Today people focus on healthy lifestyle. It is well known that probiotics have a number of beneficial health effects in humans. They play an important role in protecting the host against harmful microorganisms, reduce metabolic disorders and support immune functions. In the present study, we investigated 26 probiotic products: 16 commercially available ones and 10 from a local manufacturer. None of the commercial products contained all labelled LAB and some of them contained unacceptable microorganisms. Of 890 isolated strains only 420 met the criteria for LAB. Ninetyseven strains were investigated by phenotypic and genotypic methods. 16S-23S rDNA amplification was performed for all putative LAB isolates. Fifty-seven rod-shaped bacteria referred to genus Lactobacillus and 16 coccus bacteria, to genus Weissela. Two-step multiplex PCR identified the rodshaped strains to belong to four species: Lactobacillus delbrueckii ssp. bulgaricus (three strains), Lactobacillus acidophilus (four strains), Lactobacillus casei (three strains), Lactobacillus rhamnosus (two strains), Lactobacillus reuteri (four strains) and Lactobacillus plantarum (one strain). The transit tolerance of the isolated bacteria to in vitro simulated gastric juice (pH 2) was examined. Only four strains survived after 90 min of incubation. The antimicrobial activity of the native supernatants of the isolates was tested and 10 of them showed slight activity against three pathogenic bacteria. Our results demonstrated that only freshly produced products possessed the expected number of viable cells. The data revealed that one of the most common problems is the lower concentration of viable cells, misidentification and the presence of undesired microflora.
The ability of certain human pathogens to adapt to plants without losing their virulence toward people is a major concern today. Thus, the aim of the present work was the investigation of the presence of cross-over pathogenic bacteria in infected tomato and pepper plants. The objects of the study were 21 samples from seven different parts of the plants and three from tomato rhizosphere. In total, 26 strains were isolated, identified by MALDI-TOF, and phenotypically characterized. The PCR amplification of the rpoB gene was applied as an approach for the rapid detection of cross-over pathogens in plant samples. A great bacterial diversity was revealed from tomato samples as nine species were identified (Leclercia adecarboxylata, Pseudesherichia vulneris, Enterobacter cancerogenus, Enterobacter cloacae, Enterobacter bugandensis, Acinetobacter calcoaceticus, Pantoea agglomerans, Pantoea ananatis, and Pectobacterium carotovorum). Polymicrobial contaminations were observed in samples T2 (tomato flower) and T10 (tomato fruit). Five species were identified from pepper samples (P. agglomerans, L. adecarboxylata, Pseudomonas sp., Pseudomonas putida, and Enterococcus sp.). Antibiotic resistance patterns were assigned in accordance with EFSA recommendations. All isolates showed varying resistance to the tested antibiotics. The genetic basis for the phenotypic antibiotic resistance was not revealed. No genes for the virulence factors were found among the population. To our knowledge, this is the first overall investigation of tomato and pepper cross-over pathogenic bacterial populations in Bulgaria.
The increasing antibiotic resistance among pathogenic bacteria is a challenge that drives the development of new antibacterial substances. Marine inhabitants are excellent sources of antimicrobial proteins and considered as promising candidates for the treatment of microbial infections. In the present study, we obtained haemocyanin from Eriphia verrucosa and studied its potential to suppress the growth of some pathogenic bacteria and yeasts. The putative antibacterial molecules were isolated from the haemolymph by chromatography appropriate for producing the native haemocyanin (EvH) and its five structural units (SUs). The results showed that EvH had no antimicrobial activity unlike its glycosylated SUs. All haemocyanin SUs exhibited differential antibacterial activity depending on their grade of glycosylation. The strongest antimicrobial activity of SU1 (with highest carbohydrate content) was against Escherichia coli and Bacillus subtilis. The least glycosylated SU3 and SU4 exhibited the lowest antimicrobial activity against all strains. The fraction SU1 has the potential to be applied as a substitute for some commonly used antibiotics. It was demonstrated that the grade of haemocyanin glycosylation plays an important role in its functional antibacterial properties.
Pulsed-field gel electrophoresis (PFGE) is a highly discriminative molecular typing method that is used for epidemiological studies and investigation of outbreaks caused by different pathogens, including phytopathogenic Xanthomonas species. Bacterial spot (BS) is the most common and one of the most destructive diseases of tomato and pepper plants in Bulgaria. Several Xanthomonas species are known to cause BS, but the global distribution and genetic diversity of these species are not well understood. A collection of 100 BS-causing strains, isolated during the period of 1985-2012 from different tomato cultivars and weeds associated with tomato production areas from 11 geographic regions in Bulgaria, were screened for genetic diversity by genomic DNA restriction with rare-cutting endonucleases (XbaI and SpeI) subsequently resolved by PFGE. Two haplotypes for Xanthomonas vesicatoria and one haplotype for Xanthomonas gardneri strains were found.
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