marginal growth inhibition. The specificity of the antisense inhibition was conclusively shown in the case of auxotrophic gene ilvB. The bactericidal activity following antisense expression of ilvB was completely reversed when the growth media was supplemented with the isoleucine, leucine, valine and pantothenate. Additionally, under these conditions the expression of several genes in branched chain amino acid pathway was severely suppressed indicating targeted gene inactivation.

  14. Delineating Bacteriostatic and Bactericidal Targets in Mycobacteria Using IPTG Inducible Antisense Expression

    PubMed Central

    Kaur, Parvinder; Agarwal, Saurabh; Datta, Santanu

    2009-01-01

    In order to identify novel high value antibacterial targets it is desirable to delineate whether the inactivation of the target enzyme will lead to bacterial death or stasis. This knowledge is particularly important in slow growing organisms, like mycobacteria, where most of the viable anti-tubercular agents are bactericidal. A bactericidal target can be identified through the conditional deletion or inactivation of the target gene at a relatively high cell number and subsequently following the time course of survival for the bacteria. A simple protocol to execute conditional inactivation of a gene is by antisense expression. We have developed a mycobacteria specific IPTG inducible vector system and monitored the effect of antisense inhibition of several known essential genes in mycobacteria by following their survival kinetics. By this method, we could differentiate between genes whose down regulation lead to bacteriostatic or bactericidal effect. Targets for standard anti-tubercular drugs like inhA for isoniazid, rpoB and C for rifampicin, and gyr A/B for flouroquinolones were shown to be bactericidal. In contrast targets like FtsZ behaved in a bacteriostatic manner. Induction of antisense expression in embB and ribosomal RNA genes, viz., rplJ and rpsL showed only a marginal growth inhibition. The specificity of the antisense inhibition was conclusively shown in the case of auxotrophic gene ilvB. The bactericidal activity following antisense expression of ilvB was completely reversed when the growth media was supplemented with the isoleucine, leucine, valine and pantothenate. Additionally, under these conditions the expression of several genes in branched chain amino acid pathway was severely suppressed indicating targeted gene inactivation. PMID:19526063

  15. Nontuberculous mycobacteria: Reports of clinical laboratory isolation in a three county area, North Carolina, 2006 -2010

    EPA Science Inventory

    Background: Laboratory reports of mycobacteria isolation and identification are created during the clinical diagnostic process to differentiate Mycobacterium tuberculosis from nontuberculous mycobacteria (NTM). NTM isolation rates are expected to exceed rates of true NTM infectio...

  16. Nontuberculous mycobacteria: Reports of clinical laboratory isolation in a three county area, North Carolina, 2006 -2010

    EPA Science Inventory

    Background: Laboratory reports of mycobacteria isolation and identification are created during the clinical diagnostic process to differentiate Mycobacterium tuberculosis from nontuberculous mycobacteria (NTM). NTM isolation rates are expected to exceed rates of true NTM infectio...

  17. Epidemiology of nontuberculous mycobacteria isolations among central North Carolina residents, 2006-2010

    EPA Science Inventory

    BACKGROUND: Nontuberculous mycobacteria (NTM) are environmental mycobacteria associated with a range of infections. Reports of NTM epidemiology have primarily focused on pulmonary infections and isolations, however extrapulmonary infections of the skin, soft tissues and sterile s...

  18. Epidemiology of nontuberculous mycobacteria isolations among central North Carolina residents, 2006-2010

    EPA Science Inventory

    BACKGROUND: Nontuberculous mycobacteria (NTM) are environmental mycobacteria associated with a range of infections. Reports of NTM epidemiology have primarily focused on pulmonary infections and isolations, however extrapulmonary infections of the skin, soft tissues and sterile s...

  19. Evaluation of capillary and myofiber density in the pectoralis major muscles of rapidly growing, high-yield broiler chickens during increased heat stress.

    PubMed

    Joiner, K S; Hamlin, G A; Lien, A R J; Bilgili, S F

    2014-09-01

    Skeletal muscle development proceeds from early embryogenesis through marketing age in broiler chickens. While myofiber formation is essentially complete at hatching, myofiber hypertrophy can increase after hatch by assimilation of satellite cell nuclei into myofibers. As the diameter of the myofibers increases, capillary density peripheral to the myofiber is marginalized, limiting oxygen supply and subsequent diffusion into the myofiber, inducing microischemia. The superficial and deep pectoralis muscles constitute 25% of the total body weight in a market-age bird; thus compromise of those muscle groups can have profound economic impact on broiler production. We hypothesized that marginal capillary support relative to the hypertrophic myofibers increases the incidence of microischemia, especially in contemporary high-yield broilers under stressing conditions such as high environmental temperatures. We evaluated the following parameters in four different broiler strains at 39 and 53 days of age when reared under thermoneutral (20 to 25 C) versus hot (30 to 35 C) environmental conditions: capillary density, myofiber density and diameter, and degree of myodegeneration. Our data demonstrate that myofiber diameter significantly increased with age (P > or = 0.0001), while the absolute numbers of capillaries, blood vessels, and myofibers visible in five 400 x microscopic fields decreased (P > or = 0.0001). This is concomitant with marginalization of vascular support in rapidly growing myofibers. The myofiber diameter was significantly lower with hot environmental temperatures (P > or = 0.001); therefore, the absolute number of myofibers visible in five 400X microscopic fields was significantly higher. The incidence and subjective degree of myodegeneration characterized by loss of cross-striations, myocyte hyperrefractility, sarcoplasmic vacuolation, and nuclear pyknosis or loss also increased in hot conditions. Differences among strains were not observed.

  20. Increasing Prevalence Rate of Nontuberculous Mycobacteria ...

    EPA Pesticide Factsheets

    Rationale: Many nontuberculous mycobacteria (NTM) are clinically significant pathogens that cause disease in a variety of different human organs and tissues. Objectives: A population-based study was undertaken to investigate the prevalence of patients with a positive specimen for NTM within five states of the United States. Methods: We determined the case and age distribution of patients with at least one specimen positive for NTM, using data submitted to the disease surveillance systems of five states (Maryland, Mississippi, Missouri, Ohio, and Wisconsin) between 2008 and 2013. Crude, age-specific, and age-adjusted prevalence rates per 100,000 persons were calculated for each state. Measurements and Main Results: From 2008 to 2013, a total of 24,226 NTM cases were reported to the disease surveillance systems of the five states. The overall average annual age-adjusted prevalence rate rose from 8.7 to 13.9 per 100,000 persons between the beginning and end of the surveillance period. The number of cases and case rate in the 50–80+-year age group was higher than in the 0–49-year age group. Prevalence by age category differed among the five states. The highest number of NTM cases was observed in Mississippi for the 80+-year age group, whereas Wisconsin observed the highest number of NTM cases in the 60- to 69-year age group. Conclusions: From 2008 to 2013, the number of patients with positive specimens for NTM rose. This trend is likely to continue in the co

  1. Effect of iron concentration on the expression and activity of catalase-peroxidases in mycobacteria.

    PubMed

    Yeruva, Veena C; Sundaram, C A S Sivagami; Sritharan, Manjula

    2005-02-01

    Mycobacterial catalases are known to exist in different isoforms. We studied the influence of iron concentration on the expression and activity of the different isoforms in Mycobacterium bovis BCG, M. smegmatis, M. fortuitum, M. kansasii and M. vaccae by growing them under iron-sufficient (4 microg Fe/mL) and iron-deficient (0.02 microg Fe/ml) conditions. Upon iron deprivation, significant differences were observed in the catalase/peroxidase activities in both quantitative spectrophotometric assays and in the activity staining in native gels. Notable feature was that the peroxidase activity showed a significant decrease upon iron deprivation in all the mycobacteria, except M. vaccae. Peroxidase activity in all the mycobacteria, irrespective of the iron status was susceptible to heat inactivation. However, the isoforms of catalase showed differences in their heat stability, indicating possible structural differences in these proteins. For example, M. bovis BCG expressed a heat labile catalase under iron-sufficient conditions, while a heat stable catalase band of similar mobility was expressed under iron-deprivation conditions. The study clearly indicates that iron plays an important role in the regulation of expression of the different isoforms of the catalase-peroxidases.

  2. The ESX-5 System of Pathogenic Mycobacteria Is Involved In Capsule Integrity and Virulence through Its Substrate PPE10

    PubMed Central

    Ates, Louis S.; van der Woude, Aniek D.; Bestebroer, Jovanka; van Stempvoort, Gunny; Musters, René J. P.; Garcia-Vallejo, Juan J.; Picavet, Daisy I.; van de Weerd, Robert; Maletta, Massimiliano; Kuijl, Coenraad P.; van der Wel, Nicole N.; Bitter, Wilbert

    2016-01-01

    Mycobacteria produce a capsule layer, which consists of glycan-like polysaccharides and a number of specific proteins. In this study, we show that, in slow-growing mycobacteria, the type VII secretion system ESX-5 plays a major role in the integrity and stability of the capsule. We have identified PPE10 as the ESX-5 substrate responsible for this effect. Mutants in esx-5 and ppe10 both have impaired capsule integrity as well as reduced surface hydrophobicity. Electron microscopy, immunoblot and flow cytometry analyses demonstrated reduced amounts of surface localized proteins and glycolipids, and morphological differences in the capsular layer. Since capsular proteins secreted by the ESX-1 system are important virulence factors, we tested the effect of the mutations that cause capsular defects on virulence mechanisms. Both esx-5 and ppe10 mutants of Mycobacterium marinum were shown to be impaired in ESX-1-dependent hemolysis. In agreement with this, the ppe10 and esx5 mutants showed reduced recruitment of ubiquitin in early macrophage infection and intermediate attenuation in zebrafish embryos. These results provide a pivotal role for the ESX-5 secretion system and its substrate PPE10, in the capsular integrity of pathogenic mycobacteria. These findings open up new roads for research on the mycobacterial capsule and its role in virulence and immune modulation. PMID:27280885

  3. Flood hazard and a rapidly growing capital in the floodplain: Social response on major 18th-century Danube floods in Pest (East-Budapest)

    NASA Astrophysics Data System (ADS)

    Kiss, Andrea

    2014-05-01

    Due to its floodplain location, Pest was especially prone to damages caused by great flood events. Before water regulation works, the greatest flood events, and the highest rate of destruction occurred during ice jam floods. Whereas in the first half of the 18th century Pest is restricted to the medieval downtown located on a higher terrain (Danube terrace), from the mid 18th century onwards the rapidly growing population established suburbs around the downtown in the lower-lying flood plain. Thus, while in the first half of the century floods were more dangerous for the harvest in the agricultural lands, in the second half of the century at the same place suburbs, urban areas with thousands of inhabitants were prone to the same danger. In the first half of the century at least three particularly large flood events, in 1712, 1732 and 1744, caused increasing problems in the close vicinity of the town (and its lands), the second half of the century - as part of a climatic anomaly (Maldá) famous of its weather extremes - was characterised by two extreme (in 1775 and 1799), at least two larger (1789 and 1795) and some more, medium-sized ice jam floods. While in terms of damaged houses the loss was only some dozens in the early part of the century, several hundreds of houses - actually, complete suburbs were erased by floods in 1775 and 1799. In the poster presentation a series of known damaging 18th-century floods, occurred at Pest, is presented, the short-term impacts (e.g. damages), and medium-, long-term administrative responses as well as related long-term landscape changes influenced by floods and flood protection are discussed. Another important aim of the poster is to present the main reasons why in the 18th century these great ice jam floods caused much greater damages (e.g. percentage of collapsed houses in suburbs) in Pest protected by dams than, for example, in the Buda suburbs with no dams, partly also located in high flood-risk areas, in the immediate

  4. Identification of mycobacteria in solid-culture media by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    PubMed

    Saleeb, Paul G; Drake, Steven K; Murray, Patrick R; Zelazny, Adrian M

    2011-05-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been introduced into the clinical microbiology laboratory as a rapid and accurate method to identify bacteria and yeasts. In this paper we describe our work on the use of MALDI-TOF MS for the identification of mycobacterial isolates. We developed a protocol for protein extraction from mycobacteria and utilized it to construct a database containing 42 clinically relevant type and reference strains of mycobacteria. The database was used to identify 104 clinical isolates of mycobacteria. All members of the Mycobacterium tuberculosis complex were identified accurately at the complex level but could not be separated at the species level. All other organisms were identified at the species level, with the exception of one strain of M. kansasii (accurately identified but with a low spectral score) and three pairs of closely related strains: M. abscessus and M. massiliense, M. mucogenicum and M. phocaicum, and M. chimaera and M. intracellulare. These pairs of organisms can currently be identified only by multilocus gene sequence analysis. We conclude that MALDI-TOF MS analysis can be incorporated into the work flow of the microbiology laboratory for rapid and accurate identification of most strains of mycobacteria isolated from solid growth media.

  5. Phosphorylation of the Peptidoglycan Synthase PonA1 Governs the Rate of Polar Elongation in Mycobacteria

    PubMed Central

    Kieser, Karen J.; Baer, Christina E.; Barczak, Amy K.; Meniche, Xavier; Chao, Michael C.; Rego, E. Hesper; Sassetti, Christopher M.; Fortune, Sarah M.; Rubin, Eric J.

    2015-01-01

    Cell growth and division are required for the progression of bacterial infections. Most rod-shaped bacteria grow by inserting new cell wall along their mid-section. However, mycobacteria, including the human pathogen Mycobacterium tuberculosis, produce new cell wall material at their poles. How mycobacteria control this different mode of growth is incompletely understood. Here we find that PonA1, a penicillin binding protein (PBP) capable of transglycosylation and transpeptidation of cell wall peptidoglycan (PG), is a major governor of polar growth in mycobacteria. PonA1 is required for growth of Mycobacterium smegmatis and is critical for M. tuberculosis during infection. In both cases, PonA1’s catalytic activities are both required for normal cell length, though loss of transglycosylase activity has a more pronounced effect than transpeptidation. Mutations that alter the amount or the activity of PonA1 result in abnormal formation of cell poles and changes in cell length. Moreover, altered PonA1 activity results in dramatic differences in antibiotic susceptibility, suggesting that a balance between the two enzymatic activities of PonA1 is critical for survival. We also find that phosphorylation of a cytoplasmic region of PonA1 is required for normal activity. Mutations in a critical phosphorylated residue affect transglycosylase activity and result in abnormal rates of cell elongation. Together, our data indicate that PonA1 is a central determinant of polar growth in mycobacteria, and its governance of cell elongation is required for robust cell fitness during both host-induced and antibiotic stress. PMID:26114871

  6. Mycobacteria as Environmental Portent in Chesapeake Bay Fish Species

    PubMed Central

    Stine, Cynthia B.; Hungerford, Laura; Matsche, Mark; Driscoll, Cindy; Baya, Ana M.

    2007-01-01

    Infection with environmental mycobacteria is increasing among many Chesapeake Bay fish species. Prevalence in juvenile Atlantic menhaden differed between tributaries and ranged from 2% to 57%. Mycobacterial infection may be a syndromic sentinel of altered environmental conditions that threaten aquatic animal health. PMID:17479905

  7. Detection of mycobacteria by radiometric and standard plate procedures.

    PubMed Central

    Damato, J J; Collins, M T; Rothlauf, M V; McClatchy, J K

    1983-01-01

    A group of 89 smear-positive sputum specimens were evaluated by radiometric and standard plate procedures to determine the methodology which would provide the earliest detection of mycobacteria and maximum test sensitivity. Digested non-decontaminated specimens were concentrated and inoculated into modified selective BACTEC radiometric 7H12 broth and Mitchison selective 7H10 agar. Sodium hydroxide (1.5% final concentration) was then used to decontaminate these specimens. They were then concentrated and inoculated into both selective and nonselective 7H12 radiometric broths and into selective 7H10 and nonselective Middlebrook 7H11 agar media. The specimen processing and media combinations providing the earliest detection were non-decontaminated specimens with modified selective 7H12 BACTEC broth and decontaminated specimens with 7H12 BACTEC broths. Maximum sensitivity (percent positive) was obtained by using non-decontaminated specimens on Mitchison selective 7H10 Agar (98%) or decontaminated specimens in 7H12 BACTEC broth (95%). The decontamination process was found to reduce significantly the number of mycobacteria in clinical specimens, particularly the mycobacteria other than Mycobacterium tuberculosis. The specimen processing-media combinations providing the earliest detection and maximum recovery of mycobacteria (100%) were non-decontaminated specimens with modified selective 7H12 BACTEC broth or Mitchison selective agar and decontaminated specimens with 7H12 BACTEC broth or 7H11 agar. PMID:6348076

  8. Prospecting Environmental Mycobacteria: combined molecular approaches reveal unprecedented diversity

    USDA-ARS?s Scientific Manuscript database

    Environmental mycobacteria (EM) include species commonly found in a variety of terrestrial and aquatic environments and encompass animal and human pathogens in addition to saprophytes. Approximately 150 EM species can be separated into fast and slow growers based on sequence and copy number differen...

  9. The water environment as a source of potentially pathogenic mycobacteria.

    PubMed

    Makovcova, Jitka; Slany, Michal; Babak, Vladimir; Slana, Iva; Kralik, Petr

    2014-06-01

    Nontuberculous mycobacteria (NTM) are ubiquitous organisms of a wide variety of environmental reservoirs, including natural and municipal water, soil, aerosols, protozoans, animals and humans. Several of these species are potential pathogens which affect human health. The aim of this study was to determine the occurrence of NTM in the water environment. Samples were taken from 13 water-related facilities including fish ponds, storage ponds, drinking water reservoirs and an experimental recirculation system. Altogether, 396 samples of water, sediment and aquatic plants were collected and analysed. All samples were examined using conventional culture methods. Suspected microbial isolates were subjected to polymerase chain reaction analysis and identified using partial sequence analysis of the 16S rDNA gene. The culture revealed 94/396 samples (23.7%) that contained mycobacteria. Among known NTM we identified potentially pathogenic mycobacteria isolated from the fresh water environment for the first time: Mycobacterium asiaticum, M. chimaera, M. interjectum, M. kumamotonense, M. lentiflavum, M. montefiorense, M. nebraskense, M. paraffinicum and M. simiae. Epidemiologic studies suggest that the natural water environment is the principal source of human exposure. Our results indicate that besides the well-known potentially pathogenic mycobacteria it is important to observe occurrence, proliferation and persistence of newly discovered mycobacterial species.

  10. Evaluation of practical chromatographic procedures for identification of clinical isolates of mycobacteria.

    PubMed Central

    Luquin, M; Ausina, V; López Calahorra, F; Belda, F; García Barceló, M; Celma, C; Prats, G

    1991-01-01

    After experimental conditions were established, 366 strains of mycobacteria belonging to 23 different species were studied for fatty acids, secondary alcohols, and mycolic acid cleavage products by capillary gas-liquid chromatography. Additionally, the mycolic acid pattern was studied by thin-layer chromatography. Capillary gas-liquid chromatography allowed direct identification of the following Mycobacterium spp.: M. kansasii, M. marinum, M. szulgai, M. xenopi, M. malmoense, and M. gordonae. The patterns of mycolic acid methyl esters recorded for the test strains of M. chelonae and M. agri may be of value in the identification of these species. Moreover, the combined use of the two chromatographic techniques provided precise identification of the M. tuberculosis complex, M. simiae, M. fallax, M. triviale, and M. chelonae-like organisms. A minimal set of biochemical tests is usually required to obtain identification to the species level when chromatographic procedures alone are not sufficient. Under the reported experimental conditions, thin-layer chromatography and capillary gas-liquid chromatography are rapid and very useful techniques for the identification of mycobacteria. Images PMID:1993746

  11. Prevalence of Nontuberculous Mycobacteria among Extrapulmonary Tuberculosis Cases in Tertiary Care Centers in Northern India

    PubMed Central

    Maurya, A. K.; Nag, V. L.; Kant, S.; Kushwaha, R. A. S.; Kumar, M.; Singh, A. K.; Dhole, T. N.

    2015-01-01

    The reports of nontuberculous mycobacteria (NTM) associated with extrapulmonary diseases are increasing in tertiary care hospitals. Despite a significant increase in knowledge about NTM infections, they still represent a diagnostic and therapeutic challenge. The aim of this study is to know the prevalence of NTN among extrapulmonary tuberculosis cases in tertiary care centers in Northern India. A total of 227 culture positive isolates from 756 cases were tested for niacin production and catalase assay. BIO-LINE SD Ag MPT64 TB test and final identification and differentiation between MTBC and different species of NTM were further confirmed by GenoType Mycobacterium CM/AS assay. 71 cases (9.3%) were positive for AFB by ZN staining and 227 cases (30.1%) were positive for mycobacteria by culture. Niacin production and catalase activity were negative in 62/227 (27.4%) strains and after using a panel of different biochemicals and final confirmation by GenoType Mycobacterium CM assay. Out of 227 cultures tested, 165 (72.6%) strains were confirmed as M. tuberculosis complex, and 62 (27.4%) were confirmed as NTM. The most common NTM species identified were M. fortuitum 17 (27.5%) and M. intracellulare 13 (20.9%). The rapid identification of NTM species may help in targeted therapy and management of the diseases. PMID:25883962

  12. [Inactivation of Mycobacteria mucogenicum in drinking water: chlorine resistance and mechanism analysis].

    PubMed

    Zheng, Qi; Chen, Chao; Zhang, Xiao-Jian; Lu, Pin-Pin; Liu, Yuan-Yuan; Chen, Yu-Qiao

    2013-02-01

    In recent years, chlorine-resistant bacteria were detected in drinking water distribution systems which threatened the drinking water safety. Our group detected one strain named Mycobacteria mucogenicum from the drinking water distribution system of a city in south China. This paper studied chlorine resistance and mechanism of Mycobacteria mucogenicum. Inactivation experiments of one strain Mycobacteria mucogenicum were conducted with free chlorine, monochloramind and chlorine dioxide. The CT values of 99.9% inactivation by free chlorine, monochloramine and chlorine dioxide were detected as (76.25 +/- 47.55)mg.min.L-1, (1396 +/-382)mg.min.L-1, (13.5 +/- 4.9) mg.min L-1. Using transmission electronmicroscopy (TEM) observed the inactivation process of Mycobacteria mucogenicum. The bacteria surface hydrophobic of Mycobacteria mucogenicum was 37.2%. Mycobacteria mucogenicum has a higher hydrophobicity than other bacteria which prevented the diffusion of chlorine into cells. Mycobacteria mucogenicum is more resistant to chorine than other bacteria.

  13. Distribution of Environmental Mycobacteria in Karonga District, Northern Malawi

    PubMed Central

    Chilima, Benson Z.; Clark, Ian M.; Floyd, Sian; Fine, Paul E. M.; Hirsch, Penny R.

    2006-01-01

    The genus Mycobacterium includes many species that are commonly found in the environment (in soil and water or associated with plants and animals), as well as species that are responsible for two major human diseases, tuberculosis (Mycobacterium tuberculosis) and leprosy (Mycobacterium leprae). The distribution of environmental mycobacteria was investigated in the context of a long-term study of leprosy, tuberculosis, Mycobacterium bovis BCG vaccination, and the responses of individuals to various mycobacterial antigens in Karonga District, northern Malawi, where epidemiological studies had indicated previously that people may be exposed to different mycobacterial species in the northern and southern parts of the district. A total of 148 soil samples and 24 water samples were collected from various locations and examined to determine the presence of mycobacteria. The detection method involved semiselective culturing and acid-fast staining, following decontamination of samples to enrich mycobacteria and reduce the numbers of other microorganisms, or PCR with primers specific for the mycobacterial 16S rRNA gene, using DNA extracted directly from soil and water samples. Mycobacteria were detected in the majority of the samples, and subsequent sequence analysis of PCR products amplified directly from soil DNA indicated that most of the products were related to known environmental mycobacteria. For both methods the rates of recovery were consistently higher for dry season samples than for wet season samples. All isolates cultured from soil appeared to be strains of Mycobacterium fortuitum. This study revealed a complex pattern for the environmental mycobacterial flora but identified no clear differences between the northern and southern parts of Karonga District. PMID:16597928

  14. [Anti-mycobacteria drugs therapy for periductal mastitis with fistula].

    PubMed

    Yu, Hai-jing; Wang, Qi; Yang, Jian-min; Lian, Zhen-qiang; Zhang, An-qin; Li, Wen-ping; Xu, Juan; Zhu, Cai-xia; Gao, Hong-yi; Lai, You-xng

    2012-11-01

    To study the etiology, clinical and pathologic characteristics of periductal mastitis with fistula and estimate the effect of anti-mycobacterial agents for periductal mastitis with fistula. Totally 27 patients of periductal mastitis with fistula received anti-mycobacteria drugs therapy from December 2008 to September 2011 were analyzed retrospectively. All of the patients were female. The mean age at onset was 28 years (range 15 to 40 years old). The main clinical manifestation of the 27 patients was breast fistula, including 21 patients with single fistula and 6 patients with multiple fistula. Three patients manifested with pure fistula, 14 patients with both fistula and lump, 10 patients with fistula, lump and abscess. The samples including pus or tissues of all patients were underwent bacteria culture and all patients core needle biopsy. All patients were given primary anti-mycobacteria drugs therapy, parts of patients received surgery based on the evaluation of medical treatment. The common bacteria culture of all patients failed to demonstrate any causative microorganism. Four cases were selected randomly to undergo PCR of mycobacteria, only one case was identified as Massiliense in bacteria culture of mycobacteria. Twenty-seven patients with periductal mastitis with fistula were treated with anti-mycobacterial agents (isoniazid, rifampicin and ethambutol or pyrazinamide of triple oral drugs) for 1 to 3 months, the fistula of all 27 patients were closed well. Sixteen patients were treated with the agents only and cured. Eleven patients received surgical treatment after treated with the medical agents. None of the patients were given mastectomy. All patients had no reccurence until now. The periductal mastitis with fistula has a closely relationship with the infection of nontuberculosis mycobacteria. Those patients could be treated with triple anti-mycobacterial agents and could also avoided mastectomy.

  15. The Heater Cooler as a Source of Infection from Nontuberculous Mycobacteria

    PubMed Central

    Stammers, Alfred H.; Riley, Jeffrey B.

    2016-01-01

    Abstract: Nosocomial infections acquired during the course of cardiac surgery and hospitalization can have devastating patient consequences. The source of these infections is often difficult to determine which complicates eradication efforts. Recently it has become apparent that the heater-cooler devices used in conjunction with cardiopulmonary bypass may become contaminated with bacteria that are normally found in hospital water sources. The culprit organisms are nontuberculous mycobacteria which coat the intrinsic surfaces found within the circuits of the heater-coolers. Aerosolization of the bacteria occurs during normal heater-cooler operation which can disperse the organisms throughout the operating room. The bacteria are slow-growing and may not present for months, or years, following exposure which makes epidemiological determination a challenge. The ensuing report summarizes a recent outbreak in these infections that have been reported both in Europe and the United States, along with efforts to reduce the risk for patient infection. PMID:27578894

  16. Ecology of nontuberculous mycobacteria--where do human infections come from?

    PubMed

    Falkinham, Joseph O

    2013-02-01

    Nontuberculous mycobacteria (NTM) are environmental, opportunistic human pathogens whose reservoirs include peat-rich potting soil and drinking water in buildings and households. In fact, humans are likely surrounded by NTM. NTM are ideally adapted for residence in drinking water distribution systems and household and building plumbing as they are disinfectant-resistant, surface adherent, and able to grow on low concentrations of organic matter. For individuals at risk for NTM infection, measures can be taken to reduce NTM exposure. These include avoiding inhalation of dusts from peat-rich potting soil and aerosols from showers, hot tubs, and humidifiers. A riskanalysis of the presence of NTM in drinking water has not been initiated because the virulence of independent isolates of even single NTM species (e.g., Mycobacterium avium) is quite broad, and virulence determinants have not been identified. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  17. Essential Role of the ESX-5 Secretion System in Outer Membrane Permeability of Pathogenic Mycobacteria.

    PubMed

    Ates, Louis S; Ummels, Roy; Commandeur, Susanna; van de Weerd, Robert; van der Weerd, Robert; Sparrius, Marion; Weerdenburg, Eveline; Alber, Marina; Kalscheuer, Rainer; Piersma, Sander R; Abdallah, Abdallah M; Abd El Ghany, Moataz; Abdel-Haleem, Alyaa M; Pain, Arnab; Jiménez, Connie R; Bitter, Wilbert; Houben, Edith N G

    2015-05-01

    Mycobacteria possess different type VII secretion (T7S) systems to secrete proteins across their unusual cell envelope. One of these systems, ESX-5, is only present in slow-growing mycobacteria and responsible for the secretion of multiple substrates. However, the role of ESX-5 substrates in growth and/or virulence is largely unknown. In this study, we show that esx-5 is essential for growth of both Mycobacterium marinum and Mycobacterium bovis. Remarkably, this essentiality can be rescued by increasing the permeability of the outer membrane, either by altering its lipid composition or by the introduction of the heterologous porin MspA. Mutagenesis of the first nucleotide-binding domain of the membrane ATPase EccC5 prevented both ESX-5-dependent secretion and bacterial growth, but did not affect ESX-5 complex assembly. This suggests that the rescuing effect is not due to pores formed by the ESX-5 membrane complex, but caused by ESX-5 activity. Subsequent proteomic analysis to identify crucial ESX-5 substrates confirmed that all detectable PE and PPE proteins in the cell surface and cell envelope fractions were routed through ESX-5. Additionally, saturated transposon-directed insertion-site sequencing (TraDIS) was applied to both wild-type M. marinum cells and cells expressing mspA to identify genes that are not essential anymore in the presence of MspA. This analysis confirmed the importance of esx-5, but we could not identify essential ESX-5 substrates, indicating that multiple of these substrates are together responsible for the essentiality. Finally, examination of phenotypes on defined carbon sources revealed that an esx-5 mutant is strongly impaired in the uptake and utilization of hydrophobic carbon sources. Based on these data, we propose a model in which the ESX-5 system is responsible for the transport of cell envelope proteins that are required for nutrient uptake. These proteins might in this way compensate for the lack of MspA-like porins in slow-growing

  18. Multi-centre evaluation of the speed-oligo Mycobacteria assay for differentiation of Mycobacterium spp. in clinical isolates.

    PubMed

    Hofmann-Thiel, Sabine; Turaev, Laziz; Alnour, Tarig; Drath, Lore; Müllerova, Maria; Hoffmann, Harald

    2011-12-19

    A new DNA line probe assay (Speed-oligo Mycobacteria, Vircell) has been launched for rapid differentiation of Mycobacterium spp. from cultures. Compared to other line-probe assays, Speed-oligo Mycobacteria covers a relatively limited spectrum of species but uses a simpler and faster dip-stick technique. The present multi-centre, multi-country study aimed at evaluating the utility and usability of Speed-oligo Mycobacteria in routine mycobacteriology diagnostics. Results from Speed-oligo Myobacteria were compared to those from Genotype CM (HAIN lifescience, Nehren, Germany), another line-probe assay. Speed-oligo Mycobacteria assay was performed in three main steps: 1) DNA extraction from cultured material 2) PCR amplification of the target gene and an internal control and 3) hybridization of the PCR products to specific probes by means of a dip-stick. Two hundred forty-two clinical isolates were recovered from consecutive positive mycobacterial cultures at two German (IML Gauting, Bioscientia Ingelheim), one Czech (KLINLAB Prague), and at a Sudanese (Khartoum) laboratory. All Mycobacterium species covered by the assay were reliably recognized. The rate of false positive results was 1.2% and concerned only the species M. marinum and M. peregrinum. The identification rate, i.e. the proportion of isolates which was correctly differentiated to the level of species or complex by the assay, differed significantly among laboratories being 94.9%, 90.7%, and 75.0% at the study sites IML Gauting, KLINLAB Prague and Bioscientia Ingelheim, respectively. This difference was caused by different spectra of NTM species encountered by the laboratory centres in daily routine diagnostics. Speed-oligo Mycobacteria assay was proved a rapid and easy-to-perform alternative to conventional line-probe assays. The assay showed excellent sensitivity with regard to identification of genus Mycobacterium and species/complexes covered by the test. However, due to its relatively limited spectrum of

  19. Obesity reduces bone density through activation of PPAR gamma and suppression of Wnt/Beta-Catenin in rapidly growing male rats

    USDA-ARS?s Scientific Manuscript database

    The relationship between obesity and skeletal development remains largely ambiguous. In this report, total enteral nutrition (TEN) was used to feed growing male rats intragastrically, with a high 45% fat diet (HFD) to induce obesity. We found that fat mass was increased (P<0.05) compared to rats fed...

  20. Identification of Mycobacteria from Solid and Liquid Media by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry in the Clinical Laboratory

    PubMed Central

    Kamboj, Kamal; Pancholi, Preeti

    2013-01-01

    Mycobacteria cause significant morbidity in humans. Rapid and accurate mycobacterial identification is important for improvement of patient outcomes. However, identification may be challenging due to the slow and fastidious growth of mycobacteria. Several diagnostic methods, such as biochemical, sequencing, and probe methods, are used for mycobacterial identification. We compared the matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) Biotyper system (Bruker Daltonics) to 16S rRNA/hsp65 sequencing and/or DNA probes (Gen-Probe) for mycobacterial identification. One hundred seventy-eight mycobacterial isolates grown on solid and/or broth medium were included in the study. MALDI-TOF MS identified 93.8% of the mycobacteria isolates accurately to the species level and 98.3% to the genus level, independent of the type of medium used for isolation. The identification of mycobacteria directly from cultures using MALDI-TOF MS allows for precise identification in an hour compared to traditional biochemical and phenotypic methods that can take weeks or probes and sequencing that may take a few hours. Identification by MALDI-TOF MS potentially reduces the turnaround time and cost, thereby saving resources within the health care system. PMID:23804379

  1. The use of quaternary ammonium disinfectants selects for persisters at high frequency from some species of non-tuberculous mycobacteria and may be associated with outbreaks of soft tissue infections

    PubMed Central

    Cortesia, Claudia; Lopez, Gustavo J.; de Waard, Jacobus H.; Takiff, Howard E.

