Streptomyces sp. TR1341 was isolated from the sputum of a man with a history of lung and kidney tuberculosis, recurrent respiratory infections, and COPD. It produces secondary metabolites associated with cytotoxicity and immune response modulation. In this study, we complement our previous results by identifying the genetic features associated with the production of these secondary metabolites and other characteristics that could benefit the strain during its colonization of human tissues (virulence factors, modification of the host immune response, or the production of siderophores). We performed a comparative phylogenetic analysis to identify the genetic features that are shared by environmental isolates and human respiratory pathogens. The results showed a high genomic similarity of Streptomyces sp. TR1341 to the plant-associated Streptomyces sp. endophyte_N2, inferring a soil origin of the strain. Putative virulence genes, such as mammalian cell entry (mce) genes were not detected in the TR1341's genome. The presence of a type VII secretion system, distinct from the ones found in Mycobacterium species, suggests a different colonization strategy than the one used by other actinomycete lung pathogens. We identified a higher diversity of genes related to iron acquisition and demonstrated that the strain produces ferrioxamine B in vitro. These results indicate that TR1341 may have an advantage in colonizing environments that are low in iron, such as human tissue.
- MeSH
- fylogeneze MeSH
- genetické techniky MeSH
- genom genetika MeSH
- geny genetika MeSH
- lidé MeSH
- plíce mikrobiologie MeSH
- počet mikrobiálních kolonií metody MeSH
- Streptomyces * genetika izolace a purifikace MeSH
- tkáně mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Accurate enumeration of Paenibacillus mucilaginosus (formerly Bacillus mucilaginosus) bacterium from environmental samples on solid medium is challenging owing to its extensive extracellular polysaccharides (EPS) excretion. In the present study, P. mucilaginosus enumeration has been facilitated by a simple modification: addition of triphenyl tetrazolium chloride (TTC) to growth medium and application of a second soft agar layer. Results show distinctively better and accurate colonies' count. This method can be applied to all bacterial species that produce excessive EPS that may interfere with direct count.
AIM: Raman spectroscopy is an analytical method with a broad range of applications across multiple scientific fields. We report on a possibility to differentiate between two important Gram-positive species commonly found in clinical material - Staphylococcus aureus and Staphylococcus epidermidis - using this rapid noninvasive technique. MATERIALS & METHODS: For this, we tested 87 strains, 41 of S. aureus and 46 of S. epidermidis, directly from colonies grown on a Mueller-Hinton agar plate using Raman spectroscopy. DISCUSSION & CONCLUSION: The method paves a way for separation of these two species even on high number of samples and therefore, it can be potentially used in clinical diagnostics.
- MeSH
- bakteriologické techniky MeSH
- lidé MeSH
- počet mikrobiálních kolonií metody MeSH
- Ramanova spektroskopie metody MeSH
- stafylokokové infekce diagnóza MeSH
- Staphylococcus aureus izolace a purifikace patogenita MeSH
- Staphylococcus epidermidis izolace a purifikace patogenita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
An international standard already exists for the selective enumeration of bifidobacteria in milk products. This standard uses Transgalactosylated oligosaccharides (TOS) propionate agar supplemented with mupirocin. However, no such standard method has been described for the selective enumeration of bifidobacteria in probiotic supplements, where the presence of bifidobacteria is much more variable than in milk products. Therefore, we enumerated bifidobacteria by colony count technique in 13 probiotic supplements using three media supplemented with mupirocin (Mup; 100mg/l): TOS, Bifidobacteria selective medium (BSM) and modified Wilkins-Chalgren anaerobe agar with soya peptone (WSP). Moreover, the potential growth of bifidobacterial strains often used in probiotic products was performed in these media. All 13 products contained members of the genus Bifidobacterium, and tested mupirocin media were found to be fully selective for bifidobacteria. However, the type strain Bifidobacterium bifidum DSM 20456 and collection strain B. bifidum DSM 20239 showed statistically significant lower counts on TOS Mup media, compared to BSM Mup and WSP Mup media. Therefore, the TOS Mup medium recommended by the ISO standard cannot be regarded as a fully selective and suitable medium for the genus Bifidobacterium. In contrast, the BSM Mup and WSP Mup media supported the growth of all bifidobacterial species.
