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Streptomyces are of great interest in the pharmaceutical industry as they produce a plethora of secondary metabolites that act as antibacterial and antifungal agents. They may thrive on their own in the soil, or associate with other organisms, such as plants or invertebrates. Some soil-derived strains exhibit hemolytic properties when cultivated on blood agar, raising the question of whether hemolysis could be a virulence factor of the bacteria. In this work we examined hemolytic compound production in 23 β-hemolytic Streptomyces isolates; of these 12 were soil-derived, 10 were arthropod-associated, and 1 was plant-associated. An additional human-associated S. sp. TR1341 served as a control. Mass spectrometry analysis suggested synthesis of polyene molecules responsible for the hemolysis: candicidins, filipins, strevertene A, tetrafungin, and tetrin A, as well as four novel polyene compounds (denoted here as polyene A, B, C, and D) in individual liquid cultures or paired co-cultures. The non-polyene antifungal compounds actiphenol and surugamide A were also identified. The findings indicate that the ability of Streptomyces to produce cytolytic compounds (here manifested by hemolysis on blood agar) is an intrinsic feature of the bacteria in the soil environment and could even serve as a virulence factor when colonizing available host organisms. Additionally, a literature review of polyenes and non-polyene hemolytic metabolites produced by Streptomyces is presented.
- Klíčová slova
- Actinomycetales, Streptomyces, hemolysis, polyene antibiotics, secondary metabolites, soil ecosystem, symbiosis,
- MeSH
- antibakteriální látky farmakologie metabolismus MeSH
- antifungální látky farmakologie chemie MeSH
- faktory virulence metabolismus MeSH
- hemolýza MeSH
- lidé MeSH
- polyeny farmakologie chemie MeSH
- Streptomyces * chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- antibakteriální látky MeSH
- antifungální látky MeSH
- faktory virulence MeSH
- polyeny MeSH
BACKGROUND: Larval toxocarosis is a zoonosis caused by larvae of Toxocara canis and T. cati, a gastrointestinal nematode of canids and felids, respectively. Diagnosis is usually performed by ELISA IgG using Toxocara excretory-secretory products as an antigen. Due to laboriousness of isolation of the products and subsequent process of standardization of antigenic compounds, routine use of this method is limited and can produce inaccurate diagnostical results. The purpose of this study was to discover new specific antigenic proteins that could be used in routine serological methods of larval toxocarosis. MATERIALS AND METHODS: Toxocara excretory-secretory products were collected and separated by SDS-PAGE. Proteins from the gel were electro-transferred to a membrane and incubated with mouse sera. Antigenic proteins were analyzed using the liquid chromatography-tandem mass spectrometry approach. Selected proteins were prepared in recombinant form and tested with mice and human sera by ELISA and Western blot. RESULTS: A total of four recombinant protein antigens were prepared (rTc-TES-26, rTc-ASA, rTc-PDP, and rTc-ASP). They were analyzed by ELISA and Western blot using mice and human sera. For all sera, three of the four recombinant antigens correlated with Toxocara excretory-secretory products in ELISA analysis. By Western blot, the infection was confirmed in all experimentally infected mice and two out of seven human patients. CONCLUSION: Combination of the presented methods and analyses represents a possible method of effective identification of Toxocara protein antigens for the purpose of routine serodiagnosis.
- Klíčová slova
- Antigen, Diagnostics, Recombinant protein, Toxocara canis, Toxocariasis, Toxocarosis,
- MeSH
- antigeny helmintové MeSH
- ELISA MeSH
- larva MeSH
- lidé MeSH
- myši MeSH
- Toxocara canis * MeSH
- Toxocara MeSH
- toxokaróza * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny helmintové MeSH
Regulatory RNAs control a number of physiological processes in bacterial cells. Here we report on a 6S-like RNA transcript (scr3559) that affects both development and antibiotic production in Streptomyces coelicolor. Its expression is enhanced during the transition to stationary phase. Strains that over-expressed the scr3559 gene region exhibited a shortened exponential growth phase in comparison with a control strain; accelerated aerial mycelium formation and spore maturation; alongside an elevated production of actinorhodin and undecylprodigiosin. These observations were supported by LC-MS analyses of other produced metabolites, including: germicidins, desferrioxamines, and coelimycin. A subsequent microarray differential analysis revealed increased expression of genes associated with the described morphological and physiological changes. Structural and functional similarities between the scr3559 transcript and 6S RNA, and its possible employment in regulating secondary metabolite production are discussed.
- Klíčová slova
- 6S RNA, Streptomyces, antibiotics, secondary metabolism, small RNA,
- Publikační typ
- časopisecké články MeSH
In our study we present an overview of the use of Oxford Nanopore Technologies (ONT) sequencing technology on the background of Enteric fever. Unlike traditional methods (e.g., qPCR, serological tests), the nanopore sequencing technology enables virtually real-time data generation and highly accurate pathogen identification and characterization. Blood cultures were obtained from a 48-year-old female patient suffering from a high fever, headache and diarrhea. Nevertheless, both the initial serological tests and stool culture appeared to be negative. Therefore, the bacterial isolate from blood culture was used for nanopore sequencing (ONT). This technique in combination with subsequent bioinformatic analyses allowed for prompt identification of the disease-causative agent as Salmonella enterica subsp. enterica serovar Paratyphi A. The National Reference Laboratory for Salmonella (NIPH) independently reported this isolate also as serovar Paratyphi A on the basis of results of biochemical and agglutination tests. Therefore, our results are in concordance with certified standards. Furthermore, the data enabled us to assess some basic questions concerning the comparative genomics, i.e., to describe whether the isolated strain differs from the formerly published ones or not. Quite surprisingly, these results indicate that we have detected a novel and so far, unknown variety of this bacteria.
- Klíčová slova
- Enteric fever, Nanopore sequencing, Paratyphoid fever, Pathogen identification, Salmonella,
- MeSH
- břišní tyfus * MeSH
- lidé středního věku MeSH
- lidé MeSH
- nanopórové sekvenování * MeSH
- Salmonella paratyphi A genetika MeSH
- Salmonella MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- přehledy MeSH