    2010-01-01

    Background Non-tuberculous mycobacteria (NTM) are increasingly important as opportunistic infections after major and minor surgical procedures, likely because they are ubiquitous and not effectively killed by many commonly used disinfectants. Outbreaks of soft tissue infections with NTM appeared related to the use of commercial disinfectants based on quaternary ammonium compounds (QACs). Methods We studied the survival of clinical and environmental isolates of Mycobacterium abscessus, Mycobacterium massiliense, Mycobacterium chelonae and Mycobacterium fortuitum after 20 min, 60 min or 24 h exposures to different QACs, and the surviving bacteria were then re-exposed to QACs to see if the percentage of surviving bacteria had increased. The bacteria were labelled with a dnaA–gfp fusion and their level of QAC resistance monitored as increasing fluorescence. The QAC-resistant bacteria were then serially restreaked onto non-selective medium and retested for QAC survival. Results The frequency of survivors was <1 in 105 bacteria with Mycobacterium smegmatis, but >1 in 100 with the other mycobacteria studied. Different environmental and clinical isolates had similar QAC MICs, but QAC survivors of each strain were resistant. The QAC-surviving strains reverted to the original, non-resistant phenotype after several passages on non-selective medium. Conclusions QACs should not be used in settings where even minimally invasive procedures are performed, as they select for a non-genetically determined reversible resistant phenotype that appears at high frequency with several rapidly growing mycobacterial species associated with healthcare-related infections. M. smegmatis behaves differently and is not an adequate model for testing the activity of disinfectants against NTM. PMID:20926395

  2. Identification of nontuberculous mycobacteria in clinical samples using molecular methods: a 3-year study.

    PubMed

    Couto, I; Machado, D; Viveiros, M; Rodrigues, L; Amaral, L

    2010-08-01

    Nontuberculous mycobacteria (NTM) are being increasingly isolated in clinical laboratories and present technical and therapeutic challenges. In the present study, we report our experience with the identification of NTM received from 12 Lisbon hospitals over a 3-year period using GenoType Mycobacterium (CM/AS) assays (HAIN Lifescience GmbH, Nehren, Germany). Together, the two kits identified 96.6% of all NTM isolates tested. Among the 18 NTM species identified, Mycobacterium avium complex was the most frequent, although it accounted for only 34% of all NTM. Introducing these methods for the rapid identification of NTM highlights the importance of NTM as potential pathogens and assisted the selection of adequate therapy.

  3. Comparison of three isolation systems for the culture of mycobacteria from respiratory and non-respiratory samples

    PubMed Central

    Harris, G; Rayner, A; Blair, J; Watt, B

    2000-01-01

    Aims—To compare the recovery of mycobacteria from clinical samples using the MB/BacT rapid culture system with that obtained using egg medium or the Bactec radiometric method. Methods—The three methods were compared using 681 clinical samples (462 respiratory and 219 non-respiratory samples) and eight external quality control strains. Culture media were incubated at 35–37°C for six weeks in the MB/BacT system and for 12 weeks in the Bactec system and on egg medium. Solid media were examined macroscopically once a week and the Bactec vials were read six times in the first two weeks, and then weekly for the next 10 weeks (a growth index > 50 indicated a positive vial). The MB/BacT system positive vials were unloaded from the machine as soon as possible after detection. Confirmation of growth for all systems was by Ziehl-Neelson stained smears. Isolates were identified by a combination of phenotypic and molecular methods. Results—Of the 681 clinical samples, 59 (8.7%) were positive on culture, including 23 strains of Mycobacterium tuberculosis. None of the three systems recovered all of the isolates, but each recovered mycobacteria not detected by either of the other two systems. After six weeks incubation, isolation rates were 87%, 78%, and 90%, and mean times to detection were 13, 19, and nine days for the MB/BacT, egg medium, and Bactec systems, respectively. Although the MB/BacT system was slightly slower than the Bactec system, the biomass was greater, allowing earlier use of molecular probes and earlier inoculation of susceptibility tests. Conclusions—The MB/BacT system provides comparable performance to the Bactec radiometric system, without the problems of disposal of radioactive waste. Optimal recovery is obtained when culture on egg medium is used in conjunction with a rapid culture system. Key Words: mycobacteriarapid culture • solid media PMID:11002766

  4. Innate immunity to mycobacteria: vitamin D and autophagy.

    PubMed

    Jo, Eun-Kyeong

    2010-08-01

    Autophagy is an ancient mechanism of protein degradation and a novel antimicrobial strategy. With respect to host defences against mycobacteria, autophagy plays a crucial role in antimycobacterial resistance, and contributes to immune surveillance of intracellular pathogens and vaccine efficacy. Vitamin D3 contributes to host immune responses against Mycobacterium tuberculosis through LL-37/hCAP-18, which is the only cathelicidin identified to date in humans. In this review, we discuss recent advances in our understanding of host immune strategies against mycobacteria, including vitamin D-mediated innate immunity and autophagy activation. This review also addresses our current understanding regarding the autophagy connection to principal innate machinery, such as ubiquitin- or inflammasome-involved pathways. Integrated dialog between autophagy and innate immunity may contribute to adequate host immune defences against mycobacterial infection.

  5. Environmental reservoirs of pathogenic mycobacteria across the Ethiopian biogeographical landscape.

    PubMed

    King, Hayley C; Khera-Butler, Tanya; James, Phillip; Oakley, Brian B; Erenso, Girume; Aseffa, Abraham; Knight, Rob; Wellington, Elizabeth M; Courtenay, Orin

    2017-01-01

    The Mycobacterium genus comprises over one-hundred-and-fifty recognised species, the majority of which reside in the environment and many of which can be pathogenic to mammals. Some species of environmental mycobacteria may interfere with BCG vaccination efficacy and in tuberculin test interpretation. Examining biogeographic trends in the distribution of members of the mycobacteria across a number of physicochemical and spatial gradients in soil and water environments across Ethiopia using oligotyping identified differential distributions of pathogenic and significant species. The tuberculosis complex was identified in more than 90% of water samples and taxonomic groups implicated in lower BCG vaccine efficiency were core in both soil and water Mycobacterium communities. A reservoir of Mycobacterium bovis was identified in water, with up to 7.3×102 genome equivalents per ml. Elevation, temperature, habitat and vegetation type were important predictors of both soil and water Mycobacterium communities. These results represent the first step in understanding the potential risk of exposure to environmental mycobacteria that may undermine efforts to reduce disease incidence.

  6. Resuscitation factors from mycobacteria: homologs of Micrococcus luteus proteins.

    PubMed

    Zhu, Wenming; Plikaytis, Bonnie B; Shinnick, Thomas M

    2003-01-01

    Resuscitation promoting factors (Rpfs) are proteins, originally identified in Micrococcus luteus, that promote recovery of bacteria from a viable but non-replicating phase (e.g., stationary phase or latency) to a replicating phase. Purified M. luteus Rpf can stimulate growth and increase recovery of M. luteus bacteria as well as Mycobacterium tuberculosis bacteria from prolonged stationary cultures. To clone and characterize Rpfs from mycobacteria. We cloned one M. avium subsp. paratuberculosis rpf gene and one M. tuberculosis rpf gene into the pET19b or pET21a vector for expression in Escherichia coli. The His-tag recombinant proteins were purified and characterized. When the purified recombinant proteins were added to Sauton medium (a relatively minimal medium) at 100-500 pM, lag phase for mycobacteria from non-replicating cultures was shortened and there was a 10- to 100-fold increase in colony-forming units compared with control samples. In most probable number assays, the mycobacterial Rpfs increased recovery of mycobacteria from late stationary culture by about 10-fold. The Rpfs also promoted recovery of extensively washed Mycobacterium smegmatis bacteria inoculated into Sauton medium. Rpfs had only minor effects on growth of M. tuberculosis in BACTEC 12B broth, a rich medium. The mycobacterial Rpfs demonstrate resuscitation activities similar to those of the M. luteus Rpf.

  7. Environmental reservoirs of pathogenic mycobacteria across the Ethiopian biogeographical landscape

    PubMed Central

    James, Phillip; Oakley, Brian B.; Erenso, Girume; Aseffa, Abraham; Knight, Rob; Wellington, Elizabeth M.; Courtenay, Orin

    2017-01-01

    The Mycobacterium genus comprises over one-hundred-and-fifty recognised species, the majority of which reside in the environment and many of which can be pathogenic to mammals. Some species of environmental mycobacteria may interfere with BCG vaccination efficacy and in tuberculin test interpretation. Examining biogeographic trends in the distribution of members of the mycobacteria across a number of physicochemical and spatial gradients in soil and water environments across Ethiopia using oligotyping identified differential distributions of pathogenic and significant species. The tuberculosis complex was identified in more than 90% of water samples and taxonomic groups implicated in lower BCG vaccine efficiency were core in both soil and water Mycobacterium communities. A reservoir of Mycobacterium bovis was identified in water, with up to 7.3×102 genome equivalents per ml. Elevation, temperature, habitat and vegetation type were important predictors of both soil and water Mycobacterium communities. These results represent the first step in understanding the potential risk of exposure to environmental mycobacteria that may undermine efforts to reduce disease incidence. PMID:28333945

  8. UTILIZATION OF NITRATE BY PATHOGENIC AND SAPROPHYTIC MYCOBACTERIA

    PubMed Central

    Hedgecock, L. W.; Costello, R. L.

    1962-01-01

    Hedgecock, L. W. (Veterans Administration Hospital, Kansas City, Mo.) and R. L. Costello. Utilization of nitrate by pathogenic and saprophytic mycobacteria. J. Bacteriol. 84:195–205. 1962—The ability of mycobacteria to utilize nitrate as a sole source of nitrogen was examined. Nitrate-nitrogen was readily utilized by Mycobacterium butyricum, M. smegmatis, and M. tuberculosis H37Ra. Nitrate and nitrite were both utilized as sole sources of nitrogen by M. tuberculosis H37Rv. The saprophytes and drug-resistant strains of tubercle bacilli failed to reduce nitrate after having been cultured repeatedly and stored in Kirchner medium. It was necessary to add molybdenum to Kirchner medium for continued nitrate reduction by the saprophytic mycobacteria, but not for M. tuberculosis. Nitrate reductase of M. tuberculosis was inhibited by tungstate; the inhibition was reversed by molybdate at a concentration of 1:10,000. Optimal conditions for the reduction of nitrate by washed whole cells and by a particulate sonictreated extract of M. tuberculosis were studied. Reduced diphosphopyridine nucleotide was required for reduction of nitrate by the cell extract. Nitrate reduction was inhibited by isoniazid but not by p-aminosalicylic acid or streptomycin. PMID:13905880

  9. An inducible expression system for high-level expression of recombinant proteins in slow growing mycobacteria.

    PubMed

    Leotta, Lisa; Spratt, Joanne M; Kong, Carlyn U; Triccas, James A

    2015-09-01

    A novel protein expression vector utilising the inducible hspX promoter of Mycobacterium tuberculosis was constructed and evaluated in this study. High-level induction of three mycobacterial antigens, comprising up to 9% of bacterial sonicate, was demonstrated in recombinant Mycobacterium bovis BCG when grown under low-oxygen tension, which serves to enhance hspX promoter activity. Recombinant proteins were efficiently purified from bacterial lysates in a soluble form by virtue of a C-terminal 6-histidine tag. Purification of the immunodominant M. tuberculosis Ag85B antigen using this system resulted in a recombinant protein that stimulated significant IFN-γ release from Ag85B-reactive T cells generated after vaccination of mice with an Ag85B-expressing vaccine. Further, the M. tuberculosis L-alanine dehydrogenase (Ald) protein purified from recombinant BCG displayed strong enzymatic activity in recombinant form. This study demonstrated that high levels of native-like recombinant mycobacterial proteins can be produced in mycobacterial hosts, and this may aid the analysis of mycobacterial protein function and the development of new treatments.

  10. Sulfate Reducing Bacteria and Mycobacteria Dominate the Biofilm Communities in a Chloraminated Drinking Water Distribution System.

    PubMed

    Gomez-Smith, C Kimloi; LaPara, Timothy M; Hozalski, Raymond M

    2015-07-21

    The quantity and composition of bacterial biofilms growing on 10 water mains from a full-scale chloraminated water distribution system were analyzed using real-time PCR targeting the 16S rRNA gene and next-generation, high-throughput Illumina sequencing. Water mains with corrosion tubercles supported the greatest amount of bacterial biomass (n = 25; geometric mean = 2.5 × 10(7) copies cm(-2)), which was significantly higher (P = 0.04) than cement-lined cast-iron mains (n = 6; geometric mean = 2.0 × 10(6) copies cm(-2)). Despite spatial variation of community composition and bacterial abundance in water main biofilms, the communities on the interior main surfaces were surprisingly similar, containing a core group of operational taxonomic units (OTUs) assigned to only 17 different genera. Bacteria from the genus Mycobacterium dominated all communities at the main wall-bulk water interface (25-78% of the community), regardless of main age, estimated water age, main material, and the presence of corrosion products. Further sequencing of the mycobacterial heat shock protein gene (hsp65) provided species-level taxonomic resolution of mycobacteria. The two dominant Mycobacteria present, M. frederiksbergense (arithmetic mean = 85.7% of hsp65 sequences) and M. aurum (arithmetic mean = 6.5% of hsp65 sequences), are generally considered to be nonpathogenic. Two opportunistic pathogens, however, were detected at low numbers: M. hemophilum (arithmetic mean = 1.5% of hsp65 sequences) and M. abscessus (arithmetic mean = 0.006% of hsp65 sequences). Sulfate-reducing bacteria from the genus Desulfovibrio, which have been implicated in microbially influenced corrosion, dominated all communities located underneath corrosion tubercules (arithmetic mean = 67.5% of the community). This research provides novel insights into the quantity and composition of biofilms in full-scale drinking water distribution systems, which is critical for assessing the risks to public health and to the

  11. A microfluidic system for long-term time-lapse microscopy studies of mycobacteria.

    PubMed

    Golchin, Solmaz A; Stratford, James; Curry, Richard J; McFadden, Johnjoe

    2012-11-01

    Phenotypic heterogeneity in bacterial populations is thought to contribute to a number of important phenomena including sporulation and persistence. The latter has clinical implications in many diseases such as tuberculosis, where persistence of Mycobacterium tuberculosis within the human host is believed to be the root cause of latent tuberculosis and the ability of a minority population of cells to survive antibiotic exposure, despite being genetically identical to the bulk population that are killed. However, phenotypic variations caused by non-genetic mechanisms are difficult to study because of the transient nature of the persistent state and thereby the requirement to observe individual cells in real-time. Recently, microfluidics, combined with time-lapse microscopy, has become a powerful tool for studying population heterogeneity in bacteria. However, growth and replication of mycobacterial cells provide particular problems for the development of microfluidic systems due to their tendency to grow in three dimensions. We here describe a novel microfluidic device for the observation of growth and antibiotic killing in individual mycobacterial cells. We constructed a microfluidic device suitable for studying single cell behavior in mycobacteria. The growth of single cells of Mycobacterium smegmatis expressing green fluorescent protein was monitored using a confocal laser scanning microscope. Within the device M. smegmatis cells were tightly confined within a hydrogel matrix thus promoting planar growth. Cell growth and killing was observed in the device with dead cells highlighted by uptake of propidium iodide. Conclusions/Significance. We demonstrate that our device allows real-time analysis and long-term culture of single cells of mycobacteria, and is able to support the study of cell death during the application of antibiotics. The device will allow observation of individual cells' cell genealogy to be determined and direct observation of rare states, such

  12. OPG-Fc treatment in growing pigs leads to rapid reductions in bone resorption markers, serum calcium, and bone formation markers.

    PubMed

    Sipos, W; Zysset, P; Kostenuik, P; Mayrhofer, E; Bogdan, C; Rauner, M; Stolina, M; Dwyer, D; Sommerfeld-Stur, I; Pendl, G; Resch, H; Dall'Ara, E; Varga, P; Pietschmann, P

    2011-12-01

    Inhibition of the receptor activator of NF-κB ligand (RANKL) is a novel therapeutic option in the treatment of osteoporosis and related diseases. The aim of this study was to evaluate bone metabolism and structure in pigs after RANKL inhibition. 12 growing pigs were assigned to 2 groups with 6 animals each. The OPG group received recombinant human OPG-Fc (5 mg/kg IV) at day 0, the control group was given 0.9% NaCl solution. Serum levels of OPG-Fc, calcium (Ca), phosphorus (P), and bone turnover markers were evaluated every 5 days, and pigs were euthanized on day 20. Serum OPG-Fc concentration peaked at day 5 and coincided with significantly decreased Ca, P, and bone turnover markers. By day 15, measureable OPG-Fc serum levels could only be detected in 2/6 animals. With OPG-Fc clearance starting at day 10, serum Ca and P concentrations were not different between the 2 groups. TRACP5b, P1CP, and BAP levels significantly decreased by 40-70% relative to vehicle controls in the OPG-Fc group between days 5 and 10, indicating that pharmacologic concentration of OPG-Fc led to systemic concomitant inhibition of bone formation and resorption in young growing pigs. Dual X-ray absorptiometry data derived from the proximal femur did not differ between the 2 groups. μCT analysis of selected bone sites demonstrated an OPG-Fc-induced improvement of specific bone architectural indices and bone mineralization.

  13. Acute Pulmonary Artery Obstruction as the Primary Manifestation of a Rapidly Growing Intimal Sarcoma in a 54-Year-Old Patient.

    PubMed

    Westhofen, Sumi; Kugler, Christian; Reichenspurner, Hermann; Deuse, Tobias

    2016-12-01

    Pulmonary artery sarcoma is a rare malignant neoplasm that is often misdiagnosed and most often only recognized postmortem during the autopsy. We present the case of a male patient with a rapidly progressive pulmonary tumor who underwent urgent pneumonectomy for increasing symptoms of chest pain and septic clinical picture. Histological analysis revealed the diagnosis of a pulmonary artery sarcoma. Despite an R1-resection and adjuvant chemotherapy, the patient is in good clinical health and free of tumor relapse 1 year after the surgery.

  14. Acute Pulmonary Artery Obstruction as the Primary Manifestation of a Rapidly Growing Intimal Sarcoma in a 54-Year-Old Patient

    PubMed Central

    Westhofen, Sumi; Kugler, Christian; Reichenspurner, Hermann; Deuse, Tobias

    2016-01-01

    Pulmonary artery sarcoma is a rare malignant neoplasm that is often misdiagnosed and most often only recognized postmortem during the autopsy. We present the case of a male patient with a rapidly progressive pulmonary tumor who underwent urgent pneumonectomy for increasing symptoms of chest pain and septic clinical picture. Histological analysis revealed the diagnosis of a pulmonary artery sarcoma. Despite an R1-resection and adjuvant chemotherapy, the patient is in good clinical health and free of tumor relapse 1 year after the surgery. PMID:28018820

  15. Two-laboratory collaborative study on identification of mycobacteria: molecular versus phenotypic methods.

    PubMed Central

    Springer, B; Stockman, L; Teschner, K; Roberts, G D; Böttger, E C

    1996-01-01

    Previous studies have indicated that the conventional tests used for the identification of mycobacteria may (i) frequently result in erroneous identification and (ii) underestimate the diversity within the genus Mycobacterium. To address this issue in a more systematic fashion, a study comparing phenotypic and molecular methods for the identification of mycobacteria was initiated. Focus was given to isolates which were difficult to identify to species level and which yielded inconclusive results by conventional tests performed under day-to-day routine laboratory conditions. Traditional methods included growth rate, colonial morphology, pigmentation, biochemical profiles, and gas-liquid chromatography of short-chain fatty acids. Molecular identification was done by PCR-mediated partial sequence analysis of the gene encoding the 16S rRNA. A total of 34 isolates was included in this study; 13 of the isolates corresponded to established species, and 21 isolates corresponded to previously uncharacterized taxa. For five isolates, phenotypic and molecular analyses gave identical results. For five isolates, minor discrepancies were present; four isolates remained unidentified after biochemical testing. For 20 isolates, major discrepancies between traditional and molecular typing methods were observed. Retrospective analysis of the data revealed that the discrepant results were without exception due to erroneous biochemical test results or interpretations. In particular, phenotypic identification schemes were compromised with regard to the recognition of previously undescribed taxa. We conclude that molecular typing by 16S rRNA sequence determination is not only more rapid (12 to 36 h versus 4 to 8 weeks) but also more accurate than traditional typing. PMID:8789004

  16. Crystal growing

    NASA Technical Reports Server (NTRS)

    Neville, J. P.

    1990-01-01

    One objective is to demonstrate the way crystals grow and how they affect the behavior of material. Another objective is to compare the growth of crystals in metals and nonmetals. The procedures, which involve a supersaturated solution of a salt that will separate into crystals on cooling and the pouring off of an eutectic solution to expose the crystals formed by a solid solution when an alloy of two metals forms a solid and eutectic solution on cooling, are described.

  17. Nuclear DNA methylation and chromatin condensation phenotypes are distinct between normally proliferating/aging, rapidly growing/immortal, and senescent cells.

    PubMed

    Oh, Jin Ho; Gertych, Arkadiusz; Tajbakhsh, Jian

    2013-03-01

    This study reports on probing the utility of in situ chromatin texture features such as nuclear DNA methylation and chromatin condensation patterns - visualized by fluorescent staining and evaluated by dedicated three-dimensional (3D) quantitative and high-throughput cell-by-cell image analysis - in assessing the proliferative capacity, i.e. growth behavior of cells: to provide a more dynamic picture of a cell population with potential implications in basic science, cancer diagnostics/prognostics and therapeutic drug development. Two types of primary cells and four different cancer cell lines were propagated and subjected to cell-counting, flow cytometry, confocal imaging, and 3D image analysis at various points in culture. Additionally a subset of primary and cancer cells was accelerated into senescence by oxidative stress. DNA methylation and chromatin condensation levels decreased with declining doubling times when primary cells aged in culture with the lowest levels reached at the stage of proliferative senescence. In comparison, immortal cancer cells with constant but higher doubling times mostly displayed lower and constant levels of the two in situ-derived features. However, stress-induced senescent primary and cancer cells showed similar levels of these features compared with primary cells that had reached natural growth arrest. With regards to global DNA methylation and chromatin condensation levels, aggressively growing cancer cells seem to take an intermediate level between normally proliferating and senescent cells. Thus, normal cells apparently reach cancer-cell equivalent stages of the two parameters at some point in aging, which might challenge phenotypic distinction between these two types of cells. Companion high-resolution molecular profiling could provide information on possible underlying differences that would explain benign versus malign cell growth behaviors.

  18. An IPTG Inducible Conditional Expression System for Mycobacteria

    PubMed Central

    Ravishankar, Sudha; Ambady, Anisha; Ramu, Haripriya; Mudugal, Naina Vinay; Tunduguru, Ragadeepthi; Anbarasu, Anand; Sharma, Umender K.; Sambandamurthy, Vasan K.; Ramaiah, Sudha

    2015-01-01

    Conditional expression strains serve as a valuable tool to study the essentiality and to establish the vulnerability of a target under investigation in a drug discovery program. While essentiality implies an absolute requirement of a target function, vulnerability provides valuable information on the extent to which a target function needs to be depleted to achieve bacterial growth inhibition followed by cell death. The critical feature of an ideal conditional expression system is its ability to tightly regulate gene expression to achieve the full spectrum spanning from a high level of expression in order to support growth and near zero level of expression to mimic conditions of gene knockout. A number of bacterial conditional expression systems have been reported for use in mycobacteria. The utility of an isopropylthiogalactoside (IPTG) inducible system in mycobacteria has been reported for protein overexpression and anti-sense gene expression from a replicating multi-copy plasmid. Herein, we report the development of a versatile set of non-replicating IPTG inducible vectors for mycobacteria which can be used for generation of conditional expression strains through homologous recombination. The role of a single lac operator versus a double lac operator to regulate gene expression was evaluated by monitoring the expression levels of β-galactosidase in Mycobacterium smegmatis. These studies indicated a significant level of leaky expression from the vector with a single lac operator but none from the vector with double lac operator. The significance of the double lac operator vector for target validation was established by monitoring the growth kinetics of an inhA, a rpoB and a ftsZ conditional expression strain grown in the presence of different concentrations of IPTG. The utility of this inducible system in identifying target specific inhibitors was established by screening a focussed library of small molecules using an inhA and a rpoB conditional expression

  19. Isolation of Mycobacteria from clinical samples collected in the United States from 2004 to 2011

    USDA-ARS?s Scientific Manuscript database

    Background: Mycobacteria other than M. bovis (i.e. atypical mycobacteria) may interfere with current bovine tuberculosis diagnostic tests resulting in false positive test results. In populations with low prevalence of M. bovis (i.e., as detected within the United States), interference from atypical ...

  20. Development of an in vitro Assay, Based on the BioFilm Ring Test®, for Rapid Profiling of Biofilm-Growing Bacteria

    PubMed Central

    Di Domenico, Enea G.; Toma, Luigi; Provot, Christian; Ascenzioni, Fiorentina; Sperduti, Isabella; Prignano, Grazia; Gallo, Maria T.; Pimpinelli, Fulvia; Bordignon, Valentina; Bernardi, Thierry; Ensoli, Fabrizio

    2016-01-01

    Microbial biofilm represents a major virulence factor associated with chronic and recurrent infections. Pathogenic bacteria embedded in biofilms are highly resistant to environmental and chemical agents, including antibiotics and therefore difficult to eradicate. Thus, reliable tests to assess biofilm formation by bacterial strains as well as the impact of chemicals or antibiotics on biofilm formation represent desirable tools for a most effective therapeutic management and microbiological risk control. Current methods to evaluate biofilm formation are usually time-consuming, costly, and hardly applicable in the clinical setting. The aim of the present study was to develop and assess a simple and reliable in vitro procedure for the characterization of biofilm-producing bacterial strains for future clinical applications based on the BioFilm Ring Test® (BRT) technology. The procedure developed for clinical testing (cBRT) can provide an accurate and timely (5 h) measurement of biofilm formation for the most common pathogenic bacteria seen in clinical practice. The results gathered by the cBRT assay were in agreement with the traditional crystal violet (CV) staining test, according to the κ coefficient test (κ = 0.623). However, the cBRT assay showed higher levels of specificity (92.2%) and accuracy (88.1%) as compared to CV. The results indicate that this procedure offers an easy, rapid and robust assay to test microbial biofilm and a promising tool for clinical microbiology. PMID:27708625

  1. Development of an in vitro Assay, Based on the BioFilm Ring Test(®), for Rapid Profiling of Biofilm-Growing Bacteria.

    PubMed

    Di Domenico, Enea G; Toma, Luigi; Provot, Christian; Ascenzioni, Fiorentina; Sperduti, Isabella; Prignano, Grazia; Gallo, Maria T; Pimpinelli, Fulvia; Bordignon, Valentina; Bernardi, Thierry; Ensoli, Fabrizio

    2016-01-01

    Microbial biofilm represents a major virulence factor associated with chronic and recurrent infections. Pathogenic bacteria embedded in biofilms are highly resistant to environmental and chemical agents, including antibiotics and therefore difficult to eradicate. Thus, reliable tests to assess biofilm formation by bacterial strains as well as the impact of chemicals or antibiotics on biofilm formation represent desirable tools for a most effective therapeutic management and microbiological risk control. Current methods to evaluate biofilm formation are usually time-consuming, costly, and hardly applicable in the clinical setting. The aim of the present study was to develop and assess a simple and reliable in vitro procedure for the characterization of biofilm-producing bacterial strains for future clinical applications based on the BioFilm Ring Test® (BRT) technology. The procedure developed for clinical testing (cBRT) can provide an accurate and timely (5 h) measurement of biofilm formation for the most common pathogenic bacteria seen in clinical practice. The results gathered by the cBRT assay were in agreement with the traditional crystal violet (CV) staining test, according to the κ coefficient test (κ = 0.623). However, the cBRT assay showed higher levels of specificity (92.2%) and accuracy (88.1%) as compared to CV. The results indicate that this procedure offers an easy, rapid and robust assay to test microbial biofilm and a promising tool for clinical microbiology.

  2. Compartmental analysis of roots in intact rapidly-growing Spergularia marina and Lactuca sativa: partial characterization of the symplasms functional in the radial transport of Na/sup +/ and K/sup +/

    SciTech Connect

    Lazof, D.B.

    1987-01-01

    Techniques of compartmental analysis were adapted to the study of intact roots of rapidly-growing Spergularia marine and Lactuca sativa. Using large numbers of plants short time-courses of uptake and chase, /sup 42/K/sup +/ and /sup 22/Na/sup +/ transport could be resolved, even during a chase following a brief 10 minute labeling period. The use of intact plant systems allowed distinction of that portion of the isotope flux into the root, associated with the ion-conducting symplasms. A small compartment, which rapidly (t/sub .5/ < 1 min) exchanges with the external medium was implicated in the radial transport of N/sup +/, accounting for the observed obtention of linear translocation rates within minutes of transferring to labeled solution. The ion contents of this compartment varied in proportion to the external ion concentration. When K/sup +/ was at a high external concentration, labeled K/sup +/ exchanged into this same symplasm, but chasing a short pulse indicated that K/sup +/ transport to the xylem was not through a rapidly-exchanging compartment. At physiological concentrations of K/sup +/ the evidence indicated that transport of K/sup +/ across the root proceeded through a compartment which was not exchanging rapidly with the external medium. The rise to a linear rate of isotope translocation was gradual and translocation during a chase, following a brief pulse,was prolonged, indicating that this compartment retained its specific activity for a considerable period.

  3. Hydraulic Signals from the Roots and Rapid Cell-Wall Hardening in Growing Maize (Zea mays L.) Leaves Are Primary Responses to Polyethylene Glycol-Induced Water Deficits.

    PubMed

    Chazen, O.; Neumann, P. M.

    1994-04-01

    We investigated mechanisms involved in inhibition of maize (Zea mays L.) leaf-elongation growth following addition of non-penetrating osmolyte to the root medium. The elongation rate of the first true leaf remained inhibited for 4 h after addition of polyethylene glycol 6000 (PEG; -0.5 MPa water potential), despite progressive osmotic adjustment in the growing leaf tissues. Thus, inhibition of leaf growth did not appear to be directly related to loss of leaf capacity to maintain osmotic potential gradients. Comparative cell-wall-extension capacities of immature (still expanding) leaf tissues were measured by creep extensiometry using whole plants. Reductions in irreversible (plastic) extension capacity (i.e. wall hardening) were detected minutes and hours after addition of PEG to the roots, by both in vivo and in vitro assay. The onset of the wall-hardening response could be detected by in vitro assay only 2 min after addition of PEG. Thus, initiation of wall hardening appeared to precede transcription-regulated responses. The inhibition of both leaf growth and wall-extension capacity was reversed by removal of PEG after 4 h. Moreover, wall hardening could be induced by other osmolytes (mannitol, NaCl). Thus, the leaf responses did not appear to be related to any specific (toxic) effect of PEG. We conclude that hardening of leaf cell walls is a primary event in the chain of growth regulatory responses to PEG-induced water deficits in maize. The signaling processes by which PEG, which is not expected to penetrate root cell walls or membranes, might cause cell-wall hardening in relatively distant leaves was also investigated. Plants with live or killed roots were exposed to PEG. The killed roots were presumed to be unable to produce hormonal or electrical signals in response to addition of PEG; however, inhibition of leaf elongation and hardening of leaf cell walls were detected with both live and killed roots. Thus, neither hormonal signaling nor signaling via

  4. Risk of encountering ticks and tick-borne pathogens in a rapidly growing metropolitan area in the U.S. Great Plains.

    PubMed

    Noden, Bruce H; Loss, Scott R; Maichak, Courtney; Williams, Faithful

    2017-01-01

    The prevalence of tick-borne diseases has increased dramatically in many urban areas of the U.S., yet little is known about the ecology of ticks and tick-borne pathogens in relation to characteristics of North American urban and suburban landscapes. This study aimed to begin identification of the risk of encountering ticks and tick-borne pathogens within a rapidly expanding metropolitan area in the U.S. Great Plains region. Ten sites across Oklahoma City, Oklahoma were selected for tick sampling based on presence of tick habitat and level of urbanization intensity. Sampling was conducted using CO2 traps and flagging in June, July and October 2015. A total of 552 ticks were collected from eight of the ten sampled greenspaces. The majority of ticks collected in summer were Amblyomma americanum (N=534 (97.8%)), followed by Dermacentor variabilis (N=10 (1.8%)) and Amblyomma maculatum (N=2 (0.3%)). Ixodes scapularis adult females (N=4) and nymphal A. americanum (N=2) were also collected in October 2015. Tick species diversity was highest in sites with >15% of the surrounding landscape composed of undeveloped land. Rickettsia sp. (including R. amblyommii and 'Candidatus R. andeanae'), Ehrlichia chaffeensis and/or E. ewingii were detected in tick pools from all eight sites where ticks were found. Our data suggest that the risk of encountering ticks and tick-borne pathogens exists throughout the Oklahoma City metropolitan area and that tick populations are likely influenced by urbanization intensity. Continued research is needed to clarify the full range of abiotic and biotic features of urban landscapes that influence the risk of encountering ticks and transmitting tick-borne diseases.

  5. Mycobacteria in water and loose deposits of drinking water distribution systems in Finland.

    PubMed

    Torvinen, Eila; Suomalainen, Sini; Lehtola, Markku J; Miettinen, Ilkka T; Zacheus, Outi; Paulin, Lars; Katila, Marja-Leena; Martikainen, Pertti J

    2004-04-01

    Drinking water distribution systems were analyzed for viable counts of mycobacteria by sampling water from waterworks and in different parts of the systems. In addition, loose deposits collected during mechanical cleaning of the main pipelines were similarly analyzed. The study covered 16 systems at eight localities in Finland. In an experimental study, mycobacterial colonization of biofilms on polyvinyl chloride tubes in a system was studied. The isolation frequency of mycobacteria increased from 35% at the waterworks to 80% in the system, and the number of mycobacteria in the positive samples increased from 15 to 140 CFU/liter, respectively. Mycobacteria were isolated from all 11 deposits with an accumulation time of tens of years and from all 4 deposits which had accumulated during a 1-year follow-up time. The numbers of mycobacteria were high in both old and young deposits (medians, 1.8 x 10(5) and 3.9 x 10(5) CFU/g [dry weight], respectively). Both water and deposit samples yielded the highest numbers of mycobacteria in the systems using surface water and applying ozonation as an intermediate treatment or posttreatment. The number and growth of mycobacteria in system waters correlated strongly with the concentration of assimilable organic carbon in the water leaving the waterworks. The densities of mycobacteria in the developing biofilms were highest at the distal sites of the systems. Over 90% of the mycobacteria isolated from water and deposits belonged to Mycobacterium lentiflavum, M. tusciae, M. gordonae, and a previously unclassified group of mycobacteria. Our results indicate that drinking water systems may be a source for recently discovered new mycobacterial species.

