ABSTRACT
One of the best-known Hungarian products on world wine market is Aszú, which belongs to the family of Tokaj wine specialties and is made from aszú berries. An important condition for the formation of aszú berries is the noble rot of technologically mature grapes, which is caused by Botrytis cinerea. At the same time botrytized sweet wines are produced not only in Hungary, but in many locations of wine-producing areas of Europe as well as in certain wine growing regions of other continents. The determination of botrytization is mostly based on sensory evaluations, which is a highly subjective procedure and largely depends on the training and experience of the evaluator. Currently, the classification of aszú berries (class I and class II) is based only on visual inspection and determination of sugar content. Based on these facts the primary goal of our work was to develop a qPCR assay capable for objective rating and classification of aszú berries. The developed qPCR is highly specific and sensitive as can clearly distinguish between B. cinerea and other filamentous fungi and yeast species occur on grapes. Moreover, it is suitable for categorizing berries colonized by B. cinerea to varying degrees. Thus, the developed qPCR method can be a useful technique for classification of the grape berries into four quality groups: healthy, semi-shrivelled, Aszú Class II and Aszú Class I.
Subject(s)
Botrytis , Fruit , Vitis , Wine , Vitis/microbiology , Wine/microbiology , Wine/analysis , Fruit/microbiology , Botrytis/genetics , Botrytis/classification , Botrytis/isolation & purification , Hungary , Real-Time Polymerase Chain Reaction/methods , Plant Diseases/microbiologyABSTRACT
The spread of superbugs in dairy products can jeopardize global public health. To date, information on the incidence rates of virulent and ß-lactams-resistant (BLR) Salmonella in cheeses from rural areas of Egypt has been lacking. Biochemical, serological, antibiotic susceptibility, and multiplex PCR (M-PCR) tests were performed to identify and characterize Salmonella isolates. In this study, 44 (15.71%) Salmonella isolates of eight different serotypes were recovered from 280 samples of Rumi and pasteurized Kariesh cheeses across the Nile Delta region of Egypt. The most predominant serotypes were S. Typhimurium, S. Enteritidis, and S. Infantis. The virulence genes (invA, stn, and hilA) were identified in all isolates. However, spvC was only detected in S. Typhimurium. The highest resistance was developed against Erythromycin and Clindamycin (90.91%), followed by Ceftazidime and Cephalothin (84.09%). Meropenem and colistin were the most effective antibiotics. A high proportion (79.55%) of multi-drug resistance (MDR) isolates carried narrow spectrum (NS), extended-spectrum (ES), and AmpC-BLR genes. The blaOXA-1, blaOXA-2, blaTEM-1, blaCTX-M, blaCMY-1, and blaCMY-2 BLR genes were positive in 37.04%, 29.63%, 25.93%, 14.81%, 37.04%, and 3.70% of isolates, respectively. In conclusion, a high prevalence of virulence and BLR genes harboring Salmonella strains in Egyptian cheeses is considered a great threat to public health.
ABSTRACT
Serratia marcescens strains from a dairy-producing environment were tested for their inhibitory effect on Listeria monocytogenes, Salmonella Hartford, Yersinia enterocolitica and Escherichia coli. Inhibition of foodborne pathogens was observed in the case of a non-pigmented Serratia strain, while the pigment-producing isolate was able to inhibit only Y. enterocolitica. The co-culturing study in tryptone soya broth (TSB) and milk showed that the growth of Salmonella was inhibited in the first 24 h, but later the pathogen could grow in the presence of the Serratia strain even if its cell concentration was 1000 times higher than that of Salmonella. However, we found that (1) concentrated cell-free supernatants had stronger inhibitory activity, which confirms the extracellular nature of the antagonistic compound(s). We proved that (2) protease and chitinase enzymes can take part in this mechanism, but they are not the main inhibitory compounds. The presence of prodigiosin was observed only in the case of the pigmented strain; thus, (3) we hypothesized that prodigiosin does not take part in the inhibition of the pathogens. However, (4) the combined effect of different extracellular metabolites might be attributed to the inhibitory property. Application of concentrated S. marcescens cell-free supernatant can be an effective antibacterial strategy in the food industry, mainly in the form of a bio-disinfectant on surfaces of food-processing areas.