- MeSH
- antibakteriální látky metabolismus MeSH
- bakteriální nálož metody MeSH
- Bifidobacterium účinky léků růst a vývoj MeSH
- kultivační média chemie MeSH
- mupirocin metabolismus MeSH
- počet mikrobiálních kolonií metody MeSH
- probiotika analýza MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Klíčová slova
- Littorinidae, eusocialita, redie, infrapopulace,
- MeSH
- Echinostomatidae * cytologie embryologie růst a vývoj ultrastruktura MeSH
- plži parazitologie MeSH
- počet mikrobiálních kolonií metody přístrojové vybavení MeSH
- přijímání potravy MeSH
- rozmnožování MeSH
- statistika jako téma MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
This study provides information on the occurrence of Arcobacter in several types of water and food products of animal origin in the Czech Republic. We processed 190 samples using the modified method, and the occurrence of Arcobacter spp. was confirmed in 36.8 % of these. This total incidence consisted of Arcobacter butzleri (27.3 %), Arcobacter cryaerophilus (8.4 %) and Arcobacter skirrowii (1.1 %). We newly described the common presence of Arcobacter spp. in sewage water in the Czech Republic that is released into waterways after processing in water treatment plants (86.7 %). All the acquired isolates were subject to detailed confirmation with subsequent species classification using multiplex PCR and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In this study, we used a modification of a method using passive filtration of an enriched sample, which could be suitable for the isolation of Arcobacter, especially in combination with Campylobacter selective agar chromogenic medium. Our studies have shown this agar to be quite suited to the isolation of Arcobacter and that it can be an appropriate instrument for accelerating culture diagnostics.
- MeSH
- Arcobacter genetika růst a vývoj izolace a purifikace metabolismus MeSH
- kultivační média metabolismus MeSH
- odpadní vody mikrobiologie MeSH
- počet mikrobiálních kolonií přístrojové vybavení metody MeSH
- potravinářská mikrobiologie * MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
An agar selective enumeration of necrotoxigenic Escherichia coli O55 (NTEC2) and probiotic E. coli Nissle 1917, using modified MacConkey agar, was developed to study bacterial interference between these E. coli strains in a gnotobiotic piglet model. Replacement of lactose with saccharose in the agar enables the direct visual enumeration of red colonies of E. coli O55 and yellow colonies of E. coli Nissle 1917 that are co-cultured in the same Petri dish. A total of 336 colonies (168 for each color) were subjected to strain-specific PCR identification with LNA probes. Sensitivity, specificity, and positive and negative predictive values were 96.43%, 95.83%, 95.86% and 96.41% respectively in E. coli O55, and 98.21%, 97.02%, 97.06% and 98.19% respectively in E. coli Nissle 1917. Color-based enumeration of both E. coli strains in colonic contents and mesenteric lymph nodes homogenates of gnotobiotic piglets demonstrated the applicability of this method for the gnotobiotic piglet model of enteric diseases.
