count-plate method
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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
External fixators of serious fractures could be an attractive substrate on which microorganisms can accumulate. Therefore, this study aimed to develop a suitable method for enabling the simulation of a real situation when osteosynthetic fixation material is open for the potential threat of bacterial attack. Agar-based media represented human tissue, and the metallic pin characterized the screw in the fixation. Various types of agar, supplements, and contamination strategy by Staphylococcus aureus were tested. The influence of the initial bacterial concentration was also examined. Surfaces were observed by scanning electron microscopy (SEM), and all results were compared. Brain Heart Infusion Agar with the Egg Yolk Tellurite Emulsion was established in a transparent test tube as a suitable system for enabling the good interpretability of bacterial contamination in the pin's surroundings. Pin contamination has been found to be an appropriate approach for testing microbial growth, rather than agar surface contamination, which distorted obtained results. A lower initial colony forming units (CFU) provided better clarity of the test. SEM observation of the pin surface was comparable with the visual evaluations in the test tubes. Results were assembled for positive and negative control samples as well. Screening method for the most common bacteria S. aureus has been standardized and developed. This experimental setup could also be a useful tool for surface modification with antibacterial properties testing.
- MeSH
- antibakteriální látky farmakologie MeSH
- biofilmy účinky léků růst a vývoj MeSH
- externí fixátory mikrobiologie MeSH
- kontaminace zdravotnického vybavení * MeSH
- kultivační média MeSH
- lidé MeSH
- mikroskopie elektronová rastrovací MeSH
- počet mikrobiálních kolonií MeSH
- Staphylococcus aureus účinky léků růst a vývoj ultrastruktura MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
OBJECTIVES: The objective of the present study was to evaluate and compare the efficiency of a filter based sampling method and a high volume sampling method for sampling airborne culturable fungi present in waste sorting facilities. MATERIAL AND METHODS: Membrane filters method was compared with surface air system method. The selected sampling methods were modified and tested in 2 plastic waste sorting facilities. RESULTS: The total number of colony-forming units (CFU)/m3 of airborne fungi was dependent on the type of sampling device, on the time of sampling, which was carried out every hour from the beginning of the work shift, and on the type of cultivation medium (p < 0.001). Detected concentrations of airborne fungi ranged 2×102-1.7×106 CFU/m3 when using the membrane filters (MF) method, and 3×102-6.4×104 CFU/m3 when using the surface air system (SAS) method. CONCLUSIONS: Both methods showed comparable sensitivity to the fluctuations of the concentrations of airborne fungi during the work shifts. The SAS method is adequate for a fast indicative determination of concentration of airborne fungi. The MF method is suitable for thorough assessment of working environment contamination by airborne fungi. Therefore we recommend the MF method for the implementation of a uniform standard methodology of airborne fungi sampling in working environments of waste treatment facilities.
- MeSH
- houby izolace a purifikace MeSH
- látky znečišťující vzduch v pracovním prostředí izolace a purifikace MeSH
- mikrobiologie vzduchu * MeSH
- monitorování životního prostředí přístrojové vybavení metody MeSH
- počet mikrobiálních kolonií MeSH
- skládková zařízení * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Pneumonie způsobená Pneumocystis jirovecii (PCP) je život ohrožující mykotickou infekcí, která se vyskytuje zejména u HIV-pozitivních a jinak imunokompromitovaných jedinců. Specifickou skupinu nemocných tvoří imunokompromitovaní pacienti s hematologickými malignitami, u kterých bylo prokázáno, že konvenčně používané metody, zejména mikroskopická analýza, jsou velmi specifické, ale nejsou dostatečně citlivé. U těchto pacientů je metodou volby detekce DNA Pneumocystis jirovecii v klinických vzorcích (nejčastěji v indukovaném sputu nebo v tekutině získané bronchoalveolární laváží) metodami založenými na PCR. První metody umožňující záchyt P. jirovecii v klinických vzorcích byly publikovány v 90. letech minulého století(1) a od té doby následovaly desítky prací, které využívaly různé PCR platformy (jednokolová, nested, semi-nested, real-time PCR) a byly zaměřeny do různých cílových genů. Cílem tohoto přehledového článku je popsat přednosti a úskalí PCR diagnostiky PCP u pacientů s hematologickými malignitami.