  6. Mycobacteria in Water and Loose Deposits of Drinking Water Distribution Systems in Finland

    PubMed Central

    Torvinen, Eila; Suomalainen, Sini; Lehtola, Markku J.; Miettinen, Ilkka T.; Zacheus, Outi; Paulin, Lars; Katila, Marja-Leena; Martikainen, Pertti J.

    2004-01-01

    Drinking water distribution systems were analyzed for viable counts of mycobacteria by sampling water from waterworks and in different parts of the systems. In addition, loose deposits collected during mechanical cleaning of the main pipelines were similarly analyzed. The study covered 16 systems at eight localities in Finland. In an experimental study, mycobacterial colonization of biofilms on polyvinyl chloride tubes in a system was studied. The isolation frequency of mycobacteria increased from 35% at the waterworks to 80% in the system, and the number of mycobacteria in the positive samples increased from 15 to 140 CFU/liter, respectively. Mycobacteria were isolated from all 11 deposits with an accumulation time of tens of years and from all 4 deposits which had accumulated during a 1-year follow-up time. The numbers of mycobacteria were high in both old and young deposits (medians, 1.8 × 105 and 3.9 × 105 CFU/g [dry weight], respectively). Both water and deposit samples yielded the highest numbers of mycobacteria in the systems using surface water and applying ozonation as an intermediate treatment or posttreatment. The number and growth of mycobacteria in system waters correlated strongly with the concentration of assimilable organic carbon in the water leaving the waterworks. The densities of mycobacteria in the developing biofilms were highest at the distal sites of the systems. Over 90% of the mycobacteria isolated from water and deposits belonged to Mycobacterium lentiflavum, M. tusciae, M. gordonae, and a previously unclassified group of mycobacteria. Our results indicate that drinking water systems may be a source for recently discovered new mycobacterial species. PMID:15066787

  7. Rapid Water Uptake and Limited Storage Capacity at Height of Growing Season in Four Temperate Tree Species in a Central Pennsylvania Catchment

    NASA Astrophysics Data System (ADS)

    Gaines, K.; Meinzer, F. C.; Duffy, C.; Thomas, E.; Eissenstat, D. M.

    2014-12-01

    rapid water uptake and tree water storage limited to about a month in duration. These findings are necessary for modeling of hydrologic parameters that are influenced by tree water age. They also indicate that trees on shallow soil in this catchment may be at risk if droughts lasting over a month occur more frequently in future years.

  8. Effect of disinfectant, water age, and pipe material on occurrence and persistence of Legionella, mycobacteria, Pseudomonas aeruginosa, and two amoebas.

    PubMed

    Wang, Hong; Masters, Sheldon; Hong, Yanjuan; Stallings, Jonathan; Falkinham, Joseph O; Edwards, Marc A; Pruden, Amy

    2012-11-06

    Opportunistic pathogens represent a unique challenge because they establish and grow within drinking water systems, yet the factors stimulating their proliferation are largely unknown. The purpose of this study was to examine the influence of pipe materials, disinfectant type, and water age on occurrence and persistence of three opportunistic pathogens (Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa), broader genera (Legionella and mycobacteria), and two amoeba hosts (Acanthamoeba spp. and Hartmanella vermiformis). Triplicate simulated distribution systems (SDSs) compared iron, cement, and PVC pipe materials fed either chlorinated or chloraminated tap water and were sampled at water ages ranging from 1 day to 5.7 days. Quantitative polymerase chain reaction quantified gene copies of target microorganisms in both biofilm and bulk water. Legionella, mycobacteria, P. aeruginosa, and both amoebas naturally colonized the six SDSs, but L. pneumophila and M. avium were not detected. Disinfectant type and dose was observed to have the strongest influence on the microbiota. Disinfectant decay was noted with water age, particularly in chloraminated SDSs (due to nitrification), generally resulting in increased microbial detection frequencies and densities with water age. The influence of pipe material became apparent at water ages corresponding to low disinfectant residual. Each target microbe appeared to display a distinct response to disinfectant type, pipe materials, water age, and their interactions. Differences between the first and the second samplings (e.g., appearance of Legionella, reduction in P. aeruginosa and Acanthamoeba) suggest a temporally dynamic drinking water microbial community.

  9. Functional characterization delineates that a Mycobacterium tuberculosis specific protein kinase (Rv3080c) is responsible for the growth, phagocytosis and intracellular survival of avirulent mycobacteria.

    PubMed

    Kumari, Ruma; Singh, Susmita K; Singh, Diwakar K; Singh, Pramod K; Chaurasiya, Shivendra K; Srivastava, Kishore K

    2012-10-01

    Serine/threonine protein kinases (STPKs) are predominantly involved in growth, development, division, differentiation, and in regulating immune responses in mycobacteria. A wide variety of functions of mycobacterial STPKs persuade mycobacterial growth and further its survival in the hosts. The polymorphic studies have shown that a full length gene of Rv3080c (pknK) is present in the slow growing mycobacteria. The wild type Mycobacterium smegmatis containing only vector (M. smegmatis) and M. smegmatis containing Rv3080c (pknK) cloned in pMV261 vector (M. smegmatis::K) were cultured in different growth media. The studies have shown that M. smegmatis did not differ in the growth and in survival while a substantial reduction in the growth (four-ten-folds) and a significant delay in the colony formation were observed in M. smegmatis::K. In order to look for the stage specific and modulated expression of PknK, the study was comprehended to quantitate pknK transcripts at different phases of cultures. The mycobacterium, containing high copy number of pknK specific RNA was unable to multiply. The study thus highlights that Rv3080c is largely accountable for changing the fate of avirulent mycobacteria and hence the protein can be utilized as an important molecule to target pathogenesis.

  10. Cooccurrence of free-living amoebae and nontuberculous Mycobacteria in hospital water networks, and preferential growth of Mycobacterium avium in Acanthamoeba lenticulata.

    PubMed

    Ovrutsky, Alida R; Chan, Edward D; Kartalija, Marinka; Bai, Xiyuan; Jackson, Mary; Gibbs, Sara; Falkinham, Joseph O; Iseman, Michael D; Reynolds, Paul R; McDonnell, Gerald; Thomas, Vincent

    2013-05-01

    The incidence of lung and other diseases due to nontuberculous mycobacteria (NTM) is increasing. NTM sources include potable water, especially in households where NTM populate pipes, taps, and showerheads. NTM share habitats with free-living amoebae (FLA) and can grow in FLA as parasites or as endosymbionts. FLA containing NTM may form cysts that protect mycobacteria from disinfectants and antibiotics. We first assessed the presence of FLA and NTM in water and biofilm samples collected from a hospital, confirming the high prevalence of NTM and FLA in potable water systems, particularly in biofilms. Acanthamoeba spp. (genotype T4) were mainly recovered (8/17), followed by Hartmannella vermiformis (7/17) as well as one isolate closely related to the genus Flamella and one isolate only distantly related to previously described species. Concerning mycobacteria, Mycobacterium gordonae was the most frequently found isolate (9/17), followed by Mycobacterium peregrinum (4/17), Mycobacterium chelonae (2/17), Mycobacterium mucogenicum (1/17), and Mycobacterium avium (1/17). The propensity of Mycobacterium avium hospital isolate H87 and M. avium collection strain 104 to survive and replicate within various FLA was also evaluated, demonstrating survival of both strains in all amoebal species tested but high replication rates only in Acanthamoeba lenticulata. As A. lenticulata was frequently recovered from environmental samples, including drinking water samples, these results could have important consequences for the ecology of M. avium in drinking water networks and the epidemiology of disease due to this species.

  11. Cooccurrence of Free-Living Amoebae and Nontuberculous Mycobacteria in Hospital Water Networks, and Preferential Growth of Mycobacterium avium in Acanthamoeba lenticulata

    PubMed Central

    Ovrutsky, Alida R.; Kartalija, Marinka; Bai, Xiyuan; Jackson, Mary; Gibbs, Sara; Falkinham, Joseph O.; Iseman, Michael D.; Reynolds, Paul R.; McDonnell, Gerald

    2013-01-01

    The incidence of lung and other diseases due to nontuberculous mycobacteria (NTM) is increasing. NTM sources include potable water, especially in households where NTM populate pipes, taps, and showerheads. NTM share habitats with free-living amoebae (FLA) and can grow in FLA as parasites or as endosymbionts. FLA containing NTM may form cysts that protect mycobacteria from disinfectants and antibiotics. We first assessed the presence of FLA and NTM in water and biofilm samples collected from a hospital, confirming the high prevalence of NTM and FLA in potable water systems, particularly in biofilms. Acanthamoeba spp. (genotype T4) were mainly recovered (8/17), followed by Hartmannella vermiformis (7/17) as well as one isolate closely related to the genus Flamella and one isolate only distantly related to previously described species. Concerning mycobacteria, Mycobacterium gordonae was the most frequently found isolate (9/17), followed by Mycobacterium peregrinum (4/17), Mycobacterium chelonae (2/17), Mycobacterium mucogenicum (1/17), and Mycobacterium avium (1/17). The propensity of Mycobacterium avium hospital isolate H87 and M. avium collection strain 104 to survive and replicate within various FLA was also evaluated, demonstrating survival of both strains in all amoebal species tested but high replication rates only in Acanthamoeba lenticulata. As A. lenticulata was frequently recovered from environmental samples, including drinking water samples, these results could have important consequences for the ecology of M. avium in drinking water networks and the epidemiology of disease due to this species. PMID:23475613

  12. Mycobacteria and allograft heart valve banking: an international survey.

    PubMed

    Warwick, R M; Magee, J G; Leeming, J P; Graham, J C; Hannan, M M; Chadwick, M; Crook, D W; Yearsley, C P; Rayner, A; Parker, R

    2008-03-01

    Since the 1970s many tissue banks have been testing allograft heart valves (HVs) for Mycobacterium tuberculosis (MTB). Donor selection for low risk of tuberculosis (TB) was introduced in the 1980s and appears to have reduced the risk of TB transmission. Regulatory guidance does not specify testing for TB, but does exclude donors with a recent history of TB. This survey of HV international bank practices revealed variations in donor selection, testing and processing of valves. Participant banks (from Europe and the USA) reported that over a period of 15 years, HV tissues from 38,413 donors were banked and 32,289 donors were tested for TB, none being positive. HV-associated tissue from 27,840 donors was stained and underwent microscopy; none of these were positive for acid-fast bacilli (AFB). Non-tuberculosis mycobacteria (NTBM) were detected by culture on 24 HVs. It is recommended that HV banks employ donor selection to exclude donors at risk of TB, to culture material for mycobacteria, and to investigate potential sources when clusters of NTBM are found to facilitate corrective and preventative actions.

  13. Gene replacement and expression of foreign DNA in mycobacteria.

    PubMed Central

    Husson, R N; James, B E; Young, R A

    1990-01-01

    A system that permits molecular genetic manipulation of mycobacteria was developed on the basis of the yeast paradigm of gene replacement by homologous recombination. A shuttle vector that can replicate autonomously at a high copy number in Escherichia coli but must integrate into homologous DNA for survival in Mycobacterium smegmatis was constructed. The vector contains a ColE1 origin of replication, antibiotic resistance markers for ampicillin and kanamycin, a nutritional marker (pyrF) that allows both positive and negative selection in E. coli and M. smegmatis, and unique restriction sites that permit insertion of foreign DNA. Transformation of mycobacteria with this vector results in integration of its DNA into the genomic pyrF locus by either a single or a double homologous recombination event. With this system, the 65-kilodalton Mycobacterium leprae stress protein antigen was inserted into the M. smegmatis genome and expressed. This gene replacement technology, together with a uniquely useful pyrF marker, should be valuable for investigating mycobacterial pathobiology, for the development of candidate mycobacterial vaccine vehicles, and as a model for the development of molecular genetic systems in other pathogenic microorganisms. Images FIG. 2 FIG. 3 PMID:2153655

  14. Specific Proteins in Nontuberculous Mycobacteria: New Potential Tools

    PubMed Central

    Orduña, Patricia; Castillo-Rodal, Antonia I.; Mercado, Martha E.; Ponce de León, Samuel; López-Vidal, Yolanda

    2015-01-01

    Nontuberculous mycobacteria (NTM) have been isolated from water, soil, air, food, protozoa, plants, animals, and humans. Although most NTM are saprophytes, approximately one-third of NTM have been associated with human diseases. In this study, we did a comparative proteomic analysis among five NTM strains isolated from several sources. There were different numbers of protein spots from M. gordonae (1,264), M. nonchromogenicum type I (894), M. nonchromogenicum type II (935), M. peregrinum (806), and M. scrofulaceum/Mycobacterium mantenii (1,486) strains, respectively. We identified 141 proteins common to all strains and specific proteins to each NTM strain. A total of 23 proteins were selected for its identification. Two of the common proteins identified (short-chain dehydrogenase/reductase SDR and diguanylate cyclase) did not align with M. tuberculosis complex protein sequences, which suggest that these proteins are found only in the NTM strains. Some of the proteins identified as common to all strains can be used as markers of NTM exposure and for the development of new diagnostic tools. Additionally, the specific proteins to NTM strains identified may represent potential candidates for the diagnosis of diseases caused by these mycobacteria. PMID:26106621

  15. Specific Proteins in Nontuberculous Mycobacteria: New Potential Tools.

    PubMed

    Orduña, Patricia; Castillo-Rodal, Antonia I; Mercado, Martha E; Ponce de León, Samuel; López-Vidal, Yolanda

    2015-01-01

    Nontuberculous mycobacteria (NTM) have been isolated from water, soil, air, food, protozoa, plants, animals, and humans. Although most NTM are saprophytes, approximately one-third of NTM have been associated with human diseases. In this study, we did a comparative proteomic analysis among five NTM strains isolated from several sources. There were different numbers of protein spots from M. gordonae (1,264), M. nonchromogenicum type I (894), M. nonchromogenicum type II (935), M. peregrinum (806), and M. scrofulaceum/Mycobacterium mantenii (1,486) strains, respectively. We identified 141 proteins common to all strains and specific proteins to each NTM strain. A total of 23 proteins were selected for its identification. Two of the common proteins identified (short-chain dehydrogenase/reductase SDR and diguanylate cyclase) did not align with M. tuberculosis complex protein sequences, which suggest that these proteins are found only in the NTM strains. Some of the proteins identified as common to all strains can be used as markers of NTM exposure and for the development of new diagnostic tools. Additionally, the specific proteins to NTM strains identified may represent potential candidates for the diagnosis of diseases caused by these mycobacteria.

  16. Synthesis of N-substituted 2-[(1E)-alkenyl]-4-(1H)-quinolone derivatives as antimycobacterial agents against non-tubercular mycobacteria

    PubMed Central

    Wube, Abraham A.; Bucar, Franz; Hochfellner, Christina; Blunder, Martina; Bauer, Rudolf; Hüfner, Antje

    2011-01-01

    In an effort to improve biological activities and to examine antimycobacterial-lipophilicity relationships of 2-[(1E)-alkenyl)]-4-(1H)-quinolones, we have synthesized a series of 30 quinolones by introducing several alkyl groups, an alkenyl and an alkynyl group at N-1. All synthetic compounds were first tested in vitro against Mycobacterium smegmatis and the most active compounds (MIC values ∼3.0–7.0 μM) were further examined against three other rapidly growing strains of mycobacteria using a microtiter broth dilution assay. The Clog P values of the synthetic compounds were calculated to provide an estimate of their lipophilicity. Compounds 18e, 19a and 19b displayed the most potent inhibitory effect against M. smegmatis mc2155 with an MIC value of ∼1.5 μM, which was twenty fold and thirteen fold more potent than isoniazid and ethambutol, respectively. On the other hand, compounds 17e, 18e and 19a were most active against Mycobacterium fortuitum and Mycobacterium phlei with an MIC value of ∼3.0 μM. In the human diploid embryonic lung cell line MRC-5 cytotoxicity assay, the derivatives showed moderate to strong cytotoxic activity. Although the antimycobacterial activity of our synthetic compounds could not be correlated with the calculated log P values, an increase in lipophilicity enhances the antimycobacterial activity and C13–C15 total chain length at positions 1 and 2 is required to achieve optimal inhibitory effect against the test strains. PMID:21429630

  17. MPT 64 Antigen detection for Rapid confirmation of M.tuberculosis isolates

    PubMed Central

    2011-01-01

    Background A new rapid Immunochromatographic test kit(SD MPT64TB Ag Kit) for detection of MPT 64 Antigen in M. tuberculosis isolates using mouse monoclonal MPT 64 Antibody developed by SD Bioline, South Korea was evaluated for rapid identification of M. tuberculosis isolates. We also assessed the sensitivity, specificity and predictive values of this kit. The test kit has an excellent sensitivity, specificity, negative predictive value & positive predictive value. This rapid method is found to be a reliable, rapid and cheaper method for confirming MTB culture isolates in resource poor laboratories. Material/methods: 54 culture isolates of M. tuberculosis in broth & on LJ medium, 12 Non mycobacterial isolates, 10 Non tubercular (NTM) rapidly growing Mycobacteria isolated from pus & 5 smear positive sputum samples were tested for detection of MPT64 antigen using the SD Bioline immunochromatography (ICT)test kit. H37 RV strain was employed as the positive reference control. Findings H37 RV strain showed the presence of MPT64 antigen band. Similar band was formed in all the 54 MTB isolates tested proving 100% sensitivity. MPT64 band formation was not detected in any of the other test isolates which proved the 100% specificity of the test kit. Both PPV & NPV were 100%. Conclusion Tuberculosis is a global pandemic. Rapid identification of MTB culture isolate is very important for drug susceptibility testing. MPT 64 TB Ag detection ICT kit is a rapid, reliable method; it can be a substitute for the molecular identification methods. PMID:21429231

  18. Concurrent Nontuberculous Mycobacteria Infection and High-Grade Anterior Mediastinal Extraskeletal Osteosarcoma (ESOS): Is There a Connection?

    PubMed Central

    Faz, Gabriel T.; Eltorky, Mahmoud; Karnath, Bernard

    2016-01-01

    Patient: Male, 59 Final Diagnosis: High-grade anterior mediastinal extraskeletal osteosarcoma Symptoms: Dyspnea • hemoptysis Medication: — Clinical Procedure: Biopsy Specialty: Oncology Objective: Rare disease Background: Extraskeletal osteosarcomas (ESOS) of the mediastinum are extremely rare and may present with concurrent nontuberculous mycobacteria infection. Case Report: We present the second documented case of high-grade anterior mediastinal extraskeletal osteosarcoma in a 59-year-old man with a history of treated, latent tuberculosis (TB). Sputum samples grew Mycoplasma avium complex and Mycobacterium fortuitum. Imaging showed a right-sided 7.6 cm mass with compression of the main bronchus. Subsequent biopsy with vimentin staining established the diagnosis of ESOS. Due to the patient’s rapidly declining performance status, he was not deemed a candidate for surgery or chemotherapy. He subsequently expired within one month of presentation. Conclusions: We present a unique case of high-grade anterior mediastinum ESOS and a review of the literature regarding all documented cases of ESOS to date. We suggest there is a possible link between mediastinal masses and nontuberculous mycobacteria infection. PMID:27539718

  19. A COMPARISON OF THE GROWTH OF SELECTED MYCOBACTERIA IN HELA, MONKEY KIDNEY, AND HUMAN AMNION CELLS IN TISSUE CULTURE

    PubMed Central

    Shepard, Charles C.

    1958-01-01

    HeLa, monkey kidney, and human amnion cells in tissue cultures were compared as sites for the multiplication of strains of tubercle bacilli or original and reduced pathogenicity, and for several other species of mycobacteria capable of causing disease in humans. The arrangement of the pathogenic species inorder of their growth rates in HeLa cells was Mycobacterium fortuitum, Mycobacterium balnei, and the "yellow bacillus," followed closely by the tubercle bacillus. This order was also correct for these species in monkey kidney and human amnion cells, and is the same as that seen in bacteriological media. The arrangement of the strains of tubercle bacilli in order of their growth rates in all three types of cells was: H37Rv, then R1Rv, and lastly H37Ra, which multiplied about as slowly as BCG. An INH-resistant strain grew about as rapidly as H37Rv. Growth of the pathogenic species occurred at about the same rates in HeLa and monkey kidney cells, but was distinctly slower in human amnion cells, which are less active metabolically. Irradiation of the cells in doses up to 5000 r did not affect the subsequent growth of mycobacteria in them. Preliminary experiments with human leprosy bacilli indicate that they can be introduced into these cells in high numbers and that the bacilli then persist for the life of the cells. PMID:13491759

  20. Clinical findings in relation to mortality in non-tuberculous mycobacterial infections: patients with Mycobacterium avium complex have better survival than patients with other mycobacteria.

    PubMed

    Kotilainen, H; Valtonen, V; Tukiainen, P; Poussa, T; Eskola, J; Järvinen, A

    2015-09-01

    We compared the clinical findings and survival in patients with Mycobacterium avium complex (MAC) and other non-tuberculous mycobacteria (NTM). A total of 167 adult non-human immunodeficiency virus (HIV) patients with at least one positive culture for NTM were included. Medical records were reviewed. The patients were categorised according to the 2007 American Thoracic Society (ATS) criteria. MAC comprised 59 % of all NTM findings. MAC patients were more often female (70 % vs. 34 %, p < 0.001) and had less fatal underlying diseases (23 % vs. 47 %, p = 0.001) as compared to other NTM patients. Symptoms compatible with NTM infection had lasted for less than a year in 34 % of MAC patients but in 54 % of other NTM patients (p = 0.037). Pulmonary MAC patients had a significantly lower risk of death compared to pulmonary other NTM (hazard ratio [HR] 0.50, 95 % confidence interval [CI] 0.33-0.77, p = 0.002) or subgroup of other slowly growing NTM (HR 0.55, 95 % CI 0.31-0.99, p = 0.048) or as rapidly growing NTM (HR 0.47, 95 % CI 0.25-0.87, p = 0.02). The median survival time was 13.0 years (95 % CI 5.9-20.1) for pulmonary MAC but 4.6 years (95 % CI 3.4-5.9) for pulmonary other NTM. Serious underlying diseases (HR 3.21, 95 % CI 2.05-5.01, p < 0.001) and age (HR 1.07, 95 % CI 1.04-1.09, p < 0.001) were the significant predictors of mortality and female sex was a predictor of survival (HR 0.38, 95 % CI 0.24-0.59, p < 0.001) in the multivariate analysis. Pulmonary MAC patients had better prognosis than pulmonary other NTM patients. The symptom onset suggests a fairly rapid disease course.

  1. Diagnostic value of the strand displacement amplification method compared to those of Roche Amplicor PCR and culture for detecting mycobacteria in sputum samples.

    PubMed Central

    Ichiyama, S; Ito, Y; Sugiura, F; Iinuma, Y; Yamori, S; Shimojima, M; Hasegawa, Y; Shimokata, K; Nakashima, N

    1997-01-01

    We compared the ability of the semiautomated BDProbeTec-SDA system, which uses the strand displacement amplification (SDA) method, with that of the Roche Amplicor-PCR system and the Septi-Chek AFB culture system to directly detect Mycobacterium tuberculosis complex (MTB) and other mycobacteria in sputum samples. A total of 530 sputum samples from 299 patients were examined in this study. Of the 530 samples, 129 were culture positive for acid-fast bacilli with the Septi-Chek AFB system; 95 for MTB, 29 for M. avium-M. intracellulare complex (MAC), and 5 for other mycobacteria. The BDProbeTec-SDA system detected 90 of the 95 samples culture positive for MTB (sensitivity, 94.7%), and the Amplicor-PCR system detected 85 of the 95 samples culture positive for MTB (sensitivity, 89.5%). The specificity of each system, based on the clinical diagnosis, was 99.8% for SDA and 100% for PCR, respectively. Among the 29 samples culture positive for MAC, the BDProbeTec-SDA system detected MAC in 24 samples (sensitivity, 82.8%), whereas the Amplicor-PCR system detected MAC in 23 samples (sensitivity, 79.3%). The specificities of the systems were 98.3 and 100%, respectively. The high degrees of sensitivity and specificity of the BDProbeTec-SDA system suggest that it should be very useful in clinical laboratories for the rapid detection of mycobacteria in sputum samples. PMID:9399498

  2. Large multifocal cardiac myxoma causing the sudden unexpected death of a 2-month-old infant--a rapidly growing, acquired lesion versus a congenital process?: a case report.

    PubMed

    Kure, Kiyoe; Lingamfelter, Daniel; Taboada, Eugenio

    2011-06-01

    We report the occurrence of a clinically undiagnosed biatrial myxoma with left ventricular involvement in a 2-month-old male infant, resulting in sudden death. During a routine well-baby examination, a grade (34) holosystolic murmur was detected at the left sternal border with radiation to the axilla and back. On the following day, the patient collapsed and died suddenly. An autopsy revealed a large multifocal neoplasm diffusely involving the aortic valve while displaying mitral, tricuspid, and left ventricular extensions. The ensuing histopathologic and immunohistochemical studies were diagnostic for myxoma. We discuss the occurrence of cardiac myxoma within the pediatric population and review the literature as to theorize whether this lesion was a congenital process versus a rapidly growing tumor that developed after the child was born. Lastly, we address the potential for sudden death in patients with such tumors.

  3. Antimicrobial Susceptibility Testing, Drug Resistance Mechanisms, and Therapy of Infections with Nontuberculous Mycobacteria

    PubMed Central

    Nash, Kevin A.; Wallace, Richard J.

    2012-01-01

    Summary: Within the past 10 years, treatment and diagnostic guidelines for nontuberculous mycobacteria have been recommended by the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA). Moreover, the Clinical and Laboratory Standards Institute (CLSI) has published and recently (in 2011) updated recommendations including suggested antimicrobial and susceptibility breakpoints. The CLSI has also recommended the broth microdilution method as the gold standard for laboratories performing antimicrobial susceptibility testing of nontuberculous mycobacteria. This article reviews the laboratory, diagnostic, and treatment guidelines together with established and probable drug resistance mechanisms of the nontuberculous mycobacteria. PMID:22763637

  4. Three-Dimensional Evaluation of the Upper Airway Morphological Changes in Growing Patients with Skeletal Class III Malocclusion Treated by Protraction Headgear and Rapid Palatal Expansion: A Comparative Research

    PubMed Central

    Liu, Ju; Wu, Zizhong; Xie, Yongtao; Li, Liang; Liu, Hong; Guo, Tiantian; Chen, Chen; Zhang, Shijie

    2015-01-01

    Objective The aim of this study was to evaluate the morphological changes of upper airway after protraction headgear and rapid maxillary expansion (PE) treatment in growing patients with Class III malocclusion and maxillary skeletal deficiency compared with untreated Class III patients by cone-beam computed tomography (CBCT). Methods Thirty growing patients who have completed PE therapy were included in PE group. The control group (n = 30) was selected from the growing untreated patients with the same diagnosis. The CBCT scans of the pre-treatment (T1) and post-treatment (T2) of PE group and the control group were collected. Reconstruction and registration of the 3D models of T1 and T2 were completed. By comparing the data obtained from T1, T2 and control group, the morphological changes of the upper airway during the PE treatment were evaluated. Results Comparing with the data from T1 group, the subspinale (A) of maxilla and the upper incisor (UI) of the T2 group were moved in the anterior direction. The gnathion (Gn) of mandible was moved in the posterior-inferior direction. The displacement of the hyoid bone as well as the length and width of dental arch showed significant difference. The volume and mean cross-sectional area of nasopharynx, velopharynx and glossopharynx region showed significant difference. The largest anteroposterior/the largest lateral (AP/LR) ratios of the velopharynx and glossopharynx were increased, but the AP/LR ratio of the hypopharynx was decreased. In addition, the length and width of the maxillary dental arch, the displacement of the hyoid bone, the volume of nasopharynx and velopharynx, and the AP/LR ratio of the hypopharynx and velopharynx showed significant difference between the data from control and T2 group. Conclusion The PE treatment of Class Ⅲ malocclusion with maxillary skeletal hypoplasia leads to a significant increase in the volume of nasopharynx and velopharynx. PMID:26252015

  5. In silico identification of common epitopes from pathogenic mycobacteria

    PubMed Central

    2013-01-01

    An in silico study was carried out to identify antigens for their possible collective use as vaccine candidates against diseases caused by different classes of pathogenic mycobacteria with significant clinical relevance. The genome sequences of the relevant causative agents were used in order to search for orthologous genes among them. Bioinformatics tools permitted us to identify several conserved sequences with 100% identity with no possibility of cross-reactivity to the normal flora and human proteins. Nine different proteins were characterized using the strain H37Rv as reference and taking into account their functional category, their in vivo expression and subcellular location. T and B cell epitopes were identified in the selected sequences. Theoretical prediction of population coverage was calculated for individual epitopes as well as their combinations. Several identical sequences, belonging to six proteins containing T and B cell epitopes which are not present in selected microorganisms of the normal microbial flora or in human proteins were obtained. PMID:23458668

  6. Identification of a Copper-Binding Metallothionein in Pathogenic Mycobacteria

    PubMed Central

    Gold, Ben; Deng, Haiteng; Bryk, Ruslana; Vargas, Diana; Eliezer, David; Roberts, Julia; Jiang, Xiuju; Nathan, Carl

    2009-01-01

    A screen of a genomic library from Mycobacterium tuberculosis (Mtb) identified a small, unannotated open reading frame (MT0196) that encodes a 4.9-kDa, cysteine-rich protein. Despite extensive nucleotide divergence, the amino acid sequence is highly conserved among mycobacteria that are pathogenic in vertebrate hosts. We synthesized the protein and found that it preferentially bound up to 6 Cu(I) ions in a solvent-shielded core. Copper, cadmium and compounds that generate nitric oxide or superoxide induced the gene’s expression in Mtb up to a thousand-fold. The native protein bound copper within Mtb and partially protected Mtb from copper toxicity. We propose that the product of the MT0196 gene be named mycobacterial metallothionien (MymT). To our knowledge, MymT is the first metallothionein of a Gram-positive bacterium with a demonstrated function. PMID:18724363

  7. Current Epidemiologic Trends of the Nontuberculous Mycobacteria (NTM).

    PubMed

    Falkinham, Joseph O

    2016-06-01

    The nontuberculous mycobacteria (NTM) are waterborne opportunistic pathogens of humans. They are normal inhabitants of premise plumbing, found, for example, in household and hospital shower heads, water taps, aerators, and hot tubs. The hydrophobic NTM are readily aerosolized, and pulmonary infections and hypersensitivity pneumonitis have been traced to the presence of NTM in shower heads. Hypersensitivity pneumonitis in automotive workers was traced to the presence of NTM in metal recovery fluid used in grinding operations. Recently, NTM bacteremia in heart transplant patients has been traced to the presence of NTM in water reservoirs of instruments employed in operating rooms to heat and cool patient blood during periods of mechanical circulation. Although NTM are difficult to eradicate from premise plumbing as a consequence of their disinfectant-resistance and formation of biofilms, measures such as reduction of turbidity and reduction in carbon and nitrogen for growth and the installation of microbiological filters can reduce exposure of NTM to susceptible individuals.

  8. Benzoic Acid-Inducible Gene Expression in Mycobacteria

    PubMed Central

    Dragset, Marte S.; Barczak, Amy K.; Kannan, Nisha; Mærk, Mali; Flo, Trude H.; Valla, Svein; Rubin, Eric J.; Steigedal, Magnus

    2015-01-01

    Conditional expression is a powerful tool to investigate the role of bacterial genes. Here, we adapt the Pseudomonas putida-derived positively regulated XylS/Pm expression system to control inducible gene expression in Mycobacterium smegmatis and Mycobacterium tuberculosis, the causative agent of human tuberculosis. By making simple changes to a Gram-negative broad-host-range XylS/Pm-regulated gene expression vector, we prove that it is possible to adapt this well-studied expression system to non-Gram-negative species. With the benzoic acid-derived inducer m-toluate, we achieve a robust, time- and dose-dependent reversible induction of Pm-mediated expression in mycobacteria, with low background expression levels. XylS/Pm is thus an important addition to existing mycobacterial expression tools, especially when low basal expression is of particular importance. PMID:26348349

  9. In silico identification of common epitopes from pathogenic mycobacteria.

    PubMed

    de la Caridad Addine Ramírez, Bárbara; Marrón, Reynel; Calero, Rommel; Mirabal, Mayelin; Ramírez, Juan Carlos; Sarmiento, María E; Norazmi, Mohd Nor; Acosta, Armando

    2013-01-01

    An in silico study was carried out to identify antigens for their possible collective use as vaccine candidates against diseases caused by different classes of pathogenic mycobacteria with significant clinical relevance. The genome sequences of the relevant causative agents were used in order to search for orthologous genes among them. Bioinformatics tools permitted us to identify several conserved sequences with 100% identity with no possibility of cross-reactivity to the normal flora and human proteins. Nine different proteins were characterized using the strain H37Rv as reference and taking into account their functional category, their in vivo expression and subcellular location. T and B cell epitopes were identified in the selected sequences. Theoretical prediction of population coverage was calculated for individual epitopes as well as their combinations. Several identical sequences, belonging to six proteins containing T and B cell epitopes which are not present in selected microorganisms of the normal microbial flora or in human proteins were obtained.

