DOCSLIB.ORG

And Mycobacterium Gordonae by High-Performance Liquid Chromatography of Their Mycolic Acids W

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
And Mycobacterium Gordonae by High-Performance Liquid Chromatography of Their Mycolic Acids W JOURNAL OF CLINICAL MICROBIOLOGY, Jan. 1988, p. 50-53 Vol. 26, No. 1 0095-1137/88/010050-04$02.00/0 Copyright © 1988, American Society for Microbiology Identification of Major Slowly Growing Pathogenic Mycobacteria and Mycobacterium gordonae by High-Performance Liquid Chromatography of Their Mycolic Acids W. RAY BUTLER* AND JAMES 0. KILBURN Mycobacteriology Laboratory, Respiratory Diseases Branch, Division of Bacterial Diseases, Center for Infectious Diseases, Centers for Disease Control, Atlanta, Georgia 30333 Received 15 June 1987/Accepted 9 October 1987 A rapid, reverse-phase high-performance liquid chromatography method was used to detect p-bromophe- nacyl mycolic acid ester patterns for strains of four major pathogenic Mycobacteriunt species and for the most commonly encountered saprophytic species, Mycobacterium gordonae. Mycobacteria in low numbers (2.5 x 106 CFU) were detected and identified to the species level. Standard chromatographic patterns characteristic of each species were established. Simple pattern recognition enabled rapid identification of M. tuberculosis, M. kansasii, M. avium, M. intracellulare, and M. gordonae. Differential identification of Mycobacterium species has together account for nearly 85% of all mycobacteria recov- traditionally been based upon biochemical reactions (7). The ered in state health laboratories. time required to perform the battery of biochemical tests usually makes traditional taxonomic identification a time- MATERIALS AND METHODS consuming process. More rapid methods of differential iden- tification are needed for the various Mycobacterium species. Microorganisms and growth conditions. Mycobacterium Chemical of cellular fatty acids, especially the isolates were either from the Trudeau Mycobacterial Collec- analysis and high-molecular-weight, a-branched mycolic ac- tion (TMC), formerly at the National Jewish Hospital P-hydroxy or from the ids, has been used as a rapid test in mycobacterial system- Research Center, Denver, Colo., Mycobacteri- atics (10). Extensive studies using two-dimensional thin- ology Laboratory, Centers for Disease Control (CDC), At- were as M. TMC layer chromatography and mass spectrometry with mycolic lanta, Ga., and follows: tuberculosis 102, and 321 and acid methyl esters have revealed a variable species-related 106, 107, 110, 111, 112, 117, 119, 124, 205, 320, distribution of mycobacterial mycolic acid structural types CDC 3900, 3901, 3902, 3903, 3904, 3905, 3906, 3907, 3908 a resistance i.e., isolates (9-13). Recently, a study using thin-layer chromatography and 3909 (from primary drug study, revealed the clinical usefulness of mycolic acid analysis for with no prior exposure to drug therapy); M. kansasii TMC separating Mycobacterium simiae from M. avium-M. intra- 1203, 1204, 1214, and 1217 and CDC 769, 830, 946, 961, 983, were sub- cellulare in a mixed mycobacterial infection (8). 1010, 1069, 1087, 1210, and 1253 (these isolates Other methods of mycolic acid analysis have combined mitted from state health departments to the CDC reference absorption chromatography with high-performance liquid laboratory for study); M. gordonae TMC 1319, 1324, 1325, chromatography (HPLC) and demonstrated the chemical and 1327 and CDC 911, 946, 986, 1196, 1215, 1231, 1245, and uniqueness of the cell wall-bound mycolic acids (15, 18). We 1251 (these isolates were submitted from state health depart- previously used HPLC to separate the different mycolic ments to the CDC reference laboratory for study); M. avium acid-containing genera (1). In this study, we describe a TMC 701, 702, 706, 714, 716, 724, 801, 1461, 1462, 1463, simple, single-step HPLC analysis of the p-bromophenacyl 1464, 1468, 1472, and 1479 and CDC 82-307, 82-905, and esters of mycolic acids extracted from saponified whole cells 82-993; M. intracellulare TMC 1403, 1405, 1406, 1419, 1466, of several species of mycobacteria and demonstrate the 1469, and 1473 and CDC 81-641, 82-421, 82-625, 82-759, distinctly different mycolic acid "fingerprint" patterns pro- 82-1128, 82-1129, and 83-764. All of the CDC isolates were duced by the different species. We examined four strictly or fresh clinical specimens. The pathogenic species included