As an important source of human food, milk can be a carrier of human pathogenic bacteria, including tuberculous and nontuberculous mycobacteria (NTM), in its raw and unpasteurized state. In this research, 175 raw milk samples and 175 traditional cheese samples were collected from traditional dairy stores in 22 regions of Tehran in a 9- month period from August 2019 to May 2020. Samples were prepared and transferred to a specialized laboratory, where they were inoculated in Lowenstein-Jensen (LJ) medium containing glycerol or sodium pyruvate, as well as Herrold's egg-yolk with and without Mycobactin J. to determine the sample's identity of samples. The recommended 16S rRNA (1436 bp) and hsp65 (644 bp) gene fragments from the positive isolates identified in Ziehl-Neelsen (Z-N) staining were amplified and sequenced using PCR and compared with the sequences of the gene fragments of reference strains available in the global GenBank database. No mycobacterial species were isolated from traditional cheese samples in microbial culture. In case of raw milk samples, a total of four bacteria were collected, all of which were found in the genetic differential testing to be NTM, including n = 1 Mycobacterium heraklionense, n = 2 Mycolicibacterium fortuitum, and n = 1 Mycobacterium thermoresistibile. The analysis of the results obtained by isolate sequencing using the 16S rRNA gene showed higher discriminatory power and percentage similarities in the identification of the isolates than the hsp65 gene.
- MeSH
- atypické mykobakteriální infekce * mikrobiologie MeSH
- lidé MeSH
- mléko mikrobiologie MeSH
- netuberkulózní mykobakterie genetika MeSH
- RNA ribozomální 16S genetika MeSH
- sýr * mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Írán MeSH
The Mycobacterium marinum group (MMG) is a class of mycobacteria that includes M. marinum, the cause of chronic systemic infections in fish. This species occasionally causes granulomatous skin lesions in humans. Other members of MMG are mycolactone-producing mycobacteria (MPM; M. ulcerans, M. shottsii and M. pseudoshottsii). The cultivation-independent approach presented in this study brings a fast and reliable alternative to classically used cultivation methods. The developed triplex erp/IS2404 qPCR assay is based on a primary species-specific erp detection, which allows enumeration of MMG in analysed samples, and secondary IS2404 detection is suitable for the differentiation of M. marinum from MPM. The detection of M. marinum in clinical specimens and in artificially contaminated tissue samples has proven its applicability for diagnostic purposes.
- MeSH
- atypické mykobakteriální infekce diagnóza mikrobiologie MeSH
- bakteriální proteiny genetika MeSH
- bakteriologické techniky metody normy MeSH
- diagnostické techniky molekulární metody normy MeSH
- druhová specificita MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- multiplexová polymerázová řetězová reakce MeSH
- Mycobacterium marinum genetika izolace a purifikace MeSH
- nemoci ryb diagnóza mikrobiologie MeSH
- netuberkulózní mykobakterie genetika izolace a purifikace MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Mycobacteriosis is a progressive disease of a wide range of wild and captive, marine and freshwater fish species. Conventional detection of fish Mycobacteria is based on histopathology, culture, and biochemical characteristics. The present study analyzed the occurrence of Mycobacteria in clinically ill ornamental fish of different species, from different places of India. In first group, 60 fish were examined for presence of granulomatous inflammation and acid-fast bacteria. Thirty-eight (63.34 %) fish were positive for granulomatous inflammations. Presences of acid-fast bacteria were detected in 27 (45 %) fish having granulomatous inflammation and in two (3.33 %) fish without granulomatous inflammation. In total, AFB were found in 29 (48.34 %) of the 60 fish examined. In second group, 20 fish having granulomatous inflammation, 12 (60 %) samples were positive using Ziehl-Neelsen (Z-N) staining and 11 (55 %) of them were culture positive. Eight (40 %) samples were Z-N negative but two (10 %) of them were culture positive. In total, 13 (65 %) of the 20 examined fish were culture positive. On the basis of biochemical tests and 16S rRNA sequencing, 13 isolates were identified: five as Mycobacterium fortuitum, five as Mycobacterium gordonae, and three as Mycobacterium chelonae. In comparison of two decontamination methods, 2 % HCl treatment was better than 4 % NaOH treatment. Mycobacteria recovery from decontaminated samples was significantly high on Lowenstein-Jensen medium compared to Middlebrook 7H11 agar and Stonebrink (SB) media. The disease is transmissible from fish to fish and also from fish to human, so the significance of Mycobacteria in ornamental fish should not be overlooked.
- MeSH
- DNA bakterií chemie genetika MeSH
- histocytochemie MeSH
- mikroskopie MeSH
- Mycobacterium chelonae klasifikace genetika izolace a purifikace fyziologie MeSH
- Mycobacterium fortuitum klasifikace genetika izolace a purifikace fyziologie MeSH
- mykobakteriózy mikrobiologie patologie veterinární MeSH
- nemoci ryb mikrobiologie patologie MeSH
- netuberkulózní mykobakterie klasifikace genetika izolace a purifikace fyziologie MeSH
- ribozomální DNA chemie genetika MeSH
- RNA ribozomální 16S genetika MeSH
- ryby mikrobiologie MeSH
- sekvenční analýza DNA MeSH
- techniky typizace bakterií MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Indie MeSH
The isolation of nontuberculous mycobacteria (NTM) from clinical specimens has become very common in recent years. Such organisms are typically environmental and occasionally pathogenic for humans and animals. Standard diagnosis of mycobacterial infections relies on direct examination and culture. However, molecular tools are now available which allow quicker and more accurate diagnosis. Detection of NTM can be performed directly from clinical samples, although identification is mostly carried out after isolation. Sequencing of genomic targets (such as 16S rRNA, ITS, rpoB or hsp65) allows accurate and rapid identification, but has some technical limitations. A brief summary of the molecular methods available for NTM identification and a discussion of the problems associated with the use of sequencing analysis together with a description of available algorithms for NTM identification are the major objectives of this review.
- MeSH
- algoritmy MeSH
- bakteriální RNA analýza genetika MeSH
- DNA bakterií analýza genetika MeSH
- genom bakteriální MeSH
- lidé MeSH
- mykobakteriózy diagnóza mikrobiologie MeSH
- netuberkulózní mykobakterie klasifikace genetika izolace a purifikace MeSH
- polymerázová řetězová reakce metody MeSH
- reprodukovatelnost výsledků MeSH
- RNA ribozomální 16S analýza genetika MeSH
- sekvenční analýza DNA metody MeSH
- senzitivita a specificita MeSH
- techniky typizace bakterií metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- MeSH
- Bacteria MeSH
- elektroforéza metody přístrojové vybavení využití MeSH
- genetický výzkum MeSH
- imunohistochemie metody využití MeSH
- imunologické techniky metody využití MeSH
- izoenzymy diagnostické užití izolace a purifikace MeSH
- lidé MeSH
- Mycobacterium kansasii genetika izolace a purifikace MeSH
- Mycobacterium * enzymologie genetika izolace a purifikace MeSH
- netuberkulózní mykobakterie enzymologie genetika izolace a purifikace MeSH
- oxidoreduktasy * izolace a purifikace klasifikace MeSH
- statistika jako téma MeSH
- tetrazoliové soli * diagnostické užití MeSH
- Check Tag
- lidé MeSH
