Isolation Of Mycobacteria Research Articles

Comparative Evaluation between Detection of Mycobacterium Tuberculosis Complex in Samples of Extra Pulmonary Tuberculosis using Gene Xpert MTB/RIF Assay and Ziehl-Neelsen Staining in a Tertiary Care Hospital

Tuberculosis is a global problem and incidence of extra pulmonary tuberculosis is increasing day by day. Diagnosis of extra pulmonary tuberculosis is a little bit difficult than pulmonary tuberculosis.Aim and Objective: The aim of this study is to correlate between isolates of mycobacterium in GeneXpert and ZiehlNeelsen (ZN) staining. It also detects the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of GeneXpert assay and ZN staining.This prospective study was carried out in the Department of Microbiology, RIMS, Ranchi, Jharkhand. Seventy two samples from suspected patients of Extra pulmonary tuberculosis were taken. These samples were processed for test in GeneXpert assay, ZN staining and MGIT culture. Mycobacterium tuberculosis complex isolated from culture was taken as gold standard and compared with result of GeneXpert and ZN staining. Result: Out of 72 samples, detection rate of GeneXpert, MGIT and ZN staining were 27.77%, 26.38% and 18%. The sensitivity, specificity, PPV and NPV of ZN staining and GeneXpert were 63.15%, 98.11%, 92.3%, 88.13% and 94.73%, 96.22%, 90%, 98% respectively. GeneXpert is a rapid and easy method for extra pulmonary tuberculosis diagnosis. It not only detect the bacilli but also diagnose rifampicin drug resistance. This method prompts in diagnosis and treatment.

Prevalence of nontuberculous mycobacteria in Gomel region

Objective: to assess the species diversity and frequency of detection of nontuberculous mycobacteria (NTM) in patients of antituberculous organizations in Gomel region. Material and methods. 1057 isolates of mycobacteria were analyzed. To identify the species of the mycobacteria, phenotypic and molecular genetic studies of the isolates were performed. Results. A total of 13 NTM types have been identified. The most common were M. avium and M. intracellulare (MAC complex) - 63.7 % (60.79-66.67), followed by M. gordonae and M. fortuitum (7.66 %; 6.13-9.43 and 7.57 %; 6.05-9.33, respectively). Multiple isolation of NTMs of one species (from 2 or more samples) was observed in 190 patients, which amounted to 37.4 % of all the patients. In 75.3 % of the cases, MAC was obtained (χ2 = 97.01; p <0.001). The second place in terms of etiological significance was occupied by M. xenopi (6.43 %), the third one - by M. gordonae (5.3 %). Conclusion. Out of 13 identified NTMs, the most common causes of the development of mycobacteriosis were MAС complex, M. xenopi , M. gordonae .

Typing and classification of non-tuberculous mycobacteria isolates

Background: There are a large and growing number of non-tuberculous mycobacteria (NTM) species that have been isolated, identified, and described in the literature, yet there are many clinical isolates which are not assignable to known species even when the genome has been sequenced. Additionally, a recent manuscript has proposed the reclassification of the Mycobacterium genus into five distinct genera. Methods: We describe using a community standard fast average nucleotide identity (ANI) approximation method, MASH, for classifying NTM genomes by comparison to a resource of type strain genomes and proxy genomes. We evaluate the genus reclassification proposal in light of our ANI, MLST, and pan-genome work. Results: We describe here a sequencing study of hundreds of clinical NTM isolates. To aid in characterizing these isolates we defined a multi-locus sequence typing (MLST) schema for NTMs which can differentiate strains at the species and subspecies level using eight ribosomal protein genes. We determined and deposited the allele profiles for 2,802 NTM and Mycobacterium tuberculosis complex strains in PubMLST. Conclusions: The MLST schema and our pan-genome analysis of Mycobacteria can help inform the design of marker-gene diagnostics. The ANI comparisons likewise can assist in the classification of unknown genomes, even from previously unknown species.

