Survey of Domestic Refrigerator Storage Temperatures in Poland for Use as a QMRA Tool for Exposure Assessment.

In the framework of Quantitative Microbiological Risk Assessment, the estimation of the ingested dose of a hazard by the consumer is of paramount importance. This may be calculated by means of predictive modeling of growth/inactivation of the pathogen studied. For products that spend the majority of their shelf life in the domestic refrigerator, storage temperature will significantly impact the microbial population dynamics. To describe the variability of domestic storage temperatures in Poland, a survey including 77 participants, was carried out in Lodz, Poland. Participants were provided with temperature data loggers, which measured their refrigerator temperature for 24 h in 5-min intervals. The temperature-time profiles were used to calculate the mean working temperature, standard deviation, minimum and maximum values, and the data were statistically analyzed to find the best fitting probability distribution using R programming language. Out of the tested refrigerators, 49.35% had a mean working temperature of over 5 °C and 3.9% exceeded 10 °C. Distribution fitting scenarios were tested for goodness of fit, and the final selected distribution was a truncated normal distribution. This study can prove useful in Monte Carlo simulation analysis for stochastic quantitative food risk assessment in Poland.

Quantitative microbiological risk assessment of traditional food of animal origin produced in short supply chains in Poland.

Polish raw-milk cheeses produced in short supply chains may pose a threat to consumer safety due to pathogen presence. Listeria monocytogenes is a bacterium of great importance for the food safety of refrigerated RTE foods due to its ability to grow at refrigeration temperatures. During the EU-FORA fellowship, a stochastic risk assessment was designed and executed to estimate the risk for consumers from L. monocytogenes in these products. The aim was to develop a probabilistic QMRA model that would incorporate the variability and uncertainty of the model's inputs such as prevalence, initial concentration levels, product intrinsic factors, domestic storage temperature and consumer behaviour. The project involved data collection and analysis, growth model selection, mathematical modelling and Monte Carlo analysis in R programming language. Microbiological and physicochemical testing were carried out throughout the year on two types of cheeses in combination with a domestic refrigerator temperature survey and accompanying consumption questionnaire. Collected data were fitted to probability distributions using R. The appropriate growth model for the pathogen was selected based on an inoculation study performed on one of the raw-milk cheeses and the chosen mathematical model was written into the R script developed for the QMRA. The dose-response model used the ingested dose calculated from the modelled concentration of L. monocytogenes at the time of consumption and the single serving size from the questionnaire to estimate the probability of illness. The final risk was expressed as probability of listeriosis for Polish consumers per serving of raw-milk cheese.

Microbiological Quality and Safety of Traditional Raw Milk Cheeses Manufactured on a Small Scale by Polish Dairy Farms.

Polish raw milk artisanal cheese may pose a threat to consumer safety due to pathogen presence. The aim of this study was to assess the microbiological safety, quality and physicochemical composition of cow's and goat's milk fresh cheeses produced by farmers on a small scale. A total of 62 samples of six cheese types were analyzed for Listeria monocytogenes, Salmonella spp., lactic acid bacteria and coliform presence and concentration levels. The physicochemical analysis estimated energy, water, protein, fat, carbohydrate, ash and salt content. The cheeses were also tested for heavy metal contamination. Listeria monocytogenes and Salmonella spp. were not detected in any of the samples. Coliforms were present in all the goat's milk cheeses and only in two of the cow's milk cheeses. Low levels of cadmium, below 0.008 ppm, were detected in three of the cows' milk samples. The raw milk cheeses studied were free of the pathogens examined and were of high nutritional value.

The Prevalence of Campylobacter spp. in Polish Poultry Meat.

The prevalence, count and molecular identification of Campylobacter spp. in Polish poultry meat were analysed. 181 samples of meat from chicken (70), turkey (47), duck (54) and goose (10) were studied. Campylobacter spp. was found in 64% of meat samples. The highest prevalence of this pathogen was detected for duck meat. On average 80% of duck samples were contaminated with Campylobacter spp. The counts of Campylobacter spp. in positive samples remained under ten colony forming units per gram of product in 59% of poultry meat. C. jejuni was more frequently detected in poultry meat than C. coli.

[Elaboration of chemotaxonomical detection method of presence Legionella pneumophila cultures in water samples by GC-MS technique]. / Opracowanie chemiotaksonomicznej metody wykrywania obecnosci bakterii z rodzaju Legionella pneumophila w próbkach wody technika GC-MS.