  10. Extrapulmonary Infections Associated with Nontuberculous Mycobacteria in Immunocompetent Persons

    PubMed Central

    Scarparo, Claudio

    2009-01-01

    Over the past several years, the prevalence of human disease caused by nontuberculous mycobacteria (NTM) has increased. Whether the increase in cases is real or whether more cases are being recognized remains unclear. Despite a considerable increase in knowledge about NTM infections, they still represent a diagnostic and therapeutic challenge for several reasons: 1) pathogenic isolates may be indistinguishable from contaminant or saprophytic isolates; 2) timely and reliable identification of isolates may depend on proper communication between clinicians and laboratory staff; 3) lack of standardized susceptibility testing makes adoption of tailored therapies unrealistic; and 4) lack of treatment guidelines exposes patients to toxic drugs and disappointing outcomes. Laboratory research and multicenter controlled trials are needed to improve diagnosis and treatment of these infections. PMID:19788801

  11. Efficacy of selected disinfectants against mycobacteria and cryptosporidia.

    PubMed

    Holton, J; Nye, P; McDonald, V

    1994-06-01

    We have tested the in-vitro efficacy of various disinfectants against clinical isolates of Mycobacterium tuberculosis, Mycobacterium avium-intracellulare, and against the protozoon Cryptosporidium parvum. The disinfectants were tested both with and without an organic load. 'Sactimed sinald' (a quaternary ammonium compound), 'Steris 20' (a peracetic acid compound) and 'Pentapon DC1' (a beta-ene compound) were all equally as effective as 'Cidex' (glutaraldehyde) against all the mycobacteria tested, as well as C. parvum. 'Virkon' (a per-oxygen compound) and 'Phoraid' (an iodine compound) were less effective than glutaraldehyde. 'Pentapon DHY' (a beta-ene compound) was ineffective against any of the organisms tested. We have also assessed a method of determining disinfectant efficacy using [35S]-methionine labelling of proteins, which appears promising, although it requires further evaluation.

  12. Mycolactones: immunosuppressive and cytotoxic polyketides produced by aquatic mycobacteria

    PubMed Central

    Hong, Hui; Demangel, Caroline; Pidot, Sacha J.; Leadlay, Peter F.

    2008-01-01

    Mycolactones are a family of highly related macrocyclic polyketides that exhibit immunosuppressive and cytotoxic properties. First discovered in 1999, they are the primary virulence factors produced by the environmental human pathogen Mycobacterium ulcerans, the causative agent of Buruli ulcer, and by some closely-related aquatic mycobacteria that cause disease in fish and frogs. Mycolactones are characterized by a common 12-membered lactone core to which is appended an unsaturated fatty acyl side-chain of variable length and oxidation state. This Highlight summarizes recent progress in understanding the structural diversity of the mycolactones, their biological activity and mode of action in mammalian cells, and the genetics, evolution, and enzymology of their biosynthesis. PMID:18497894

  13. Increased Lytic Efficiency of Bovine Macrophages Trained with Killed Mycobacteria.

    PubMed

    Juste, Ramon A; Alonso-Hearn, Marta; Garrido, Joseba M; Abendaño, Naiara; Sevilla, Iker A; Gortazar, Christian; de la Fuente, José; Dominguez, Lucas

    2016-01-01

    Innate immunity is evolutionarily conserved in multicellular organisms and was considered to lack memory until very recently. One of its more characteristic mechanisms is phagocytosis, the ability of cells to engulf, process and eventually destroy any injuring agent. We report the results of an ex vivo experiment in bovine macrophages in which improved clearance of Mycobacterium bovis (M. bovis) was induced by pre-exposure to a heat killed M. bovis preparation. The effects were independent of humoral and cellular adaptive immune responses and lasted up to six months. Specifically, our results demonstrate the existence of a training effect in the lytic phase of phagocytosis that can be activated by killed mycobacteria, thus suggesting a new mechanism of vaccine protection. These findings are compatible with the recently proposed concept of trained immunity, which was developed to explain the observation that innate immune responses provide unspecific protection against pathogens including other than those that originally triggered the immune response.

  14. Comparative Genomics of Mycobacteria: Some Answers, Yet More New Questions

    PubMed Central

    Behr, Marcel A.

    2015-01-01

    Comparative genomic studies permit a genus-level perspective on the distinction between environmental mycobacteria and Mycobacterium tuberculosis, as well as a species-level assessment of genetic variability within M. tuberculosis. Both of these strata of evolutionary analysis serve to generate hypotheses regarding the genomic basis of M. tuberculosis virulence. In contrasting lessons from macroevolutionary study and microevolutionary study, one can form predictions about which segments of the genome are likely to be essential for or dispensable for the pathogenesis of tuberculosis. Although some of these predictions have been experimentally verified, notable exceptions challenge the direct link between these virulence factors and the capacity of M. tuberculosis to successfully cause disease and propagate between human hosts. These unexpected findings serve as the stimulus for further studies, using genomic comparisons and other approaches, to better define the remarkable success of this recalcitrant pathogen. PMID:25395374

  15. Natural disasters and nontuberculous mycobacteria: a recipe for increased disease?

    PubMed

    Honda, Jennifer R; Bernhard, Jon N; Chan, Edward D

    2015-02-01

    Infectious diseases acquired by survivors of large-scale natural disasters complicate the recovery process. During events such as tsunamis, hurricanes, earthquakes, and tornados and well into the recovery period, victims often are exposed to water-soil mixtures that have relocated with indigenous microbes. Because nontuberculous mycobacteria (NTM) are ubiquitous in water and soil, there is potential for increased exposure to these organisms during natural disasters. In this hypothesis-driven commentary, we discuss the rise in NTM lung disease and natural disasters and examine the geographic overlap of NTM infections and disaster frequencies in the United States. Moreover, we show an increased number of positive NTM cultures from Louisiana residents in the years following three of the relatively recent epic hurricanes and posit that such natural disasters may help to drive the increased number of NTM infections. Finally, we advocate for increased environmental studies and surveillance of NTM infections before and after natural disasters.

  16. Increased Lytic Efficiency of Bovine Macrophages Trained with Killed Mycobacteria

    PubMed Central

    Juste, Ramon A.; Alonso-Hearn, Marta; Garrido, Joseba M.; Abendaño, Naiara; Sevilla, Iker A.; Gortazar, Christian; de la Fuente, José; Dominguez, Lucas

    2016-01-01

    Innate immunity is evolutionarily conserved in multicellular organisms and was considered to lack memory until very recently. One of its more characteristic mechanisms is phagocytosis, the ability of cells to engulf, process and eventually destroy any injuring agent. We report the results of an ex vivo experiment in bovine macrophages in which improved clearance of Mycobacterium bovis (M. bovis) was induced by pre-exposure to a heat killed M. bovis preparation. The effects were independent of humoral and cellular adaptive immune responses and lasted up to six months. Specifically, our results demonstrate the existence of a training effect in the lytic phase of phagocytosis that can be activated by killed mycobacteria, thus suggesting a new mechanism of vaccine protection. These findings are compatible with the recently proposed concept of trained immunity, which was developed to explain the observation that innate immune responses provide unspecific protection against pathogens including other than those that originally triggered the immune response. PMID:27820836

  17. Nontuberculous Mycobacteria: An Underestimated Cause of Bioprosthetic Valve Infective Endocarditis

    PubMed Central

    Bouchiat, Coralie; Saison, Julien; Boisset, Sandrine; Flandrois, Jean-Pierre; Issartel, Bertrand; Dauwalder, Olivier; Benito, Yvonne; Jarraud, Sophie; Grando, Jacqueline; Boibieux, Andre; Dumitrescu, Oana; Delahaye, François; Farhat, Fadi; Thivolet-Bejui, Françoise; Frieh, Jean-Philippe; Vandenesch, François

    2015-01-01

    Background. Atypical mycobacteria, or nontuberculous mycobacteria (NTM), have been barely reported as infective endocarditis (IE) agents. Methods. From January 2010 to December 2013, cardiac valve samples sent to our laboratory as cases of blood culture-negative suspected IE were analyzed by 16S rDNA polymerase chain reaction (PCR). When positive for NTM, hsp PCR allowed species identification. Demographic, clinical, echocardiographic, histopathological, and Ziehl-Neelsen staining data were then collected. Results. Over the study period, 6 of 370 cardiac valves (belonging to 5 patients in 3 hospitals) were positive for Mycobacterium chelonae (n = 5) and Mycobacterium lentiflavum (n = 1) exclusively on bioprosthetic material. The 5 patients presented to the hospital for heart failure without fever 7.1–18.9 months (median 13.1 months) after biological prosthetic valve implantation. Echocardiography revealed paravalvular regurgitation due to prosthesis dehiscence in all patients. Histopathological examination of the explanted material revealed inflammatory infiltrates in all specimens, 3 of which were associated with giant cells. Gram staining and conventional cultures remained negative, whereas Ziehl-Neelsen staining showed acid-fast bacilli in all patients. Allergic etiology was ruled out by antiporcine immunoglobulin E dosages. These 5 cases occurred exclusively on porcine bioprosthetic material, revealing a statistically significant association between bioprosthetic valves and NTM IE (P < .001). Conclusions. The body of evidence confirmed the diagnosis of prosthetic IE. The statistically significant association between bioprosthetic valves and NTM IE encourages systematic Ziehl-Neelsen staining of explanted bioprosthetic valves in case of early bioprosthesis dysfunction, even without an obvious sign of IE. In addition, we strongly question the cardiac bioprosthesis conditioning process after animal sacrifice. PMID:26213691

  18. Dual Analysis for Mycobacteria and Propionibacteria in Sarcoidosis BAL

    PubMed Central

    Oswald-Richter, Kyra A.; Beachboard, Dia C.; Seeley, Erin H.; Abraham, Susamma; Shepherd, Bryan E.; Jenkins, Cathy A.; Culver, Daniel A.; Caprioli, Richard M.; Drake, Wonder P.

    2012-01-01

    Purpose Sarcoidosis is a non-caseating granulomatous disease for which a role for infectious antigens continues to strengthen. Recent studies have reported molecular evidence of mycobacteria or propionibacteria. We assessed for immune responses against mycobacterial and propionibacterial antigens in sarcoidosis bronchoalveolar lavage (BAL) using flow cytometry, and localized signals consistent with microbial antigens with sarcoidosis specimens, using matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS). Methods BAL cells from 27 sarcoidosis, 14 PPD- controls, and 9 subjects with nontuberculosis mycobacterial (NTM) infections were analyzed for production of IFN-γ after stimulation with mycobacterial ESAT-6 and Propionibacterium acnes proteins. To complement the immunological data, MALDI-IMS was performed to localize ESAT-6 and Propionibacterium acnes signals within sarcoidosis and control specimens. Results CD4+ immunologic analysis for mycobacteria was positive in 17/27 sarcoidosis subjects, compared to 2/14 PPD-subjects, and 5/9 NTM subjects (p=00.008 and p=00.71 respectively, Fisher's exact test). There was no significant difference for recognition of P. acnes, which occurred only in sarcoidosis subjects that also recognized ESAT-6. Similar results were also observed for the CD8+ immunologic analysis. MALDI-IMS localized signals consistent with ESAT-6 only within sites of granulomatous inflammation, whereas P. acnes signals were distributed throughout the specimen. Conclusions MALDI-IMS localizes signals consistent with ESAT-6 to sarcoidosis granulomas, whereas no specific localization of P. acnes signals is detected. Immune responses against both mycobacterial and P. acnes are present within sarcoidosis BAL, but only mycobacterial signals are distinct from disease controls. These immunologic and molecular investigations support further investigation of the microbial community within sarcoidosis granulomas. PMID:22552860

  19. Nontuberculous mycobacteria isolations from residents of three counties in North Carolina, 2006 – 2010

    EPA Science Inventory

    Background: Nontuberculous mycobacteria (NTM) are emerging infections among the elderly and immunocompromised but the epidemiology is poorly characterized. Reports of NTM isolation from clinical specimens is a readily available, if imperfect surrogate for disease prevalence. Meth...

  20. Occurrence of Opportunistic Pathogens Legionella pneumophila and non-tuberculous mycobacteria in hospital plumbing systems

    EPA Science Inventory

    Occurrence of Opportunistic Pathogens Legionella pneumophila and non-tuberculous mycobacteria in hospital plumbing systems Jill Hoelle, Michael Coughlin, Elizabeth Sotkiewicz, Jingrang Lu, Stacy Pfaller, Mark Rodgers, and Hodon Ryu U.S. Environmental Protection Agency, Cincinnati...

  1. Series of Case Patients with Nontuberculous Mycobacteria Isolation, Central North Carolina, 2006-2010

    EPA Science Inventory

    Nontuberculous mycobacteria (NTM) infection/colonization, associated with human morbidity/mortality, is linked to drinking water and drinking water distribution systems. To characterize rates and distribution of NTM isolation among residents living in three North Carolina countie...

  2. Occurrence of Opportunistic Pathogens Legionella pneumophila and non-tuberculous mycobacteria in hospital plumbing systems

    EPA Science Inventory

    Occurrence of Opportunistic Pathogens Legionella pneumophila and non-tuberculous mycobacteria in hospital plumbing systems Jill Hoelle, Michael Coughlin, Elizabeth Sotkiewicz, Jingrang Lu, Stacy Pfaller, Mark Rodgers, and Hodon Ryu U.S. Environmental Protection Agency, Cincinnati...

  3. Nontuberculous mycobacteria isolations from residents of three counties in North Carolina, 2006 – 2010

    EPA Science Inventory

    Background: Nontuberculous mycobacteria (NTM) are emerging infections among the elderly and immunocompromised but the epidemiology is poorly characterized. Reports of NTM isolation from clinical specimens is a readily available, if imperfect surrogate for disease prevalence. Meth...

  4. Increasing Prevalence Rate of Nontuberculous Mycobacteria Infections in Five States, 2008–2013

    EPA Science Inventory

    Rationale: Many nontuberculous mycobacteria (NTM) are clinically significant pathogens that cause disease in a variety of different human organs and tissues. Objectives: A population-based study was undertaken to investigate the prevalence of patients with a positive specimen fo...

  5. Series of Case Patients with Nontuberculous Mycobacteria Isolation, Central North Carolina, 2006-2010

    EPA Science Inventory

    Nontuberculous mycobacteria (NTM) infection/colonization, associated with human morbidity/mortality, is linked to drinking water and drinking water distribution systems. To characterize rates and distribution of NTM isolation among residents living in three North Carolina countie...

  6. Mycobacterial growth and bacterial contamination in the mycobacteria growth indicator tube and BACTEC 460 culture systems.

    PubMed Central

    Cornfield, D B; Beavis, K G; Greene, J A; Bojak, M; Bondi, J

    1997-01-01

    The BACTEC 460 system currently provides the most rapid detection of mycobacterial growth, but the system is radiometric and requires needles to inoculate specimens through the bottle's septum. The Mycobacteria Growth Indicator Tube (MGIT) system has a liquid medium, like the BACTEC system, and does not require needles when inoculating specimens. We compared mycobacterial growth from 510 specimens in the two systems. Average time to acid-fast bacillus (AFB) detection and identification to the species level was less with the BACTEC system, but this result was statistically significant only for AFB detection in specimens containing Mycobacterium avium-M. intracellulare complex. The contamination rate with MGIT was 29%; the BACTEC rate was 5%. To investigate MGIT contamination, we initiated a second study with changes in specimen processing. The MGIT contamination rate was reduced to 12%; the BACTEC rate was not significantly affected (5.5%). The most likely explanation for the contamination in MGIT is the richness of its medium compared to the BACTEC medium. Cost analysis for the two systems in a laboratory that processes 4,500 specimens a year is presented. The data suggest that the BACTEC 460 and the MGIT systems are approximately equivalent in cost and ability to support the growth of AFB. The MGIT system appears safer and easier to use and was preferred by laboratory personnel, but it cannot currently be used for blood specimens or antituberculosis susceptibility testing. PMID:9230383

  7. Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform.

    PubMed

    Rock, Jeremy M; Hopkins, Forrest F; Chavez, Alejandro; Diallo, Marieme; Chase, Michael R; Gerrick, Elias R; Pritchard, Justin R; Church, George M; Rubin, Eric J; Sassetti, Christopher M; Schnappinger, Dirk; Fortune, Sarah M

    2017-02-06

    The development of new drug regimens that allow rapid, sterilizing treatment of tuberculosis has been limited by the complexity and time required for genetic manipulations in Mycobacterium tuberculosis. CRISPR interference (CRISPRi) promises to be a robust, easily engineered and scalable platform for regulated gene silencing. However, in M. tuberculosis, the existing Streptococcus pyogenes Cas9-based CRISPRi system is of limited utility because of relatively poor knockdown efficiency and proteotoxicity. To address these limitations, we screened eleven diverse Cas9 orthologues and identified four that are broadly functional for targeted gene knockdown in mycobacteria. The most efficacious of these proteins, the CRISPR1 Cas9 from Streptococcus thermophilus (dCas9Sth1), typically achieves 20- to 100-fold knockdown of endogenous gene expression with minimal proteotoxicity. In contrast to other CRISPRi systems, dCas9Sth1-mediated gene knockdown is robust when targeted far from the transcriptional start site, thereby allowing high-resolution dissection of gene function in the context of bacterial operons. We demonstrate the utility of this system by addressing persistent controversies regarding drug synergies in the mycobacterial folate biosynthesis pathway. We anticipate that the dCas9Sth1 CRISPRi system will have broad utility for functional genomics, genetic interaction mapping and drug-target profiling in M. tuberculosis.

  8. Growth characteristics of atypical mycobacteria in water and their comparative resistance to disinfectants.

    PubMed Central

    Carson, L A; Petersen, N J; Favero, M S; Aguero, S M

    1978-01-01

    With the increasing significance of group IV atypical mycobacteria as etiological agents in a variety of infections, studies were conducted to determine their growth capabilities in water and their comparative resistance to disinfectants used to decontaminate hospital equipment. Isolates of Mycobaterium chelonei (TM strains) from peritoneal fluids of patients and peritoneal dialysis machines were able to multiply in commercial distilled water, with generation times at 25 degrees C ranging from 8 to 15 h. Levels of 10(5) to 10(6) cells per ml were attained, and these stationary-phase populations declined only slightly over a 1-year period. Results of studies to determine resistance to disinfectants showed the following. (i) TM strains of M. chelonei cultured in commercial distilled water showed survivors in 2% aqueous formaldehyde (HCHO) solutions up to 24 h; in 8% HCHO, only a 2-log reduction in viable counts was observed over a 2-h sampling period. Reference ATCC strains of M. chelonei and M. fortuitum were rapidly inactivated, with no survivors after 2 h of exposure to 2% HCHO or 15 min of exposure to 8% HCHO. (ii) In 2% alkaline glutaraldehyde, TM strains survived 60 min. whereas ATCC strains showed no survivors after 2 min of contact time. (iii) All M. chelonei and M. fortuitum strains survived 60 min of exposure to concentrations of 0.3 and 0.7 microgram of free chlorine per ml at pH 7. PMID:104656

  9. [Contamination of a bronchial fiberscope by mycobacteria linked to an automated bronchoscope disinfection machine].

    PubMed

    Nomura, K; Ogawa, M; Chang, B; Miyamoto, H; Tanabe, T; Taniguchi, H; Matsumoto, T

    2000-06-01

    Mycobacteria are being isolated with increasing frequency from automated bronchoscope disinfection machines. This has led to misdiagnosis and nosocomial infections. In this study, we isolated Mycobacterium chelonae from a bronchoscope disinfection machine and found one strain to be resistant to 2% glutaraldehyde and sensitive to 70% ethanol. Since we began cleaning the sink of the machine with 70% ethanol, no mycobacteria has been seen throughout the machine.

  10. Fine-tuning the space, time, and host distribution of mycobacteria in wildlife

    PubMed Central

    2011-01-01

    Background We describe the diversity of two kinds of mycobacteria isolates, environmental mycobacteria and Mycobacterium bovis collected from wild boar, fallow deer, red deer and cattle in Doñana National Park (DNP, Spain), analyzing their association with temporal, spatial and environmental factors. Results High diversity of environmental mycobacteria species and M. bovis typing patterns (TPs) were found. When assessing the factors underlying the presence of the most common types of both environmental mycobacteria and M. bovis TPs in DNP, we evidenced (i) host species differences in the occurrence, (ii) spatial structuration and (iii) differences in the degree of spatial association of specific types between host species. Co-infection of a single host by two M. bovis TPs occurred in all three wild ungulate species. In wild boar and red deer, isolation of one group of mycobacteria occurred more frequently in individuals not infected by the other group. While only three TPs were detected in wildlife between 1998 and 2003, up to 8 different ones were found during 2006-2007. The opposite was observed in cattle. Belonging to an M. bovis-infected social group was a significant risk factor for mycobacterial infection in red deer and wild boar, but not for fallow deer. M. bovis TPs were usually found closer to water marshland than MOTT. Conclusions The diversity of mycobacteria described herein is indicative of multiple introduction events and a complex multi-host and multi-pathogen epidemiology in DNP. Significant changes in the mycobacterial isolate community may have taken place, even in a short time period (1998 to 2007). Aspects of host social organization should be taken into account in wildlife epidemiology. Wildlife in DNP is frequently exposed to different species of non-tuberculous, environmental mycobacteria, which could interact with the immune response to pathogenic mycobacteria, although the effects are unknown. This research highlights the suitability of

  11. Disseminated Nontuberculous Mycobacteria in HIV-Infected Patients, Oregon, USA, 2007–2012

    PubMed Central

    Ku, Jennifer H.; Henkle, Emily; Schafer, Sean D.; Winthrop, Kevin L.

    2017-01-01

    We determined disseminated nontuberculous mycobacteria incidence in the HIV-infected population of Oregon, USA, during 2007–2012 by using statewide laboratory surveillance. We identified 37 disseminated nontuberculous mycobacteria cases among 7,349 patients with median annual incidence of 110/100,000 HIV person-years and the highest incidence in those with CD4 counts <50 cells/mm3 (5,300/100,000 person-years). PMID:28221103

  12. Effect of relative centrifugal force and centrifugation time on sedimentation of mycobacteria in clinical specimens.

    PubMed

    Ratnam, S; March, S B

    1986-03-01

    Optimum relative centrifugal force (RCF) and centrifugation time to concentrate mycobacteria in clinical specimens were determined by processing split samples of sputa and urines containing mycobacteria with combinations of different RCFs and centrifugation times. Although individual test results showed considerable variation in the recovery rates of mycobacteria in the sediment, the data indicated that higher recovery rates occurred as centrifugation speed and time were increased. With a 15- to 20-min centrifugation time, on the average, 67 to 71% of mycobacteria were recovered at an RCF of 2,074 X g, and 76 to 80% were recovered at 3,005 or 3,895 X g at maximum radius. The remainder of mycobacteria was mostly recovered from the supernatant, but culturing of supernatant was not profitable. Increasing RCF had a negligible effect on acid-fast bacillus smear sensitivity. The smear sensitivity for about 25,000 clinical specimens processed with an RCF of 3,800 X g for 20 min was 71% compared with 69% as determined for over 30,000 specimens processed in a similar manner but an RCF of 2,000 X g. An RCF of 3,000 X g applied for 15 min, or an RCF of about 2,000 to 2,500 X g applied for 20 min, is considered adequate to concentrate mycobacteria in clinical specimens.

  13. Effect of relative centrifugal force and centrifugation time on sedimentation of mycobacteria in clinical specimens.

    PubMed Central

    Ratnam, S; March, S B

    1986-01-01

    Optimum relative centrifugal force (RCF) and centrifugation time to concentrate mycobacteria in clinical specimens were determined by processing split samples of sputa and urines containing mycobacteria with combinations of different RCFs and centrifugation times. Although individual test results showed considerable variation in the recovery rates of mycobacteria in the sediment, the data indicated that higher recovery rates occurred as centrifugation speed and time were increased. With a 15- to 20-min centrifugation time, on the average, 67 to 71% of mycobacteria were recovered at an RCF of 2,074 X g, and 76 to 80% were recovered at 3,005 or 3,895 X g at maximum radius. The remainder of mycobacteria was mostly recovered from the supernatant, but culturing of supernatant was not profitable. Increasing RCF had a negligible effect on acid-fast bacillus smear sensitivity. The smear sensitivity for about 25,000 clinical specimens processed with an RCF of 3,800 X g for 20 min was 71% compared with 69% as determined for over 30,000 specimens processed in a similar manner but an RCF of 2,000 X g. An RCF of 3,000 X g applied for 15 min, or an RCF of about 2,000 to 2,500 X g applied for 20 min, is considered adequate to concentrate mycobacteria in clinical specimens. PMID:3082926

  14. Microscopy, culture, and quantitative real-time PCR examination confirm internalization of mycobacteria in plants.

    PubMed

    Kaevska, M; Lvoncik, S; Slana, I; Kulich, P; Kralik, P

    2014-07-01

    The environment is a reservoir of nontuberculous mycobacteria and is considered a source of infection for animals and humans. Mycobacteria can persist in different types of environments for a relatively long time. We have studied their possible internalization into plant tissue through intact, as well as damaged, root systems of different types of plants grown in vitro and under field conditions. The substrate into which plants were seeded was previously contaminated with different strains of Mycobacterium avium (10(8) to 10(10) cells/g of soil) and feces from animals with paratuberculosis. We detected M. avium subsp. avium, hominissuis, and paratuberculosis in the stems and leaves of the plants by both culture and real-time quantitative PCR. The presence of mycobacteria in the plant tissues was confirmed by microscopy. The concentration of mycobacteria found inside plant tissue was several orders of magnitude lower (up to 10(4) cells/g of tissue) than the initial concentration of mycobacteria present in the culture medium or substrate. These findings led us to the hypothesis that plants may play a role in the spread and transmission of mycobacteria to other organisms in the environment.

  15. Mycobacterium shottsii sp. nov., a slowly growing species isolated from Chesapeake Bay striped bass (Morone saxatilis)

    USGS Publications Warehouse

    Rhodes, M.W.; Kator, H.; Kotob, S.; van Berkum, P.; Kaattari, I.; Vogelbein, W.; Quinn, F.; Floyd, M.M.; Butler, W.R.; Ottinger, C.A.

    2003-01-01

    Slowly growing, non-pigmented mycobacteria were isolated from striped bass (Morone saxatilis) during an epizootic of mycobacteriosis in the Chesapeake Bay. Growth characteristics, acid-fastness and results of 16S rRNA gene sequencing were consistent with those of the genus Mycobacterium. A unique profile of biochemical reactions was observed among the 21 isolates. A single cluster of eight peaks identified by analysis of mycolic acids (HPLC) resembled those of reference patterns but differed in peak elution times from profiles of reference species of the Mycobacterium tuberculosis complex. One isolate (M175T) was placed within the slowly growing mycobacteria by analysis of aligned 16S rRNA gene sequences and was proximate in phylogeny to Mycobacterium ulcerans and Mycobacterium marinum. However, distinct nucleotide differences were detected in the 16S rRNA gene sequence among M175T, M. ulcerans and M. marinum (99.2% similarity). Isolate M175T could be differentiated from other slowly growing, non-pigmented mycobacteria by its inability to grow at 37??C, production of niacin and urease, absence of nitrate reductase and resistance to isoniazid (1 ??g ml-1), thiacetazone and thiophene-2-carboxylic hydrazide. Based upon these genetic and phenotypic differences, isolate M175T (= ATCC 700981T = NCTC 13215T) is proposed as the type strain of a novel species, Mycobacterium shottsii sp. nov.

  16. Multicenter Evaluation of the Mycobacteria Growth Indicator Tube for Testing Susceptibility of Mycobacterium tuberculosis to First-Line Drugs

    PubMed Central

    Rüsch-Gerdes, Sabine; Domehl, Cornelia; Nardi, Giampietro; Gismondo, Maria Rita; Welscher, Hans-Martin; Pfyffer, Gaby E.

    1999-01-01

    In a multicenter study involving three reference centers for mycobacteria, the reliability of the Mycobacteria Growth Indicator Tube (MGIT) for rapid antimicrobial susceptibility testing (AST) of Mycobacterium tuberculosis was evaluated and compared to the radiometric method (BACTEC 460TB). Test cultures for which the results of the MGIT and BACTEC 460TB tests were discordant were checked by the conventional proportion method on solid medium. Four hundred forty-one isolates have been tested for susceptibility to isoniazid (INH), rifampin (RMP), ethambutol (EMB), and streptomycin (SM). Discrepant results were obtained for three isolates (0.7%) with INH (susceptible by MGIT, resistant by BACTEC 460TB), for four isolates (0.9%) with RMP (susceptible by MGIT, resistant by BACTEC 460TB), for six isolates (1.9%) with EMB (four susceptible by MGIT, resistant by BACTEC 460TB; two resistant by MGIT, susceptible by BACTEC 460TB), and for four isolates (0.9%) with SM (two susceptible by MGIT, resistant by BACTEC 460TB; two resistant by MGIT, susceptible by BACTEC 460TB). When cultures with discordant results were tested by the conventional proportion method, about half of the cultures yielded results similar to the BACTEC 460TB results, while the other half yielded results similar to the MGIT results. Turnaround times were 3 to 14 days (median, 8.8 days) for MGIT and 3 to 15 days (median, 7.8 days) for BACTEC 460TB. There was no statistically significant difference between the susceptibility testing results of the two methods (P > 0.05). These data demonstrate that the MGIT system is an accurate, nonradiometric alternative to the BACTEC 460TB method for rapid susceptibility testing of M. tuberculosis. PMID:9854062

  17. Multifork chromosome replication in slow-growing bacteria

    PubMed Central

    Trojanowski, Damian; Hołówka, Joanna; Ginda, Katarzyna; Jakimowicz, Dagmara; Zakrzewska-Czerwińska, Jolanta

    2017-01-01

    The growth rates of bacteria must be coordinated with major cell cycle events, including chromosome replication. When the doubling time (Td) is shorter than the duration of chromosome replication (C period), a new round of replication begins before the previous round terminates. Thus, newborn cells inherit partially duplicated chromosomes. This phenomenon, which is termed multifork replication, occurs among fast-growing bacteria such as Escherichia coli and Bacillus subtilis. In contrast, it was historically believed that slow-growing bacteria (including mycobacteria) do not reinitiate chromosome replication until the previous round has been completed. Here, we use single-cell time-lapse analyses to reveal that mycobacterial cell populations exhibit heterogeneity in their DNA replication dynamics. In addition to cells with non-overlapping replication rounds, we observed cells in which the next replication round was initiated before completion of the previous replication round. We speculate that this heterogeneity may reflect a relaxation of cell cycle checkpoints, possibly increasing the ability of slow-growing mycobacteria to adapt to environmental conditions. PMID:28262767

  18. Multifork chromosome replication in slow-growing bacteria.

    PubMed

    Trojanowski, Damian; Hołówka, Joanna; Ginda, Katarzyna; Jakimowicz, Dagmara; Zakrzewska-Czerwińska, Jolanta

    2017-03-06

    The growth rates of bacteria must be coordinated with major cell cycle events, including chromosome replication. When the doubling time (Td) is shorter than the duration of chromosome replication (C period), a new round of replication begins before the previous round terminates. Thus, newborn cells inherit partially duplicated chromosomes. This phenomenon, which is termed multifork replication, occurs among fast-growing bacteria such as Escherichia coli and Bacillus subtilis. In contrast, it was historically believed that slow-growing bacteria (including mycobacteria) do not reinitiate chromosome replication until the previous round has been completed. Here, we use single-cell time-lapse analyses to reveal that mycobacterial cell populations exhibit heterogeneity in their DNA replication dynamics. In addition to cells with non-overlapping replication rounds, we observed cells in which the next replication round was initiated before completion of the previous replication round. We speculate that this heterogeneity may reflect a relaxation of cell cycle checkpoints, possibly increasing the ability of slow-growing mycobacteria to adapt to environmental conditions.

  19. Multiphasic strain differentiation of atypical mycobacteria from elephant trunk wash.

    PubMed

    Chan, Kok-Gan; Loke, Mun Fai; Ong, Bee Lee; Wong, Yan Ling; Hong, Kar Wai; Tan, Kian Hin; Kaur, Sargit; Ng, Hien Fuh; Abdul Razak, Mfa; Ngeow, Yun Fong

    2015-01-01

    Background. Two non-tuberculous mycobacterial strains, UM_3 and UM_11, were isolated from the trunk wash of captive elephants in Malaysia. As they appeared to be identical phenotypes, they were investigated further by conventional and whole genome sequence-based methods of strain differentiation. Methods. Multiphasic investigations on the isolates included species identification with hsp65 PCR-sequencing, conventional biochemical tests, rapid biochemical profiling using API strips and the Biolog Phenotype Microarray analysis, protein profiling with liquid chromatography-mass spectrometry, repetitive sequence-based PCR typing and whole genome sequencing followed by phylogenomic analyses. Results. The isolates were shown to be possibly novel slow-growing schotochromogens with highly similar biological and genotypic characteristics. Both strains have a genome size of 5.2 Mbp, G+C content of 68.8%, one rRNA operon and 52 tRNAs each. They qualified for classification into the same species with their average nucleotide identity of 99.98% and tetranucleotide correlation coefficient of 0.99999. At the subspecies level, both strains showed 98.8% band similarity in the Diversilab automated repetitive sequence-based PCR typing system, 96.2% similarity in protein profiles obtained by liquid chromatography mass spectrometry, and a genomic distance that is close to zero in the phylogenomic tree constructed with conserved orthologs. Detailed epidemiological tracking revealed that the elephants shared a common habitat eight years apart, thus, strengthening the possibility of a clonal relationship between the two strains.

  20. Unveiling unusual features of formation of septal partition and constriction in mycobacteria--an ultrastructural study.