Whole genome sequencing of Nontuberculous Mycobacterium (NTM) isolates from sputum specimens of co-habiting patients with NTM pulmonary disease and NTM isolates from their environment

BackgroundNontuberculous mycobacterium (NTM) species are ubiquitous microorganisms. NTM pulmonary disease (NTM-PD) is thought to be caused not by human-to-human transmission but by independent environmental acquisition. However, recent studies using next-generation sequencing (NGS) have reported trans-continental spread of Mycobacterium abscessus among patients with cystic fibrosis.ResultsWe investigated NTM genomes through NGS to examine transmission patterns in three pairs of co-habiting patients with NTM-PD who were suspected of patient-to-patient transmission. Three pairs of patients with NTM-PD co-habiting for at least 15 years were enrolled: a mother and a daughter with M. avium-PD, a couple with M. intracellulare-PD, and a second couple, one of whom was infected with M. intracellulare and the other of whom was infected with M. abscessus. Whole genome sequencing was performed using patients’ NTM isolates as well as environmental specimens. Genetic distances were estimated based on single nucleotide polymorphisms (SNPs). By comparison with the genetic distances among 78 publicly available NTM genomes, NTM isolates derived from the two pairs of patients infected with the same NTM species were not closely related to each other. In phylogenetic analysis, the NTM isolates from patients with M. avium-PD clustered with isolates from different environmental sources.ConclusionsIn conclusion, considering the genetic distances between NTM strains, the likelihood of patient-to-patient transmission in pairs of co-habiting NTM-PD patients without overt immune deficiency is minimal.

Gene mutations related to rifampin resistance of tuberculosis in northwest of Iran

BackgroundMycobacterium tuberculosis causes tuberculosis disease, is a communicable disease and multidrug-resistant or rifampicin-resistant M. tuberculosis continuous to be a public health threat worldwide. Therefore, the aim of this study was the assessment of the mutations profile of rifampin-resistant and MDR M. tuberculosis in the north-west of Iran. Material and methodsThe specimens of suspicious individuals were collected and processed by direct microscopy and culture. Drug susceptibility testing was assessed using the conventional proportional method, and the rpoB genes of rifampin-resistant isolates were amplified and sequence analyses were performed. ResultsA total of 6294 individuals referred, 295 (4.68%) isolates of Mycobacterium spp. were isolated, which only 5 (1.69%) isolates were found resistant to rifampin. Three of the rifampin-resistant isolates were multidrug-resistant, originated from Azerbaijani patients, and two from Iran that were monoresistant to rifampin. Three different types of mutations were identified in the rpoB gene, involving four codons Gln510His, Asp516Val, His526Tyr and Ser531Leu. The most frequent codons affected by point mutation were codons 531 and 526. The katG315 and emb306 mutations were responsible for isoniazid- and ethambutol-resistant isolates. ConclusionThe results of this study revealed that the frequency of the mutations in the RRDR region of the rpoB gene were low, but most of the isolates were collected from patients originated from Rep. of Azerbaijan, which have a high potential for transmission of MDR tuberculosis. These results indicate the importance of policies to restrict infection distribution in the region by patients from neighbor countries. The rate of mutations in the isolates of the present study was lower than the rest of the world.

Typing and classification of non-tuberculous mycobacteria isolates

Background: There are a large and growing number of non-tuberculous mycobacteria (NTM) species that have been isolated, identified, and described in the literature, yet there are many clinical isolates which are not assignable to known species even when the genome has been sequenced. Additionally, a recent manuscript has proposed the reclassification of the Mycobacterium genus into five distinct genera. Methods: We describe using a fast average nucleotide identity (ANI) approximation method, MASH, for classifying NTM genomes by comparison to a resource of type strain genomes and proxy genomes. We evaluate the genus reclassification proposal in light of our ANI, MLST, and pan-genome work. Results: We describe here a sequencing study of hundreds of clinical NTM isolates. To aid in characterizing these isolates we defined a multi-locus sequence typing (MLST) schema for NTMs which can differentiate strains at the species and subspecies level using eight ribosomal protein genes. We determined and deposited the allele profiles for 2,802 NTM and Mycobacterium tuberculosis complex strains in PubMLST. Conclusions: The MLST schema and our pan-genome analysis of Mycobacteria can help inform the design of marker-gene diagnostics. The ANI comparisons likewise can assist in the classification of unknown genomes, even from previously unknown species.