The aim of this study was elaboration of chemotaxonomical detection method of presence Legionella pneumophila cultures in water samples. In research, the profile of ester-linked fatty acids were specified, which are situated in the cell wall of the model bacteria cultures Legionella pneumophila 33152, which originate from ATCC collection Philadelphia 1 type. The profile were applied as a standard to detection L. pneumophila presence in water supply network. During the research, water samples were esterified and extracted and then specific taxonomic markers of this bacteria were indicated by gas chromatography coupled with mass spectrometry technique (GC-MS). The limit of detection were determined to 100 jtk/100 ml water. Innovative chemotaxonomic method may be used in preliminary selection of water samples in routine analyses. Its application cuts down on time of analysis, by limitation on number of samples being diagnosed by classical microbiological methods.

Structure of a glucosyl phosphate-containing O-polysaccharide of Proteus vulgaris O42.

An O-polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of Proteus vulgaris O42 and studied by sugar and methylation analyses along with 1H, 13C and 31P NMR spectroscopy. The following structure of the polysaccharide having a linear pentasaccharide phosphate repeating unit was established: -->3)-alpha-L-FucpNAc4Ac-(1-->4)-alpha-D-Glcp-1-P-(O-->4)-alpha-D-GlcpNAc-(1-->3)-alpha-L-FucpNAc4Ac-(1-->3))-alpha-D-GlcpNAc6Ac-(1--> where the degree of O-acetylation is approximately 80% on GlcNAc and approximately 40% on each of the FucNAc residues. A weak serological cross-reaction of anti-P. vulgaris O42 serum with the lipopolysaccharide of P. vulgaris O39 was observed and accounted for by the sharing of a disaccharide fragment of the O-polysaccharides.

Structure of the O-polysaccharide of Proteus mirabilis O19 and reclassification of certain Proteus strains that were formerly classified in serogroup O19.

INTRODUCTION: Bacteria of the genus Proteus are a common cause of urinary tract infections. The O-polysaccharide chain of their LPS (O-antigen) defines the serological specificity of these bacteria. Based on the immunospecificity of the O-antigens, two species, P. mirabilis and P. vulgaris, were classified into 49 O-serogroups, and more O-serogroups for strains of these species and P. penneri have been subsequently proposed. MATERIAL AND METHODS: The lipopolysaccharide of P.mirabilis CCUG 19011 from serogroup O19 was degraded under mildly acidic and mildly alkaline conditions. Polysaccharides thus obtained were studied by chemical methods, including O -deacetylation, sugar and methylation analyses, and 1H- and 13C-NMR spectroscopy. Antisera were obtained by immunization of New Zealand white rabbits with heat-killed bacteria. In serological studies, enzyme immunosorbent assay, passive hemolysis test, and inhibition of passive hemolysis were used. RESULTS: The following structure of the O-polysaccharide repeating unit was established:-->3)- beta-D-GlcrhoNAc-(1-->3)- alpha-D-GalrhoNAc4,6(R-Pyr)-(1-->4)- a-D-GalrhoA-(1-->3) alpha-L-Rhap2Ac-(1-->where R-Pyr is (R)-1-carboxyethylidene (an acetal-linked pyruvic acid). This structure is significantly different from the O-polysaccharide structures of P. vulgaris, P.hauseri and P. penneri strains from the same Proteus serogroup O19. CONCLUSIONS: Based on immunochemical studies of the lipopolysaccharides, it is suggested 1) to keep P. vulgaris CCUG 4654 and P. penneri 31 in serogroup O19 as two subgroups, 2) to reclassify P. mirabilis CCUG 19011 into a new Proteus serogroup, O51, and 3) to classify serologically related strains, including P. vulgaris ATCC 49990, P. hauseri> 1732-80 and 1086-80, P. penneri 15, and some other P. penneri strains, in yet another Proteus serogroup, O52.

Structure of the O-polysaccharide of Proteus penneri 28 and Proteus vulgaris O31 and classification of P. penneri 26 and 28 in Proteus serogroup O31.

