• Biomedical Science Letters
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• v.16 no.3
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• pp.133-138
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• 2010

Despite of the increasing importance of environmental mycobacteria, detection and identification of mycobacteria from environmental sources including water have been fraught with technical difficulties. Although, several protocols to optiruize isolation of mycobacteria from water sources have been reported, standard method has not yet been established. In this study, usefulness of cetylpyridinium chloride (CPC), a cationic quaternary ammonium compound, for the isolation of environmental mycobacteria from diverse water samples was evaluated. For this, water samples from diverse water sources such as effluent water, lake water, and underground water were collected, treated with diverse concentrations of CPC, and plated on the solid agar plates. Subsequently individual colonies grown on the plates were sequence analyzed for identification of each colony. In brief, the results from this study showed that the growth of mycobacteria was enhanced by use of CPC as a pre-treatment reagent to water samples by inhibiting growth of other non-mycobacteria in water. In fact, the effect of CPC to decontaminate non-mycobacteria for isolation of mycobacteria was better than 1~4% of NaOH, which is a routinely used decontaminating reagent widely employed for culturing mycobactera from sputum specimens. Therefore, the results from this study seems to support that the CPC pre-treatment may be useful for isolation of mycobacteria from diverse sources including clinical specimens which are often contaminated with other bacteria.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1207-1215
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• 2008

To investigate the occurrence and species diversity of mycobacteria in waters, surface water samples were collected monthly from the Han River and tap water samples at the terminal sites of the distribution system. Mycobacteria in each water sample were isolated by decontamination using cetylpyridinium chloride (CPC) and cultivation on Middlebrook 7H10 agar, and then identified by polymerase chain reaction-restriction fragment length polymorphism analysis (PRA) and sequencing of the 65-kDa heat-shock protein gene (hsp65 gene). Mycobacteria were detected in 59% of the surface water samples and 26% of the tap water samples. Over half of the 158 isolates could not be identified by hsp65 PRA and gene sequencing, and several identification discrepancies were observed between the two methods. The most frequently isolated species was Mycobacterium gordonae in surface water and M. lentiflavum in tap water. M. avium complex (MAC), the most important pathogen among environmental mycobacteria, was detected in the surface water samples but not found in the tap water samples. The result demonstrated that water is an important environmental source of mycobacteria and the combined application of hsp65 PRA and sequencing was more reliable than hsp65 PRA alone to accurately identify mycobacteria present in water.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.33 no.3
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• pp.355-364
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• 2023

Objectives: The objective of the present study was to identify mycobacterial infection in retired dusty workers who were ineligible for medical care benefits for work-related pneumoconiosis. Methods: Sputum samples were collected from 170 retired dusty workers living in Gangwon-do. The mycobacterial culture was grown in 2% Ogawa medium and Mycobacteria Growth Indicator Tube(MGIT). Mycobacterial species were identified using MolecuTech REBA Myco-ID^Ⓡ. Results: Thirty-one(18.2%) out of 170 sputum samples were identified as positive for culture. Among the positive culture samples, eleven(6.5%) were identified as mycobacterial species. The proportion of mycobacteria was M. avium 2.3%(4/170), M. fortuitum complex 1.2%(2/170), M. intracellulare 1.2%(2/170), M. abscessus 0.6%(1/170), M. tuberculosis(MTB) complex 0.6%(1/170), and MYC(NTM except 19 species) 0.6%(1/170). Conclusions: In comparison with previous studies, the incidence rate of tuberculosis(TB) in retired dusty workers who were ineligible for medical care benefits for work-related pneumoconiosis was higher than in close contact with TB patients, workers exposed to silica, and patients with silicosis. And the proportion of non-tuberculosis mycobacteria(NTM) was higher than that of MTB.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.1
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• pp.1-5
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• 2015

The nontuberculous mycobacteria (NTM) have been found in different environmental sources. They tend to colonize different body surfaces and secretions. The purpose of this study is to evaluate the presence of NTM in the theater environment. Fifty of Theater environment sample were examined using acid-fast stain, Lowenstein-Jensen medium culture, PCR and DNA-Sequencing. 4 of 50 samples were detected as NFB in AFB stain, L-J medium culture, PCR. and then, All of 4 NTM stains identified as Mycobacterium fortitum type in DNA-sequencing result.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.23 no.3
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• pp.169-176
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• 2013