Generation of mycobacterial lipoarabinomannan-specific monoclonal antibodies and their ability to identify mycobacterium isolates.

The World Health Organization has recommended commercial urine-sourced lipoarabinomannan (LAM) detection as a tool for screening HIV patients with suspected TB, but more sensitive immunodetection assays would help to identify HIV-negative TB patients. Here, we aimed to develop novel rabbit monoclonal antibodies (mAbs) against LAM for immunodetection purposes. Rabbits were immunized with cell-wall components from the Mycobacterium tuberculosis (Mtb) H37Rv strain. An immune single-chain fragment variable (scFv) phage display library was generated. The scFv mAbs to LAM were identified through ELISA screening. The light and heavy chain variable region genes from the selected clones were sequenced. Vectors containing the full-length light and heavy chains were constructed and co-expressed in 293T cells to generate whole IgG antibodies. The performances and binding characteristics of the mAbs against purified LAM from M.tb H37Rv, multiple mycobacteria species (M.tb H37Rv, M. bovis and non-tuberculous mycobacteria (NTM) strains), and mycobacteria clinical isolates (Mtb and NTM isolates) were determined using various immunoassay methods. We obtained five rabbit mAbs against LAM, four of which had high sensitivities (100pg/ml) and affinities (1.16-1.73×10-9M) toward LAM. They reacted with M.tb H37Rv, M. bovis, and slow-growing NTM, but not with rapid-growing NTM. Similar results were obtained with mycobacterium isolates, where 96% of the Mtb isolates and 90% of the M. avium-intracellulare isolates were successfully identified. The novel rabbit LAM-specific mAbs performed well at recognizing LAM from slow-growing pathogenic mycobacteria, which support their future clinical application.

Nontuberculous mycobacterial infections in a Brazilian pediatric population: a seven-year survey

ABSTRACTThe aim of this study was to describe the frequency of nontuberculous mycobacteria (NTM) isolation and their related outcomes among pediatric patients of a Brazilian university hospital from 2012 to 2019. NTM were identified in different clinical samples by microbiological culture and molecular-based methods. NTM were isolated from 14 patients, out of whom four (27%) were infected and were treated accordingly. Two were infected with Mycobacterium avium complex (MAC), two with M. abscessus complex (MABSC) and one with M. intracellulare. Two patients had cystic fibrosis-related lung disease and improved after successful NTM eradication. One patient was HIV-positive and died. One patient had severe combined immunodeficiency (SCID)-related pneumonia and is currently being followed-up. We conclude that NTM frequency in our center was low among pediatric patients. Whether this is inherent to Brazilian patients, due to the broad coverage of the Bacille Calmette-Guérin (BCG) vaccine in Brazil, or a result of underdiagnosis remains to be elucidated.

In Vitro Susceptibility Testing of GSK656 against Mycobacterium Species.

