Identification of Three Strains of <i>Mycobacterium</i> Species Isolated from Clinical Samples Using Fatty Acid Methyl Ester Profiling

Özbek, Ahmet; Aktaş, Osman

doi:10.1177/147323000303100210

Cited by 19 publications

(20 citation statements)

References 12 publications

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“…During the last decades, fatty acid profiles have been widely used to characterize microbial community [26]. GC analysis of fatty acids, in particular, derived from phospholipid, has proven to be highly applicable method for the bacteria species identification [27-30].…”

Section: Resultsmentioning

confidence: 99%

Fatty acid variability in three medicinal herbs of Panaxspecies

Zhang

Huang

Cai

et al. 2013

Chemistry Central Journal

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BackgroundFatty acid profiling has been widely used in the bacteria species identification, we hypothesized that fatty acid characteristics might discriminate the Panax herbs according to species. To test the hypothesis, fatty acids of Panax species, including Panax ginseng, Panax notoginseng and Panax quinquefolius, were characterized and compared using gas chromatography–mass spectrometry (GC-MS) followed by multivariate statistical analysis.ResultsThe content of investigated 11 fatty acids, including myristic acid, pentadecanoic acid, palmitic acid, palmitoleic acid, heptadecanoic acid, stearic acid, oleic acid, linoleic acid, α-linolenic acid, arachidic acid and eicosadienoic acid, obviously varied among three species, suggesting each species has its own fatty acid pattern. Principal component analysis and hierarchical clustering analysis according to the absolute and relative contents of fatty acids, showed that 30 tested samples could be clearly differentiated according to the species.ConclusionsThese findings demonstrated that GC-MS-based fatty acid profiling coupled with multivariate statistical analysis provides reliable platform to classify these three Panax species, which is helpful for ensuring their safety and efficacy.

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Section: Resultsmentioning

confidence: 99%

Fatty acid variability in three medicinal herbs of Panaxspecies

Zhang

Huang

Cai

et al. 2013

Chemistry Central Journal

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“…Chemotaxonomic investigations by FAME analysis have often played a cardinal role in resolving inadvertencies in case of taxonomic investigations in bacteria including the ones belonging to Mycobacterium as it examines the features at whole organism level [23], [24]. FAME (fatty acid methyl ester) analysis is a very sensitive approach, which proficiently reflects on biochemical and physiological attributes of an organism to correctly define its precise taxonomic position [25]–[27].…”

Section: Resultsmentioning

confidence: 99%

“…Hence, as a next step, the FAME analysis of MIP was carried out. It offered the obvious advantages that: i) FAME analysis reflects on the biochemical and physiological attributes of the associated organisms rather than on the mutations in the genes encoding the candidate orthologues in order to correctly define their precise taxonomic position; and ii) it offers a highly reproducible value based on the comparisons with other members of the genus [23], [24]. Its significance became apparent, when the FAME profile of MIP was compared with its counterparts from several other members of family Mycobacteriaceae .…”

Section: Discussionmentioning

confidence: 99%

Polyphasic Taxonomic Analysis Establishes Mycobacterium indicus pranii as a Distinct Species

Saini

Raghuvanshi

Talwar³

et al. 2009

PLoS ONE

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Background Mycobacterium indicus pranii (MIP), popularly known as Mw, is a cultivable, non-pathogenic organism, which, based on its growth and metabolic properties, is classified in Runyon Group IV along with M. fortuitum, M. smegmatis and M. vaccae. The novelty of this bacterium was accredited to its immunological ability to undergo antigen driven blast transformation of leukocytes and delayed hypersensitivity skin test in leprosy patients, a disease endemic in the Indian sub-continent. Consequently, MIP has been extensively evaluated for its biochemical and immunological properties leading to its usage as an immunomodulator in leprosy and tuberculosis patients. However, owing to advances in sequencing and culture techniques, the citing of new strains with almost 100% similarity in the sequences of marker genes like 16S rRNA, has compromised the identity of MIP as a novel species. Hence, to define its precise taxonomic position, we have carried out polyphasic taxonomic studies on MIP that integrate its phenotypic, chemotaxonomic and molecular phylogenetic attributes.Methodology/Principal FindingsThe comparative analysis of 16S rRNA sequence of MIP by using BLAST algorithm at NCBI (nr database) revealed a similarity of ≥99% with M. intracellulare, M. arosiense, M. chimaera, M. seoulense, M. avium subsp. hominissuis, M. avium subsp. paratuberculosis and M. bohemicum. Further analysis with other widely used markers like rpoB and hsp65 could resolve the phylogenetic relationship between MIP and other closely related mycobacteria apart from M. intracellulare and M. chimaera, which shares ≥99% similarity with corresponding MIP orthologues. Molecular phylogenetic analysis, based on the concatenation of candidate orthologues of 16S rRNA, hsp65 and rpoB, also substantiated its distinctiveness from all the related organisms used in the analysis excluding M. intracellulare and M. chimaera with which it exhibited a close proximity. This necessitated further analysis of MIP with more sensitive and segregating parameters to ascertain its precise taxonomic position as a new species. The analysis of MIP and its comparison with other mycobacterial reference strains based on cellular and biochemical features, growth characteristics and chemotaxonomic studies like FAME profiling confirmed that MIP is uniquely endowed with diverse metabolic attributes that effectively distinguishes it from all the closely related mycobacteria including M. intracellulare and M. chimaera.ConclusionThe results presented in this study coupled with the non-pathogenic nature and different biochemical and immunomodulatory properties of MIP affirm it as a distinct species belonging to M. avium complex (MAC). It is further proposed to use an earlier suggested name Mycobacterium indicus pranii for this newly established mycobacterial species. This study also exemplifies the growing need for a uniform, consensus based broader polyphasic frame work for the purpose of taxonomy and speciation, particularly in the genus Mycobacterium.