- MeSH
- agar chemie MeSH
- barva MeSH
- Escherichia coli klasifikace genetika růst a vývoj metabolismus MeSH
- infekce vyvolané Escherichia coli farmakoterapie mikrobiologie MeSH
- kultivační média chemie metabolismus MeSH
- lidé MeSH
- počet mikrobiálních kolonií přístrojové vybavení metody MeSH
- prasata MeSH
- probiotika aplikace a dávkování chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Real-time PCR in nuclear ribosomal DNA (nrDNA) is becoming a well-established tool for the quantification of arbuscular mycorrhizal (AM) fungi, but this genomic region does not allow the specific amplification of closely related genotypes. The large subunit of mitochondrial DNA (mtDNA) has a higher-resolution power, but mtDNA-based quantification has not been previously explored in AM fungi. We applied real-time PCR assays targeting the large subunit of mtDNA to monitor the DNA dynamics of two isolates of Glomus intraradices sensu lato coexisting in the roots of medic (Medicago sativa). The mtDNA-based quantification was compared to quantification in nrDNA. The ratio of copy numbers determined by the nrDNA- and mtDNA-based assays consistently differed between the two isolates. Within an isolate, copy numbers of the nuclear and the mitochondrial genes were closely correlated. The two quantification approaches revealed similar trends in the dynamics of both isolates, depending on whether they were inoculated alone or together. After 12 weeks of cultivation, competition between the two isolates was observed as a decrease in the mtDNA copy numbers of one of them. The coexistence of two closely related isolates, which cannot be discriminated by nrDNA-based assays, was thus identified as a factor influencing the dynamics of AM fungal DNA in roots. Taken together, the results of this study show that real-time PCR assays targeted to the large subunit of mtDNA may become useful tools for the study of coexisting AM fungi.
- MeSH
- DNA fungální chemie genetika MeSH
- Glomeromycota genetika růst a vývoj MeSH
- kořeny rostlin mikrobiologie MeSH
- kvantitativní polymerázová řetězová reakce metody MeSH
- Medicago sativa mikrobiologie MeSH
- mikrobiální interakce MeSH
- mitochondriální DNA genetika MeSH
- molekulární sekvence - údaje MeSH
- počet mikrobiálních kolonií metody MeSH
- sekvenční analýza DNA MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
We developed and assessed the diagnostic value of a novel quantitative nested real-time (QNRT) PCR assay targeting the internal transcribed spacer region of ribosomal DNA (rDNA) in a guinea pig model of invasive pulmonary aspergillosis. Groups of 5 immunosuppressed animals that were infected using an aerosol chamber with Aspergillus fumigatus conidia were humanely terminated 1 h postinoculation and at days 3, 5, 7, and 11 postchallenge, and lung tissue, bronchoalveolar lavage (BAL) fluid, whole blood, and serum samples were collected. The QNRT PCR results obtained with the serum and BAL fluid were compared to those achieved with galactomannan and (1→3)-β-d-glucan assays. High fungal burden levels were detected by QNRT PCR in both lung tissue and BAL fluid in all infected animals at each time point, and the sensitivity of each assay in BAL fluid was 100% by day 3 and remained so through the remainder of the study. The sensitivity of detection of fungi in whole blood and serum samples was significantly lower, and some samples remained negative by all three assays despite the advanced stage of the infection. From these data, we can conclude that this novel QNRT PCR method was highly sensitive for the detection of A. fumigatus from different types of samples in this model. In addition, BAL fluid samples appeared to be the most suitable for the early diagnosis of invasive pulmonary aspergillosis. When testing serum, the use of a combination of available assays may increase the possibility of early detection of this opportunistic mycosis.
- MeSH
- Aspergillus fumigatus izolace a purifikace MeSH
- beta-glukany analýza MeSH
- bronchoalveolární lavážní tekutina mikrobiologie MeSH
- DNA fungální genetika MeSH
- invazivní plicní aspergilóza mikrobiologie patologie MeSH
- kvantitativní polymerázová řetězová reakce metody MeSH
- mannany analýza MeSH
- mezerníky ribozomální DNA genetika MeSH
- modely nemocí na zvířatech MeSH
- morčata MeSH
- mykologie metody MeSH
- plíce mikrobiologie MeSH
- počet mikrobiálních kolonií metody MeSH
- polymerázová řetězová reakce metody MeSH
- senzitivita a specificita MeSH
- zvířata MeSH
- Check Tag
- morčata MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- srovnávací studie MeSH