The lipopolysaccharides (LPS) of Proteus penneri 28 and Proteus vulgaris O31 (PrK 55/57) were degraded with dilute acetic acid and structurally identical high-molecular-mass O-polysaccharides were isolated by gel-permeation chromatography. Sugar analysis and nuclear magnetic resonance (NMR) spectroscopic studies showed that both polysaccharides contain D-GlcNAc, 2-acetamido-2,6-dideoxy-L-glucose (L-2-acetamido-2,6-dideoxyglucose (N-acetylquinovosamine)) and 2-acetamido-3-O-[(S)-1-carboxyethyl]-2-deoxy-D-glucose (N-acetylisomuramic acid) and have the following structure: [carbohydrate structure: see text] where (S)-1-carboxyethyl [a residue of (S)-lactic acid] (S-Lac) is an ether-linked residue of (S)-lactic acid. The O-polysaccharide studied is structurally similar to that of P. penneri 26, which differs only in the absence of S-Lac from the GlcNAc residue. Based on the O-polysaccharide structures and serological data of the LPS, it was suggested classifying these strains in one Proteus serogroup, O31, as two subgroups: O(31a), 31b for P. penneri 28 and P. vulgaris PrK 55/57 and O31a for P. penneri 26. A serological relatedness of the LPS of Proteus O(31a), 31b and P. penneri 62 was revealed and substantiated by sharing epitope O31b, which is associated with N-acetylisomuramic acid. It was suggested that a cross-reactivity of P. penneri 28 O-antiserum with the LPS of several other P. penneri strains is due to a common epitope(s) on the LPS core.

Structure of the O-polysaccharide of Proteus vulgaris O44: a new O-antigen that contains an amide of D-glucuronic acid with L-alanine.

The O-polysaccharide of Proteus vulgaris O44, strain PrK 67/57 was studied by 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, ROESY, H-detected 1H, 13C HMQC, HMQC-TOCSY and HMBC experiments. The polysaccharide was found to contain an amide of D-glucuronic acid with L-alanine [D-GlcA6(L-Ala)], and the following structure of the linear pentasaccharide repeating unit was established: [structure: see text]. The structural data of the O-polysaccharide and the results of serological studies with P. vulgaris O44 O-antiserum showed that the strain studied is unique among Proteus bacteria, which is in agreement with its classification in a separate Proteus serogroup, O44.

Immunochemical studies on the O-antigens of Proteus mirabilis O23 and Proteus vulgaris O23.

Analysis by 1H and 13C nuclear magnetic resonance (NMR) spectroscopy demonstrated that the O-specific polysaccharides of Proteus mirabilis PrK 42/57 and P. vulgaris PrK 43/57 are structurally similar to that of P. vulgaris PrK 44/57 and different from the polysaccharide of P. mirabilis PrK 41/57 studied earlier. The lipopolysaccharides of these strains were tested using enzyme immunosorbent assay, passive hemolysis and Western blot with O-antisera against P. mirabilis 42/57 and P. vulgaris 43/57 and 44/57, as well as with cross-absorbed O-antisera. The chemical and serological data revealed the basis for combining the four strains into Proteus serogroup O23 and division of this serogroup to three subgroups, one for P. vulgaris 43/57 and 44/57 and two others for P. mirabilis 41/57 and 42/57.

Structure of the O-specific polysaccharide of Proteus vulgaris O45 containing 3-acetamido-3,6-dideoxy-D-galactose.

An O-specific polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of Proteus vulgaris O45 and studied by sugar and methylation analyses along with 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, ROESY, H-detected 1H,13C HSQC and HMBC experiments. The following structure of the pentasaccharide repeating unit of the polysaccharide was established:-->6)-alpha-D-GlcpNAc-(1-->4)-alpha-D-GalpNAc-(1-->4)-alpha-D-GalpA-(1-->3)-beta-D-GlcpNAc-(1-->2)-beta-D-Fucp3NAc4Ac-(1-->where Fuc3NAc4Ac is 3-acetamido-4-O-acetyl-3,6-dideoxygalactose. A cross-reactivity of anti-P. vulgaris O45 serum was observed with several other Proteus lipopolysaccharides, which contains Fuc3N derivatives.

[Investigation of hydrophobicity of Proteus vulgaris strains and ability of Proteus vulgaris and Proteus penneri strains to penetrate bladder membrane HCV T-29 cells ]. / Badania hydrofobowosci szczepów Proteus vulgaris oraz zdolnosci szczepów Proteus vulgaris i Proteus penneri do penetracji komórek nablonka pecherza moczowego HCV T-29.

Proteus bacilli play a particularly important role in urinary tract infections (UTI). Fimbriae and adherence ability and hemolysins production (HpmA, HlyA) are one of the factors of pathogenicity of these bacteria. In this paper we describe the invasion of HCV T-29 transitional bladder urothelial cells carcinoma strains of P. penneri, as well as P. vulgaris strains belonging to different serogroups. The cytotoxic effect was observed at 8 hour of incubation of the tested cells with P. vulgaris O21 and the same effect (complete lysis) at 6 hours by P. vulgaris O4 (this strain manifests maximal activity in the production of HlyA hemolysin). P. penneri strains, produce different types of fimbriae, expressed similar bacterial invasiveness. The hydrophobic properties of 25 P. vulgaris strains were also tested and only 3 strains occur to have hydrophobic cell surface.