Detection of Salmonella contamination in food samples by dot-ELISA, DNA amplification and bacterial culture.
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A dot-blot enzyme-linked immunosorbent assay (dot-ELISA) employing a genus Salmonella specific monoclonal antibody (MAb) was used for detection of the bacteria in food samples in comparison with the conventional culture method and the DNA amplification. Among the 200 chicken and pork samples (100 each) tested, 9% and 33%, 7% and 20% and 7 and 23% were positive for salmonellae by the dot-ELISA, the culture method and the DNA amplification, respectively. Statistical analyses revealed that the sensitivity, specificity, efficacy, and positive and negative predictive values of the detection of Salmonella in the food samples by dot-ELISA compared with the culture method were 93.33%, 91.76%, 92%, 66.66% and 98.73%, respectively. Comparison of the DNA amplification and the culture method revealed the sensitivity, specificity, efficacy, and positive and negative predictive values of 100%, 91.58%, 92%, 65.21% and 100%, respectively. The dot-ELISA and the DNA amplification results were in a better agreement when the two assays were compared. The sensitivity, specificity, efficacy, positive and negative predictive values of the dot-ELISA compared to the DNA amplification were 91.3%, 100%, 98%, 100% and 97.5%, respectively. From this study, the dot-ELISA is rapid, simple, sensitive, specific at low cost with limited amount of infectious waste to be disposed and offers another advantage in that it detects only the smooth LPS of Salmonella which implies the possible presence of the virulent organisms. (+info)
Abiotrophia elegans strains comprise 8% of the nutritionally variant streptococci isolated from the human mouth.
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Ninety-one isolates of nutritionally variant streptococci (NVS) that were previously isolated from the human mouth were regarded as consisting of 7 Streptococcus defectivus isolates, 78 Streptococcus adjacens isolates, and 6 Gemella morbillorum isolates. However, recent references to the taxonomic reclassification of NVS, from S. defectivus to Abiotrophia defectiva and from S. adjacens to Abiotrophia adiacens, and the newly introduced species Abiotrophia elegans as a third Abiotrophia species, emphasize the need for genetic analyses for identification of NVS. When PCR-restriction fragment length polymorphism (RFLP) and phylogenetic distances were examined based on 16S rRNA gene sequences, the results indicated that 7 of the 91 NVS isolates were closely related to A. elegans. These seven isolates consisted of four isolates previously identified as G. morbillorum and three isolates previously identified as S. adjacens. Two isolates previously identified as G. morbillorum were related to A. adiacens. In biochemical tests, A. elegans and the seven isolates related to it possessed arginine dihydrolase (ADH) activity but the other Abiotrophia species did not. As a result, A. elegans strains comprised 8% of the 91 NVS isolates. Our findings suggest that A. elegans, A. adiacens, and A. defectiva exist in the human mouth in proportions of about 1:11:1 and that A. elegans can be genetically distinguished from the other two Abiotrophia species by PCR-RFLP analysis of 16S rRNA gene sequences and can be biochemically distinguished by ADH activity. (+info)
A predictive model that describes the effect of prolonged heating at 70 to 90 degrees C and subsequent incubation at refrigeration temperatures on growth from spores and toxigenesis by nonproteolytic Clostridium botulinum in the presence of lysozyme.
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Refrigerated processed foods of extended durability such as cook-chill and sous-vide foods rely on a minimal heat treatment at 70 to 95 degrees C and then storage at a refrigeration temperature for safety and preservation. These foods are not sterile and are intended to have an extended shelf life, often up to 42 days. The principal microbiological hazard in foods of this type is growth of and toxin production by nonproteolytic Clostridium botulinum. Lysozyme has been shown to increase the measured heat resistance of nonproteolytic C. botulinum spores. However, the heat treatment guidelines for prevention of risk of botulism in these products have not taken into consideration the effect of lysozyme, which can be present in many foods. In order to assess the botulism hazard, the effect of heat treatments at 70, 75, 80, 85, and 90 degrees C combined with refrigerated storage for up to 90 days on growth from 10(6) spores of nonproteolytic C. botulinum (types B, E, and F) in an anaerobic meat medium containing 2,400 U of lysozyme per ml (50 microg per ml) was studied. Provided that the storage temperature was no higher than 8 degrees C, the following heat treatments each prevented growth and toxin production during 90 days; 70 degrees C for >/=2,545 min, 75 degrees C for >/=463 min, 80 degrees C for >/=230 min, 85 degrees C for >/=84 min, and 90 degrees C for >/=33.5 min. A factorial experimental design allowed development of a predictive model that described the incubation time required before the first sample showed growth, as a function of heating temperature (70 to 90 degrees C), period of heat treatment (up to 2,545 min), and incubation temperature (5 to 25 degrees C). Predictions from the model provided a valid description of the data used to generate the model and agreed with observations made previously. (+info)
Production of acylated homoserine lactones by psychrotrophic members of the Enterobacteriaceae isolated from foods.