In this study, we aimed to assess the in vitro susceptibility to GSK656 among multiple mycobacterial species and to investigate the correlation between leucyl-tRNA synthetase (LeuRS) sequence variations and in vitro susceptibility to GSK656 among mycobacterial species. A total of 187 mycobacterial isolates, comprising 105 Mycobacterium tuberculosis isolates and 82 nontuberculous mycobacteria (NTM) isolates, were randomly selected for the determination of in vitro susceptibility. For M. tuberculosis, 102 of 105 isolates had MICs of ≤0.5 mg/liter, demonstrating a MIC50 of 0.063 mg/liter and a MIC90 of 0.25 mg/liter. An epidemiological cutoff value of 0.5 mg/liter was proposed for identification of GSK656-resistant M. tuberculosis strains. For NTM, the MIC50 and MIC90 values were >8.0 mg/liter for both Mycobacterium intracellulare and Mycobacterium avium In contrast, all Mycobacterium abscessus isolates had MICs of ≤0.25 mg/liter, yielding a MIC90 of 0.063 mg/liter. LeuRS from M. abscessus showed greater sequence similarity to M. tuberculosis LeuRS than to LeuRSs from M. avium and M. intracellulare Sequence alignment revealed 28 residues differing between LeuRSs from M. avium and M. intracellulare and LeuRSs from M. tuberculosis and M. abscessus; among them, 15 residues were in the drug binding domain. Structure modeling revealed that several different residues were close to the tRNA-LeuRS interface or the entrance of the drug-tRNA binding pocket. In conclusion, our data demonstrate significant species diversity in in vitro susceptibility to GSK656 among various mycobacterial species. GSK656 has potent efficacy against M. tuberculosis and M. abscessus, whereas inherent resistance was noted for M. intracellulare and M. avium.

A rare case of spinal tuberculosis due to Mycobacterium bovis. Is zoonotic tuberculosis underdiagnosed?

We report a rare presentation of Pott's disease caused by M. bovis, suggesting transmission from infected cattle, and only the second case described so far in scientific reports. Noteworthy of this case was that the strain was only isolated on Stonebrink medium, a sodium pyruvate-containing culture medium for the isolation of mycobacteria. This medium is frequently ignored in diagnostic laboratories and in the laboratory manuals of most international health organizations. In general laboratories use a culture medium that contains glycerol, a carbon substrate considered inhibitory for the growth of M. bovis. The use of glycerol-containing medium therefore likely contributes towards underestimating zoonotic tuberculosis. Our case suggests that, in order to improved surveillance efforts for zoonotic TB and increase the notification rate for M. bovis to human TB, the use of pyruvate-containing media should be promoted, particularly in developing countries with a high prevalence of bovine TB, but also through the World Health Organization' (WHO) End TB Strategy and the Roadmap for Zoonotic TB.

An improved simple method for the identification of Mycobacteria by MALDI-TOF MS (Matrix-Assisted Laser Desorption- Ionization mass spectrometry)

The aim of this study was to establish a simple method for the rapid identification of Mycobacteria species by MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass spectrometry) using the Bruker MALDI-TOF Biotyper system (Bruker Daltonik, Bremen, Germany). A multicentre, prospective, and single blind study was performed in three European Hospitals, two Spanish and one UK hospital from May to August 2018. The BD BACTEC MGIT (Becton Dickinson, Berks, UK) liquid culture system was used in all three centres for the growth of Mycobacteria. When signal positive, tubes were removed from the analyser and in addition to standard laboratory procedures were subcultured on blood agar plates for MALDI-TOF analysis. Plates were incubated aerobically for 1 to 7 days at 37 °C and inspected every day. Once any growth was visible, it was transferred to the steel target plate, overlaid with 1 μl of neat formic acid and 1 μl HCCA matrix (alpha hydroxyl 4 cinnamic acid), and analysed in a Bruker Biotyper MALDI-TOF. Results given by MALDI-TOF were compared with the reference methods used for identification in the different centres. At two Spanish hospitals, identification by MALDI-TOF was only attempted on presumptive non-tuberculosis mycobacteria (NTM) and the results were initially compared with the results obtained by a commercial reverse hybridisation assay, GenoType CM/AS (Hain Lifescience, Tübingen, Germany). At the UK Hospital, identification of any presumptive mycobacteria was attempted and compared with the results obtained by whole genome sequencing (WGS). Overall in 142/167 (85%) of cases the identifications obtained were concordant; all Mycobacterium tuberculosis (MTB) isolates 43/43 (100%), 57/76 (75%) of the rapid growing nontuberculous mycobacteria (NTM), and 42/48 (85%) slow growing NTM tested were identified correctly. We report a new, easy, cheap and quick method for isolation and identification of Mycobacterium spp. without the need for additional steps or equipment and this method is in routine used in all three centres.