in potentially pathogenic species (M. tuberculosis, M. avium, this study were M. tuberculosis, M. kansasii, M. avium, and M. intracellulare, and M. kansasii) that account for almost M. intracellulare. M. gordonae, a commonly nonpathogenic 90% of the potentially pathogenic mycobacteria and 71% of species, was also included. Isolates were grown on commer- the total (both pathogenic and saprophytic) mycobacteria cially prepared Lowenstein-Jensen slants (Carr-Scarborough recovered in state health laboratories in the United States Microbiologicals, Atlanta, Ga.) at 35°C for 3 weeks before (5). We compared the mycolic acid patterns of both refer- analysis. ence culture strains and clinical isolates to establish standard Identification of mycobacteria. Mycobacteria were identi- species patterns. It was of special interest to determine if the fied by conventional laboratory methods (6, 7). mycolic acid patterns could be used as a rapid identification Mycolic acid extraction and derivatization procedures. My- method for these slowly growing pathogenic mycobacteria. cobacteria were harvested either by washing the Lowen- Although M. gordonae is not pathogenic, it was included in stein-Jensen slants with 2 ml of saponification reagent (25% the study because it represents about 15% of the isolates KOH in 50% ethanol) if growth was scant or by suspending recovered in public health laboratories (5). The five species 1 loopful of cells from Lowenstein-Jensen slants with abun- dant growth in 2 ml of alcoholic KOH. ]3iomass saponifica- tion was performed at 85°C for 18 h in 13-mm glass tubes * Corresponding author. with Teflon-lined screw caps. Saponified cells were cooled to 50 VOL. 26, 1988 IDENTIFICATION OF MYCOBACTERIAL MYCOLIC ACIDS BY HPLC 51 k M. tuberculosis (TMC 102) M. avium (TMC 724) w (n z Oo- (n w k r wcc h w a m 15 20 24 15 20 24 RETENTION TIME (MIN) RETENTION TIME (MIN) M. intracellulare (TMC 1466) w (n M. kansasii (TMC 1214) z O o- h k (n w O Ulc-c w c e 1--w 1 1 h 9D 15 20 24 15 20 24 RETENTION TIME (MIN) RETENTION TIME (MIN) M. gordonae (TMC 1325) w (n 9 z k CLo- (n w Cr: cc O F- w F- w a il1 M 15 20 24 RETENTION TIME (MIN) FIG. 1. High-performance liquid chromatograms of the p-bromophenacyl mycolic acid esters from saponified whole cells of mycobacteria. 52 BUTLER AND KILBURN J. CLIN. MICROBIOL. TABLE 1. Reproductibility of mycolic acid patterns of mycobacteria Species (no. of strains) At (±SEM)a of peak: a b c d e f M. tuberculosis (22) M. kansasii (14) -0.62 (0.006) M. gordonae (12) -0.69 (0.009) M. aviumb (17) -5.07 (0.02) -4.78 (0.02) -4.27 (0.03) -3.75 (0.02) -3.21 (0.01) -0.62 (0.008) M. intracellulareb (14) -5.00 (0.04) -4.69 (0.04) -4.27 (0.04) -3.68 (0.03) -3.18 (0.03) -0.64 (0.008) room temperature and acidified to pH 1 with 6 N HCI. The RESULTS AND DISCUSSION mycolic acids were extracted by adding 2 ml of chloroform, mixing for 20 s on a test tube mixer, and centrifuging to assist The elution patterns of the p-bromophenacyl esters of the separation of the chloroform layer. The extracted mycolic mycolic acids were determined by HPLC for 85 isolates acids were evaporated to dryness and resuspended in chlo- representing five Mycobacterium species (Fig. 1). Each roform for derivatization. The mycolic acids were deriva- analysis was completed in 34 min, from the initial column tized to p-bromophenacyl esters by the method of Durst et equilibration to the end of the program run. The mycolic acid al. (4). Briefly, a mixture containing the extracted mycolic esters of these mycobacteria eluted between 15 and 23 min. acids in 0.5 ml of chloroform-2 mg of potassium car- The p-bromophenacyl esters of "standard," purified my- bonate-25 ,ul ofp-bromophenacyl bromide in acetonitrile-25 colic acids from M. tuberculosis and M. smegmatis corre- pul of the catalyst dicyclohexyl-18-crown-6 ether in acetoni- sponded directly in retention times and overall patterns to trile was heated for 45 min at 85°C in Teflon-capped 5-ml the derivatized mycolic acids that we obtained from whole Reacti-Vials (Pierce Chemical Co., Rockford, Ill.) with organisms, and these standards were used as a presumptive constant stirring with Teflon-coated magnetic stirrers. The identification of the mycolic acids. We found that the deri- reagents were included a kit obtained from Alltech Associ- vatized products were stable for months at an ambient ates, Inc., Applied Science Div., State College, Pa. Chloro- temperature. Components