Objectives: The aim of this study is to critically review the health effects of not only direct exposure to biocide, but also indirect exposure to by-product hazardous agents generated through the use of biocide in metalworking operations. Methods: An extensive literature review was conducted of studies reporting on respiratory disease cases, particularly hypersensitivity pneumonitis (HP), in environments using water-soluble metalworking fluids (MWFs). Keyword search terms included 'metalworking fluids', 'machining fluids', 'metalworking operation' 'machining operation' and 'biocide', which were also used in combination. Additional articles were identified in references cited in the articles reviewed. Results: Several of the field, epidemiological and experimental studies reviewed assumed that the symptoms and signs typical of HP developed in machinists who handled water-soluble MWF could be caused by inhalation exposure to nontuberculous mycobacteria (NTM). Most NTM are known to be not only resistant to both biocide and disinfectant, but also to have acid-fast cell walls that are highly antigenic. The presence or persistence of the Mycobacterium species, referred to as NTM, in metalworking fluid-using operations may be caused by NTM contamination in either the natural water or tap water that is used to dilute the base oil and additives for water-soluble MWFs. This hypothesis that NTM contamination in water-soluble MWFs is a causative agent of HP has high biologic plausibility, such as antigenic property, hydrophobicity and small diameter (< 5 um). Conclusions: Aerosolized mycobacteria colonized from MWF are likely to be causing the HP. Inhalation exposure to mycobacteria should be considered as a possible cause for the development of HP.

• Tuberculosis and Respiratory Diseases
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• v.82 no.2
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• pp.94-101
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• 2019

Nontuberculous mycobacteria (NTM) are ubiquitous organisms that are generally found not only in the natural environment but also in the human engineered environment, including water, soil, and dust. These organisms can form biofilms and can be readily aerosolized because they are hydrophobic owing to the presence of the lipid-rich outer membrane. Aerosolization and subsequent inhalation were the major route of NTM lung disease. Water distribution systems and household plumbing are ideal habit for NTM and the main transmission route from natural water to household. NTM have been isolated from drinking water, faucets, pipelines, and water tanks. Studies that used genotyping have shown that NTM isolates from patients are identical to those in the environment, that is, from shower water, showerheads, tap water, and gardening soil. Humans are likely to be exposed to NTM in their homes through simple and daily activities, such as drinking, showering, or gardening. In addition to environmental factors, host factors play an important role in the development of NTM lung disease. The incidence and prevalence of NTM lung disease are increasing worldwide, and this disease is rapidly becoming a major public health problem. NTM lung disease is associated with substantially impaired quality of life, increased morbidity and mortality, and high medical costs. A more comprehensive understanding of the infection source and epidemiology of NTM is essential for the development of new strategies that can prevent and control NTM infection.

• Journal of Environmental Health Sciences
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• v.39 no.2
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• pp.105-116
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• 2013

Background: In Korea, there is low awareness of the respiratory health problems caused by the use of humidifiers, leading to a lack of governmental measures. Objectives: The objectives of this study were to review cases of varying degrees of humidifier lung and fever in connection with the use of humidifiers and to summarize the probable environmental agents causing these cases. Methods: We searched all articles reporting on humidifier lung, humidifier fever, and humidifier diseases caused by the use of a humidifier both at home and in the workplace. Results: We summarized a number of cases of varying degrees of respiratory diseases resulting from inhalation of water mist of humidifiers containing various species of bacteria and fungi and their toxins. Type of respiratory disease connected with humidifier lung includes interstitial pneumonitis, hypersensitivity pneumonitis, fever and several respiratory symptoms. Non-tuberculous mycobacteria (NTM), Actinomycetes, endotoxins and contaminated humidifier water were the most commonly suspected probable environmental agents causing humidifier lung. In Korea, the use of humidifier biocide is suspected as a likely cause of fatal lung injury including death and lung transplantation. Conclusion: Governmental policy should be devised and measures including a national surveillance system should be taken to prevent humidifier lung caused by the use of humidifiers.