Optimized Method for Isolation of Nontuberculous Mycobacteria from Hospital Aquatic Sources

Optimized Method for Isolation of Nontuberculous Mycobacteria from Hospital Aquatic Sources

Predictors of radiographic progression for NTM-pulmonary disease diagnosed by bronchoscopy.

A single isolate of nontuberculous mycobacterium (NTM) from bronchoscopic samples satisfies the microbiological criterion for diagnosing NTM-pulmonary disease (PD). Studies investigating patients with NTM-PD and multiple culture-negative sputum samples but culture-positive bronchoscopic samples are lacking. We investigated the clinical characteristics, outcome, and predictors of radiographic progression in this special population. Patients with negative NTM culture from ≥2 expectorated sputum samples within the 3 months prior to bronchoscopy diagnosis of NTM-PD between 2009 and 2017 were included. Patient characteristics and clinical course were described. Predictors for radiographic progression of NTM-PD within 2 years were analysed by using multivariate logistic regression. Among 66 patients with bronchoscopy-diagnosed NTM-PD, radiographic progression occurred within 2 years in 17 (26%). Of the 60 patients not initially treated, radiographic progression occurred in 17 (28%). Among them, 10 never received treatment, with 6 deteriorating and 3 dying. Of the 6 and 7 patients who received treatment immediately after NTM-PD diagnosis and after radiographic progression, respectively, none had further radiographic progression. The independent predictors of radiographic progression were male sex, body mass index <18.5kg/m2, use of inhaled corticosteroids, and acid-fast smear grade ≥2 of index bronchoscopic samples. Among patients with bronchoscopy-diagnosed NTM-PD, one fourth experienced radiographic progression within 2 years. The risk was even higher in those with the aforementioned predictors, immediate treatment or close monitoring is recommended. For others, conservative management by regular microbiological monitoring for sputum samples and image follow-up may be the optimal choice.

Whole-Genome Sequencing of a Mycobacterium orygis Strain Isolated from Cattle in Chennai, India.

Here, we report the isolation of Mycobacterium orygis from dairy cattle in Chennai, India. Spoligotyping assigned the isolate to spoligotype 587 (ST587), which belongs to M. orygis This species was confirmed as M. orygis using whole-genome sequencing.

Nontuberculous mycobacteria isolated from specimens of pulmonary tuberculosis suspects, Northern Tunisia: 2002\u20132016

BackgroundReports on the worldwide ascending trend of pulmonary nontuberculous mycobacteria (NTM) isolation rates and their effective role in respiratory tract infections are compelling. However, as yet, there are no such data relating to Tunisia.MethodsHere we carried out a retrospective review of mycobacterial cultures originating from Northern Tunisia, which have been processed in the laboratory of mycobacteria of the Institut Pasteur de Tunis, during the time period 2002–2016. All pulmonary NTM (PNTM) isolates available for culture were characterized phenotypically and their taxonomic status was further established based on polymorphisms in rpoB, 16S rRNA, hsp65, and sodA DNA gene sequences.ResultsOf the 10,466 specimens collected from HIV-negative Tunisian patients with presumptive clinical pulmonary TB, 60 (0.6%) yielded PNTM isolates. An overall annual PNTM isolation prevalence of 0.2/100,000 was estimated. As far as could be ascertained, this isolation rate accounts amongst the lowest reported hitherto throughout the world. Among the 30 NTM isolates that were available for culture, 27 (90.0%) have been identified to the species level. The most commonly encountered species was Mycobacterium kansasii (23.3%) subtype 1. Strikingly, all M. kansasii cases were male patients originating from Bizerte, an industrialized region particularly known for iron industry. The remaining NTM species were M. fortuitum (16.6%), M. novocastrense (16.6%), M. chelonae (10.0%), M. gordonae (6.6%), M. gadium (6.6%), M. peregrinum (3.3%), M. porcinum (3.3%), and M. flavescens (3.3%). There were no bacteria of the M. avium complex, the most frequently isolated NTM globally, and the main driver of the rise of NTM-lung diseases.ConclusionsThis study uncovered an exceptional low prevalence of PNTM isolation among HIV-negative TB suspects in Northern Tunisia, suggesting a very low burden of NTM pulmonary disease. However, the frequent isolation of M. kansasii subtype 1, the most pathogenic subtype, particularly from the industrialized region of Bizerte, strongly suggests its effective involvement in a typical pulmonary disease.