of Lowenstein-Jensen medium, form was used instead of the acetonitrile recommended for excess derivatization reagents, acetonitrile (from the reagent shorter-chain fatty acid derivatization because the long- kit), and short-chain cellular fatty acids (C18 or less) all chain, high-molecular-weight mycolic acids were more sol- eluted within 5 min and did not interfere with the identifica- uble in chloroform. After derivatization, the p-bromophe- tion of the mycolic acids. Quantitation of the method by use nacyl esters of the mycolic acids were filtered through of an M. kansasii culture revealed that definitive mycolic 0.45-,um-pore nylon 66 membrane filters, evaporated to acid fingerprint patterns could be achieved with as few as 2.5 dryness, and resuspended in 100 pil of chloroform for anal- x 106 CFU. ysis. M. tuberculosis had the least complicated mycolic acid HPLC conditions. HPLC of the p-bromophenacyl esters of pattern, with a cluster of seven peaks that eluted between 21 the mycolic acids was done with an HPLC model 450 data and 23 min and one major, central peak (designated k). M. system controller (Beckman Instruments, Inc., Fullerton, kansasii and M.
Load more

Recommended publications
  • S1 Sulfate Reducing Bacteria and Mycobacteria Dominate the Biofilm
    Sulfate Reducing Bacteria and Mycobacteria Dominate the Biofilm Communities in a Chloraminated Drinking Water Distribution System C. Kimloi Gomez-Smith 1,2 , Timothy M. LaPara 1, 3, Raymond M. Hozalski 1,3* 1Department of Civil, Environmental, and Geo- Engineering, University of Minnesota, Minneapolis, Minnesota 55455 United States 2Water Resources Sciences Graduate Program, University of Minnesota, St. Paul, Minnesota 55108, United States 3BioTechnology Institute, University of Minnesota, St. Paul, Minnesota 55108, United States Pages: 9 Figures: 2 Tables: 3 Inquiries to: Raymond M. Hozalski, Department of Civil, Environmental, and Geo- Engineering, 500 Pillsbury Drive SE, Minneapolis, MN 554555, Tel: (612) 626-9650. Fax: (612) 626-7750. E-mail: [email protected] S1 Table S1. Reference sequences used in the newly created alignment and taxonomy databases for hsp65 Illumina sequencing. Sequences were obtained from the National Center for Biotechnology Information Genbank database. Accession Accession Organism name Organism name Number Number Arthrobacter ureafaciens DQ007457 Mycobacterium koreense JF271827 Corynebacterium afermentans EF107157 Mycobacterium kubicae AY373458 Mycobacterium abscessus JX154122 Mycobacterium kumamotonense JX154126 Mycobacterium aemonae AM902964 Mycobacterium kyorinense JN974461 Mycobacterium africanum AF547803 Mycobacterium lacticola HM030495 Mycobacterium agri AY438080 Mycobacterium lacticola HM030495 Mycobacterium aichiense AJ310218 Mycobacterium lacus AY438090 Mycobacterium aichiense AF547804 Mycobacterium
    [Show full text]
  • Opportunist Mycobacteria
    Thorax: first published as 10.1136/thx.44.6.449 on 1 June 1989. Downloaded from Thorax 1989;44:449454 Editorial Treatment of pulmonary disease caused by opportunist mycobacteria During the early 1950s it was recognised that recommended that surgical treatment for opportunist mycobacteria other than Mycobacterium tuberculosis mycobacterial infection should be given to those could cause pulmonary disease in man.' Over 30 years patients who are suitable surgical candidates.'7 The later there is no general agreement about the treatment failure of chemotherapy was often attributed to drug of patients with these mycobacterial infections. The resistance,'2 18 but the importance of prolonging the greatest controversy surrounds the treatment of in- duration of chemotherapy in opportunist mycobac- fection caused by the M aviwn-intracellulare- terial disease beyond that which would normally be scrofulaceum complex (MAIS), for which various required in tuberculosis was not appreciated. In treatments have been advocated, including chemo- several surgically treated series preoperative chemo- therapy with three23 or more' drugs or, alternatively, therapy was given on average for only four to seven surgical resection of the affected lung.78 Although the months,'3 19-22 some patients receiving as little as eight treatment of infection caused by M kansasii is less weeks of treatment before surgery.2324 In contrast, controversial there is no uniform approach to treat- chemotherapy alone, with isoniazid, p-aminosalicyclic ment. Disease caused by M xenopi has been described acid, and streptomycin for 24 months, produced by some as easy to treat with chemotherapy,9 whereas successful results in 80-100% of patients with M others have found the response to drug treatment to be kansasii infection despite reports of in vitro resistance unpredictable.'' " to these agents.2125 This diversity ofopinion and approach to treatment has arisen for two reasons.