• Animal cells and systems
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• v.4 no.3
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• pp.299-304
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• 2000

In the present paper, the immunostimulatory effects of the extracellular products (ECP) from Mycobacterium spp. and various adjuvants on the non-specific immune responses of Nile tilapia, Oreochromis nilotica, were examined. Nile tilapia were immunized by injecting ECP of Mycobacterium spp. (strain TB40, TB267 or the type strain Mycobacterium marinum) into their swim bladders. A variety of adjuvants like as Freund s complete adjuvant (FCA), Freund's incomplete adjuvant (FIA) and Titremax were similarly injected into additional groups of tilapia. The number of nitroblue tetrazolium (NBT)-positive cells observed in the swim bladder of the immunized fish was signigicantly increased by the fourth day post-immunization. By day 8, the numbers of NBT-positive cells were fewer in fish immunized with ECP from mycobacteria strains TB40 or TB267 than those immunized with ECP from M. marinum or fish injected with FCA or FIA. The level of Iysozyme activity detected in the serum of fish 40 alter immunization with ECP from various Mycobacterium spp. was also significantly higher than that found in the serum of the control fish. Head kidney macrophages showed enhanced reduction of NBT when cultured in vitro with 1 $\mu$ g/ml of ECP. Concentrations greater than this (10 or 100 $\mu$g/ml) were found to suppress the reduction of NBT by the macrophages. ECP from Mycobacterium spp. and the various adjuvants used in the study all appear to be good activators of the non-specific immune responses of Nile tilapia.

• Biomedical Science Letters
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• v.19 no.2
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• pp.153-157
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• 2013

Although culture is the gold standard method to identify mycobacteria, its use in tuberculous lymphadenitis (TBL) is limited due to formalin fixation of the submitted specimens. We evaluated the performance of quantitative real-time PCR (q-PCR) for Mycobacterium Tuberculosis (MTB) in granulomatous lymphadenitis using formalin-fixed paraffin-embedded (FFPE) tissues. From 2000 to 2010, a total number of 117 cases of lymph node samples with granulomatous inflammation which were surgically removed and fixed in formalin were studied. Hematoxylin & Eosin (H&E) and Ziehl-Neelsen-stained (ZN) slides were reviewed. qPCR using Real TB-Taq$^{(R)}$ was performed for all cases to identify Mycobacterium tuberculosis. Thirteen non-tuberculous lymphadenopathy cases were used as negative control. Cervical lymph nodes were more frequently affected (60%, 70/117) than other sites. ZN stain for acid fast bacilli was positive in 19 (16.24%) cases. qPCR for tuberculosis was positive in 92 (78.63%) cases. Caseous necrosis was found in 103 (88.03%) cases. While the ZN stain and qPCR were both negative in all control cases, the qPCR showed a significantly higher positive rate (78.63% vs. 16.24%) compared to ZN stain in histologically diagnosed TBL. Quantitative real-time PCR proves to be more sensitive than ZN stain for diagnosis of tuberculous lymphadenitis.

• Biomedical Science Letters
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• v.24 no.2
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• pp.138-142
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• 2018

The incidence of tuberculosis (TB) in the Republic of Korea remains high when compared to the incidence in other Organization for Economic Cooperation and Development (OECD) countries. The prompt diagnosis and effective treatment of latent TB infection (LTBI) are very important in terms of controlling the burden of TB. The tuberculin skin test (TST) has long been the "gold standard" assay for the diagnosis of LTBI. However, it can show false positive results due to Bacille Calmette-$Gu{\acute{e}}rin$ (BCG) vaccination and infection with many environmental nontuberculous mycobacteria (NTM). The interferon gamma release assay (IGRA) using Mycobacterium tuberculosis (MTB)-specific antigens, was developed for the detection of LTBI. The QuantiFERON-TB Gold In-Tube assay is one of the most commonly used forms of the IGRA. In order to compare the diagnostic efficacy of the TST and IGRA in relation to LTBI among BCG-vaccinated healthy donors, whole blood samples were collected from 51 participants, and the results of the TST and IGRA were compared. Of the 51 cases, 18 cases (35.3%) were positive and 33 cases (64.7%) were negative when using the TST, while four cases (7.8%) were positive and 47 cases (92.2%) negative when using the IGRA. There was no correlation between the size of the induration in the TST and the $IFN-{\gamma}$ protein level. In conclusion, the TST showed higher cross-reactivity among the BCG-vaccinated healthy participants, therefore, the IGRA might be the most suitable assay for the rapid screening of LTBI in BCG-vaccinated healthy population, or for TB contact investigation.