Resistance To First-Line Antituberculosis Drugs And Prevalence Of pncA Mutations In Clinical Isolates Of Mycobacterium tuberculosis From Zunyi, Guizhou Province Of China.

BackgroundChina is one of the high-burden countries for multidrug-resistant tuberculosis (MDR-TB), and pyrazinamide is one of the anti-TB drugs used for the shorter MDR-TB treatment regimen. The aim of this study was to determine the correlation between pncA gene mutations and resistance to four first-line anti-TB drugs as well as treatment history in clinical isolates of Mycobacterium tuberculosis.Patients and methodsM. tuberculosis clinical isolates were collected from 318 in-patients with smear-positive TB between October 2008 and September 2016 at a major hospital in Zunyi, Guizhou Province of China, and used for drug susceptibility testing against four first-line anti-TB drugs. Genomic DNA extracted from clinical isolates was used for PCR amplification and DNA sequencing of the pncA gene.ResultsAmong 318 clinical isolates, 129 (40.6%), 170 (53.5%), 66 (20.8%) and 109 (34.3%) were resistant to rifampicin, isoniazid, ethambutol and streptomycin respectively. In addition, 124 clinical isolates were MDR-TB and 71.8% of them were previously treated cases. Sequencing results showed that 46.8% of MDR-TB and 2.2% of drug susceptible isolates harbored a pncA mutation, and 52 types of pncA mutations were detected from 64 isolates. The prevalence of pncA mutations in isolates resistant to first-line anti-TB drugs and previously treated TB cases was significantly higher than that in drug-susceptible isolates and new cases of TB.ConclusionHigh prevalence of pncA mutations in clinical isolates of M. tuberculosis from Zunyi, Guizhou Province of China, is correlated with resistance to four first-line anti-TB drugs, MDR-TB and previously treated TB cases.

Phenotypic characterization of mycobacteria isolates from tuberculosis patients in Kaduna State, Nigeria

Background: Tuberculosis (TB) remains one of the leading public health challenges in Nigeria and the burden is still high. There is hence a need for continuous characterization of mycobacteria to obtain current data that will aid the ongoing TB prevention and control programme. The aim of this study was to phenotypically characterize mycobacteria isolates recovered from clinical specimens of patients with tuberculosis in Kaduna State, Nigeria.Methods: Two thousand, two hundred and twelve (2212) sputum samples were collected from patients clinically suspected to have TB in three different zones of Kaduna State, Nigeria, between May 2017 and October, 2018. Samples were processed by decontaminating with NaOH-Citrate N-acetyl-L-Cystein method for Ziehl Neelsen (ZN) AFB microscopy and culture on Lowenstein Jensen (LJ) slants which were incubated at 37ᵒC for 8 weeks. Positive LJ cultures were further analyzed with a rapid TB antigen assay (SD-Bioline) to differentiate Mycobacterium tuberculosis complex (MTBC) from Non Tuberculous Mycobacteria (NTM).Results: Out of the 2212 patients with suspected TB, 300 (13.6%) were positive for AFB by microscopy with Zone A (Kaduna North) having the highest AFB positive cases of 169 (15.2%). Of the 300 AFB positive samples, 272 (91.0%) were culture positive on LJ medium, 18 (6.0%) were culture negative and 10 (3.0%) were culture contaminated. Result of the distribution of mycobacteria among infected patients within the study area revealed that 219 (80.5%) were infected with MTBC, 42 (15.4%) with NTM and 11 (4.0%) with both MTBC and NTM.Conclusion: A relatively high number of TB in the study area was caused by NTM. There is need for advanced diagnostic tools that can differentiate MTBC and NTM strains among TB patients in all TB Reference Laboratories in Nigeria.Keywords: Phenotypic, Characterization, Tuberculosis, Mycobacteria