    [Show full text]
  • Mycobacterium Goodii Endocarditis Following Mitral Valve Ring Annuloplasty Rohan B
    Parikh and Grant Ann Clin Microbiol Antimicrob (2017) 16:14 DOI 10.1186/s12941-017-0190-4 Annals of Clinical Microbiology and Antimicrobials CASE REPORT Open Access Mycobacterium goodii endocarditis following mitral valve ring annuloplasty Rohan B. Parikh1 and Matthew Grant2* Abstract Background: Mycobacterium goodii is an infrequent human pathogen which has been implicated in prosthesis related infections and penetrating injuries. It is often initially misidentified as a gram-positive rod by clinical microbio- logic laboratories and should be considered in the differential diagnosis. Case presentation: We describe here the second reported case of M. goodii endocarditis. Species level identification was performed by 16S rDNA (ribosomal deoxyribonucleic acid) gene sequencing. The patient was successfully treated with mitral valve replacement and a prolonged combination of ciprofloxacin and trimethoprim/sulfamethoxazole. Conclusion: Confirmation of the diagnosis utilizing molecular techniques and drug susceptibility testing allowed for successful treatment of this prosthetic infection. Keywords: Mycobacterium goodii, Endocarditis, Gene sequencing, Prostheses related infections Background appreciated at the apex, and a drain was in place for a Mycobacterium goodii is a rapidly growing non-tubercu- groin seroma related to recent left heart catheterization. lous mycobacterium (NTM) belonging to the Mycobac- He had an unsteady gait and exhibited mild left lower terium smegmatis [1] group. Its importance has become extremity weakness (4/5). His brain magnetic resonance increasingly appreciated as a pathogen over the last imaging showed multiple ring-enhancing lesions in the 20 years, with a predilection towards infecting tissues at pons and posterior fossa suggestive of septic emboli. the site of penetrating injuries. Antibacterial treatment Transthoracic echocardiography showed moderate strategies against this pathogen are diverse but reported mitral regurgitation without any vegetation.
    [Show full text]
  • An Abstract of the Dissertation of Melanie J. Harriff
    AN ABSTRACT OF THE DISSERTATION OF MELANIE J. HARRIFF for the degree of Doctor of Philosophy in Molecular and Cellular Biology presented on June 8, 2007. Title: Mechanisms for the Interaction of Environmental Mycobacteria with Host Cells Abstract approved: Luiz E. Bermudez Michael L. Kent Environmental mycobacteria are important opportunistic pathogens for many hosts, including humans, cattle, and fish. Two well-studied species are Mycobacterium avium subsp. avium, a significant cause of disseminated bacterial disease in patients with AIDS, and Mycobacterium avium subsp. paratuberculosis, the cause of Johne’s disease in cattle. Many other species that are considerable sources of infections in fish, such as Mycobacterium chelonae and Mycobacterium marinum, also have zoonotic potential. To gain knowledge about the invasion of epithelial cells by environmental mycobacteria, selected genes and proteins involved in the uptake of M. avium by HEp-2 cells were analyzed by a variety of methods. Two transcriptional regulators (MAV_3679 and MAV_5138) were identified as being involved in invasion. A mycobacterial protein (CipA) with an amino acid sequence suggestive of an ability to be a part of the scaffolding complex that forms during cell signaling leading to actin polymerization was found to putatively interact with host cell Cdc42. Fusion of CipA to GFP, expressed in Mycobacterium smegmatis, revealed that CipA localizes to a structure on the surface of bacteria approaching HEp-2 cells. To establish whether species of environmental mycobacteria isolated from different hosts use similar mechanisms to M. avium for interaction with the mucosa, and for survival in macrophages, assays to determine invasion and replication were performed in different cell types, and a custom DNA microarray containing probes for known mycobacterial virulence determinants was developed.