    PubMed

    Vijay, Srinivasan; Anand, Deepak; Ajitkumar, Parthasarathi

    2012-02-01

    The ultrastructural functions of the electron-dense glycopeptidolipid-containing outermost layer (OL), the arabinogalactan-mycolic acid-containing electron-transparent layer (ETL), and the electron-dense peptidoglycan layer (PGL) of the mycobacterial cell wall in septal growth and constriction are not clear. Therefore, using transmission electron microscopy, we studied the participation of the three layers in septal growth and constriction in the fast-growing saprophytic species Mycobacterium smegmatis and the slow-growing pathogenic species Mycobacterium xenopi and Mycobacterium tuberculosis in order to document the processes in a comprehensive and comparative manner and to find out whether the processes are conserved across different mycobacterial species. A complete septal partition is formed first by the fresh synthesis of the septal PGL (S-PGL) and septal ETL (S-ETL) from the envelope PGL (E-PGL) in M. smegmatis and M. xenopi. The S-ETL is not continuous with the envelope ETL (E-ETL) due to the presence of the E-PGL between them. The E-PGL disappears, and the S-ETL becomes continuous with the E-ETL, when the OL begins to grow and invaginate into the S-ETL for constriction. However, in M. tuberculosis, the S-PGL and S-ETL grow from the E-PGL and E-ETL, respectively, without a separation between the E-ETL and S-ETL by the E-PGL, in contrast to the process in M. smegmatis and M. xenopi. Subsequent growth and invagination of the OL into the S-ETL of the septal partition initiates and completes septal constriction in M. tuberculosis. A model for the conserved sequential process of mycobacterial septation, in which the formation of a complete septal partition is followed by constriction, is presented. The probable physiological significance of the process is discussed. The ultrastructural features of septation and constriction in mycobacteria are unusually different from those in the well-studied organisms Escherichia coli and Bacillus subtilis.

  1. Radiometric measurement of differential metabolism of fatty acid by mycobacteria.

    PubMed

    Camargo, E E; Kertcher, J A; Larson, S M; Tepper, B S; Wagner, H N

    1982-06-01

    An assay system has been developed based on automated radiometric quantification of 14CO2 produced through oxidation of [1-14C] fatty acids by mycobacteria. Two stains of M. tuberculosis (H37Rv and Erdman) and one of M. bovis (BCG) in 7H9 medium (ADC) with 1.0 microCi of one of the fatty acids (butyric, hexanoic, octanoic, decanoic, lauric, myristic, palmitic, stearic, oleic, linoleic and linolenic) were studied. Results previously published on M. lepraemurium (Hawaiian) were also included for comparison. Both strains of M. tuberculosis had maximum 14CO2 production from hexanoic acid. Oxidation of butyric and avid oxidation of lauric acids were also found with the H37Rv strain but not with Erdman. In contrast, 14CO2 production by M. bovis was greatest from lauric and somewhat less from decanoic acid. M. lepraemurium showed increasing oxidation rates from myristic, decanoic and lauric acids. Assimilation studies of M. tuberculosis H37Rv confirmed that most of the oxidized substrates were converted into by-products with no change in those from which no oxidation was found. These data suggest that the radiometric measurement of differential fatty acid metabolism may provide a basis of strain identification of the genus Mycobacterium.

  2. Review: Environmental mycobacteria as a cause of human infection.

    PubMed

    Halstrom, Samuel; Price, Patricia; Thomson, Rachel

    2015-06-01

    Pulmonary infections with nontuberculous mycobacteria (NTM) are recognized as a problem in immunodeficient individuals and are increasingly common in older people with no known immune defects. NTM are found in soil and water, but factors influencing transmission from the environment to humans are mostly unknown. Studies of the epidemiology of NTM disease have matched some clinical isolates of NTM with isolates from the patient's local environment. Definitive matching requires strain level differentiation based on molecular analyses, including partial sequencing, PCR-restriction fragment length polymorphism (RFLP) analysis, random amplified polymorphic DNA (RAPD) PCR, repetitive element (rep-) PCR and pulsed field gel electrophoresis (PFGE) of large restriction fragments. These approaches have identified hospital and residential showers and faucets, hot-tubs and garden soil as sources of transmissible pathogenic NTM. However, gaps exist in the literature, with many clinical isolates remaining unidentified within environments that have been tested, and few studies investigating NTM transmission in developing countries. To understand the environmental reservoirs and transmission routes of pathogenic NTM, different environments, countries and climates must be investigated. Copyright © 2015 Asian African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.

  3. Nontuberculous Mycobacteria Isolated from Tuberculosis Suspects in Ibadan, Nigeria

    PubMed Central

    Cadmus, Simeon Idowu; Diarra, Bassirou; Traore, Brehima; Maiga, Mamoudou; Siddiqui, Sophia; Tounkara, Anatole; Falodun, Olutayo; Lawal, Wole; Adewole, Isaac Folurunso; Murphy, Rob; van Soolingen, Dick; Taiwo, Babafemi

    2016-01-01

    In Nigeria, one of the highest tuberculosis (TB) burdened nations, sputum smear microscopy is routinely employed for TB diagnosis at Directly Observed Treatment Short-Course (DOTS) Centers. This diagnostic algorithm does not differentiate Mycobacterium tuberculosis complex (MTC) from nontuberculous mycobacteria (NTM). Between December 2008 and January 2009, consecutive patients diagnosed with TB were screened for inclusion at 10 DOTS centers in Ibadan, Nigeria. To verify Mycobacterium species in patients diagnosed, we cultured and identified mycobacterial isolates using PCR, line probe assay, and spoligotyping techniques. From 48 patients screened, 23 met the inclusion criteria for the study. All the 23 study patients had a positive culture. Overall, we identified 11/23 patients (48%) with MTC only, 9/23 (39%) with NTM only, and 3/23 (13%) with evidence of both MTC and NTM. Strains of MTC identified were Latin American Mediterranean (LAM) genotype (n = 12), M. africanum (n = 1), and the genotype family T (n = 1). Four M. avium-intracellulare-M. scrofulaceum complexes, one M. chelonae complex, one M. abscessus, and one M. intracellulare were identified. Our findings underscore the need to incorporate molecular techniques for more precise diagnosis of TB at DOTS centers to improve clinical outcomes and safe guard public health, particularly in TB endemic countries. PMID:27099795

  4. Mycobacteria manipulate macrophage recruitment through coordinated use of membrane lipids.

    PubMed

    Cambier, C J; Takaki, Kevin K; Larson, Ryan P; Hernandez, Rafael E; Tobin, David M; Urdahl, Kevin B; Cosma, Christine L; Ramakrishnan, Lalita

    2014-01-09

    The evolutionary survival of Mycobacterium tuberculosis, the cause of human tuberculosis, depends on its ability to invade the host, replicate, and transmit infection. At its initial peripheral infection site in the distal lung airways, M. tuberculosis infects macrophages, which transport it to deeper tissues. How mycobacteria survive in these broadly microbicidal cells is an important question. Here we show in mice and zebrafish that M. tuberculosis, and its close pathogenic relative Mycobacterium marinum, preferentially recruit and infect permissive macrophages while evading microbicidal ones. This immune evasion is accomplished by using cell-surface-associated phthiocerol dimycoceroserate (PDIM) lipids to mask underlying pathogen-associated molecular patterns (PAMPs). In the absence of PDIM, these PAMPs signal a Toll-like receptor (TLR)-dependent recruitment of macrophages that produce microbicidal reactive nitrogen species. Concordantly, the related phenolic glycolipids (PGLs) promote the recruitment of permissive macrophages through a host chemokine receptor 2 (CCR2)-mediated pathway. Thus, we have identified coordinated roles for PDIM, known to be essential for mycobacterial virulence, and PGL, which (along with CCR2) is known to be associated with human tuberculosis. Our findings also suggest an explanation for the longstanding observation that M. tuberculosis initiates infection in the relatively sterile environment of the lower respiratory tract, rather than in the upper respiratory tract, where resident microflora and inhaled environmental microbes may continually recruit microbicidal macrophages through TLR-dependent signalling.

  5. Transformation of the Antibacterial Agent Norfloxacin by Environmental Mycobacteria

    PubMed Central

    Adjei, Michael D.; Heinze, Thomas M.; Deck, Joanna; Freeman, James P.; Williams, Anna J.; Sutherland, John B.

    2006-01-01

    Because fluoroquinolone antimicrobial agents may be released into the environment, the potential for environmental bacteria to biotransform these drugs was investigated. Eight Mycobacterium sp. cultures in a sorbitol-yeast extract medium were dosed with 100 μg ml−1 of norfloxacin and incubated for 7 days. The MICs of norfloxacin for these strains, tested by an agar dilution method, were 1.6 to 25 μg ml−1. Cultures were extracted with ethyl acetate, and potential metabolites in the extracts were purified by high-performance liquid chromatography. The metabolites were identified using mass spectrometry and nuclear magnetic resonance spectroscopy. N-Acetylnorfloxacin (5 to 50% of the total absorbance at 280 nm) was produced by the eight Mycobacterium strains. N-Nitrosonorfloxacin (5 to 30% of the total absorbance) was also produced by Mycobacterium sp. strain PYR100 and Mycobacterium gilvum PYR-GCK. The MICs of N-nitrosonorfloxacin and N-acetylnorfloxacin were 2- to 38- and 4- to 1,000-fold higher, respectively, than those of norfloxacin for several different bacteria, including the two strains that produced both metabolites. Although N-nitrosonorfloxacin had less antibacterial activity, nitrosamines are potentially carcinogenic. The biotransformation of fluoroquinolones by mycobacteria may serve as a resistance mechanism. PMID:16957195

  6. Iron Deprivation Affects Drug Susceptibilities of Mycobacteria Targeting Membrane Integrity

    PubMed Central

    Pal, Rahul; Hameed, Saif; Fatima, Zeeshan

    2015-01-01

    Multidrug resistance (MDR) acquired by Mycobacterium tuberculosis (MTB) through continuous deployment of antitubercular drugs warrants immediate search for novel targets and mechanisms. The ability of MTB to sense and become accustomed to changes in the host is essential for survival and confers the basis of infection. A crucial condition that MTB must surmount is iron limitation, during the establishment of infection, since iron is required by both bacteria and humans. This study focuses on how iron deprivation affects drug susceptibilities of known anti-TB drugs in Mycobacterium smegmatis, a “surrogate of MTB.” We showed that iron deprivation leads to enhanced potency of most commonly used first line anti-TB drugs that could be reverted upon iron supplementation. We explored that membrane homeostasis is disrupted upon iron deprivation as revealed by enhanced membrane permeability and hypersensitivity to membrane perturbing agent leading to increased passive diffusion of drug and TEM images showing detectable differences in cell envelope thickness. Furthermore, iron seems to be indispensable to sustain genotoxic stress suggesting its possible role in DNA repair machinery. Taken together, we for the first time established a link between cellular iron and drug susceptibility of mycobacteria suggesting iron as novel determinant to combat MDR. PMID:26779346

  7. Pathogenic mycobacteria achieve cellular persistence by inhibiting the Niemann-Pick Type C disease cellular pathway

    PubMed Central

    2016-01-01

    Background. Tuberculosis remains a major global health concern. The ability to prevent phagosome-lysosome fusion is a key mechanism by which intracellular mycobacteria, including Mycobacterium tuberculosis, achieve long-term persistence within host cells. The mechanisms underpinning this key intracellular pro-survival strategy remain incompletely understood. Host macrophages infected with persistent mycobacteria share phenotypic similarities with cells taken from patients suffering from Niemann-Pick Disease Type C (NPC), a rare lysosomal storage disease in which endocytic trafficking defects and lipid accumulation within the lysosome lead to cell dysfunction and cell death. We investigated whether these shared phenotypes reflected an underlying mechanistic connection between mycobacterial intracellular persistence and the host cell pathway dysfunctional in NPC. Methods. The induction of NPC phenotypes in macrophages from wild-type mice or obtained from healthy human donors was assessed via infection with mycobacteria and subsequent measurement of lipid levels and intracellular calcium homeostasis. The effect of NPC therapeutics on intracellular mycobacterial load was also assessed. Results. Macrophages infected with persistent intracellular mycobacteria phenocopied NPC cells, exhibiting accumulation of multiple lipid types, reduced lysosomal Ca2+ levels, and defects in intracellular trafficking. These NPC phenotypes could also be induced using only lipids/glycomycolates from the mycobacterial cell wall. These data suggest that persistent intracellular mycobacteria inhibit the NPC pathway, likely via inhibition of the NPC1 protein, and subsequently induce altered acidic store Ca2+ homeostasis. Reduced lysosomal calcium levels may provide a mechanistic explanation for the reduced levels of phagosome-lysosome fusion in mycobacterial infection. Treatments capable of correcting defects in NPC mutant cells via modulation of host cell calcium were of benefit in promoting

  8. MALDI-TOF MS Andromas strategy for the routine identification of bacteria, mycobacteria, yeasts, Aspergillus spp. and positive blood cultures.

    PubMed

    Bille, E; Dauphin, B; Leto, J; Bougnoux, M-E; Beretti, J-L; Lotz, A; Suarez, S; Meyer, J; Join-Lambert, O; Descamps, P; Grall, N; Mory, F; Dubreuil, L; Berche, P; Nassif, X; Ferroni, A

    2012-11-01

    All organisms usually isolated in our laboratory are now routinely identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) using the Andromas software. The aim of this study was to describe the use of this strategy in a routine clinical microbiology laboratory. The microorganisms identified included bacteria, mycobacteria, yeasts and Aspergillus spp. isolated on solid media or extracted directly from blood cultures. MALDI-TOF MS was performed on 2665 bacteria isolated on solid media, corresponding to all bacteria isolated during this period except Escherichia coli grown on chromogenic media. All acquisitions were performed without extraction. After a single acquisition, 93.1% of bacteria grown on solid media were correctly identified. When the first acquisition was not contributory, a second acquisition was performed either the same day or the next day. After two acquisitions, the rate of bacteria identified increased to 99.2%. The failures reported on 21 strains were due to an unknown profile attributed to new species (9) or an insufficient quality of the spectrum (12). MALDI-TOF MS has been applied to 162 positive blood cultures. The identification rate was 91.4%. All mycobacteria isolated during this period (22) were correctly identified by MALDI-TOF MS without any extraction. For 96.3% and 92.2% of yeasts and Aspergillus spp., respectively, the identification was obtained with a single acquisition. After a second acquisition, the overall identification rate was 98.8% for yeasts (160/162) and 98.4% (63/64) for Aspergillus spp. In conclusion, the MALDI-TOF MS strategy used in this work allows a rapid and efficient identification of all microorganisms isolated routinely.

  9. Control of the Bcg gene of early resistance in mice to infections with BCG substrains and atypical mycobacteria.

    PubMed Central

    Denis, M; Forget, A; Pelletier, M; Turcotte, R; Skamene, E

    1986-01-01

    The effect of the Bcg gene on the early host response to intravenous infection with a variety of BCG substrains and some atypical mycobacteria was investigated. The numbers of live bacilli of BCG Pasteur and BCG Tice recovered from the spleens of Bcgs mice (C57BL/6, B10.A and BALB/c) at 3 weeks following infection exceeded the bacterial dose injected, whereas the number of CFU recovered from the spleens of Bcgr mice (A/J, DBA/2 and C3H/HeN) did not exceed the number of CFU injected, thus following the pattern observed in Bcgr mice and Bcgs infected with BCG Montreal. BCG Russia failed to multiply in both test groups; however, the number of CFU recovered in Bcgr mice was significantly lower than in Bcgs mice. On the other hand, the presence of live bacilli in the spleens of either Bcgr or Bcgs mice injected with BCG Japan was undetectable in most cases. Involvement of the Bcg gene in the early resistance to infection with BCG Pasteur, BCG Russia, Mycobacterium kansasii and M. intracellulare was documented by the significant differences in the kinetics of infections in mice of the C.D2 (BALB/c-Bcgr) and BALB/c (Bcgs) congenic lines. In BCG Russia, M. intracellulare and M. fortuitum infections, the phenotypic expression of the Bcg gene resulted in a more rapid elimination of the bacteria in the spleens of Bcgr when compared with Bcgs mice. On the other hand, the hepatic granuloma formation correlated with bacterial load except when C.D2 mice were infected with a small dose of BCG Pasteur or M. kansasii where extensive granulomatous hepatitis developed although no bacterial multiplication occurred in the spleen. It is suggested that granuloma formation could depend of the properties of the mycobacteria as well as the genetic background of the host without implicating the bacterial burden. PMID:3086001

  10. Role of the horizontal gene exchange in evolution of pathogenic Mycobacteria

    PubMed Central

    2015-01-01

    Background Mycobacterium tuberculosis is one of the most dangerous human pathogens, the causative agent of tuberculosis. While this pathogen is considered as extremely clonal and resistant to horizontal gene exchange, there are many facts supporting the hypothesis that on the early stages of evolution the development of pathogenicity of ancestral Mtb has started with a horizontal acquisition of virulence factors. Episodes of infections caused by non-tuberculosis Mycobacteria reported worldwide may suggest a potential for new pathogens to appear. If so, what is the role of horizontal gene transfer in this process? Results Availing of accessibility of complete genomes sequences of multiple pathogenic, conditionally pathogenic and saprophytic Mycobacteria, a genome comparative study was performed to investigate the distribution of genomic islands among bacteria and identify ontological links between these mobile elements. It was shown that the ancient genomic islands from M. tuberculosis still may be rooted to the pool of mobile genetic vectors distributed among Mycobacteria. A frequent exchange of genes was observed between M. marinum and several saprophytic and conditionally pathogenic species. Among them M. avium was the most promiscuous species acquiring genetic materials from diverse origins. Conclusions Recent activation of genetic vectors circulating among Mycobacteria potentially may lead to emergence of new pathogens from environmental and conditionally pathogenic Mycobacteria. The species which require monitoring are M. marinum and M. avium as they eagerly acquire genes from different sources and may become donors of virulence gene cassettes to other micro-organisms. PMID:25708825

  11. Transcriptome analysis of mycobacteria in sputum samples of pulmonary tuberculosis patients

    PubMed Central

    Sharma, Sumedha; Ryndak, Michelle B.; Aggarwal, Ashutosh N.; Yadav, Rakesh; Sethi, Sunil; Masih, Shet; Laal, Suman; Verma, Indu

    2017-01-01

    Pulmonary tuberculosis, the disease caused by Mycobacterium tuberculosis, still retains a top rank among the deadliest communicable diseases. Sputum expectorated during the disease continues to be a primary diagnostic specimen and also serves as a reservoir of bacteria. The expression pattern of mycobacteria in sputum will lead to an insight into bacterial adaptation at the most highly transmissible stage of infection and can also help in identifying newer diagnostic as well as drug targets. Thus, in the present study, a whole genome microarray of Mycobacterium tuberculosis was used to elucidate the transcriptional profile of mycobacteria in the sputum samples of smear positive pulmonary tuberculosis patients. Overall, the mycobacteria in sputum appeared to be in a low energy and low replicative state as compared to in vitro grown log phase M. tb with downregulation of genes involved in ATP synthesis, aerobic respiration and translational machinery. Simultaneously, downregulation was also seen in the genes involved in secretion machinery of mycobacteria along with the downregulation of genes involved in the synthesis of phthiocerol dimycocerosate and phenol glycolipids. In contrast, the majority of the genes which showed an upregulation in sputum mycobacteria were of unknown function. Further identification of these genes may provide new insights into the mycobacterial behavior during this phase of infection and may help in deciphering candidates for development of better diagnostic and drug candidates. PMID:28282458

  12. First evidence of amoebae-mycobacteria association in drinking water network.

    PubMed

    Delafont, Vincent; Mougari, Faïza; Cambau, Emmanuelle; Joyeux, Michel; Bouchon, Didier; Héchard, Yann; Moulin, Laurent

    2014-10-21

    Free-living amoebae are protozoa ubiquitously found in water systems. They mainly feed on bacteria by phagocytosis, but some bacterial species are able to resist or even escape this lethal process. Among these amoeba resistant bacteria are numerous members of the genus Mycobacterium. Nontuberculous Mycobacteria (NTM) are opportunistic pathogens that share the same ecological niches as amoebae. While several studies have demonstrated the ability of these bacteria to colonise and persist within drinking water networks, there is also strong suspicion that mycobacteria could use amoebae as a vehicle for protection and even replication. We investigated here the presence of NTM and FLA on a drinking water network during an all year round sampling campaign. We observed that 87.6% of recovered amoebal cultures carried high numbers of NTM. Identification of these amoeba and mycobacteria strains indicated that the main genera found in drinking water networks, that is, Acanthamoeba, Vermamoeba, Echinamoeba, and Protacanthamoeba are able to carry and likely to allow replication of several environmental and potentially pathogenic mycobacteria including M. llatzerense and M. chelonae. Direct Sanger sequencing as well as pyrosequencing of environmental isolates demonstrated the frequent association of mycobacteria and FLA, as they are part of the most represented genera composing amoebae's microbiome. This is the first time that an association between FLA and NTM is observed in water networks, highlighting the importance of FLA in the ecology of NTM.

  13. Effects of protein concentration and degradability on performance, ruminal fermentation, and nitrogen metabolism in rapidly growing heifers fed high-concentrate diets from 100 to 230 kg body weight.

    PubMed

    Devant, M; Ferret, A; Gasa, J; Calsamiglia, S; Casals, R

    2000-06-01

    Twenty crossbred heifers (101 +/- 4.5 kg BW) were used to examine the effects of protein concentration and degradability on performance, ruminal fermentation, nutrient digestion, N balance, and urinary excretion of purine derivatives. Heifers were offered concentrate and barley straw for ad libitum consumption. Two protein concentrations (17 vs 14%, DM basis) and two protein sources differing in ruminal degradability (58 vs 42% of CP for soybean meal and treated soybean meal, respectively) were tested. The experiment was divided into four consecutive 28-d periods to evaluate the age (period) effect. Increasing protein concentration and degradability did not improve ADG or intake (P > .05). The increase in urinary N excretion (P < .001) in heifers fed 17% CP suggests that N was in excess of requirements. When the low-degradable protein source was supplemented and(or) CP concentration was low, ruminal NH3 N concentrations fell below 5 mg/100 mL. Urinary excretion of purine derivatives was not affected (P > .05) by protein concentration and degradability, suggesting that in high-concentrate diets NH3 N concentration was not limiting microbial growth. Total VFA concentration decreased (P < .001) and the acetate:propionate ratio increased (P < .01) with advancing period, suggesting an increase in ruminal absorption capacity and an increase in fiber fermentation. The decrease in ruminal NH3 N concentration in the last period suggests a greater use of NH3 N by microorganisms. This hypothesis is supported by the increase (P < .001) in urinary excretion of allantoin and estimated duodenal flows of purine bases and microbial protein with advancing period. Reducing CP concentration and increasing ruminal undegradable protein supply did not affect animal performance or estimated duodenal flow of microbial protein in rapidly growing heifers fed high-concentrate diets.

  14. How Does Your Garlic Grow?

    ERIC Educational Resources Information Center

    Shimabukuro, Mary A.; Fearing, Vickie

    1993-01-01

    Garlic is an ideal plant for the elementary classroom. It grows rapidly in water without aeration for several weeks and remains relatively free of microbial contamination. Simple experiments with garlic purchased at grocery stores can illustrate various aspects of plant growth. (PR)

  15. How Does Your Garlic Grow?

    ERIC Educational Resources Information Center

    Shimabukuro, Mary A.; Fearing, Vickie

    1993-01-01

    Garlic is an ideal plant for the elementary classroom. It grows rapidly in water without aeration for several weeks and remains relatively free of microbial contamination. Simple experiments with garlic purchased at grocery stores can illustrate various aspects of plant growth. (PR)

  16. [Distribution and drug resistance of nontuberculous Mycobacteria in Beijing].

    PubMed

    Zhang, J; Su, J R; Ding, B C; Liu, J W; Yi, J L; Yang, X Y; Wang, N H; Wang, S M

    2017-03-12

    Objective: To analyze the distribution and drug resistance of nontuberculous mycobacteria(NTM) in Beijing. Methods: Using PCR-fluorescence probe method we identified 1 552 mycobacterial isolates in 2009 and 1 553 mycobacterial isolates in 2013, which were stored by Beijing Research Institute for Tuberculosis Control.All identified NTM strains were confirmed by 16S rRNA gene sequencing, and drug sensitivity testing was performed by using 1% ratio method.SPSS 13.0 was used for statistical analysis. Results: The isolation rate for NTM in 2009 and 2013 was 3.8%(59/1 552), and 4.6%(71/1 553) respectively. A total of 130 NTM strains were identified to 13 species by 16S rRNA gene sequencing, including M. intracellulare strains 39.2%(51/130), M. kansasii strains 37.7%(49/130), M. avium strains 6.9%(9/130), M. abscessus strains 5.4%(7/130), M. fortuitum strains 3.0%(4/130), M. gordonae strains 1.5%(2/130), M. xenopi strains 1.5%(2/130), M. scrofulaceum, M. Phlei, M. smegmatis, M. vaccae, M. neoaurum, M. kumamotonense 1 strain each. For the patients infected with NTM, 87 were male and 43 were female, with an average age of 55 years. The results of drug sensitivity test from 97 strains of NTM showed that isoniazid and p-aminosalicylic acid showed the highest drug resistant rate of 98%(95/97), followed by streptomycin 94.8%(92/97), capreomycin 81.4%(79/97), amikacin 69.1%(67/97), levofloxacin 56.7%(55/97), rifampicin 54.6%(53/97), prothionamide 51.5%(50/97), and ethambutol 50.5%(49/97). Conclusions:Mycobacterium intracellulare and Mycobacterium kansasii were the main strains isolated from patients infected with NTM in Beijing. Patients infected with NTM were mostly males. NTM showed high resistance to anti-tuberculosis drugs.

  17. Multidrug-resistant nontuberculous mycobacteria isolated from cystic fibrosis patients.

    PubMed

    Cândido, Pedro Henrique Campanini; Nunes, Luciana de Souza; Marques, Elizabeth Andrade; Folescu, Tânia Wrobel; Coelho, Fábrice Santana; de Moura, Vinicius Calado Nogueira; da Silva, Marlei Gomes; Gomes, Karen Machado; Lourenço, Maria Cristina da Silva; Aguiar, Fábio Silva; Chitolina, Fernanda; Armstrong, Derek T; Leão, Sylvia Cardoso; Neves, Felipe Piedade Gonçalves; Mello, Fernanda Carvalho de Queiroz; Duarte, Rafael Silva

    2014-08-01

    Worldwide, nontuberculous mycobacteria (NTM) have become emergent pathogens of pulmonary infections in cystic fibrosis (CF) patients, with an estimated prevalence ranging from 5 to 20%. This work investigated the presence of NTM in sputum samples of 129 CF patients (2 to 18 years old) submitted to longitudinal clinical supervision at a regional reference center in Rio de Janeiro, Brazil. From June 2009 to March 2012, 36 NTM isolates recovered from 10 (7.75%) out of 129 children were obtained. Molecular identification of NTM was performed by using PCR restriction analysis targeting the hsp65 gene (PRA-hsp65) and sequencing of the rpoB gene, and susceptibility tests were performed that followed Clinical and Laboratory Standards Institute recommendations. For evaluating the genotypic diversity, pulsed-field gel electrophoresis (PFGE) and/or enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) was performed. The species identified were Mycobacterium abscessus subsp. bolletii (n = 24), M. abscessus subsp. abscessus (n = 6), Mycobacterium fortuitum (n = 3), Mycobacterium marseillense (n = 2), and Mycobacterium timonense (n = 1). Most of the isolates presented resistance to five or more of the antimicrobials tested. Typing profiles were mainly patient specific. The PFGE profiles indicated the presence of two clonal groups for M. abscessus subsp. abscessus and five clonal groups for M. abscesssus subsp. bolletii, with just one clone detected in two patients. Given the observed multidrug resistance patterns and the possibility of transmission between patients, we suggest the implementation of continuous and routine investigation of NTM infection or colonization in CF patients, including countries with a high burden of tuberculosis disease.

  18. Effects of storage and transport on the cultivability of mycobacteria

    PubMed Central

    Šula, Ladislav; Sundaresan, T. K.; Langerová, M.

    1960-01-01

    In the course of WHO/UNICEF-assisted tuberculosis surveys carried out in a number of African territories, sputa were microscopically examined for the presence of acid-fast bacilli. Since adequate facilities for performing cultures to confirm the diagnosis of tuberculosis and other mycobacterial infections were not available in these territories, it was necessary to despatch sputum specimens to certain European laboratories for culturing and typing of mycobacteria. However, it was noticed that the number of positive cultures from such sputa was very low, being limited largely to specimens in which acid-fast bacilli were easily demonstrable by microscopy. Specimens containing scanty acid-fast bacilli, or microscopically negative specimens, usually failed to exhibit growth on culture, contrary to the usual observations made with European sputum specimens cultured in European laboratories. As the sputa were mostly taken from new cases with lung pathology, previous treatment could not have been responsible for these poor culture results, and it was thought that the conditions in which the specimens were transported, and possibly also the chemical composition of the containers in which they were stored, might be the cause. In an experiment carried out by the WHO Tuberculosis Research Office, in collaboration with a WHO field team in Africa and the Tuberculosis Research Institute in Prague, pure cultures of the H37 Rv strain and sputa were sent from Prague to East Africa and book in conditions simulating those in which specimens collected by African field teams are routinely sent to European laboratories. The results show that the cultivability of tubercle bacilli is adversely affected by storage and transport. PMID:20604080

  19. Multidrug-Resistant Nontuberculous Mycobacteria Isolated from Cystic Fibrosis Patients

    PubMed Central

    Cândido, Pedro Henrique Campanini; Nunes, Luciana de Souza; Marques, Elizabeth Andrade; Folescu, Tânia Wrobel; Coelho, Fábrice Santana; de Moura, Vinicius Calado Nogueira; da Silva, Marlei Gomes; Gomes, Karen Machado; Lourenço, Maria Cristina da Silva; Aguiar, Fábio Silva; Chitolina, Fernanda; Armstrong, Derek T.; Leão, Sylvia Cardoso; Neves, Felipe Piedade Gonçalves; Mello, Fernanda Carvalho de Queiroz

    2014-01-01

    Worldwide, nontuberculous mycobacteria (NTM) have become emergent pathogens of pulmonary infections in cystic fibrosis (CF) patients, with an estimated prevalence ranging from 5 to 20%. This work investigated the presence of NTM in sputum samples of 129 CF patients (2 to 18 years old) submitted to longitudinal clinical supervision at a regional reference center in Rio de Janeiro, Brazil. From June 2009 to March 2012, 36 NTM isolates recovered from 10 (7.75%) out of 129 children were obtained. Molecular identification of NTM was performed by using PCR restriction analysis targeting the hsp65 gene (PRA-hsp65) and sequencing of the rpoB gene, and susceptibility tests were performed that followed Clinical and Laboratory Standards Institute recommendations. For evaluating the genotypic diversity, pulsed-field gel electrophoresis (PFGE) and/or enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) was performed. The species identified were Mycobacterium abscessus subsp. bolletii (n = 24), M. abscessus subsp. abscessus (n = 6), Mycobacterium fortuitum (n = 3), Mycobacterium marseillense (n = 2), and Mycobacterium timonense (n = 1). Most of the isolates presented resistance to five or more of the antimicrobials tested. Typing profiles were mainly patient specific. The PFGE profiles indicated the presence of two clonal groups for M. abscessus subsp. abscessus and five clonal groups for M. abscesssus subsp. bolletii, with just one clone detected in two patients. Given the observed multidrug resistance patterns and the possibility of transmission between patients, we suggest the implementation of continuous and routine investigation of NTM infection or colonization in CF patients, including countries with a high burden of tuberculosis disease. PMID:24920766

  20. Inventory study of non-tuberculous mycobacteria in the European Union

    PubMed Central

    2014-01-01

    Background Since non-tuberculous mycobacteria (NTM) disease is not notifiable in most European Union (EU) and European Economic Area (EEA) countries, the epidemiological situation of the >150 NTM species is largely unknown. We aimed to collect data on the frequency of NTM detection and NTM species types in EU/EEA countries. Methods Officially nominated national tuberculosis reference laboratories of all EU/EEA countries were asked to provide information on: laboratory routines for detection and identification of NTM, including drug sensitivity testing (DST) methods; data on the number and type of NTM species identified; coverage and completeness of the provided data on NTM; type and number of human specimens tested for NTM; and number of specimens tested for Mycobacterium tuberculosis complex and NTM. This information was summarized and the main results are described. Results In total, 99 different NTM species were identified with M. avium, M. gordonae, M. xenopi , M. intracellulare, and M. fortuitum identified most frequently. Seven percent of the NTM species could not be identified. NTM was cultured from between 0.4-2.0% of the specimens (data from four countries). The laboratories use culturing methods optimised for M. tuberculosis complex. Identification is mainly carried out by a commercial line probe assay supplemented with sequencing. Most laboratories carried out DST for rapid growers and only at the explicit clinical request for slow growers. Conclusion It is likely that the prevalence of NTM is underestimated because diagnostic procedures are not optimized specifically for NTM and isolates may not be referred to the national reference laboratory for identification. Due to the diagnostic challenges and the need to establish the clinical relevance of NTM, we recommend that countries should concentrate detection and identification in only few laboratories. PMID:24502462

  1. Infection by tubercular mycobacteria is spread by nonlytic ejection from their amoeba hosts

    PubMed Central

    Hagedorn, Monica; Rohde, Kyle H.; Russell, David G.; Soldati, Thierry

    2009-01-01

    To generate efficient vaccines and cures for Mycobacterium tuberculosis, we need a far better understanding of modes of infection, persistence and spreading. Host cell entry and establishment of a replication niche are well understood, but little is known about how tubercular mycobacteria exit host cells and disseminate the infection. Using the social amoeba Dictyostelium as a genetically tractable host for pathogenic mycobacteria, we discovered that M. tuberculosis and M. marinum but not M. avium are ejected from the cell through an actin-based structure, the ejectosome. This conserved nonlytic spreading mechanism requires a cytoskeleton regulator from the host and an intact mycobacterial ESX-1 secretion system. This insight offers new directions for research into the spreading of tubercular mycobacteria infections in mammalian cells. PMID:19325115

  2. Growing and Growing: Promoting Functional Thinking with Geometric Growing Patterns

    ERIC Educational Resources Information Center

    Markworth, Kimberly A.