In vitro Antibacterial and Antitubercular Activities of Leaf Extracts of Senna occidentalis

Aim: This study aimed to evaluate the antibacterial and antitubercular activities of ethylacetate and ethanol leaf extracts of Senna occidentalis.&#x0D; Study Design: Fresh leaves of Senna occidentalis collected from Suleja, Niger state were used for this study against some medically important micro-organisms viz; Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, Salmonella paratyphi, Klebsiella pneumoniae, Mycobacterium bovis and Mycobacterium smegmatis.&#x0D; Place and Duration of Study: The study was conducted in Abuja, Nigeria at the Department of Microbiology and Biotechnology, National Institute for Pharmaceutical Research and Development [NIPRD], from February 2019 to April 2019.&#x0D; Methodology: Senna occidentalis leaves were extracted successively with ethyl-acetate and ethanol. The obtained extracts were tested in vitro for antibacterial activity by agar well diffusion method, while anti-tubercular screening was carried out by broth micro-dilution method. A fixed-dose concentration of chloramphenicol was used as a control drug against the bacterial isolates while isoniazid was used as control drug against the mycobacterium isolates.&#x0D; Results: The in vitro antibacterial screening showed that the crude extracts exhibited varying activity against the different microbes with highest zone of inhibition at 12 mm, and anti-tubercular activity with MICs ranging from 97.6-390.6 μg/mL. Among these extracts, ethyl-acetate extract showed significant antibacterial activity against most of the test micro-organisms. The most susceptible micro-organism was P. aeruginosa (12mm zone in ethyl-acetate at 80 mg/mL) followed by B. subtilis (10 mm zone in ethyl-acetate extract at 80 mg/mL) and E. coli (9 mm zone in ethyl-acetate extract at 80 mg/mL). The ethanol extract was the most effective in inhibiting the growth of M. smegmatis and M. bovis with MICs of 97.6 μg/mL and 195.3 μg/mL.&#x0D; Conclusion: The activities observed could be attributed to the presence of some active metabolites contained in the extracts which could be useful in drug development for therapeutic purposes.

Isolation and identification of Nontuberculous mycobacteria from environmental water samples in the northeast of Iran

Isolation and identification of Nontuberculous mycobacteria from environmental water samples in the northeast of Iran

Experience of selection, indication and identication of non-tuberculous mycobacteria, got during I National research on study of distribution of drugresistant of tuberculosis in Ukraine