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“…FAs analysis allows us to consider not only the presence or absence of each FA but also use the data in a quantitative fashion. Previously, a set of 10 FAs (14:0, 16:1w7c, 16:1w6c, 16:0, 18:2w6,9c, 18:1w9c, 18:0, 18:0 10 Me, Summed in Feature 2 and Summed in Feature 3) were found in five Mycobacterium avium complex strains, which were differentiated from Mtb and Mycobacterium xenopi strains by the presence of Palmitoleic acid (16:1 w7c), and Summed in Feature 2 and 3 (Ozbek and Aktas, 2003). As a result, it was suggested that these three FAs could be used as markers to identify and distinguish Mycobacterium avium complex strains from other Mycobacteria.…”

Section: Discussionmentioning

confidence: 99%

Mycobacterium avium subsp. paratuberculosis (Map) Fatty Acids Profile Is Strain-Dependent and Changes Upon Host Macrophages Infection

Alonso‐Hearn

Abendaño

Ruvira

et al. 2017

Front. Cell. Infect. Microbiol.

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Johne's disease is a chronic granulomatous enteritis of ruminants caused by the intracellular bacterium Mycobacterium avium subsp. paratuberculosis (Map). We previously demonstrated that Map isolates from sheep persisted within host macrophages in lower CFUs than cattle isolates after 7 days of infection. In the current study, we hypothesize that these phenotypic differences between Map isolates may be driven be the fatty acids (FAs) present on the phosphadidyl-1-myo-inositol mannosides of the Map cell wall that mediate recognition by the mannose receptors of host macrophages. FAs modifications may influence Map's envelope fluidity ultimately affecting pathogenicity. To test this hypothesis, we investigated the responses of two Map isolates from cattle (K10 isolate) and sheep (2349/06-1) to the bovine and ovine macrophage environment by measuring the FAs content of extracellular and intracellular bacteria. For this purpose, macrophages cell lines of bovine (BOMAC) and ovine (MOCL-4) origin were infected with the two isolates of Map for 4 days at 37°C. The relative FAs composition of the two isolates recovered from infected BOMAC and MOCL-4 cells was determined by gas chromatography and compared with that of extracellular bacteria and that of bacteria grown in Middlebrook 7H9 medium. Using this approach, we demonstrated that the FAs composition of extracellular and 7H9-grown bacteria was highly conserved within each Map isolate, and statistically different from that of intracellular bacteria. Analysis of FAs composition from extracellular bacteria enabled the distinction of the two Map strains based on the presence of the tuberculostearic acid (18:0 10Me) exclusively in the K10 strain of Map. In addition, significant differences in the content of Palmitic acid and cis-7 Palmitoleic acid between both isolates harvested from the extracellular environment were observed. Once the infection established itself in BOMAC and MOCL-4 cells, the FAs profiles of both Map isolates appeared conserved. Our results suggest that the FAs composition of Map might influence its recognition by macrophages and influence the survival of the bacillus within host macrophages.

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Identification of Three Strains of Mycobacterium Species Isolated from Clinical Samples Using Fatty Acid Methyl Ester Profiling

Cited by 19 publications

References 12 publications

Fatty acid variability in three medicinal herbs of Panaxspecies

Fatty acid variability in three medicinal herbs of Panaxspecies

Polyphasic Taxonomic Analysis Establishes Mycobacterium indicus pranii as a Distinct Species

Mycobacterium avium subsp. paratuberculosis (Map) Fatty Acids Profile Is Strain-Dependent and Changes Upon Host Macrophages Infection

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