    2010-01-01

    Design research methodology is used in this study to develop an empirically-substantiated instruction theory about students' development of functional thinking in the context of geometric growing patterns. The two research questions are: (1) How does students' functional thinking develop in the context of geometric growing patterns? (2) What are…

  3. [Isolation of environmental mycobacteria from soils of Córdoba city Argentina].

    PubMed

    Ballarino, Guillermo J; Eseverri, M Verónica; Salas, Andrea V; Giayetto, Víctor O; González, Silvia; Wolff, Lidia; Pessah, Oscar

    2002-01-01

    The interest for the research on enviromental mycobacteria has risen over the last decades, in part, due to a significant incidence rate rise. Reports from all over the world address the soil as the major source for human contamination. In Argentina two documents report the prevalence of atypical mycobacteriosis at Córdoba (1997), and the isolation of enviromental mycobacteria from soils of the Province of La Pampa (1999) respectively. The aim of our study was to confirm the presence of enviromental mycobacteria in soil of the city of Córdoba. The map of the city was divided in 9 regions according to avenues and major streets distribution. A total of 120 soil samples were recollected with spatula from a 10 x 10 cm square up to 1 cm deep. Samples were kept at 4 degrees C no more than 7 days. Soil samples were homogenized with destilled water in a 1:1 proportion, and decontaminated according to Petroff's method. The cultures were made in Lowestein-Jehnsen media and incubated at 37 degrees C controlling development every 7 days for 2 months. An acid-fast-bacilli smear was made from colonies obtained. Twenty three cultures (19%) were discarded due to contamination. Twenty cultures (17%) developed acid fast bacilli (AFB). Colonies obtained were sent to the Mycobacteria Service of the Instituto Nacional de Enfermedades Infecciosas Dr. Carlos G. Malbrán, in the city of Buenos Aires, for identification. A single isolation was identified as Mycobacterium triviale. A positive correlation was observed between the frequency of positive AFB isolation and the number of samples taken from park areas. The presence of enviromental mycobacteria in soils of Córdoba was confirmed. Results suggest higher odds of isolation in parklands and soils where animals live. Extensive works are needed to asset the features that allow and contribute the proliferation of mycobacteria in soils.

  4. [Pulmonary infection caused by non-tuberculous mycobacteria in two patients with bronchiectasis].

    PubMed

    Daniels, J M A; Haitjema, T; van Altena, R; van den Aardweg, J G; Vlaspolder, F; Boersma, W G

    2005-04-30

    Two patients, a woman aged 67 years and a man aged 80 years, had chronic cough among other respiratory symptoms. In the woman, chest radiograph and CT-scan revealed partial atelectasis of the middle lobe and bronchiectasis. In the man, an interstitial pattern was seen on chest radiograph, and CT scan showed diffuse bronchiectasis. In both the man and the woman, non-tuberculous mycobacteria were identified (Mycobacterium avium complex and Mycobacterium abscessus, respectively). Treatment was successful in both patients. Non-tuberculous mycobacteria can cause considerable pulmonary infection in patients with bronchiectasis.

  5. Validation of biomarkers for distinguishing Mycobacterium tuberculosis from non-tuberculous mycobacteria using gas chromatography-mass spectrometry and chemometrics.

    PubMed

    Dang, Ngoc A; Kuijper, Sjoukje; Walters, Elisabetta; Claassens, Mareli; van Soolingen, Dick; Vivo-Truyols, Gabriel; Janssen, Hans-Gerd; Kolk, Arend H J

    2013-01-01

    Tuberculosis (TB) remains a major international health problem. Rapid differentiation of Mycobacterium tuberculosis complex (MTB) from non-tuberculous mycobacteria (NTM) is critical for decisions regarding patient management and choice of therapeutic regimen. Recently we developed a 20-compound model to distinguish between MTB and NTM. It is based on thermally assisted hydrolysis and methylation gas chromatography-mass spectrometry and partial least square discriminant analysis. Here we report the validation of this model with two independent sample sets, one consisting of 39 MTB and 17 NTM isolates from the Netherlands, the other comprising 103 isolates (91 MTB and 12 NTM) from Stellenbosch, Cape Town, South Africa. All the MTB strains in the 56 Dutch samples were correctly identified and the model had a sensitivity of 100% and a specificity of 94%. For the South African samples the model had a sensitivity of 88% and specificity of 100%. Based on our model, we have developed a new decision-tree that allows the differentiation of MTB from NTM with 100% accuracy. Encouraged by these findings we will proceed with the development of a simple, rapid, affordable, high-throughput test to identify MTB directly in sputum.

  6. Growing media [Chapter 5

    Treesearch

    Douglass F. Jacobs; Thomas D. Landis; Tara Luna

    2009-01-01

    Selecting the proper growing medium is one of the most important considerations in nursery plant production. A growing medium can be defined as a substance through which roots grow and extract water and nutrients. In native plant nurseries, a growing medium can consist of native soil but is more commonly an "artificial soil" composed of materials such as peat...

  7. Culture-Independent Detection of Nontuberculous Mycobacteria in Clinical Respiratory Samples

    PubMed Central

    Scoleri, Gianny P.; Choo, Jocelyn M.; Leong, Lex E. X.; Goddard, Thomas R.; Shephard, Lisa; Burr, Lucy D.; Bastian, Ivan; Thomson, Rachel M.

    2016-01-01

    Culture-based detection of nontuberculous Mycobacteria (NTM) in respiratory samples is time consuming and can be subject to overgrowth by nonmycobacterial bacteria. We describe a single-reaction TaqMan quantitative PCR assay for the direct detection of NTM species in clinical samples that is specific, sensitive, and robust. PMID:27413194

  8. New Insights in to the Intrinsic and Acquired Drug Resistance Mechanisms in Mycobacteria

    PubMed Central

    Nasiri, Mohammad J.; Haeili, Mehri; Ghazi, Mona; Goudarzi, Hossein; Pormohammad, Ali; Imani Fooladi, Abbas A.; Feizabadi, Mohammad M.

    2017-01-01

    Infectious diseases caused by clinically important Mycobacteria continue to be an important public health problem worldwide primarily due to emergence of drug resistance crisis. In recent years, the control of tuberculosis (TB), the disease caused by Mycobacterium tuberculosis (MTB), is hampered by the emergence of multidrug resistance (MDR), defined as resistance to at least isoniazid (INH) and rifampicin (RIF), two key drugs in the treatment of the disease. Despite the availability of curative anti-TB therapy, inappropriate and inadequate treatment has allowed MTB to acquire resistance to the most important anti-TB drugs. Likewise, for most mycobacteria other than MTB, the outcome of drug treatment is poor and is likely related to the high levels of antibiotic resistance. Thus, a better knowledge of the underlying mechanisms of drug resistance in mycobacteria could aid not only to select the best therapeutic options but also to develop novel drugs that can overwhelm the existing resistance mechanisms. In this article, we review the distinctive mechanisms of antibiotic resistance in mycobacteria. PMID:28487675

  9. Structural insights into the mycobacteria transcription initiation complex from analysis of X-ray crystal structures

    DOE PAGES

    Hubin, Elizabeth A.; Lilic, Mirjana; Darst, Seth A.; ...

    2017-07-13

    The mycobacteria RNA polymerase (RNAP) is a target for antimicrobials against tuberculosis, motivating structure/function studies. Here we report a 3.2 Å-resolution crystal structure of a Mycobacterium smegmatis (Msm) open promoter complex (RPo), along with structural analysis of the Msm RPo and a previously reported 2.76 Å-resolution crystal structure of an Msm transcription initiation complex with a promoter DNA fragment. We observe the interaction of the Msm RNAP α-subunit C-terminal domain (αCTD) with DNA, and we provide evidence that the a CTD may play a role in Mtb transcription regulation. Here, our results reveal the structure of an Actinobacteria-unique insert ofmore » the RNAP β' subunit. Finally, our analysis reveals the disposition of the N-terminal segment of Msm σA, which may comprise an intrinsically disordered protein domain unique to mycobacteria. The clade-specific features of the mycobacteria RNAP provide clues to the profound instability of mycobacteria RPo compared with E. coli.« less

  10. Modeling Human Exposure Risk to Nontuberculous Mycobacteria in Central North Carolina

    EPA Science Inventory

    Nontuberculous mycobacteria (NTM) are a broad group of soil-and water-borne bacteria. Some species are pathogenic and may cause serious infections in the lungs, soft tissues, bones and skin. Infections in humans are associated with environmental exposures to contaminated soil, ae...

  11. HUMAN INFECTION WITH NONTUBERCULOUS MYCOBACTERIA SPP. IN KING COUNTY, WASHINGTON, 1999-2002

    EPA Science Inventory

    Human infection with nontuberculous Mycobacteria spp. in King County, Washington, 1999 - 2002
    E Hilborn, T Covert, M Yakrus, G Stelma, M Schmitt
    1) US Environmental Protection Agency, Office of Research and Development, National Health and Environmental Research Laboratory,...

  12. Synthetic trehalose esters of cis-alkene and diene α'-mycolic acids of Mycobacteria.

    PubMed

    Taher, Salam G; Muzael, Maged; Al Dulayymi, Juma'a R; Baird, Mark S

    2015-07-01

    The synthesis of an α'-mycolic acid of Mycobacterium smegmatis and other mycobacteria, containing a cis,cis-diene, and of the trehalose mono- and di-mycolates of this, and of a related α'-mycolic acid containing one cis-alkene, is reported. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  13. Mild Nutrient Starvation Triggers the Development of a Small-Cell Survival Morphotype in Mycobacteria

    PubMed Central

    Wu, Mu-Lu; Gengenbacher, Martin; Dick, Thomas

    2016-01-01

    Mycobacteria, generally believed to be non-sporulating, are well known to survive shock starvation in saline for extended periods of time in a non-replicating state without any apparent morphological changes. Here, we uncover that mycobacteria can undergo cellular differentiation by exposing Mycobacterium smegmatis to mild starvation conditions. Traces of various carbon sources in saline triggered the development of a novel small resting cell (SMRC) morphotype. Development of SMRCs could also be observed for other mycobacteria, suggesting evolutionary conservation of this differentiation pathway. Fluorescence microscopic analyses showed that development of SMRCs progresses via septated, multi-nucleoided cell intermediates, which divide to generate mono-nucleoided SMRCs. Intriguingly, saline shock-starved large resting cells (LARCs), which did not show cell size or surface changes when observed by scanning electron microscopy, remodeled their internal structure to septated, multi-nucleoided cells, similar to the intermediates seen during differentiation to SMRCs. Our results suggest that mycobacteria harbor a starvation-induced differentiation program in which at first septated, multi-nucleoided cells are generated. Under zero-nutrient conditions bacteria terminate development at this stage as LARCs. In the presence of traces of a carbon source, these multi-nucleoided cells continue differentiation into mono-nucleoided SMRCs. Both SMRCs and LARCs exhibited extreme antibiotic tolerance. SMRCs showed increased long-term starvation survival, which was associated with the presence of lipid inclusion bodies. PMID:27379076

  14. Does infection with environmental mycobacteria suppress the protective response to subsequent vaccination with BCG?

    PubMed

    Smith, D; Reeser, P; Musa, S

    1985-03-01

    Using a guinea pig model of experimental airborne tuberculosis, we were unable to find evidence to support the hypothesis that infection with environmental mycobacteria (M. simiae or M. avium-intracellulare) interferes with the induction of a protective response in animals subsequently vaccinated with BCG.

  15. HUMAN INFECTION WITH NONTUBERCULOUS MYCOBACTERIA SPP. IN KING COUNTY, WASHINGTON, 1999-2002

    EPA Science Inventory

    Human infection with nontuberculous Mycobacteria spp. in King County, Washington, 1999 - 2002
    E Hilborn, T Covert, M Yakrus, G Stelma, M Schmitt
    1) US Environmental Protection Agency, Office of Research and Development, National Health and Environmental Research Laboratory,...

  16. Modeling Human Exposure Risk to Nontuberculous Mycobacteria in Central North Carolina

    EPA Science Inventory

    Nontuberculous mycobacteria (NTM) are a broad group of soil-and water-borne bacteria. Some species are pathogenic and may cause serious infections in the lungs, soft tissues, bones and skin. Infections in humans are associated with environmental exposures to contaminated soil, ae...

  17. Membrane-active antimicrobial peptides and human placental lysosomal extracts are highly active against mycobacteria.

    PubMed

    Jena, Prajna; Mishra, Bibhuti; Leippe, Matthias; Hasilik, Andrej; Griffiths, Gareth; Sonawane, Avinash

    2011-05-01

    Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, manifests discreet strategies to subvert host immune responses, which enable the pathogen to survive and multiply inside the macrophages. This problem is further worsened by the emergence of multidrug resistant mycobacterial strains, which make most of the anti-tuberculous drugs ineffective. It is thus imperative to search for and design better therapeutic strategies, including employment of new antibiotics. Recently, naturally produced antimicrobial molecules such as enzymes, peptides and their synthetic analogs have emerged as compounds with potentially significant therapeutical applications. Although, many antimicrobial peptides have been identified only very few of them have been tested against mycobacteria. A major limitation in using peptides as therapeutics is their sensitivity to enzymatic degradation or inactivity under certain physiological conditions such as relatively high salt concentration. Here, we show that NK-2, a peptide representing the cationic core region of the lymphocytic effector protein NK-lysin, and Ci-MAM-A24, a synthetic salt-tolerant peptide derived from immune cells of Ciona intestinalis, efficiently kill Mycobacterium smegmatis and Mycobacterium bovis-BCG. In addition, NK-2 and Ci-MAM-A24 showed a synergistic killing effect against M. smegmatis, no cytotoxic effect on mouse macrophages at bactericidal concentrations, and were even found to kill mycobacteria residing inside the macrophages. We also show that human placental lysosomal contents exert potent killing effect against mycobacteria under acidic and reducing growth conditions. Electron microscopic studies demonstrate that the lysosomal extract disintegrate bacterial cell membrane resulting in killing of mycobacteria.

  18. Mycobacteria-specific cytokine responses as correlates of treatment response in active and latent tuberculosis.

    PubMed

    Clifford, Vanessa; Tebruegge, Marc; Zufferey, Christel; Germano, Susie; Forbes, Ben; Cosentino, Lucy; McBryde, Emma; Eisen, Damon; Robins-Browne, Roy; Street, Alan; Denholm, Justin; Curtis, Nigel

    2017-08-01

    A biomarker indicating successful tuberculosis (TB) therapy would assist in determining appropriate length of treatment. This study aimed to determine changes in mycobacteria-specific antigen-induced cytokine biomarkers in patients receiving therapy for latent or active TB, to identify biomarkers potentially correlating with treatment success. A total of 33 adults with active TB and 36 with latent TB were followed longitudinally over therapy. Whole blood stimulation assays using mycobacteria-specific antigens (CFP-10, ESAT-6, PPD) were done on samples obtained at 0, 1, 3, 6 and 9 months. Cytokine responses (IFN-γ, IL-1ra, IL-2, IL-10, IL-13, IP-10, MIP-1β, and TNF-α) in supernatants were measured by Luminex xMAP immunoassay. In active TB cases, median IL-1ra (with CFP-10 and with PPD stimulation), IP-10 (CFP-10, ESAT-6), MIP-1β (ESAT-6, PPD), and TNF-α (ESAT-6) responses declined significantly over the course of therapy. In latent TB cases, median IL-1ra (CFP-10, ESAT-6, PPD), IL-2 (CFP-10, ESAT-6), and IP-10 (CFP-10, ESAT-6) responses declined significantly. Mycobacteria-specific cytokine responses change significantly over the course of therapy, and their kinetics in active TB differ from those observed in latent TB. In particular, mycobacteria-specific IL-1ra responses are potential correlates of successful therapy in both active and latent TB. Copyright © 2017 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

  19. Patients with nontuberculous mycobacteria: comparison of updated and previous diagnostic criteria for lung disease.

    PubMed

    Epson, Erin; Cassidy, Maureen; Marshall-Olson, Angela; Hedberg, Katrina; Winthrop, Kevin L

    2012-09-01

    We classified patients with respiratory nontuberculous mycobacteria (NTM) isolates using updated (2007) and previous (1997) American Thoracic Society/Infectious Diseases Society of America criteria for NTM lung disease. We found that a greater proportion of such patients have disease using updated criteria due to improved sensitivity of the microbiologic component of the disease definition.

  20. [The prevalence of salmonella, yersinia and mycobacteria in slaughtered pigs in Switzerland].

    PubMed

    Offermann, U; Bodmer, T; Audigé, L; Jemmi, T

    1999-01-01

    Clinically healthy food animals can be reservoirs for various foodborne pathogens. In general, such animals do not have lesions that are visible during meat inspection. Pigs are considered to be carriers of salmonella, yersinia and mycobacteria, but the risk of transmission to humans is difficult to assess. The aim of this study was to estimate the actual prevalence of the three above mentioned pathogens in the Swiss pig population and to comment on their significance. A total of 570 samples each of tonsils and mesenteric lymphnodes, were collected at two slaughterhouses from carcasses of apparently healthy pigs and analyzed for the presence of salmonella, yersinia and mycobacteria. The prevalence of salmonella (0.9%) was found to be lower than--while that of yersinia (8.1%) and mycobacteria (12.8%) about equal to--results reported from other European countries. Yersinia typing showed that serotype O:9 of Yersinia enterocolitica (2.5%) was 6 to 7 times more frequent than serotype O:3 (0.4%)--formerly the most frequent serotype. Mycobacterium avium was the most frequent isolate (90.7%) among the mycobacteria isolated. Although all three pathogens are present in the Swiss pig population, we consider the risk of transmission to humans via consumption of pork as low. Appropriate preventive measures and quality management should contribute to keep the risk under control.

  1. Simplified Automated Image Analysis for Detection and Phenotyping of Mycobacterium tuberculosis on Porous Supports by Monitoring Growing Microcolonies

    PubMed Central

    den Hertog, Alice L.; Visser, Dennis W.; Ingham, Colin J.; Fey, Frank H. A. G.; Klatser, Paul R.; Anthony, Richard M.

    2010-01-01

    Background Even with the advent of nucleic acid (NA) amplification technologies the culture of mycobacteria for diagnostic and other applications remains of critical importance. Notably microscopic observed drug susceptibility testing (MODS), as opposed to traditional culture on solid media or automated liquid culture, has shown potential to both speed up and increase the provision of mycobacterial culture in high burden settings. Methods Here we explore the growth of Mycobacterial tuberculosis microcolonies, imaged by automated digital microscopy, cultured on a porous aluminium oxide (PAO) supports. Repeated imaging during colony growth greatly simplifies “computer vision” and presumptive identification of microcolonies was achieved here using existing publically available algorithms. Our system thus allows the growth of individual microcolonies to be monitored and critically, also to change the media during the growth phase without disrupting the microcolonies. Transfer of identified microcolonies onto selective media allowed us, within 1-2 bacterial generations, to rapidly detect the drug susceptibility of individual microcolonies, eliminating the need for time consuming subculturing or the inoculation of multiple parallel cultures. Significance Monitoring the phenotype of individual microcolonies as they grow has immense potential for research, screening, and ultimately M. tuberculosis diagnostic applications. The method described is particularly appealing with respect to speed and automation. PMID:20544033

  2. Confirmation of the presence of Mycobacterium tuberculosis and other mycobacteria in mycobacterial growth indicator tubes (MGIT) by multiplex strand displacement amplification.

    PubMed Central

    Badak, F Z; Kiska, D L; O'Connell, M; Nycz, C M; Hartley, C; Setterquist, S; Hopfer, R L

    1997-01-01

    Multiplex strand displacement amplification (mSDA) is capable of amplifying three distinct DNA sequences simultaneously. These include sequences present in most genera of mycobacteria, a sequence specific for Mycobacterium tuberculosis, and an internal control. mSDA was used to detect the presence of these target sequences in 154 (72 positive, 76 negative, and 6 failed) clinical specimens cultured in the mycobacterial growth indicator tube (MGIT) system. A wide variety of specimen types were processed and cultured. Once these cultures were deemed positive by MGIT fluorescence or were deemed negative after 8 weeks of incubation, MGIT culture aliquots were processed for mSDA analyses. A chemiluminescent microwell assay was used to detect the amplified products. The procedure was relatively simple and took less than 6 h to complete. The sensitivity of mSDA for detecting acid-fast bacilli was 96.4% compared to that of MGIT culture. Sensitivity and specificity were 97.2 and 96.1%, respectively, when all clinical criteria were considered. mSDA was shown to be a rapid and effective method for confirming the presence of M. tuberculosis and other mycobacteria in positive MGIT cultures. PMID:9114414

  3. Mycobacteria in Municipal Wastewater Treatment and Reuse: Microbial Diversity for Screening the Occurrence of Clinically and Environmentally Relevant Species in Arid Regions.

    PubMed

    Amha, Yamrot M; Anwar, M Zohaib; Kumaraswamy, Rajkumari; Henschel, Andreas; Ahmad, Farrukh

    2017-03-07

    With accumulating evidence of pulmonary infection via aerosolized nontuberculous mycobacteria (NTM), it is important to characterize their persistence in wastewater treatment, especially in arid regions where treated municipal wastewater is extensively reused. To achieve this goal, microbial diversity of the genus Mycobacterium was screened for clinically and environmentally relevant species using pyrosequencing. Analysis of the postdisinfected treated wastewater showed the presence of clinically relevant slow growers like M. kansasii, M. szulgai, M. gordonae, and M. asiaticum; however, in these samples, rapid growers like M. mageritense occurred at much higher relative abundance. M. asiaticum and M. mageritense have been isolated in pulmonary samples from NTM-infected patients in the region. Diversity analysis along the treatment train found environmentally relevant organisms like M. poriferae and M. insubricum to increase in relative abundance across the chlorine disinfection step. A comparison to qPCR results across the chlorine disinfection step saw no significant change in slow grower counts at CT disinfection values ≤90 mg·min/L; only an increase to 180 mg·min/L in late May brought slow growers to below detection levels. The study confirms the occurrence of clinically and environmentally relevant mycobacteria in treated municipal wastewater, suggesting the need for vigilant monitoring of treated wastewater quality and disinfection effectiveness prior to reuse.

  4. Comparison of methods for the isolation of mycobacteria from water treatment plant sludge.

    PubMed

    Makovcova, Jitka; Babak, Vladimir; Slany, Michal; Slana, Iva

    2015-05-01

    Nontuberculous mycobacteria (NTM) are ubiquitous organisms in all natural ecosystems, including water environments. Several of these species are potential pathogens which affect human health. NTM most commonly cause pulmonary, skin or soft tissue infections. Primary sludge obtained from the water treatment plants of four drinking water reservoirs were subjected to analysis for mycobacteria. Five decontamination methods (5% oxalic acid, modified Petroff, HCl-NaOH, N-acetyl-L-cysteine-sodium hydroxide and 0.05% cetylpyridinium chloride), three growth media (Herrold's egg yolk medium with and without the antibiotic cocktail PANTA and Löwenstein-Jensen medium with sodium pyruvate) and three incubation temperatures (25, 30 and 37 °C) for isolation of mycobacteria were compared in the analysis of 18 sludge samples. To evaluate examined methods, the overall positive, negative, and contamination rate, and these rates in respect to localities are taken into account. Statistical analysis demonstrated that the best combination for the recovery of mycobacteria with the minimum number of contaminating microorganisms is 5% oxalic acid decontamination cultured on Herrold's egg yolk medium with the antibiotic cocktail PANTA at 25 °C. The least suitable is N-acetyl-L-cysteine-sodium hydroxide decontamination cultured on Löwenstein-Jensen medium with sodium pyruvate at 25 °C. From 18 sludge samples we isolated 27 mycobacterial species or groups; Mycobacterium algericum, M. arabiense, M. heraklionense, M. minnesotense, M. moriokaense, M. salmoniphilum and M. vulneris were isolated from the natural water environment for the first time. Because the natural water environment is the main source of potentially pathogenic mycobacteria for humans, it is important to direct particular focus to newly described mycobacterial species.

  5. Isolation and characterization of mycobacteria from striped bass Morone saxatilis from the Chesapeake Bay

    USGS Publications Warehouse

    Rhodes, M.W.; Kator, H.; Kaattari, I.; Gauthier, D.; Vogelbein, W.; Ottinger, C.A.

    2004-01-01

    Mycobacteriosis in striped bass Morone saxatilis of Chesapeake Bay, USA, was first diagnosed in 1997 based on the presence of granulomatous inflammation and acid-fast bacteria in skin and spleen. To confirm histopathology, bacteriological detection and identification of mycobacteria were begun using splenic tissue from fish with and without skin ulcerations. On the basis of initial studies using a variety of selective and nonselective media, decontamination, homogenization and incubation conditions, a simple and quantitative recovery method using aseptic necropsy of splenic tissue was developed. Optimal recovery was obtained by spread-plating homogenates on Middlebrook 7H10 agar with incubation for 3 mo at 23??C. Mycobacteria were recovered from 76% (n = 149/196) of fish examined. Mycobacterial densities exceeded 104 colony forming units??g tissue-1 in 38% of samples (n = 63/168) that were examined using a quantitative approach. The most frequently recovered mycobacterium, present in 57% (n = 109/192) of characterized samples, was the recently named new species Mycobacterium shottsii. Polyinfections of M. shottsii and other mycobacteria were observed in 25% of samples (n = 47/192) with densities of M. shottsii usually 1 or more orders of magnitude higher than co-isolate(s). Other mycobacteria recovered included isolates that, based on phenotypic traits, resembled M. interjectum, M. marinum, M. scrofulaceum, M. szulgai and M. triplex. M. marinum, commonly associated with fish mycobacteriosis and human disease, was recovered infrequently (3%, n = 6/192). The presence of multiple mycobacterial types occurring at high densities suggests that a variety of mycobacteria could be causative agents of mycobacteriosis in striped bass from the Chesapeake Bay. Striped bass is the major recreational fish species in the Chesapeake Bay, and the significance of the current epizootic to human health and the potential adverse effects on fish stocks are not known.

  6. Mycobacteria release active membrane vesicles that modulate immune responses in a TLR2-dependent manner in mice

    PubMed Central

    Prados-Rosales, Rafael; Baena, Andres; Martinez, Luis R.; Luque-Garcia, Jose; Kalscheuer, Rainer; Veeraraghavan, Usha; Camara, Carmen; Nosanchuk, Joshua D.; Besra, Gurdyal S.; Chen, Bing; Jimenez, Juan; Glatman-Freedman, Aharona; Jacobs, William R.; Porcelli, Steven A.; Casadevall, Arturo

    2011-01-01

    Bacteria naturally release membrane vesicles (MVs) under a variety of growth environments. Their production is associated with virulence due to their capacity to concentrate toxins and immunomodulatory molecules. In this report, we show that the 2 medically important species of mycobacteria, Mycobacterium tuberculosis and Mycobacterium bovis bacille Calmette-Guérin, release MVs when growing in both liquid culture and within murine phagocytic cells in vitro and in vivo. We documented MV production in a variety of virulent and nonvirulent mycobacterial species, indicating that release of MVs is a property conserved among mycobacterial species. Extensive proteomic analysis revealed that only MVs from the virulent strains contained TLR2 lipoprotein agonists. The interaction of MVs with macrophages isolated from mice stimulated the release of cytokines and chemokines in a TLR2-dependent fashion, and infusion of MVs into mouse lungs elicited a florid inflammatory response in WT but not TLR2-deficient mice. When MVs were administered to mice before M. tuberculosis pulmonary infection, an accelerated local inflammatory response with increased bacterial replication was seen in the lungs and spleens. Our results provide strong evidence that actively released mycobacterial vesicles are a delivery mechanism for immunologically active molecules that contribute to mycobacterial virulence. These findings may open up new horizons for understanding the pathogenesis of tuberculosis and developing vaccines. PMID:21364279

  7. THE INDUCED DEVELOPMENT OF NON-ACID-FAST FORMS OF BACILLUS TUBERCULOSIS AND OTHER MYCO-BACTERIA

    PubMed Central

    Miller, Franklin R.

    1932-01-01

    Six strains of mycobacteria,—three human strains, Saranac H-37, T. S., and No. 90, a bovine strain, B-1, a smegma strain, No. 74, and a Saranac strain of B. phlei,—have been made to grow as non-acid-fast organisms by the addition to the culture media of a filtered extract of the chromogenic H-37 strain of B. tuberculosis. The action of the extract produced acceleration of growth of the treated culture, followed by macroscopic and microscopic changes, and differentiation into non-acid-fast forms. The bacterial forms grown from these treated cultures were pleomorphic, usually consisting of cocci and small rods; but branching forms and spore-like bodies also developed. The sterility of the extract causing the changes was demonstrated by frequent control inoculations on various media, including Kendall's K medium; and autoclaved extracts had the same effects as non-autoclaved. After transfer to media suitable for acid growths four of the strains reverted not only to acid-fastness but to their original cultural characteristics, providing evidence that the non-acid-fast forms were specific for the strain. PMID:19870075

  8. A Flow Cytometry Method for Rapidly Assessing Mycobacterium tuberculosis Responses to Antibiotics with Different Modes of Action

    PubMed Central

    Hendon-Dunn, Charlotte Louise; Doris, Kathryn Sarah; Thomas, Stephen Richard; Allnutt, Jonathan Charles; Marriott, Alice Ann Neville; Hatch, Kim Alexandra; Watson, Robert James; Bottley, Graham; Marsh, Philip David; Taylor, Stephen Charles

    2016-01-01

    Current methods for assessing the drug susceptibility of Mycobacterium tuberculosis are lengthy and do not capture information about viable organisms that are not immediately culturable under standard laboratory conditions as a result of antibiotic exposure. We have developed a rapid dual-fluorescence flow cytometry method using markers for cell viability and death. We show that the fluorescent marker calcein violet with an acetoxy-methyl ester group (CV-AM) can differentiate between populations of M. tuberculosis growing at different rates, while Sytox green (SG) can differentiate between live and dead mycobacteria. M. tuberculosis was exposed to isoniazid or rifampin at different concentrations over time and either dual stained with CV-AM and SG and analyzed by flow cytometry or plated to determine the viability of the cells. Although similar trends in the loss of viability were observed when the results of flow cytometry and the plate counting methods were compared, there was a lack of correlation between these two approaches, as the flow cytometry analysis potentially captured information about cell populations that were unable to grow under standard conditions. The flow cytometry approach had an additional advantage in that it could provide insights into the mode of action of the drug: antibiotics targeting the cell wall gave a flow cytometry profile distinct from those inhibiting intracellular processes. This rapid drug susceptibility testing method could identify more effective antimycobacterials, provide information about their potential mode of action, and accelerate their progress to the clinic. PMID:26902767

  9. A Flow Cytometry Method for Rapidly Assessing Mycobacterium tuberculosis Responses to Antibiotics with Different Modes of Action.

    PubMed

    Hendon-Dunn, Charlotte Louise; Doris, Kathryn Sarah; Thomas, Stephen Richard; Allnutt, Jonathan Charles; Marriott, Alice Ann Neville; Hatch, Kim Alexandra; Watson, Robert James; Bottley, Graham; Marsh, Philip David; Taylor, Stephen Charles; Bacon, Joanna

    2016-07-01

    Current methods for assessing the drug susceptibility of Mycobacterium tuberculosis are lengthy and do not capture information about viable organisms that are not immediately culturable under standard laboratory conditions as a result of antibiotic exposure. We have developed a rapid dual-fluorescence flow cytometry method using markers for cell viability and death. We show that the fluorescent marker calcein violet with an acetoxy-methyl ester group (CV-AM) can differentiate between populations of M. tuberculosis growing at different rates, while Sytox green (SG) can differentiate between live and dead mycobacteria. M. tuberculosis was exposed to isoniazid or rifampin at different concentrations over time and either dual stained with CV-AM and SG and analyzed by flow cytometry or plated to determine the viability of the cells. Although similar trends in the loss of viability were observed when the results of flow cytometry and the plate counting methods were compared, there was a lack of correlation between these two approaches, as the flow cytometry analysis potentially captured information about cell populations that were unable to grow under standard conditions. The flow cytometry approach had an additional advantage in that it could provide insights into the mode of action of the drug: antibiotics targeting the cell wall gave a flow cytometry profile distinct from those inhibiting intracellular processes. This rapid drug susceptibility testing method could identify more effective antimycobacterials, provide information about their potential mode of action, and accelerate their progress to the clinic.

  10. Pulmonary infection with hypervirulent Mycobacteria reveals a crucial role for the P2X7 receptor in aggressive forms of tuberculosis.

    PubMed

    Amaral, Eduardo P; Ribeiro, Simone C M; Lanes, Verônica R; Almeida, Fabrício M; de Andrade, Marcelle R M; Bomfim, Caio Cesar Barbosa; Salles, Erika M; Bortoluci, Karina R; Coutinho-Silva, Robson; Hirata, Mario H; Alvarez, José M; Lasunskaia, Elena B; D'Império-Lima, Maria Regina

    2014-07-01

    The purinergic P2X7 receptor (P2X7R) is a sensor of extracellular ATP, a damage-associated molecule that is released from necrotic cells and that induces pro-inflammatory cytokine production and cell death. To investigate whether the innate immune response to damage signals could contribute to the development of pulmonary necrotic lesions in severe forms of tuberculosis, disease progression was examined in C57BL/6 and P2X7R-/- mice that were intratracheally infected with highly virulent mycobacterial strains (Mycobacterium tuberculosis strain 1471 of the Beijing genotype family and Mycobacterium bovis strain MP287/03). The low-dose infection of C57BL/6 mice with bacteria of these strains caused the rapid development of extensive granulomatous pneumonia with necrotic areas, intense bacillus dissemination and anticipated animal death. In contrast, in P2X7R-/- mice, the lung pathology presented with moderate infiltrates of mononuclear leukocytes without visible signs of necrosis; the disease attenuation was accompanied by a delay in mortality. In vitro, the hypervirulent mycobacteria grew rapidly inside macrophages and induced death by a P2X7R-dependent mechanism that facilitated the release of bacilli. Furthermore, these bacteria were resistant to the protective mechanisms elicited in macrophages following extracellular ATP stimulation. Based on this study, we propose that the rapid intracellular growth of hypervirulent mycobacteria results in massive macrophage damage. The ATP released by damaged cells engages P2X7R and accelerates the necrotic death of infected macrophages and the release of bacilli. This vicious cycle exacerbates pneumonia and lung necrosis by promoting widespread cell destruction and bacillus dissemination. These findings suggest the use of drugs that have been designed to inhibit the P2X7R as a new therapeutic approach to treat the aggressive forms of tuberculosis.