Objective — a study of presence of non-tuberculosis mycobacteriumin a circulatory mycobacterial population among patients by a white plague in Ukraine, finding out of etiologic role of non-tuberculosis mycobacterium, their selection and identification, study of some morphological and biological properties.Materials and methods. The strokes of sputum were investigational in the presence of acid proof bacteria by a direct microscopy after coloring on the method of Ziehl—Neelsen. For all standards with the positive results of microscopy all spectrum of bacteriologicexaminations was executed on diagnostics of tubercu­losis, according to the algorithm ratified by the order of MH of Ukraine N 620. Bacteriological studies were undertaken on identification of clinical isolates on standard methodologies that is expounded in the order of MH of Ukraine N 45. Specific identification of the got cultures consisted of two stages: primary and final.In process for sowing environments were used: liquid — broth of Middlebrook 7Н9 and dense egg Lowenstein—Jensen medium. Pressed treatment of clinical material was carried out by means of a 0,5 % sterile solution of N-acetyl-L-of cystein from 1.0 % gidroxid of natrium (BBL Mycoprep NALC-NAOH). Test tubes placed in the system ВАСТЕС МGIТ 960 and did incubation. Sowing was carried out in parallel on the dense environment of Lowenstein—Jensen medium. For identification of the distinguished strains ID МТВ MGIT test used and determination of ability of height of clinical isolates of mycobacterium on an environment with a salicylic sour natrium and p-nitrodracylic acid with the purpose of determination of belonging of distinguished mycobacterium to the tuberculosis complex.For realization of final identification of mycobacterium used the system GenoType® MTB DR (Нain, Laifsains), that allowed to define the type of non-tuberculosis mycobacterium.Results and discussion. During Ist of National research on the study of distribution of drug resistant of tuberculosis1598 clinical isolates of mycobacterium were got in Ukraine. The analysis of results of bacteriologicexamination educed, that 48 (2.9 %) clinical isolates had been identified as non-tuberculosis mycobacterium. A chart over of primary authentication of non-tuberculosis mycobacteriumis brought.Nonphotochromogen mycobacterium prevailed in the structure of distinguished non-tuberculosis mycobacterium– 77.1 %. Photochromogen mycobacterium distinguished at level — 16.7 %, fast-growing mycobacterium was made the least — 6.3 %.For determination of specific belonging of non-tuberculosis mycobacterium used the system GenoType® MTB DR (Нain, Laifsains). Next non-tuberculosis mycobacterium were distinguished: M. kansasii, M. xenopi, M. avium, M. fortuitum. From 48 distinguished strains 20 strains were the clean cultures of non-tuberculosis mycobacterium (41.7 %), 28 strains (58.3 %) were in mix (М. tuberculosis + non-­tuberculosis mycobacterium). The results of researches showed that more often from the sputum of patients M. avium was distinguished in mix with М. tuberculosis — 39.6 %, in the clean culture of M. avium was distinguished in 20.8 % cases. Minimum was specific gravity of selection of M. fortuitum — 6.25 %. Here found out reliable difference was between prevalence of the distinguished strains of M. avium and other strains of non-tuberculosis mycobacterium.Conclusions. During Ist of National research on study of distribution of drug resistant of tuberculosis 48 strains of mycobacterium (2.9 %) that were identified as non-tuberculosis mycobacterium were got in Ukraine. Thus in 20 cases (1.2 %) were found out the clean cultures of non-tuberculosis mycobacterium. In 28 (1.7 %) cases were discovered in mix, that contained as M. tuberculosis, so non-tuberculosis mycobac­terium. Authors are bringing a chart over of primary identification of non-tuberculosis mycobacterium. Nonphotochromogen mycobacterium prevailed in the structure of non-tuberculosis mycobacterium — 77.1 %. Photochromogen mycobacterium distinguished at level — 16.7 %, fast-growing mycobacterium was made the least — 6.3 %. For determination of specific belonging of non-tuberculosis mycobacterium used the system GenoType® MTB DR (Нain, Laifsains). Next non-tuberculosis mycobacterium were distinguished: M. kansasii, M. xenopi, M. avium, M. fortuitum. The results of researches showed that more often from the patient’s sputum M. avium was distinguished. In mix with М. tuberculosis – 39.6 %, in the clean culture of M. avium was distinguished in 20.8 % cases. Minimum was specific gravity of selection of M. fortuitum — 6.25 %. Here found out a reliable difference was between prevalence of the distinguished strains of M. avium and other strains of non-tuberculosis mycobacterium.