Diagnostics of pneumocystis pneumonia using methods of molecular biology Pneumonia caused by Pneumocystis jirovecii (PCP) is a life-threatening fungal infection that occurs especially in HIV-positive and other immunocompromised individuals. A specific group are patients with haematological malignancies in which it was shown that conventionally used methods, in particular microscopic analysis, are highly specific but not sensitive enough. In these patients, the method of choice is detection of P. jirovecii DNA in clinical specimens (mostly induced sputum or bronchoalveolar lavage fluid) using methods based on PCR. The first methods allowing detection of P. jirovecii in clinical specimens have been published in the 1990’s(1) and were followed by dozens of papers that were aimed at various target genes and used different PCR platforms (single-round, nested, semi-nested, real-time PCR). The aim of this review is to describe the advantages and pitfalls of PCR diagnosis of PCP in patients with haematological malignancies.
- MeSH
- bronchoalveolární lavážní tekutina mikrobiologie MeSH
- imunokompromitovaný pacient * imunologie MeSH
- lidé MeSH
- pneumocystová pneumonie * diagnóza MeSH
- počet mikrobiálních kolonií * klasifikace normy MeSH
- polymerázová řetězová reakce * metody MeSH
- sputum mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- 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.
Arbuscular mycorrhizal fungi (AMFs) are important plant symbionts, but we know little about the effects of plant taxonomic identity or functional group on the AMF community composition. To examine the effects of the surrounding plant community, of the host, and of the AMF pool on the AMF community in plant roots, we manipulated plant community composition in a long-term field experiment. Within four types of manipulated grassland plots, seedlings of eight grassland plant species were planted for 12 wk, and AMFs in their roots were quantified. Additionally, we characterized the AMF community of individual plots (as their AMF pool) and quantified plot abiotic conditions. The largest determinant of AMF community composition was the pool of available AMFs, varying at metre scale due to changing soil conditions. The second strongest predictor was the host functional group. The differences between grasses and dicotyledonous forbs in AMF community variation and diversity were much larger than the differences among species within those groups. High cover of forbs in the surrounding plant community had a strong positive effect on AMF colonization intensity in grass hosts. Using a manipulative field experiment enabled us to demonstrate direct causal effects of plant host and surrounding vegetation.
- MeSH
- fylogeneze MeSH
- interakce hostitele a patogenu * MeSH
- metoda Monte Carlo MeSH
- multivariační analýza MeSH
- mykobiom * MeSH
- mykorhiza růst a vývoj fyziologie MeSH
- pastviny * MeSH
- počet mikrobiálních kolonií MeSH
- pravděpodobnostní funkce MeSH
- půda chemie MeSH
- rostliny mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The presence of raffinose series oligosaccharides (RSO) was determined by an enzymic method in three commercially available chicken feed mixtures. All feed mixtures contained RSO at a concentration of 2.1-2.2%. Soya meal was identified as the exclusive source of RSO. Subsequently, the bifidogenic effect of stachyose (main soya bean RSO) was also assigned on the growth of poultry intestinal bifidobacteria. Bifidobacteria were counted in chicken intestinal tract using cultivation and FISH methods. Four out of 6 bifidobacterial strains tested grew significantly better on stachyose than on glucose. It can be thus concluded that chicken feed mixtures naturally contain prebiotic oligosaccharides in the form of RSO in higher levels (>2%) compared with the concentration (usually up to 1%) recommended for artificially added prebiotics. Our results therefore indicate that there is no reason for the supplementation of chicken feed mixtures with prebiotics with bifidogenic properties.
- MeSH
- Bifidobacterium izolace a purifikace MeSH
- dietní sacharidy MeSH
- financování organizované MeSH
- hybridizace in situ fluorescenční MeSH
- krmivo pro zvířata MeSH
- kur domácí MeSH
- oligosacharidy analýza MeSH
- počet mikrobiálních kolonií MeSH
- střeva mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