  11. Comparative quantitative prevalence of Mycobacteria and functionally abundant nidA, nahAc, and nagAc dioxygenase genes in coal tar contaminated sediments.

    PubMed

    Debruyn, Jennifer M; Chewning, Christopher S; Sayler, Gary S

    2007-08-01

    The Chattanooga Creek Superfund site is heavily contaminated with metals, pesticides, and coal tar with sediments exhibiting high concentrations of polycyclic aromatic hydrocarbons (PAHs). High molecular weight PAHs are of concern because of their toxicity and recalcitrance in the environment; as such, there is great interest in microbes, such as fast-growing Mycobacterium spp., capable of degradation of these compounds. Real-time quantitative PCR assays were developed targeting multiple dioxygenase genes to assess the ecology and functional diversity of PAH-degrading communities. These assays target the Mycobacterium nidA, beta-proteobacteria nagAc, and gamma-proteobacteria nahAc with the specific goal of testing the hypothesis that Mycobacteria catabolic genes are enriched and may be functionally associated with high molecular weight PAH biodegradation in Chattanooga Creek. Dioxygenase gene abundances were quantitatively compared to naphthalene and pyrene mineralization, and temporal and spatial PAH concentrations. nidA abundances ranged from 5.69 x 10(4) to 4.92 x 10(6) copies per gram sediment; nagAc from 2.42 x 10(3) to 1.21 x 10(7), and nahAc from below detection to 4.01 x 10(6) copies per gram sediment. There was a significantly greater abundance of nidA and nagAc at sites with the greatest concentrations of PAHs. In addition, nidA and nagAc were significantly positively correlated (r = 0.76), indicating a coexistence of organisms carrying these genes. A positive relationship was also observed between nidA and nagAc and pyrene mineralization indicating that these genes serve as biomarkers for pyrene degradation. A 16S rDNA clone library of fast-growing Mycobacteria indicated that the population is very diverse and likely plays an important role in attenuation of high molecular weight PAHs from Chattanooga Creek.

  12. Comparative quantitative prevalence of mycobacteria and functionally abundant nidA, nahAc, and nagAc Dioxygenase genes in coal tar contaminated sediments

    SciTech Connect

    Jennifer M. DeBruyn; Christopher S. Chewning; Gary S. Sayler

    2007-08-01

    The Chattanooga Creek Superfund site is heavily contaminated with metals, pesticides, and coal tar with sediments exhibiting high concentrations of polycyclic aromatic hydrocarbons (PAHs). High molecular weight PAHs are of concern because of their toxicity and recalcitrance in the environment; as such, there is great interest in microbes, such as fast-growing Mycobacterium spp., capable of degradation of these compounds. Real-time quantitative PCR assays were developed targeting multiple dioxygenase genes to assess the ecology and functional diversity of PAH-degrading communities. These assays target the Mycobacterium nidA, {beta}-proteobacteria nagAc, and {gamma}-proteobacteria nahAc with the specific goal of testing the hypothesis that Mycobacteria catabolic genes are enriched and may be functionally associated with high molecular weight PAH biodegradation in Chattanooga Creek. Dioxygenase gene abundances were quantitatively compared to naphthalene and pyrene mineralization, and temporal and spatial PAH concentrations. nidA abundances ranged from 5.69 x 10{sup 4} to 4.92 x 10{sup 6} copies per gram sediment; nagAc from 2.42 x 10{sup 3} to 1.21 x 10{sup 7}, and nahAc from below detection to 4.01 x 10{sup 6} copies per gram sediment. There was a significantly greater abundance of nidA and nagAc at sites with the greatest concentrations of PAHs. In addition, nidA and nagAc were significantly positively correlated, indicating a coexistence of organisms carrying these genes. A positive relationship was also observed between nidA and nagAc and pyrene mineralization indicating that these genes serve as biomarkers for pyrene degradation. A 16S rDNA clone library of fast-growing Mycobacteria indicated that the population is very diverse and likely plays an important role in attenuation of high molecular weight PAHs from Chattanooga Creek. 35 refs., 5 figs., 1 tab.

  13. Correlation between virulence of various strains of mycobacteria and their susceptibility to ethanolic extract of propolis (EEP).

    PubMed

    Scheller, S; Kawalski, H; Oklek, K; Dworniczak, S; Matsuno, T; Waldemar-Klimmek, K; Rajca, M; Shani, J

    1998-01-01

    Ethanol extract of propolis (EEP) has antibacterial, antiviral, antiprotozoal and antifungal properties, in addition to many biological effects. Our laboratory has demonstrated a synergistic effect of EEP and antibiotics on the growth of Staphylococcus aureus, and suggested that the bactericidal effect of EEP was expressed mainly on virulent mycobacteria rather than on non-virulent (attenuated) ones. The present study was designed to reconfirm the latter finding, by subjecting 17 different mycobacteria strains to EEP, and evaluating whether there is a correlation between the virulence of the mycobacteria strains studied and their susceptibility to EEP. Our findings demonstrate that while the four non-virulent strains studied are not susceptible to EEP, out of the 13 virulent strains studied seven are susceptible and six are resistant to it. These results suggest that while there is no full correlation between virulence of the mycobacteria tested and their susceptibility to EEP, the few strains that were resistant to EEP were non-virulent.

  14. Use of green fluorescent protein labeled non-tuberculous mycobacteria to evaluate the activity quaternary ammonium compound disinfectants and antibiotics.

    PubMed

    Cortesia, Claudia; Bello, Teresita; Lopez, Gustavo; Franzblau, Scott; de Waard, Jacobus; Takiff, Howard

    Although infections with NonTuberculous Mycobacteria have become less common in AIDS patients, they are important opportunistic infections after surgical procedures, likely because they are ubiquitous and not efficiently killed by many commonly used disinfectants. In Venezuela there have recently been many non-tuberculous mycobacteria soft tissue infections after minor surgical procedures, some apparently related to the use of a commercial disinfectant based on a Quaternary Ammonium Compound. We studied the activity of this and other quaternary ammonium compounds on different non-tuberculous mycobacteria by transforming the mycobacteria with a dnaA-gfp fusion and then monitoring fluorescence to gauge the capacity of different quaternary ammonium compounds to inhibit bacterial growth. The minimum inhibitory concentration varied for the different quaternary ammonium compounds, but M. chelonae and M. abscessus were consistently more resistant than M. smegmatis, and M. terrae more resistant than M. bovis BCG. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  15. Counting mycobacteria in infected human cells and mouse tissue: a comparison between qPCR and CFU.

    PubMed

    Pathak, Sharad; Awuh, Jane A; Leversen, Nils Anders; Flo, Trude H; Asjø, Birgitta

    2012-01-01

    Due to the slow growth rate and pathogenicity of mycobacteria, enumeration by traditional reference methods like colony counting is notoriously time-consuming, inconvenient and biohazardous. Thus, novel methods that rapidly and reliably quantify mycobacteria are warranted in experimental models to facilitate basic research, development of vaccines and anti-mycobacterial drugs. In this study we have developed quantitative polymerase chain reaction (qPCR) assays for simultaneous quantification of mycobacterial and host DNA in infected human macrophage cultures and in mouse tissues. The qPCR method cannot discriminate live from dead bacteria and found a 10- to 100-fold excess of mycobacterial genomes, relative to colony formation. However, good linear correlations were observed between viable colony counts and qPCR results from infected macrophage cultures (Pearson correlation coefficient [r] for M. tuberculosis = 0.82; M. a. avium = 0.95; M. a. paratuberculosis = 0.91). Regression models that predict colony counts from qPCR data in infected macrophages were validated empirically and showed a high degree of agreement with observed counts. Similar correlation results were also obtained in liver and spleen homogenates of M. a. avium infected mice, although the correlations were distinct for the early phase (< day 9 post-infection) and later phase (≥ day 20 post-infection) liver r = 0.94 and r = 0.91; spleen r = 0.91 and r = 0.87, respectively. Interestingly, in the mouse model the number of live bacteria as determined by colony counts constituted a much higher proportion of the total genomic qPCR count in the early phase (geometric mean ratio of 0.37 and 0.34 in spleen and liver, respectively), as compared to later phase of infection (geometric mean ratio of 0.01 in both spleen and liver). Overall, qPCR methods offer advantages in biosafety, time-saving, assay range and reproducibility compared to colony counting. Additionally, the duplex format allows enumeration of

  16. 16S rDNA-based probes for two polycyclic aromatic hydrocarbon (PAH)-degrading soil Mycobacteria

    SciTech Connect

    Govindaswami, M.; Feldhake, D.J.; Loper, J.C.

    1994-12-31

    PAHs are a class of widespread pollutants, some of which have been shown to be genotoxic, hence the fate of these compounds in the environment is of considerable interest. Research on the biodegradation of 4 and 5 ring PAHs has been limited by the general lack of microbial isolates or consortia which can completely degrade these toxicants. Heitkamp and Cerniglia have described an oxidative soil Mycobacterium-strain PYR-1 that metabolizes pyrene and fluoranthene more rapidly than the 2 and 3 ring naphthalene and phenanthrene; although some metabolites of benzo-(a)-pyrene (BaP) were detected, no mineralization of BaP was observed. In 1991 Grosser et al. reported the isolation of a Mycobacterium sp. which mineralizes pyrene and also causing some mineralization of BaP. Their study describes a comparative analysis of these two strains, which show very similar colony morphology, growth rate and yellow-orange pigmentation. Genetic differences were shown by DNA amplification fingerprinting (DAF) using two arbitrary GC-rich octanucleotide primers, and by sequence comparison of PCR amplified 16S rDNA, although both strains show similarity closest to that of the genus Mycobacteria. These 16S rDNA sequences are in use for the construction of strain-specific DNA probes to monitor the presence, survival and growth of these isolates in PAH-contaminated soils in studies of biodegradation.

  17. Prevalence and concentration of non-tuberculous mycobacteria in cooling towers by means of quantitative PCR: a prospective study.

    PubMed

    Adrados, Bárbara; Julián, Esther; Codony, Francesc; Torrents, Eduard; Luquin, Marina; Morató, Jordi

    2011-01-01

    There is an increasing level of interest in non-tuberculous mycobacteria (NTM) due to the increasing reported rates of diseases caused by them. Although it is well known that NTM are widely distributed in the environment it is necessary to identify its reservoirs to prevent possible infections. In this study, we aimed to investigate the occurrence and levels of NTM in cooling towers to provide evidences for considering these settings as possible sources of respiratory infections. In the current study, we detected and quantified the presence of NTM by means of a rapid method in water samples taken from 53 cooling towers of an urban area (Barcelona, Spain). A genus-specific quantitative PCR (Q-PCR) assay with a quantification limit (QL) of 500 cells l(-1) was used. 56% (30) of samples were positive with a concentration range from 4.6 × 10(3) to 1.79 × 10(6) cells l(-1). In some cases (9/30), samples were positive but with levels below the QL. The colonization rate confirmed that cooling towers could be considered as a potential reservoir for NTM. This study also evaluated Q-PCR as a useful method to detect and quantify NTM in samples coming from environmental sources.

  18. Rapid radiometric methods to detect and differentiate Mycobacterium tuberculosis/M. bovis from other mycobacterial species

    SciTech Connect

    Siddiqi, S.H.; Hwangbo, C.C.; Silcox, V.; Good, R.C.; Snider, D.E. Jr.; Middlebrook, G.

    1984-10-01

    Rapid methods for the differentiation of Mycobacterium tuberculosis/M. bovis (TB complex) from other mycobacteria (MOTT bacilli) were developed and evaluated in a three-phase study. In the first phase, techniques for identification of Mycobacterium species were developed by using radiometric technology and BACTEC Middlebrook 7H12 liquid medium. Based on /sup 14/CO/sub 2/ evolution, characteristic growth patterns were established for 13 commonly encountered mycobacterial species. Mycobacteria belonging to the TB complex were differentiated from other mycobacteria by cellular morphology and rate of /sup 14/CO/sub 2/ evolution. For further differentiation, radiometric tests for niacin production and inhibition by Q-nitro-alpha-acetyl amino-beta-hydroxy-propiophenone (NAP) were developed. In the second phase, 100 coded specimens on Lowenstein-Jensen medium were identified as members of the TB complex, MOTT bacilli, bacteria other than mycobacteria, or ''no viable organisms'' within 3 to 12 (average 6.4) days of receipt from the Centers for Disease Control. Isolation and identification of mycobacteria from 20 simulated sputum specimens were carried out in phase III. Out of 20 sputum specimens, 16 contained culturable mycobacteria, and all of the positives were detected by the BACTEC method in an average of 7.3 days. The positive mycobacterial cultures were isolated and identified as TB complex or MOTT bacilli in an average of 12.8 days. The radiometric NAP test was found to be highly sensitive and specific for a rapid identification of TB complex, whereas the radiometric niacin test was found to have some inherent problems. Radiometric BACTEC and conventional methodologies were in complete agreement in Phase II as well as in Phase III.

  19. Mycobacteria emulsified in olive oil-in-water trigger a robust immune response in bladder cancer treatment

    PubMed Central

    Noguera-Ortega, Estela; Blanco-Cabra, Núria; Rabanal, Rosa Maria; Sánchez-Chardi, Alejandro; Roldán, Mónica; Guallar-Garrido, Sandra; Torrents, Eduard; Luquin, Marina; Julián, Esther

    2016-01-01

    The hydrophobic composition of mycobacterial cell walls leads to the formation of clumps when attempting to resuspend mycobacteria in aqueous solutions. Such aggregation may interfere in the mycobacteria-host cells interaction and, consequently, influence their antitumor effect. To improve the immunotherapeutic activity of Mycobacterium brumae, we designed different emulsions and demonstrated their efficacy. The best formulation was initially selected based on homogeneity and stability. Both olive oil (OO)- and mineral oil-in-water emulsions better preserved the mycobacteria viability and provided higher disaggregation rates compared to the others. But, among both emulsions, the OO emulsion increased the mycobacteria capacity to induce cytokines’ production in bladder tumor cell cultures. The OO-mycobacteria emulsion properties: less hydrophobic, lower pH, more neutralized zeta potential, and increased affinity to fibronectin than non-emulsified mycobacteria, indicated favorable conditions for reaching the bladder epithelium in vivo. Finally, intravesical OO-M. brumae-treated mice showed a significantly higher systemic immune response, together with a trend toward increased tumor-bearing mouse survival rates compared to the rest of the treated mice. The physicochemical characteristics and the induction of a robust immune response in vitro and in vivo highlight the potential of the OO emulsion as a good delivery vehicle for the mycobacterial treatment of bladder cancer. PMID:27265565

  20. Mycobacteria emulsified in olive oil-in-water trigger a robust immune response in bladder cancer treatment.

    PubMed

    Noguera-Ortega, Estela; Blanco-Cabra, Núria; Rabanal, Rosa Maria; Sánchez-Chardi, Alejandro; Roldán, Mónica; Guallar-Garrido, Sandra; Torrents, Eduard; Luquin, Marina; Julián, Esther

    2016-06-06

    The hydrophobic composition of mycobacterial cell walls leads to the formation of clumps when attempting to resuspend mycobacteria in aqueous solutions. Such aggregation may interfere in the mycobacteria-host cells interaction and, consequently, influence their antitumor effect. To improve the immunotherapeutic activity of Mycobacterium brumae, we designed different emulsions and demonstrated their efficacy. The best formulation was initially selected based on homogeneity and stability. Both olive oil (OO)- and mineral oil-in-water emulsions better preserved the mycobacteria viability and provided higher disaggregation rates compared to the others. But, among both emulsions, the OO emulsion increased the mycobacteria capacity to induce cytokines' production in bladder tumor cell cultures. The OO-mycobacteria emulsion properties: less hydrophobic, lower pH, more neutralized zeta potential, and increased affinity to fibronectin than non-emulsified mycobacteria, indicated favorable conditions for reaching the bladder epithelium in vivo. Finally, intravesical OO-M. brumae-treated mice showed a significantly higher systemic immune response, together with a trend toward increased tumor-bearing mouse survival rates compared to the rest of the treated mice. The physicochemical characteristics and the induction of a robust immune response in vitro and in vivo highlight the potential of the OO emulsion as a good delivery vehicle for the mycobacterial treatment of bladder cancer.

  1. Evaluation of an immunohistochemical test with polyclonal antibodies raised against mycobacteria used in formalin-fixed tissue compared with mycobacterial specific culture.

    PubMed

    Carabias, E; Palenque, E; Serrano, R; Aguado, J M; Ballestín, C

    1998-03-01

    An immunohistochemical (IH) test (commercially available polyclonal antiserum rabbit anti-Myco-bacterium bovis; DAKO A/S) was used to detect the presence of mycobacteria in 65 formalin-fixed, paraffin-embedded tissue blocks from different organs, showing necrotizing caseous granuloma lesions on hematoxylin and eosin sections from 65 patients. These 65 samples were dyed using an acid-fast fluorescent technique and compared using the immunohistochemical method. Both results were also compared with the mycobacterial cultures. The IH test, compared with the culture, showed a sensitivity (S) of 52%, a specificity (Sp) of 76%, a positive predictive value (PV pos) of 61% and a negative predictive value (PV neg) of 69%. We analyze these data and discuss the possible causes of false-positive and -negative results of the IH test. This rapid test on paraffin embedded tissue seems valuable in the period when waiting for the culture results.

  2. Conserved Pro-Glu (PE) and Pro-Pro-Glu (PPE) protein domains target LipY lipases of pathogenic mycobacteria to the cell surface via the ESX-5 pathway.

    PubMed

    Daleke, Maria H; Cascioferro, Alessandro; de Punder, Karin; Ummels, Roy; Abdallah, Abdallah M; van der Wel, Nicole; Peters, Peter J; Luirink, Joen; Manganelli, Riccardo; Bitter, Wilbert

    2011-05-27

    The type VII secretion system ESX-5 is a major pathway for export of PE and PPE proteins in pathogenic mycobacteria. These mycobacteria-specific protein families are characterized by conserved N-terminal domains of 100 and 180 amino acids, which contain the proline-glutamic acid (PE) and proline-proline-glutamic acid (PPE) motifs after which they are named. Here we investigated secretion of the triacylglycerol lipase LipY, which in fast-growing mycobacteria contains a signal sequence, but in slow-growing species appears to have replaced the signal peptide with a PE or PPE domain. Selected LipY homologues were expressed in wild-type Mycobacterium marinum and its corresponding ESX-5 mutant, and localization of the proteins was investigated by immunoblotting and electron microscopy. Our study shows that Mycobacterium tuberculosis PE-LipY (LipY(tub)) and M. marinum PPE-LipY (LipY(mar)) are both secreted to the bacterial surface in an ESX-5-dependent fashion. After transport, the PE/PPE domains are removed by proteolytic cleavage. In contrast, Mycobacterium gilvum LipY, which has a signal sequence, is not transported to the cell surface. Furthermore, we show that LipY(tub) and LipY(mar) require their respective PE and PPE domains for ESX-5-dependent secretion. The role of the PE domain in ESX-5 secretion was confirmed in a whole cell lipase assay, in which wild-type bacteria expressing full-length LipY(tub), but not LipY(tub) lacking its PE domain, were shown to hydrolyze extracellular lipids. In conclusion, both PE and PPE domains contain a signal required for secretion of LipY by the ESX-5 system, and these domains are proteolytically removed upon translocation.

  3. Th1 – Th2 polarisation and autophagy in the control of intracellular mycobacteria by macrophages

    PubMed Central

    Harris, James; Master, Sharon S.; De Haro, Sergio A.; Delgado, Monica; Roberts, Esteban A.; Hope, Jayne C.; Keane, Joseph; Deretic, Vojo

    2009-01-01

    Autophagy is a major intracellular pathway for the lysosomal degradation of long-lived cytoplasmic macromolecules and damaged or surplus organelles. More recently, autophagy has also been linked with innate and adaptive immune responses against intracellular pathogens, including Mycobacterium tuberculosis, which can survive within macrophages by blocking fusion of the phagosome with lysosomes. Induction of autophagy by the Th1 cytokine IFN-γ enables infected macrophages to overcome this phagosome maturation block and inhibit the intracellular survival of mycobacteria. Conversely, theTh2 cytokines IL-4 and IL-13 inhibit autophagy in murine and human macrophages. We discuss how differential modulation of autophagy by Th1 and Th2 cytokines may represent an important feature of the host response to mycobacteria. PMID:19026454

  4. Non-tuberculous mycobacteria and microbial populations in drinking water distribution systems.

    PubMed

    Briancesco, Rossella; Semproni, Maurizio; Della Libera, Simonetta; Sdanganelli, Massimo; Bonadonna, Lucia

    2010-01-01

    Data on the occurrence of non-tuberculous mycobacteria (NTM), in parallel with those obtained for bacterial indicators and amoebae, are presented with the aim to collect information on the spread of NTM in drinking water distribution systems in Italy. Samples were collected from taps of hospitals and households in Central and Southern Italy. The concentration values obtained for the more traditional microbial parameters complied with the mandatory requirements for drinking water. Conversely, moderate-to-high microbial loads (till 300 CFU/L) were observed for the NTM. Positive samples were obtained from 62% of the investigated water samples. Analogous results were observed for amoebae showing a higher percentage of positive samples (76%). In terms of public health, the presence of mycobacteria in water distribution systems may represent a potential risk especially for vulnerable people such as children, the elderly or immunocompromised individuals.

  5. Current efforts and future prospects in the development of live mycobacteria as vaccines.

    PubMed

    Ng, Tony W; Saavedra-Ávila, Noemí A; Kennedy, Steven C; Carreño, Leandro J; Porcelli, Steven A

    2015-01-01

    The development of more effective vaccines against Mycobacterium tuberculosis (Mtb) remains a major goal in the effort to reduce the enormous global burden of disease caused by this pathogen. Whole-cell vaccines based on live mycobacteria with attenuated virulence represent an appealing approach, providing broad antigen exposure and intrinsic adjuvant properties to prime durable immune responses. However, designing vaccine strains with an optimal balance between attenuation and immunogenicity has proven to be extremely challenging. Recent basic and clinical research efforts have broadened our understanding of Mtb pathogenesis and created numerous new vaccine candidates that have been designed to overcome different aspects of immune evasion by Mtb. In this review, we provide an overview of the current efforts to create improved vaccines against tuberculosis based on modifications of live attenuated mycobacteria. In addition, we discuss the use of such vaccine strains as vectors for stimulating protective immunity against other infectious diseases and cancers.

  6. Current Efforts and Future Prospects in the Development of Live Mycobacteria as Vaccines

    PubMed Central

    Porcelli, Steven A.; Ng, Tony W.; Saavedra-Avila, Noemi A; Kennedy, Steven C.; Carreno, Leandro J.

    2016-01-01

    Summary The development of more effective vaccines against Mycobacterium tuberculosis (Mtb) remains a major goal in the effort to reduce the enormous global burden of disease caused by this pathogen. Whole-cell vaccines based on live mycobacteria with attenuated virulence represent an appealing approach, providing broad antigen exposure and intrinsic adjuvant properties to prime durable immune responses. However, designing vaccine strains with an optimal balance between attenuation and immunogenicity has proven to be extremely challenging. Recent basic and clinical research efforts have broadened our understanding of Mtb pathogenesis and created numerous new vaccine candidates that are designed to overcome different aspects of immune evasion by Mtb. In this review, we provide an overview of current efforts to create improved vaccines against tuberculosis based on modifications of live attenuated mycobacteria. In addition, we discuss the use of such vaccine strains as vectors for stimulating protective immunity against other infectious diseases and cancers. PMID:26366616

  7. Commercial DNA Probes for Mycobacteria Incorrectly Identify a Number of Less Frequently Encountered Species▿ †

    PubMed Central

    Tortoli, Enrico; Pecorari, Monica; Fabio, Giuliana; Messinò, Massimino; Fabio, Anna

    2010-01-01

    Although commercially available DNA probes for identification of mycobacteria have been investigated with large numbers of strains, nothing is known about the ability of these probes to identify less frequently encountered species. We analyzed, with INNO LiPA MYCOBACTERIA (Innogenetics) and with GenoType Mycobacterium (Hein), 317 strains, belonging to 136 species, 61 of which had never been assayed before. INNO LiPA misidentified 20 taxa, the majority of which cross-reacted with the probes specific for Mycobacterium fortuitum and the Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum group. GenoType misidentified 28 taxa, most of which cross-reacted with M. intracellulare and M. fortuitum probes; furthermore, eight species were not recognized as members of the genus Mycobacterium. Among 54 strains investigated with AccuProbe (Gen-Probe), cross-reactions were detected for nine species, with the probes aiming at the M. avium complex being most involved in cross-reactions. PMID:19906898

  8. Performance Assessment of New Multiplex Probe Assay for Identification of Mycobacteria

    PubMed Central

    Tortoli, Enrico; Nanetti, Anna; Piersimoni, Claudio; Cichero, Paola; Farina, Claudio; Mucignat, Giorgio; Scarparo, Claudio; Bartolini, Laura; Valentini, Roberta; Nista, Domenico; Gesu, Giampietro; Tosi, Cristiana Passerini; Crovatto, Marina; Brusarosco, Giuliana

    2001-01-01

    A new DNA probe assay (INNO LiPA Mycobacteria; Innogenetics, Ghent, Belgium) for the simultaneous identification, by means of reverse hybridization and line-probe technology, of Mycobacterium tuberculosis complex, Mycobacterium kansasii, Mycobacterium xenopi, Mycobacterium gordonae, the species of the Mycobacterium avium complex (MAC), Mycobacterium scrofulaceum, and Mycobacterium chelonae was evaluated on a panel of 238 strains including, besides representatives of all the taxa identifiable by the system, a number of other mycobacteria, some of which are known to be problematic with the only other commercial DNA probe system (AccuProbe; Gen-Probe, San Diego, Calif.), and two nocardiae. The new kit, which includes a control probe reacting with the whole genus Mycobacterium, correctly identified 99.6% of the strains tested; the one discrepancy, which remained unresolved, concerned an isolate identified as MAC intermediate by INNO LiPA Mycobacteria and as Mycobacterium intracellulare by AccuProbe. In five cases, because of an imperfect checking of hybridization temperature, a very slight, nonspecific, line was visible which was no longer evident when the test was repeated. Two strains whose DNA failed amplification at the first attempt were regularly identified when the test was repeated. Interestingly, the novel kit dodged all the pitfalls presented by the strains giving anomalous reactions with AccuProbe. A unique feature of INNO LiPA Mycobacteria is its ability to recognize different subgroups within the species M. kansasii and M. chelonae, while the declared overlapping reactivity of probe 4 with some M. kansasii and Mycobacterium gastri organisms and of probe 9 with MAC, Mycobacterium haemophilum, and Mycobacterium malmoense, may furnish a useful aid for their identification. The turnaround time of the method is approximately 6 h, including a preliminary PCR amplification. PMID:11230430

  9. Carbohydrate-Conjugated Hollow Oblate Mesoporous Silica Nanoparticles as Nanoantibiotics to Target Mycobacteria.

    PubMed

    Hao, Nanjing; Chen, Xuan; Jeon, Seaho; Yan, Mingdi

    2015-12-30

    Engineering nanomaterials with enhanced antibacterial activities remains a critical and practical challenge. Hollow oblate mesoporous silica nanoparticles (HOMSNs) are synthesized by a simple protocol of ammonia hydrothermal treatment of oblate mesoporous silica nanoparticles prepared using dibenzyl ether as a cosolvent. When conjugated with trehalose as the targeting ligand, the antibiotic-encapsulated HOMSNs exhibit high binding affinity and antibacterial efficacy toward mycobacteria. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Carbohydrate-Conjugated Hollow Oblate Mesoporous Silica Nanoparticles as Nanoantibiotics to Target Mycobacteria

    PubMed Central

    Hao, Nanjing; Chen, Xuan; Jeon, Seaho

    2015-01-01

    Engineering nanomaterials with enhanced antibacterial activities remains a critical and practical challenge. Hollow oblate mesoporous silica nanoparticles (HOMSNs) are synthesized by a simple protocol of ammonia hydrothermal treatment of oblate mesoporous silica nanoparticles prepared using dibenzyl ether as a co-solvent. When conjugate with trehalose as the targeting ligand, the antibiotic-encapsulated HOMSNs exhibit high binding affinity and antibacterial efficacy towards mycobacteria. PMID:26450697

  11. Aldehyde-Resistant Mycobacteria Associated with the Use of Endoscope Reprocessing Systems

    PubMed Central

    Fisher, Christopher W.; Fiorello, Anthony; Shaffer, Diana; Jackson, Mary

    2012-01-01

    Bacteria can develop resistance to antibiotics, but less is known about their ability to increase resistance to chemical disinfectants. This study randomly sampled three AERs in the USA using aldehydes for endoscope disinfection. Bacterial contamination was found post-disinfection in all AERs and some mycobacteria isolated demonstrated significant resistance to glutaraldehyde and OPA disinfectants. Bacteria can survive aldehyde-based disinfection and may pose a cross-contamination risk to patients. PMID:22325730

  12. Synergism between ethanolic extract of propolis (EEP) and anti-tuberculosis drugs on growth of mycobacteria.

    PubMed

    Scheller, S; Dworniczak, S; Waldemar-Klimmek, K; Rajca, M; Tomczyk, A; Shani, J

    1999-01-01

    Ethanolic extract of propolis exerts a strong anti-bacterial activity, in addition to antifungal, antiviral and antiprotozoal properties. In previous studies from these laboratories we have demonstrated that the intensity of the bactericidal activity of EEP is correlated with the virulence of the mycobacteria tested, and that EEP has a synergistic effect with antibiotics on growth of staphylococcus aureus. In the present study we investigated whether the same synergism and correlation exists between EEP and some anti-tuberculosis drugs on tuberculosis mycobacteria with different degrees of virulence. Six standard strains and 11 wild strains of mycobacteria were exposed for 30 days to EEP, with or without streptomycin, rifamycin, isoniazid or ethambutol. Out of the 17 strains, 8 were resistant to at least two standard antibiotics, and were considered "multi-resistant strains". The rest were either susceptible or resistant to only one of the antimycobacterial drugs. Antagonism was recorded only in one case, when Staphylococcus aureus were treated with a mixture of EEP and ethambutol, suggesting that a chemical bond could have been formed between this anti-tuberculosis antibiotic and one of the active components of the ethanol extract of propolis.

  13. Comparison of filtering methods, filter processing and DNA extraction kits for detection of mycobacteria in water.

    PubMed

    Kaevska, Marija; Slana, Iva

    2015-01-01

    Mycobacteria have been isolated from almost all types of natural waters, as well as from man-made water distribution systems. Detection of mycobacteria using PCR has been described in different types of water; however, currently, there is no standardised protocol for the processing of large volumes of water. In the present study, different filtering methods are tested and optimised for tap or river water filtration up to 10 L, as well as filter processing and DNA isolation using four commercially available kits. The PowerWater DNA isolation kit (MoBio, USA), together with a kit used for soil and other environmental samples (PowerSoil DNA isolation kit, MoBio), had the highest efficiency. Filtration of 10 L of water and elution of the filter in PBS with the addition of 0.05% of Tween 80 is suggested. The described protocol for filter elution is recommended, and the use of the PowerWater DNA isolation kit for the highest mycobacterial DNA yield from water samples. The described protocol is suitable for parallel detection of mycobacteria using cultivation.

  14. Detection of mycobacteria in aquarium fish in Slovenia by culture and molecular methods.

    PubMed

    Pate, M; Jencic, V; Zolnir-Dovc, M; Ocepek, M

    2005-04-06

    Thirty-five aquarium fish were investigated for the presence of mycobacteria by culture and molecular methods. The following species were examined: goldfish Carassius auratus auratus, guppy Poecilia reticulata, 4 three-spot gourami Trichogaster trichopterus, dwarf gourami Colisa lalia, Siamese fighting fish Betta splendens, freshwater angelfish Pterophyllum scalare, African cichlid fish Cichlidae spp., cichlid fish Microgeophagus altispinosus, cichlid fish Pseudotropheus lombardoi, blue streak hap Labidochromis caeruleus, sterlet Acipenser ruthenus, southern platyfish Xiphophorus maculatus, and catfish Corydoras spp. Isolates of mycobacteria were obtained in 29 cases (82.9%). Two specimens were positive using Ziehl-Neelsen (ZN) staining, but the cultivation failed. Four specimens were both ZN- and culture-negative. On the basis of GenoType Mycobacterium assay (Hain Life-science) and restriction enzyme analysis of the amplified products (PCR-RFLP), 23 isolates (79.3%) were identified: 7 as Mycobacterium fortuitum, 6 as M. gordonae, 6 as M. marinum, 3 as M. chelonae, and 1 as M. peregrinum. Five isolates remained unidentified (Mycobacterium spp.). One case probably represented a mixed infection (M. marinum/M. fortuitum). Since M. marinum infections are also detected in humans, the significance of mycobacteria in aquarium fish should not be overlooked.