ABSTRACT
Bacterial isolates derived from food or raw food materials of animal origin were screened for potential antagonistic activity against foodborne pathogenic Listeria monocytogenes. Using the agar spot method, ten out of the 94 tested bacteria showed antilisterial activity. All of the antagonistic isolates identified by sequence analysis as strains of the genus Pseudomonas were able to inhibit the growth of all the examined Listeria species including the ruminal pathogenic L. ivanovii and the opportunistic human pathogenic L. innocua. Pseudomonas sp. CMI-1 had the highest inhibitory effect on the growth of different Listeria strains. Co-culturing studies revealed that the inhibition of L. monocytogenes could not be achieved efficiently. Although the population of the Pseudomonas sp. CMI-1 strain increased by up to 10 orders of magnitude during 2 days of culturing period at 20 °C in the presence of L. monocytogenes, the cell count of the pathogen also increased by approx. 6 orders of magnitude. At the same time, appropriate inhibition of cell-free supernatants generated from 6-day-old cultures of Pseudomonas sp. CMI-1 was observed. The inhibitory compound of this antagonistic strain is presumably a chromopeptide siderophore, whose activity and production can be affected by iron supplementation, and which had an absorption maximum typical of siderophores of fluorescent Pseudomonas species. Production of the antilisterial substance was influenced by the oxygen concentration, as in static cultures the concentration of the siderophore was higher than in shake flask cultures.
ABSTRACT
Cold-active lipases have attracted attention in recent years due to their potential applications in reactions requiring lower temperatures. Both bacterial and fungal lipases have been investigated, each having distinct advantages for particular applications. Among yeasts, cold-active lipases from the genera Candida, Yarrowia, Rhodotorula, and Pichia have been reported. In this paper, biosynthesis and properties of a novel cold-active lipase from Candida zeylanoides isolated from refrigerated poultry meat are described. Heat-sterilized olive oil was found to be the best lipase biosynthesis inducer, while nonionic detergents were not effective. The enzyme was purified to homogeneity using hydrophobic chromatography and its enzymatic properties were tested. Pure enzyme activity at 7 °C was about 60% of the maximal activity at 27 °C. The enzyme had rather good activity at higher temperatures, as well. Optimal pH of pure lipase was between 7.3 and 8.2, while the enzyme from the crude extract had an optimum pH of about 9.0. The enzyme was sensitive to high ionic strength and lost most of its activity at high salt concentrations. Due to the described properties, cold-active C. zeylanoides lipase has comparative advantages to most similar enzymes with technological applications and may have potential to become an industrially important enzyme.
Subject(s)
Candida/enzymology , Lipase/chemistry , Lipase/isolation & purification , Cold Temperature , Detergents/pharmacology , Enzyme Activation , Enzyme Stability , Fungal Proteins/chemistry , Fungal Proteins/metabolism , Hydrogen-Ion Concentration , Lipase/metabolism , Meat/microbiology , Olive Oil/pharmacology , TemperatureABSTRACT
Phytochemical studies of Epilobium angustifolii herba aimed the determination of the active ingredients; antimicrobial activity of the aqueous and ethanolic extracts against human pathogenic bacteria and fungi was also examined. Six flavonoid and 4 phytosterol fractions were identified by thin layer chromatography, while tannins were present in low concentration. It has been shown that not only the ethanolic but also the aqueous extracts had inhibitory effect on certain pathogenic microorganisms, therefore E. angustifolium could be used for the development of external phytotherapic or disinfectant preparations.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Epilobium , Plant Extracts/pharmacology , Disinfectants/pharmacology , Humans , Phytochemicals , PhytotherapyABSTRACT
In this study, 22 yeast strains isolated from foods were characterized by traditional and molecular techniques. With the help of traditional identification tests, yeast strains were grouped in 12 species belonging to 11 genera as follows: Candida parapsilosis, Rhodotorula mucilaginosa, Debaryomyces hansenii, Cryptococcus humicolus, Cryptococcus albidus, Aureobasidium spp., Hanseniaspora valbyensis, Metschnikowia pulcherrima, Lachancea thermotolerans, Pichia anomala, Geotrichum candidum and Yarrowia lipolytica. The patterns obtained by the digestion of ITS-18S rRNA gene with MspI and HaeIII restriction endonucleases were similar among strains belonging to the same species. With the help of randomly amplified polymorphic DNA (RAPD) analysis performed within the same species, discrimination of M. pulcherrima strains could be achieved.