    [Show full text]
  • Identification of Mycobacterium Avium Pathogenicity Island Important For
    MICROBIOLOGY. For the article ‘‘Identification of Mycobacterium avium pathogenicity island important for macrophage and amoeba infection,’’ by Lia Danelishvili, Martin Wu, Bernadette Stang, Melanie Harriff, Stuart Cirillo, Jeffrey Cirillo, Robert Bildfell, Brian Arbogast, and Luiz E. Bermudez, which appeared in issue 26, June 26, 2007, of Proc Natl Acad Sci USA (104:11038– 11043; first published June 19, 2007; 10.1073͞pnas.0610746104), the author name Stuart Cirillo should have appeared as Suat L. G. Cirillo, and the author name Jeffrey Cirillo should have appeared as Jeffrey D. Cirillo. The online version has been corrected. The corrected author line appears below. Addition- ally, the present address for both these authors should be: Department of Microbial and Molecular Pathogenesis, Texas A&M University College of Medicine, College Station, TX 77843-1114. The authors also note that Fig. 1 did not print at high resolution. The corrected figure and its legend appear below. Lia Danelishvili, Martin Wu, Bernadette Stang, Melanie Harriff, Suat L. G. Cirillo, Jeffrey D. Cirillo, Robert Bildfell, Brian Arbogast, and Luiz E. Bermudez Fig. 1. Chromosome regions. (A) Organization of the chromosome region inactivated in the 8H8 clone of M. avium involved in the glycosylation of the lipopeptide core. (B) Organization of the chromosome region inactivated in the M. avium 9B9 clone. The M. avium gene names correspond to MAP numbers from the M. avium subsp. paratuberculosis genome sequence. (C) Genetic organization of M. avium 104 PI associated with low invasion of macrophages and virulence in mice. The M. avium 104 (b) sequence and gene organization of this region are presented in comparison with M.
    [Show full text]
  • Pathogenicity of Mycobacterium Fortuitum and Mycobacterium Smegmatis to Goldfish, Carassius Auratus Adel M
    Veterinary Microbiology 66 (1999) 151±164 Pathogenicity of Mycobacterium fortuitum and Mycobacterium smegmatis to goldfish, Carassius auratus Adel M. Talaata,b,1, Michele Trucksisa,c, Andrew S. Kaneb, Renate Reimschuesselb,* aCenter for Vaccine Development, Division of Geographic Medicine, Department of Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA bDepartment of Pathology, University of Maryland School of Medicine, Baltimore, MD 21201, USA cMedical Service, Veterans' Affairs Medical Center, Baltimore, MD 21201, USA Received 3 June 1998; accepted 22 December 1998 Abstract Despite the ubiquitous presence of atypical mycobacteria in the environment and the potential risk of infection in humans and animals, the pathogenesis of diseases caused by infection with atypical mycobacteria has been poorly characterized. In this study, goldfish, Carassius auratus were infected either with the rapidly growing fish pathogen, Mycobacterium fortuitum or with another rapidly growing mycobacteria, Mycobacterium smegmatis. Bacterial persistence and pathological host response to mycobacterial infection in the goldfish are described. Mycobacteria were recovered from a high percentage of inoculated fish that developed a characteristic chronic granulomatous response similar to that associated with natural mycobacterial infection. Both M. fortuitum and M. smegmatis were pathogenic to fish. Fish infected with M. smegmatis ATCC 19420 showed the highest level of giant cell recruitment compared to fish inoculated with M. smegmatis mc2155 and M. fortuitum. Of the three strains of mycobacteria examined, M. smegmatis ATCC 19420 was the most virulent strain to goldfish followed by M. fortuitum and M. smegmatis mc2155, respectively. # 1999 Elsevier Science B.V. All rights reserved. Keywords: Fish; Virulence; Mycobacteria; Mycobacterium fortuitum; Mycobacterium smegmatis; Pathogenesis * Corresponding author.