  15. Distribution and respiratory activity of mycobacteria in household water system of healthy volunteers in Japan.

    PubMed

    Ichijo, Tomoaki; Izumi, Yoko; Nakamoto, Sayuri; Yamaguchi, Nobuyasu; Nasu, Masao

    2014-01-01

    The primary infectious source of nontuberculous mycobacteria (NTM), which are known as opportunistic pathogens, appears to be environmental exposure, and it is important to reduce the frequency of exposure from environmental sources for preventing NTM infections. In order to achieve this, the distribution and respiratory activity of NTM in the environments must be clarified. In this study, we determined the abundance of mycobacteria and respiratory active mycobacteria in the household water system of healthy volunteers using quantitative PCR and a fluorescent staining method, because household water has been considered as one of the possible infectious sources. We chose healthy volunteer households in order to lessen the effect of possible residential contamination from an infected patient. We evaluated whether each sampling site (bathroom drain, kitchen drain, bath heater pipe and showerhead) have the potential to be the sources of NTM infections. Our results indicated that drains in the bathroom and kitchen sink are the niche for Mycobacterium spp. and M. avium cells were only detected in the bathtub inlet. Both physicochemical and biologic selective pressures may affect the preferred habitat of Mycobacterium spp. Regional differences also appear to exist as demonstrated by the presence (US) or absence (Japan) of Mycobacterium spp. on showerheads. Understanding of the country specific human activities and water usage will help to elucidate the infectious source and route of nontuberculous mycobacterial disease.

  16. Evaluation of hemostatic field dressing for bacteria, mycobacteria, or fungus contamination.

    PubMed

    Murray, Clinton K; Brunstetter, Tyson; Beckius, Miriam; Dunne, James R; Mende, Katrin

    2013-03-01

    Infectious complications have a major impact on wounded warriors. Pathogens causing infections include multidrug-resistant bacteria, fungi, and mycobacteria. The potential sources for these pathogens include nosocomial transmission, the environment (e.g., dirt), or the patients (skin flora) themselves. The purpose of this pilot study was to explore the possibility that hemostatic field dressings might act as an inoculation source of pathogens into wounds. To accomplish this, hemostatic field dressings were assessed for the presence of bacterial, fungal, or mycobacterial contamination. We evaluated two samples of QuikClot Combat Gauze and two samples of CELOX Gauze subjected to normal stresses associated with storage after receipt from the manufacturer. We then evaluated 16 samples of QuikClot Combat Gauze that were collected from personnel deployed in Afghanistan and had undergone routine mechanical stress. Samples underwent screening with Trypticase Soy Broth, blood agar plates, MacConkey agar plates, CHROMagar Staphylococcus aureus plates, chocolate agar plates, Potato Flake agar, Lowenstein-Jensen media, and Middlebrook 7H11 media. No bacteria, fungi, or mycobacteria were recovered from the dressings. It does not appear that hemostatic field dressings are contaminated, even after subjected to field conditions. Further research is needed to identify inoculation sources of fungi and mycobacteria, which cause infections.

  17. Targeting drug tolerance in mycobacteria: a perspective from mycobacterial biofilms

    PubMed Central

    Islam, Mohammad S; Richards, Jacob P; Ojha, Anil K

    2013-01-01

    Multidrug chemotherapy for 6–9-months is one of the primary treatments in effective control of tuberculosis, although the mechanisms underlying the persistence of its etiological agent, Mycobacterium tuberculosis, against antibiotics remain unclear. Ever-mounting evidence indicates that the survival of many environmental and pathogenic microbial species against antibiotics is influenced by their ability to grow as surface-associated multicellular communities called biofilms. In recent years, several mycobacterial species, including M. tuberculosis, have been found to form drug-tolerant biofilms in vitro through genetically controlled mechanisms. In this review, the authors discuss the relevance of the in vitro mycobacterial biofilms in understanding the antibiotic recalcitrance of tuberculosis infections. PMID:23106280

  18. Isolation of non-tuberculous mycobacteria from pastoral ecosystems of Uganda: public health significance.

    PubMed

    Kankya, Clovice; Muwonge, Adrian; Djønne, Berit; Munyeme, Musso; Opuda-Asibo, John; Skjerve, Eystein; Oloya, James; Edvardsen, Vigdis; Johansen, Tone B

    2011-05-16

    The importance of non-tuberculous mycobacteria (NTM) infections in humans and animals in sub-Saharan Africa at the human-environment-livestock-wildlife interface has recently received increased attention. NTM are environmental opportunistic pathogens of humans and animals. Recent studies in pastoral ecosystems of Uganda detected NTM in humans with cervical lymphadenitis and cattle with lesions compatible with bovine tuberculosis. However, little is known about the source of these mycobacteria in Uganda. The aim of this study was to isolate and identify NTM in the environment of pastoral communities in Uganda, as well as assess the potential risk factors and the public health significance of NTM in these ecosystems. A total of 310 samples (soil, water and faecal from cattle and pigs) were examined for mycobacteria. Isolates were identified by the INNO-Lipa test and by 16S rDNA sequencing. Additionally, a questionnaire survey involving 231 pastoralists was conducted during sample collection. Data were analysed using descriptive statistics followed by a multivariable logistic regression analysis. Forty-eight isolates of NTM were detected; 25.3% of soil samples, 11.8% of water and 9.1% from animal faecal samples contained mycobacteria. Soils around water sources were the most contaminated with NTM (29.8%). Of these samples, M. fortuitum-peregrinum complex, M. avium complex, M. gordonae, and M. nonchromogenicum were the most frequently detected mycobacteria. Drinking untreated compared to treated water (OR = 33), use of valley dam versus stream water for drinking and other domestic use (OR = 20), sharing of water sources with wild primates compared to antelopes (OR = 4.6), sharing of water sources with domestic animals (OR = 5.3), and close contact with cattle or other domestic animals (OR = 13.8) were the most plausible risk factors for humans to come in contact with NTM in the environment. The study detected a wide range of potentially pathogenic NTM from the

  19. Identification of Nontuberculous Mycobacteria Species Isolated from Water Samples Using Phenotypic and Molecular Methods and Determination of their Antibiotic Resistance Patterns by E- Test Method, in Isfahan, Iran.

    PubMed

    Moghim, Sharareh; Sarikhani, Ensieh; Nasr Esfahani, Bahram; Faghri, Jamshid

    2012-09-01

    Many studies have shown epidemiological links between strains isolated in tap water, and those isolated from patients. Molecular methods linked to PCR are more reliable and faster for identification of non- tuberculous mycobacteria (NTM). In this study molecular methods were used for identification and typing of NTM. Five hundred ml of 85 water samples was passed through 0.45 μm filters. The filters were transferred directly onto 7H10 Middle Brook solid media, containing 15% OADC. PCR for 16S rRNA was done and the PCR product (1500 bp) was sequenced. PRA of the hsp65 gene was investigated to identify the species of isolates. For evaluation of susceptibility of NTM to antimycobacterial agents, E-test method was used. The genus of 26 isolated NTM was confirmed by 16s rRNA sequence based method. Nineteen isolates of Mycobacteria were differentiated using hsp65 genes PRA. The dominant isolates were M. fortuitum (26.7%), M. chelonae like organism (13.3%) and M. mucogenicum (13.3%). Seventy one percent of NTM species were resistant to isoniazid, 64% to rifampin, 57% to ethambutol, 35% to tetracycline, 14 % to azithromycin and 7.1 % to amikacin. The results showed that E-test method is not a proper technique for antimycobacterial assay because some NTM species are slow in growing and have no growth on Muller Hinton agar. Regarding the 16S rRNA sequence analysis, the identification of isolates was restricted to the genus level, because 99% similarity within 16S rRNA of two isolates may or may not determine the same species.

  20. Microbiological Quality and Occurrence of Nontuberculous Mycobacteria in Fresh-Squeezed Orange Juice Samples Purchased from Street Vendors in Mexico City.

    PubMed

    Cerna-Cortes, Jorge F; Cortes-Cueto, Ana Laura; Cano-Gaona, Maria Rosalia; Leon-Montes, Nancy; Helguera-Repetto, Addy C; Rivera-Gutierrez, Sandra; Salas-Rangel, Laura P; Castro-Rosas, Javier; Gonzalez-Y-Merchand, Jorge A

    2016-12-01

    Nontuberculous mycobacteria (NTM) are potentially pathogenic agents commonly found in a natural ecosystem. For this reason, food is considered another source of NTM transmission for humans. The aims of this study were to evaluate the microbiological quality and the occurrence of NTM in fresh-squeezed orange juice samples purchased from street vendors. All 102 samples analyzed were positive for aerobic mesophilic bacteria (AMB), with limits ranging from 1.8 to 6.2 log CFU/ml. A total of 55 (54%), 25 (25%), and 13 (13%) orange juice samples were positive for total coliforms (TC), fecal coliforms (FC), and Escherichia coli , respectively. TC, FC, and E. coli were present with limits ranging from <3 to >1,100 most probable number (MPN)/ml, <3 to 460 MPN/ml, and <3 to 11 MPN/ml, respectively. Six orange juice samples harbored NTM. These NTM were identified by using three molecular markers (hsp65, rrs, and rpoB genes) and corresponded to the fast-growing mycobacteria: Mycobacterium fortuitum (n = 3), Mycobacterium rhodesiae (n = 1), Mycobacterium obuense (n = 1), and a mixture of M. fortuitum and Mycobacterium mucogenicum in an additional sample (n = 1). No correlation was found between the presence NTM in orange juice samples with the presence and concentration of the indicator microorganisms (aerobic mesophilic bacteria, TC, and FC). Overall, these results suggest that fresh-squeezed orange juice might represent a vehicle for NTM transmission in humans. Therefore, prevention of contamination by humans (proper handling and washing of oranges) during juice preparation should be recommended.

  1. Growing Pains (For Parents)

    MedlinePlus

    ... Lessons? Visit KidsHealth in the Classroom What Other Parents Are Reading Your Child's Development (Birth to 3 Years) Feeding Your 1- to 3-Month-Old Feeding Your 4- to 7-Month-Old Feeding Your 8- to 12-Month-Old Feeding Your 1- to 2-Year-Old Growing ... > For Parents > Growing Pains Print A A A What's in ...

  2. Rapid Reagentless Detection of M. tuberculosis H37Ra in Respiratory Effluents

    SciTech Connect

    Adams, K L; Steele, P T; Bogan, M J; Sadler, N M; Martin, S; Martin, A N; Frank, M

    2008-01-29

    Two similar mycobacteria, Mycobacteria tuberculosis H37Ra and Mycobacteria smegmatis are rapidly detected and identified within samples containing a complex background of respiratory effluents using Single Particle Aerosol Mass Spectrometry (SPAMS). M. tuberculosis H37Ra (TBa), an avirulent strain, is used as a surrogate for virulent tuberculosis (TBv); M. smegmatis (MSm) is utilized as a near neighbor confounder for TBa. Bovine lung surfactant and human exhaled breath condensate are used as first-order surrogates for infected human lung expirations from patients with pulmonary tuberculosis. This simulated background sputum is mixed with TBa or MSm and nebulized to produce conglomerate aerosol particles, single particles that contain a bacterium embedded within a background respiratory matrix. Mass spectra of single conglomerate particles exhibit ions associated with both respiratory effluents and mycobacteria. Spectral features distinguishing TBa from MSm in pure and conglomerate particles are shown. SPAMS pattern matching alarm algorithms are able to distinguish TBa containing particles from background matrix and MSm for >50% of the test particles, which is sufficient to enable a high probability of detection and a low false alarm rate if an adequate number of such particles are present. These results indicate the potential usefulness of SPAMS for rapid, reagentless tuberculosis screening.

  3. Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Combined Species Identification and Drug Sensitivity Testing in Mycobacteria.

    PubMed

    Ceyssens, Pieter-Jan; Soetaert, Karine; Timke, Markus; Van den Bossche, An; Sparbier, Katrin; De Cremer, Koen; Kostrzewa, Markus; Hendrickx, Marijke; Mathys, Vanessa

    2017-02-01

    Species identification and drug susceptibility testing (DST) of mycobacteria are important yet complex processes traditionally reserved for reference laboratories. Recent technical improvements in matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has started to facilitate routine mycobacterial identifications in clinical laboratories. In this paper, we investigate the possibility of performing phenotypic MALDI-based DST in mycobacteriology using the recently described MALDI Biotyper antibiotic susceptibility test rapid assay (MBT-ASTRA). We randomly selected 72 clinical Mycobacterium tuberculosis and nontuberculous mycobacterial (NTM) strains, subjected them to MBT-ASTRA methodology, and compared its results to current gold-standard methods. Drug susceptibility was tested for rifampin, isoniazid, linezolid, and ethambutol (M. tuberculosis, n = 39), and clarithromycin and rifabutin (NTM, n = 33). Combined species identification was performed using the Biotyper Mycobacteria Library 4.0. Mycobacterium-specific MBT-ASTRA parameters were derived (calculation window, m/z 5,000 to 13,000, area under the curve [AUC] of >0.015, relative growth [RG] of <0.5; see the text for details). Using these settings, MBT-ASTRA analyses returned 175/177 M. tuberculosis and 65/66 NTM drug resistance profiles which corresponded to standard testing results. Turnaround times were not significantly different in M. tuberculosis testing, but the MBT-ASTRA method delivered on average a week faster than routine DST in NTM. Databases searches returned 90.4% correct species-level identifications, which increased to 98.6% when score thresholds were lowered to 1.65. In conclusion, the MBT-ASTRA technology holds promise to facilitate and fasten mycobacterial DST and to combine it directly with high-confidence species-level identifications. Given the ease of interpretation, its application in NTM typing might be the first in finding its way to current

  4. Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Combined Species Identification and Drug Sensitivity Testing in Mycobacteria

    PubMed Central

    Ceyssens, Pieter-Jan; Soetaert, Karine; Timke, Markus; Van den Bossche, An; Sparbier, Katrin; De Cremer, Koen; Kostrzewa, Markus; Hendrickx, Marijke

    2016-01-01

    ABSTRACT Species identification and drug susceptibility testing (DST) of mycobacteria are important yet complex processes traditionally reserved for reference laboratories. Recent technical improvements in matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has started to facilitate routine mycobacterial identifications in clinical laboratories. In this paper, we investigate the possibility of performing phenotypic MALDI-based DST in mycobacteriology using the recently described MALDI Biotyper antibiotic susceptibility test rapid assay (MBT-ASTRA). We randomly selected 72 clinical Mycobacterium tuberculosis and nontuberculous mycobacterial (NTM) strains, subjected them to MBT-ASTRA methodology, and compared its results to current gold-standard methods. Drug susceptibility was tested for rifampin, isoniazid, linezolid, and ethambutol (M. tuberculosis, n = 39), and clarithromycin and rifabutin (NTM, n = 33). Combined species identification was performed using the Biotyper Mycobacteria Library 4.0. Mycobacterium-specific MBT-ASTRA parameters were derived (calculation window, m/z 5,000 to 13,000, area under the curve [AUC] of >0.015, relative growth [RG] of <0.5; see the text for details). Using these settings, MBT-ASTRA analyses returned 175/177 M. tuberculosis and 65/66 NTM drug resistance profiles which corresponded to standard testing results. Turnaround times were not significantly different in M. tuberculosis testing, but the MBT-ASTRA method delivered on average a week faster than routine DST in NTM. Databases searches returned 90.4% correct species-level identifications, which increased to 98.6% when score thresholds were lowered to 1.65. In conclusion, the MBT-ASTRA technology holds promise to facilitate and fasten mycobacterial DST and to combine it directly with high-confidence species-level identifications. Given the ease of interpretation, its application in NTM typing might be the first in finding its way to

  5. [Evaluation of the performances of AdvanSure TB/NTM real time PCR kit for detection of mycobacteria in respiratory specimens].

    PubMed

    Kim, Young Jin; Park, Mi Young; Kim, Shine Young; Cho, Son A; Hwang, Sang Hyun; Kim, Hyung Hoi; Lee, Eun Yup; Jeong, Joseph; Kim, Kyeong Hee; Chang, Chulhun L

    2008-02-01

    PCR is a widely used method for rapid and accurate diagnosis of mycobacteriosis. The sensitivity and specificity of a real time PCR kit newly developed in Korea were evaluated for detecting mycobacteria in respiratory specimens. One hundred twenty nine Mycobacterium tuberculosis (TB) culture positive respiratory specimens (82 AFB stain positive and 47 stain negative specimens) were used for evaluation of the sensitivity. Nine non-tuberculous mycobacteria (NTM) culture positive specimens were also included. For evaluation of the specificity, 48 AFB stain and culture negative respiratory specimens from patients who were initially not fully excluded from mycobacterial diseases (specificity group 1) were used. Other 51 respiratory specimens from patients who were not suspected of mycobacterial diseases were also included (specificity group 2). Real time PCR was performed by using AdvanSure TB/NTM real time PCR Kit (LG Lifescience, Korea) and SLAN real time PCR detection system (LG Lifescience). The target genes of TB and NTM were IS6110 and rpoB, respectively. Among 129 TB culture positive specimens, 82 of 82 AFB stain positive specimens (100%) and 35 of 47 (74.5%) stain negative specimens revealed real time PCR positivity for TB, resulting in sensitivity of 90.7%. Five of nine NTM culture positive specimens resulted in real time PCR positivity for NTM (55.6%). Forty seven of 48 specimens (97.9%) and all 51 specimens (100%) of the specificity group 1 and 2, respectively, were real time PCR negative for TB and NTM. AdvanSure TB/NTM real time PCR Kit should be useful for detecting TB in respiratory specimens with high sensitivity and specificity.

  6. Numerical taxonomy of mycobactin-dependent mycobacteria, emended description of Mycobacterium avium, and description of Mycobacterium avium subsp. avium subsp. nov., Mycobacterium avium subsp. paratuberculosis subsp. nov., and Mycobacterium avium subsp. silvaticum subsp. nov.

    PubMed

    Thorel, M F; Krichevsky, M; Lévy-Frébault, V V

    1990-07-01

    We performed a numerical taxonomy analysis of 38 Mycobacterium paratuberculosis and related mycobacterial strains, including wood pigeon mycobacteria; this analysis was based on 22 tests, which were selected for their potential discriminative value from a total of 51 tests studied and produced four well-defined clusters. Cluster 1 contained the M. paratuberculosis strains, including two strains isolated from Crohn's disease patients; cluster 2 contained Mycobacterium avium and Mycobacterium intracellulare reference strains; cluster 3 consisted of the wood pigeon mycobacteria; and the only strain in cluster 4 was M. paratuberculosis 316F, which is used for antigen and vaccine production. Strains in cluster 1 were mycobactin dependent even when they were subcultured, whereas strains in cluster 3 were unable to grow on egg medium and their growth was stimulated by pH 5.5. Growth stimulation by pyruvate, resistance to D-cycloserine (50 micrograms/ml), and alkaline phosphatase activity also were characteristics that were useful for discriminating between clusters 1 and 3. The results of previous DNA-DNA hybridization studies have demonstrated that M. avium Chester 1901, M. paratuberculosis Bergey et al. 1923, and the wood pigeon mycobacteria belong to a single genomic species, and we propose that the name of this species should be M. avium. On the basis of the results of previous genomic analyses based on restriction fragment length, the results of polymorphism studies, and DNA patterns determined by field inversion gel electrophoresis as well as the results of our phenotypic study, we propose that the species should be divided into subspecies which correspond to pathogenicity and host range characteristics.(ABSTRACT TRUNCATED AT 250 WORDS)

  7. Carbonaceous Matter in Growing Nanoparticles

    NASA Astrophysics Data System (ADS)

    Johnston, M. V.; Stangl, C. M.; Horan, A. J.

    2015-12-01

    Atmospheric nanoparticles constitute the greatest portion of ambient aerosol loading by number. A major source of atmospheric nanoparticles is new particle formation (NPF), a gas to particle conversion process whereby clusters nucleate from gas phase precursors to form clusters on the order of one or a few nanometers and then grow rapidly to climatically relevant sizes. A substantial fraction of cloud condensation nuclei (CCN) are thought to arise from NPF. In order to better predict the frequency, growth rates, and climatic impacts of NPF, knowledge of the chemical mechanisms by which nucleated nanoparticles grow is needed. The two main contributors to particle growth are (neutralized) sulfate and carbonaceous matter. Particle growth by sulfuric acid condensation is generally well understood, though uncertainty remains about the extent of base neutralization and the relative roles of ammonia and amines. Much less is known about carbonaceous matter, and field measurements suggest that nitrogen-containing species are important. In this presentation, recent work by our group will be described that uses a combination of ambient measurements, laboratory experiments and computational work to study carbonaceous matter in growing nanoparticles. These studies span a range of particle sizes from the initial adsorption of molecules onto a nanometer-size ammonium bisulfate seed cluster to reactions in particles that are large enough to support condensed-phase chemistry.

  8. The Growing Human Population.

    ERIC Educational Resources Information Center

    Keyfitz, Nathan

    1989-01-01

    Discusses the issue of human population. Illustrates the projections of the growing human population in terms of developed and less developed countries. Describes the family planning programs in several countries. Lists three references for further reading. (YP)

  9. Apparatus for growing crystals

    NASA Technical Reports Server (NTRS)

    Jasinski, Thomas J. (Inventor); Witt, August F. (Inventor)

    1986-01-01

    An improved apparatus and method for growing crystals from a melt employing a heat pipe, consisting of one or more sections, each section serving to control temperature and thermal gradients in the crystal as it forms inside the pipe.

  10. How Your Baby Grows

    MedlinePlus

    ... Pregnancy > Prenatal care > Pregnancy week by week Pregnancy week by week Week by week Videos Swipe to advance Learn ... grows each week during pregnancy. Pick your week. Weeks 1-2 Conception (also called fertilization) usually happens ...

  11. Crop growing practices

    USDA-ARS?s Scientific Manuscript database

    This chapter reviews the literature on two specific aspects of cotton growing practices; tillage management and nutrient management. Conservation tillage systems were developed to reduce soil erosion from agricultural fields. Besides this function, conservation tillage systems can improve the water ...

  12. The Growing Human Population.

    ERIC Educational Resources Information Center

    Keyfitz, Nathan

    1989-01-01

    Discusses the issue of human population. Illustrates the projections of the growing human population in terms of developed and less developed countries. Describes the family planning programs in several countries. Lists three references for further reading. (YP)

  13. Antimicrobial agent resistance in mycobacteria: molecular genetic insights.

    PubMed Central

    Musser, J M

    1995-01-01

    The primary theme emerging from molecular genetic work conducted with Mycobacterium tuberculosis and several other mycobacterial species is that resistance is commonly associated with simple nucleotide alterations in target chromosomal genes rather than with acquisition of new genetic elements encoding antibiotic-altering enzymes. Mutations in an 81-bp region of the gene (rpoB) encoding the beta subunit of RNA polymerase account for rifampin resistance in 96% of M. tuberculosis and many Mycobacterium leprae isolates. Streptomycin resistance in about one-half of M. tuberculosis isolates is associated with missense mutations in the rpsL gene coding for ribosomal protein S12 or nucleotide substitutions in the 16S rRNA gene (rrs). Mutations in the katG gene resulting in catalase-peroxidase amino acid alterations nad nucleotide substitutions in the presumed regulatory region of the inhA locus are repeatedly associated with isoniazid-resistant M. tuberculosis isolates. A majority of fluoroquinolone-resistant M. tuberculosis isolates have amino acid substitutions in a region of the DNA gyrase A subunit homologous to a conserved fluoroquinolone resistance-determining region. Multidrug-resistant isolates of M. tuberculosis arise as a consequence of sequential accumulation of mutations conferring resistance to single therapeutic agents. Molecular strategies show considerable promise for rapid detection of mutations associated with antimicrobial resistance. These approaches are now amenable to utilization in an appropriately equipped clinical microbiology laboratory. PMID:8665467

  14. Interdigitated array microelectrode capacitive sensor for detection of paraffinophilic mycobacteria

    NASA Astrophysics Data System (ADS)

    Sampson, Andrew M.; Peterson, Erik T. K.; Papautsky, Ian

    2008-02-01

    Mycobacterium Avium Complex (MAC) is an opportunistic pathogen that threatens public health and has high clinical relevance. While culture-based and molecular biology techniques for identification are available, these methods are prone to error and require weeks to perform. There is a critical need for improved portable lab-on-a-chip sensor technology which will enable accurate and rapid point-of-care detection of these microorganisms. In this work, a new capacitive sensing strategy is explored utilizing interdigitated array (IDA) microelectrodes and exploiting the paraffinophilic nature of MAC. In this approach, paraffin wax is deposited over IDA microelectrodes to selectively extract these microorganisms from samples. As bacteria consume the dielectric paraffin layer, the charging current of the IDA capacitor changes to facilitate detection. Several IDA geometries were designed and simulated using CFD-ACE+ modeling software and compared with mathematical models. Capacitance of fabricated devices was determined using a charge-based capacitance measurement (CBCM) technique. Modeling and experimental results were in good agreement. Detection of femto-Farad changes in capacitance is possible, making this a feasible technique for sensing small changes in the paraffin for detection of paraffinophilic MAC.

  15. Growing America's Energy Future

    SciTech Connect

    2016-06-01

    The emerging U.S. bioenergy industry provides a secure and growing supply of transportation fuels, biopower, and bioproducts produced from a range of abundant, renewable biomass resources. Bioenergy can help ensure a secure, sustainable, and economically sound future by reducing U.S. dependence on foreign oil, developing domestic clean energy sources, and generating domestic green jobs. Bioenergy can also help address growing concerns about climate change by reducing greenhouse gas emissions to create a healthier environment for current and future generations.

  16. Th1-skewed tissue responses to a mycolyl glycolipid in mycobacteria-infected rhesus macaques

    SciTech Connect

    Morita, Daisuke; Miyamoto, Ayumi; Hattori, Yuki; Komori, Takaya; Nakamura, Takashi; Igarashi, Tatsuhiko; Harashima, Hideyoshi; Sugita, Masahiko

    2013-11-08

    Highlights: •Glucose monomycolate (GMM) is a marker glycolipid for active tuberculosis. •Tissue responses to GMM involved up-regulation of Th1-attracting chemokines. •Th1-skewed local responses were mounted at the GMM-injected tissue. -- Abstract: Trehalose 6,6′-dimycolate (TDM) is a major glycolipid of the cell wall of mycobacteria with remarkable adjuvant functions. To avoid detection by the host innate immune system, invading mycobacteria down-regulate the expression of TDM by utilizing host-derived glucose as a competitive substrate for their mycolyltransferases; however, this enzymatic reaction results in the concomitant biosynthesis of glucose monomycolate (GMM) which is recognized by the acquired immune system. GMM-specific, CD1-restricted T cell responses have been detected in the peripheral blood of infected human subjects and monkeys as well as in secondary lymphoid organs of small animals, such as guinea pigs and human CD1-transgenic mice. Nevertheless, it remains to be determined how tissues respond at the site where GMM is produced. Here we found that rhesus macaques vaccinated with Mycobacterium bovis bacillus Calmette–Guerin mounted a chemokine response in GMM-challenged skin that was favorable for recruiting T helper (Th)1 T cells. Indeed, the expression of interferon-γ, but not Th2 or Th17 cytokines, was prominent in the GMM-injected tissue. The GMM-elicited tissue response was also associated with the expression of monocyte/macrophage-attracting CC chemokines, such as CCL2, CCL4 and CCL8. Furthermore, the skin response to GMM involved the up-regulated expression of granulysin and perforin. Given that GMM is produced primarily by pathogenic mycobacteria proliferating within the host, the Th1-skewed tissue response to GMM may function efficiently at the site of infection.

  17. The tracing of mycobacteria in drinking water supply systems by culture, conventional, and real time PCRs.

    PubMed

    Klanicova, Barbora; Seda, Jaromir; Slana, Iva; Slany, Michal; Pavlik, Ivo

    2013-12-01

    Mycobacteria are widely present in diverse aquatic habitats, where they can survive for months or years while some species can even proliferate. The resistance of different mycobacterial species to disinfection methods like chlorination or ozonation could result in their presence in the final tap water of consumers. In this study, the culture method, Mycobacterium tuberculosis complex conventional duplex PCR for detection of non-tuberculous mycobacteria (NTM) and quantitative real-time PCR (qPCR) to detect three subspecies of M. avium species (M. a. avium, M. a. hominissuis, and M. a. paratuberculosis) were used to trace their possible path of transmission from the watershed through the reservoir and drinking water plant to raw drinking water and finally to households. A total of 124 samples from four drinking water supply systems in the Czech Republic, 52 dam sediments, 34 water treatment plant sludge samples, and 38 tap water household sediments, were analyzed. NTM of 11 different species were isolated by culture from 42 (33.9 %) samples; the most prevalent were M. gordonae (16.7 %), M. triplex (14.3 %), M. lentiflavum (9.5 %), M. a. avium (7.1 %), M. montefiorenase (7.1 %), and M. nonchromogenicum (7.1 %). NTM DNA was detected in 92 (76.7 %) samples. By qPCR analysis a statistically significant decrease (P < 0.01) was observed along the route from the reservoir (dam sediments), through water treatment sludge and finally to household sediments. The concentrations ranged from 10(0) to 10(4) DNA cells/g. It was confirmed that drinking water supply systems (watershed-reservoir-drinking water treatment plant-household) might be a potential transmission route for mycobacteria.

  18. Direct Visualization of the Outer Membrane of Mycobacteria and Corynebacteria in Their Native State▿ †

    PubMed Central

    Zuber, Benoît; Chami, Mohamed; Houssin, Christine; Dubochet, Jacques; Griffiths, Gareth; Daffé, Mamadou

    2008-01-01

    The cell envelope of mycobacteria, which include the causative agents of tuberculosis and leprosy, is crucial for their success as pathogens. Despite a continued strong emphasis on identifying the multiple chemical components of this envelope, it has proven difficult to combine its components into a comprehensive structural model, primarily because the available ultrastructural data rely on conventional electron microscopy embedding and sectioning, which are known to induce artifacts. The existence of an outer membrane bilayer has long been postulated but has never been directly observed by electron microscopy of ultrathin sections. Here we have used cryo-electron microscopy of vitreous sections (CEMOVIS) to perform a detailed ultrastructural analysis of three species belonging to the Corynebacterineae suborder, namely, Mycobacterium bovis BCG, Mycobacterium smegmatis, and Corynebacterium glutamicum, in their native state. We provide new information that accurately describes the different layers of the mycobacterial cell envelope and challenges current models of the organization of its components. We show a direct visualization of an outer membrane, analogous to that found in gram-negative bacteria, in the three bacterial species examined. Furthermore, we demonstrate that mycolic acids, the hallmark of mycobacteria and related genera, are essential for the formation of this outer membrane. In addition, a granular layer and a low-density zone typifying the periplasmic space of gram-positive bacteria are apparent in CEMOVIS images of mycobacteria and corynebacteria. Based on our observations, a model of the organization of the lipids in the outer membrane is proposed. The architecture we describe should serve as a reference for future studies to relate the structure of the mycobacterial cell envelope to its function. PMID:18567661

  19. Distribution of mycobacteria in clinically healthy ornamental fish and their aquarium environment.

    PubMed

    Beran, V; Matlova, L; Dvorska, L; Svastova, P; Pavlik, I

    2006-07-01

    Some mycobacterial species (particularly Mycobacterium marinum) found in aquarium environments may cause chronic diseases in fish and cutaneous infections in humans, the so-called 'fish tank granuloma'. The presence and distribution of mycobacterial species in clinically healthy aquarium fish and their environment has not been adequately explored. The present study analysed the occurrence of mycobacteria in a decorative aquarium (Brno, South Moravia) and in five aquaria of a professional fish breeder (Bohumin, North Moravia). After Ziehl-Neelsen staining, acid-fast rods (AFR) were observed in six (14.3%) and mycobacteria were detected by culture in 18 (42.9%) of 42 tissue samples from 19 fish. Sixty-five samples of the aqueous environment from all six aquaria were examined; AFR were found in 16 (24.6%) and mycobacteria were detected by culture in 49 (75.4%) samples. Forty-one (70.7%) of 58 selected mycobacterial isolates were identified biochemically as follows: M. fortuitum, M. flavescens, M. chelonae, M. gordonae, M. terrae, M. triviale, M. diernhoferi, M. celatum, M. kansasii and M. intracellulare. The clinically important species for humans and fish, M. marinum, was not detected. Mycobacterium kansasii was isolated from one sample of the aquarium environment from North Moravia, which is a region of the Czech Republic with endemic incidence of M. kansasii in water. The incidence of other conditionally pathogenic mycobacterial species in healthy fish and in all investigated constituents of the aquarium environment including snails and crustaceans used for fish feeding, was quite high. Accordingly, mycobacterial species from aquarium environments may serve as a possible source of infection for both aquarium fish and immunodeficient fish handlers.

  20. Multicenter evaluation of fully automated BACTEC Mycobacteria Growth Indicator Tube 960 system for susceptibility testing of Mycobacterium tuberculosis.

    PubMed

    Bemer, Pascale; Palicova, Frantiska; Rüsch-Gerdes, Sabine; Drugeon, Henri B; Pfyffer, Gaby E

    2002-01-01

    The reliability of the BACTEC Mycobacteria Growth Indicator Tube (MGIT) 960 system for testing of Mycobacterium tuberculosis susceptibility to the three front-line drugs (isoniazid [INH], rifampin [RIF], and ethambutol [EMB]) plus streptomycin (STR) was compared to that of the BACTEC 460 TB system. The proportion method was used to resolve discrepant results by an independent arbiter. One hundred and ten strains were tested with an overall agreement of 93.5%. Discrepant results were obtained for seven strains (6.4%) with INH (resistant by BACTEC MGIT 960; susceptible by BACTEC 460 TB), for one strain (0.9%) with RIF (resistant by BACTEC MGIT 960; susceptible by BACTEC 460 TB), for seven strains (6.4%) with EMB (six resistant by BACTEC MGIT 960 and susceptible by BACTEC 460 TB; one susceptible by BACTEC MGIT 960 and resistant by BACTEC 460 TB), and for 19 strains (17.3%) with STR (resistant by BACTEC MGIT 960 and susceptible by BACTEC 460 TB). After resolution of discrepant results, the sensitivity of the BACTEC MGIT 960 system was 100% for all four drugs and specificity ranged from 89.8% for STR to 100% for RIF. Turnaround times were 4.6 to 11.7 days (median, 6.5 days) for BACTEC MGIT 960 and 4.0 to 10.0 days (median, 7.0 days) for BACTEC 460 TB. These data demonstrate that the fully automated and nonradiometric BACTEC MGIT 960 system is an accurate method for rapid susceptibility testing of M. tuberculosis.