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This review article summarizes the scientific literature that is currently available about a probiotic E. coli that is known under the name Symbioflor E. coli. The probiotic is marketed for human use and has been subjected to over 20 years of scientific research. As is presented here, the available literature not only contains multiple works to investigate and analyse the probiotic activity of this E. coli, but also describes a variety of other research experiments, dealing with a surprising and interesting range of subjects. By compiling all these works into one review article, more insights into this interesting probiotic E. coli were obtained.

Microbacterium sp. strain BR1 is a bacterial strain that recently received attention for its capability to mineralize sulfamethoxazole (SMX) and other sulfonamides. In this study, the survival of Microbacterium sp. in municipal sludge waters was tested in batch experiments to explore optimal process conditions. Inoculation of Microbacterium sp. was subsequently performed in a pilot membrane bioreactor (MBR) operated in two configurations: treating full-scale MBR permeate (post-treatment) and treating raw municipal wastewater. SMX removal by Microbacterium sp. could not be proved in any of the configurations, except for SMX concentrations far higher than the ones normally found in municipal wastewater. By use of molecular tools (fluorescence in situ hybridization analysis) a low capability to survive in activated sludge systems was assessed. After inoculation, Microbacterium sp. was reduced to a small fraction of the viable biomass. The observed growth rate appeared to be many times lower than the one of typical activated sludge micro-organisms. Possibilities of application in full-scale municipal wastewater treatment are scarce.

Conventional cultivation methods and molecular approaches were utilised to describe the filamentous bacterial population of industrial activated sludge WWTPs. In total 43 strains were isolated by micromanipulation and were affiliated with 12 different species, comprising two new species and a new genus. In particular, a new species of Microthrix, a new genus of a filamentous Alphaproteobacteria morphologically similar to Nostocoida limicola, and a new filamentous species closely related to the opportunistic pathogen Propionibacterium propionicum were obtained. Despite the high number of isolates, the cultivation approach was unable to describe the filamentous bacteria most common in industrial WWTP. A culture-independent approach, termed the cell sorting/RT-PCR method, was therefore applied to identify fastidious or non-culturable filamentous microrganisms from different industrial plants. By this method the relevant filaments were micromanipulated and their 16S rDNA genes were amplified by RT-PCR. This approach was highly efficient. In total 31 16S rRNA sequences were obtained and 16 of them were used for the design of new specific oligonucleotide probes that highlighted dominant filaments in industrial activated sludge plants.

BackgroundEnterobacter sakazakii is a foodborne pathogen that has been associated with sporadic cases and outbreaks causing meningitis, necrotizing enterocolitis and sepsis especially in neonates. The current FDA detection method includes two enrichment steps, the subculturing of the second enrichment broth on a selective agar (VRBG), a further subculturing of selected grown colonies on TSA and the subsequent biochemical identification of yellow-pigmented colonies by API20E. However, there is a strong need for simplified methods for isolation and identification of E. sakazakii. In this study, two chromogenic media, which allow to indicate presumptive E. sakazakii colonies by the alpha glucosidase activity, as well as a newly developed 1,6-alpha-glucosidase based conventional PCR assay and a rRNA oligonucleotide probe based commercial test system for identification of presumptive E. sakazakii were evaluated on 98 target and non-target strains. The methods were compared with respect to specificity aspects.ResultsA total of 75 presumptive E. sakazakii and 23 non-target strains were analysed by using chromogenic media, alpha-glucosidase based PCR assay, and the VIT assay. For most presumptive E. sakazakii strains on the chromogenic media, the PCR and VIT assay confirmed the identification. However, for a number of presumptive E. sakazakii isolates from fruit powder, the alpha-glucosidase PCR and VIT assay did not correspond to the typical E. sakazakii colonies on DFI and ESIA. Further characterization by API32E identification, phylogenetic analysis of partial 16S rRNA sequences and ribotyping strongly suggested, that these strains did not belong to the species E. sakazakii. The newly developed alpha-glucosidase based PCR assay as well as the commercially available VIT Enterobacter sakazakii identification test showed an excellent correlation with the 16S rRNA data, and are thus well suited for identification of E. sakazakii.ConclusionThe results indicate that presumptive colonies on ESIA and DFI media need further species identification. Both evaluated molecular methods, the alpha-glucosidase PCR and the 16S RNA in situ hybridisation test (VIT), although based on completely different target regions and methodologies performed equally well in terms of specificity.

Fluorescent In Situ Hybridisation (FISH) was used to monitor the presence of filamentous microorganisms in industrial wastewater treatment plants (WWTPs). Monitoring with a restricted set of FISH probes in WWTPs from potato industry showed growth and decline of Thiothrix populations that could be linked to operational procedures. In a follow up project new FISH probes were developed for filamentous bacteria in industrial WWTPs and 70 WWTPs were analysed for presence of these filaments. Several newly described species of filamentous bacteria appear to be common and dominant in industrial WWTPs. Monitoring of a WWTP from textile industry showed growth and decline of one of these organisms when operational conditions in the plant were varied. The present paper demonstrates that bulking sludge in industrial wastewater treatment plants can effectively be monitored using a combination of standard chemical analyses and the FISH technique.

Mycorrhizal ascomycetous fungi are obligate ectosymbionts that colonize the roots of gymnosperms and angiosperms. In this paper we describe a straightforward approach in which a combination of morphological and molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria associated with the ascomycetous ectomycorrhizal fungus Tuber borchii Vittad. Universal eubacterial primers specific for the 5' and 3' ends of the 16S rRNA gene (16S rDNA) were used for PCR amplification, direct sequencing, and phylogenetic analyses. The 16S rDNA was amplified directly from four pure cultures of T. borchii Vittad. mycelium. A nearly full-length sequence of the gene coding for the prokaryotic small-subunit rRNA was obtained from each T. borchii mycelium studied. The 16S rDNA sequences were almost identical (98 to 99% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belonging to the Cytophaga-Flexibacter-Bacteroides (CFB) phylogroup which had not been described previously. In situ detection of the CFB bacterium in the hyphal tissue of the fungus T. borchii was carried out by using 16S rRNA-targeted oligonucleotide probes for the eubacterial domain and the Cytophaga-Flexibacter phylum, as well as a probe specifically designed for the detection of this mycelium-associated bacterium. Fluorescent in situ hybridization showed that all three of the probes used bound to the mycelium tissue. This study provides the first direct visual evidence of a not-yet-cultured CFB bacterium associated with a mycorrhizal fungus of the genus Tuber.