    [Show full text]
  • Detection of and Efflux Pump Genes Among Clinical Isolates of Non-Pigmented Rapidly Growing Mycobacteria J
    Detection of and efflux pump genes among clinical isolates of non-pigmented rapidly growing mycobacteria J. Esteban, N.Z. Martín-De-Hijas, A. Ortiz, T.J. Kinnari, A. Bodas Sánchez, I. Gadea, R. Fernández-Roblas To cite this version: J. Esteban, N.Z. Martín-De-Hijas, A. Ortiz, T.J. Kinnari, A. Bodas Sánchez, et al.. Detection of and ef- flux pump genes among clinical isolates of non-pigmented rapidly growing mycobacteria. International Journal of Antimicrobial Agents, Elsevier, 2009, 34 (5), pp.454. 10.1016/j.ijantimicag.2009.06.026. hal-00556354 HAL Id: hal-00556354 https://hal.archives-ouvertes.fr/hal-00556354 Submitted on 16 Jan 2011 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Accepted Manuscript Title: Detection of lfrA and tap efflux pump genes among clinical isolates of non-pigmented rapidly growing mycobacteria Authors: J. Esteban, N.Z. Mart´ın-de-Hijas, A. Ortiz, T.J. Kinnari, A. Bodas Sanchez,´ I. Gadea, R. Fernandez-Roblas´ PII: S0924-8579(09)00348-3 DOI: doi:10.1016/j.ijantimicag.2009.06.026 Reference: ANTAGE 3083 To appear in: International Journal of Antimicrobial Agents Received date: 8-6-2009 Revised date: 16-6-2009 Accepted date: 19-6-2009 Please cite this article as: Esteban J, Mart´ın-de-Hijas NZ, Ortiz A, Kinnari TJ, Sanchez´ AB, Gadea I, Fernandez-Roblas´ R, Detection of lfrA and tap efflux pump genes among clinical isolates of non-pigmented rapidly growing mycobacteria, International Journal of Antimicrobial Agents (2008), doi:10.1016/j.ijantimicag.2009.06.026 This is a PDF file of an unedited manuscript that has been accepted for publication.
    [Show full text]
  • The Impact of Chlorine and Chloramine on the Detection and Quantification of Legionella Pneumophila and Mycobacterium Spp
    The impact of chlorine and chloramine on the detection and quantification of Legionella pneumophila and Mycobacterium spp. Maura J. Donohue Ph.D. Office of Research and Development Center of Environmental Response and Emergency Response (CESER): Water Infrastructure Division (WID) Small Systems Webinar January 28, 2020 Disclaimer: The views expressed in this presentation are those of the author and do not necessarily reflect the views or policies of the U.S. Environmental Protection Agency. A Tale of Two Bacterium… Legionellaceae Mycobacteriaceae • Legionella (Genus) • Mycobacterium (Genus) • Gram negative bacteria • Nontuberculous Mycobacterium (NTM) (Gammaproteobacteria) • M. avium-intracellulare complex (MAC) • Flagella rod (2-20 µm) • Slow grower (3 to 10 days) • Gram positive bacteria • Majority of species will grow in free-living • Rod shape(1-10 µm) amoebae • Non-motile, spore-forming, aerobic • Aerobic, L-cysteine and iron salts are required • Rapid to Slow grower (1 week to 8 weeks) for in vitro growth, pH: 6.8 to 7, T: 25 to 43 °C • ~156 species • ~65 species • Some species capable of causing disease • Pathogenic or potentially pathogenic for human 3 NTM from Environmental Microorganism to Opportunistic Opponent Genus 156 Species Disease NTM =Nontuberculous Mycobacteria MAC = M. avium Complex Mycobacterium Mycobacterium duvalii Mycobacterium litorale Mycobacterium pulveris Clinically Relevant Species Mycobacterium abscessus Mycobacterium elephantis Mycobacterium llatzerense. Mycobacterium pyrenivorans, Mycobacterium africanum Mycobacterium europaeum Mycobacterium madagascariense Mycobacterium rhodesiae Mycobacterium agri Mycobacterium fallax Mycobacterium mageritense, Mycobacterium riyadhense Mycobacterium aichiense Mycobacterium farcinogenes Mycobacterium malmoense Mycobacterium rufum M. avium, M. intracellulare, Mycobacterium algericum Mycobacterium flavescens Mycobacterium mantenii Mycobacterium rutilum Mycobacterium alsense Mycobacterium florentinum. Mycobacterium marinum Mycobacterium salmoniphilum ( M. fortuitum, M.
    [Show full text]
  • MYCOBACTERIOSIS in FARMED SEA BREAM (Sparus Aurata) CAUSED by Mycobacterium Frederiksbergense in TURKEY
    Acta Veterinaria-Beograd 2018, 68 (4), 391-400 UDK: 597.556.331.1-12:[616.98:579.873.2(560) Research article DOI: 10.2478/acve-2018-0032 MYCOBACTERIOSIS IN FARMED SEA BREAM (Sparus aurata) CAUSED BY Mycobacterium frederiksbergense IN TURKEY URKU Cigdem1*, GENC Gonca Erkose2, WITTWER Franziska3, ERTURAN Zayre2, PFYFFER Gaby3 1Department of Fish Disease, Faculty of Aquatic Sciences, University of Istanbul, 34470, Ordu Cad. No: 200 Laleli- Istanbul/ Turkey; 2Department of Medical Microbiology, Istanbul Faculty of Medicine, Istanbul University, Capa, 34093, Istanbul/ Turkey; 3Department of Medical Microbiology, Luzerner Kantonsspital, 6000 Luzern 16, Switzerland (Received 10 September, Accepted 19 November 2018) Over the past decades, different mycobacteriosis agents have become important fi sh pathogens. The present study describes a different mycobacteriosis in farmed sea bream (Sparus aurata) caused by Mycobacterium frederiksbergense in Turkey. Affected 15 fi sh, weighing 15 to 20 g, showed lethargy, stunted growth, pale skin, dorsal fi n necrosis, and a signifi cant level of mortality (40 %) in fi sh stocks. Internally, no multifocal white- colored granulomas in the visceral organs were observed. Inoculation of the visceral organs onto Löwenstein-Jensen medium and Tryptic Soy Agar (1.5% NaCl) slants produced only fast-growing (2-3 weeks), orange to yellow-colored, photochromogenic acid-fast colonies. Ziehl-Nielsen positive bacterial isolates were identifi ed using a commercially available line probe assay (Genotype Mycobacterium CM/AS assay) and with 16S rRNA gene sequencing analysis based on 16S rRNA gene sequencing, fi fteen isolates were identifi ed as Mycobacterium frederiksbergense. Histopathologically, epithelioid cell granulomas were not observed in any vicseral organs but acid-fast bacteria were detected in the liver, kidney, spleen, and heart tissue.
    [Show full text]
  • Maturing Mycobacterium Smegmatis Peptidoglycan Requires Non
    RESEARCH ARTICLE Maturing Mycobacterium smegmatis peptidoglycan requires non-canonical crosslinks to maintain shape Catherine Baranowski1, Michael A Welsh2, Lok-To Sham2,3, Haig A Eskandarian4,5, Hoong Chuin Lim2, Karen J Kieser1, Jeffrey C Wagner1, John D McKinney4, Georg E Fantner5, Thomas R Ioerger6, Suzanne Walker2, Thomas G Bernhardt2, Eric J Rubin1,2*, E Hesper Rego7* 1Department of Immunology and Infectious Disease, Harvard TH Chan School of Public Health, Boston, United States; 2Department of Microbiology and Immunobiology, Harvard Medical School, Boston, United States; 3Department of Microbiology and Immunology, National University of Singapore, Singapore, Singapore; 4School of Life Sciences, Swiss Federal Institute of Technology in Lausanne, Lausanne, Switzerland; 5School of Engineering, Swiss Federal Institute of Technology in Lausanne, Lausanne, Switzerland; 6Department of Computer Science and Engineering, Texas A&M University, Texas, United States; 7Department of Microbial Pathogenesis, Yale University School of Medicine, New Haven, United States Abstract In most well-studied rod-shaped bacteria, peptidoglycan is primarily crosslinked by penicillin-binding proteins (PBPs). However, in mycobacteria, crosslinks formed by L,D- transpeptidases (LDTs) are highly abundant. To elucidate the role of these unusual crosslinks, we characterized Mycobacterium smegmatis cells lacking all LDTs. We find that crosslinks generate by LDTs are required for rod shape maintenance specifically at sites of aging cell wall, a byproduct of *For correspondence: polar elongation. Asymmetric polar growth leads to a non-uniform distribution of these two types [email protected] (EJR); of crosslinks in a single cell. Consequently, in the absence of LDT-mediated crosslinks, PBP- [email protected] (EHR) catalyzed crosslinks become more important.
    [Show full text]
  • Diagnosis, Treatment, and Prevention of Nontuberculous Mycobacterial Diseases
    American Thoracic Society Documents An Official ATS/IDSA Statement: Diagnosis, Treatment, and Prevention of Nontuberculous Mycobacterial Diseases David E. Griffith, Timothy Aksamit, Barbara A. Brown-Elliott, Antonino Catanzaro, Charles Daley, Fred Gordin, Steven M. Holland, Robert Horsburgh, Gwen Huitt, Michael F. Iademarco, Michael Iseman, Kenneth Olivier, Stephen Ruoss, C. Fordham von Reyn, Richard J. Wallace, Jr., and Kevin Winthrop, on behalf of the ATS Mycobacterial Diseases Subcommittee This Official Statement of the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA) was adopted by the ATS Board Of Directors, September 2006, and by the IDSA Board of Directors, January 2007 CONTENTS Health Care– and Hygiene-associated Disease and Disease Prevention Summary NTM Species: Clinical Aspects and Treatment Guidelines Diagnostic Criteria of Nontuberculous Mycobacterial M. avium Complex (MAC) Lung Disease Key Laboratory Features of NTM M. kansasii Health Care- and Hygiene-associated M. abscessus Disease Prevention M. chelonae Prophylaxis and Treatment of NTM Disease M. fortuitum Introduction M. genavense Methods M. gordonae Taxonomy M. haemophilum Epidemiology M. immunogenum Pathogenesis M. malmoense Host Defense and Immune Defects M. marinum Pulmonary Disease M. mucogenicum Body Morphotype M. nonchromogenicum Tumor Necrosis Factor Inhibition M. scrofulaceum Laboratory Procedures M. simiae Collection, Digestion, Decontamination, and Staining M. smegmatis of Specimens M. szulgai Respiratory Specimens M. terrae
    [Show full text]
  • Mycobacterium Smegmatis Bacteremia in an Immunocompetent Host
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by IUPUIScholarWorks IDCases 15 (2019) xxx–xxx Contents lists available at ScienceDirect IDCases journal homepage: www.elsevier.com/locate/idcr Mycobacterium smegmatis bacteremia in an immunocompetent host a, b Saira Butt *, Amir Tirmizi a Indiana University School of Medicine, Department of Medicine, Division of Infectious Diseases, United States b Infectious Diseases Consultant, Columbus Regional Hospital, Columbus, IN, United States A R T I C L E I N F O A B S T R A C T Article history: Non-tuberculous mycobacteria can cause catheter associated blood stream infections. The causative Received 24 February 2019 agents are generally rapid growers that belong to the Mycobacterium fortuitum and Mycobacterium Received in revised form 7 March 2019 mucogenicum groups. A 65 year hospitalized patient with temporary central venous catheter who Accepted 7 March 2019 developed Mycobacterium smegmatis bacteremia. Bacteremia cleared after removal of the catheter. Patient was treated initially with 4 weeks of intravenous amikacin, intravenous meropenem, oral Keywords: doxycycline and oral ethambutol and then deescalated to oral doxycycline and oral ciprofloxacin for 8 Mycobacterium weeks. He improved clinically and remained stable. A literature search identified total of 22 articles that Atypical mycobacteria reported 47 unique cases of Mycobacterium smegmatis infection. To our knowledge, this is the first case CVC infection of Mycobacterium smegmatis central venous catheter associated bacteremia in an immunocompetent M smegmatis Line infection host. © 2019 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
    [Show full text]