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Bacteria are able to communicate and gene regulation can be mediated through the production of acylated homoserine lactone (AHL) signal molecules. These signals play important roles in several pathogenic and symbiotic bacteria. The following study was undertaken to investigate whether AHLs are produced by bacteria found in food at temperatures and NaCl conditions commercially used for food preservation and storage. A minimum of 116 of 154 psychrotrophic Enterobacteriaceae strains isolated from cold-smoked salmon or vacuum-packed chilled meat produced AHLs. Analysis by thin-layer chromatography indicated that N-3-oxo-hexanoyl homoserine lactone was the major AHL of several of the strains isolated from cold-smoked salmon and meat. AHL-positive strains cultured at 5 degrees C in medium supplemented with 4% NaCl produced detectable amounts of AHL(s) at cell densities of 10(6) CFU/ml. AHLs were detected in cold-smoked salmon inoculated with strains of Enterobacteriaceae stored at 5 degrees C under an N(2) atmosphere when mean cell densities increased to 10(6) CFU/g and above. Similarly, AHLs were detected in uninoculated samples of commercially produced cold-smoked salmon when the level of indigenous Enterobacteriaceae reached 10(6) CFU/g. This level of Enterobacteriaceae is often found in lightly preserved foods, and AHL-mediated gene regulation may play a role in bacteria associated with food spoilage or food toxicity. (+info)
Rapid fluorescence assessment of the viability of stressed Lactococcus lactis.
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The aim of this study was to establish the use of the fluorescent probes carboxyfluorescein (cF) and propidium iodide (PI) for rapid assessment of viability, using Lactococcus lactis subsp. lactis ML3 exposed to different stress treatments. The cF labeling indicated the reproductive capacity of mixtures of nontreated cells and cells killed at 70 degrees C very well. However, after treatment up to 60 degrees C the fraction of cF-labeled cells remained high, whereas the survival decreased for cells treated at above 50 degrees C and was completely lost for those treated at 60 degrees C. In an extended series of experiments, cell suspensions were exposed to heating, freezing, low pH, or bile salts, after which the colony counts, acidification capacity, glycolytic activity, PI exclusion, cF labeling, and cF efflux were measured and compared. The acidification capacity corresponded with the number of CFU. The glycolytic activity, which is an indicator of vitality, was more sensitive to the stress conditions than the reproduction, acidification, and fluorescence parameters. The cF labeling depended on membrane integrity, as was confirmed by PI exclusion. The fraction of cF-labeled cells was not a general indicator of reproduction or acidification, nor was PI exclusion or cF labeling capacity (the internal cF concentration). When the cells were labeled by cF, a subsequent lactose-energized efflux assay was needed for decisive viability assessment. This novel assay proved to be a good and rapid indicator of the reproduction and acidification capacities of stressed L. lactis and has potential for physiological research and dairy applications related to lactic acid bacteria. (+info)
Health risks associated with unpasteurized goats' and ewes' milk on retail sale in England and Wales. A PHLS Dairy Products Working Group Study.
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A pilot study to determine the microbiological quality of unpasteurized milk from goats and ewes sampled from farm shops, health food shops, and other retail premises found that 47%, (47/100) of goats' and 50% (13/26) of ewes' milk samples failed the standards prescribed by the Dairy Products (Hygiene) Regulations 1995. In addition, Staphylococcus aureus, haemolytic streptococci or enterococci, were present in excess of 10(2) c.f.u./ml in 9 (7 %) 2 (2 %) and 19 (15%) samples, respectively. Salmonella, campylobacter, verocytotoxin-producing Escherichia coli O157:H7 and Listeria monocytogenes were not detected in the samples. At the time of purchase approximately half of the milk samples (58 %) were frozen, the rest were liquid. Farm outlets sold predominantly liquid milk, other retail premises sold a frozen product. The microbiological quality of goats' and ewes' milk, whether frozen or liquid, was not significantly different. Milk sold from farm shops was of lower quality than that from health food shops and other retail premises. In this pilot study most producers (92 %) supplied, and most retailers (76 %) sold unpasteurized goats' and ewes' milk that contained unacceptable levels of indicator organisms. The study was carried out during the winter when goats' milk production is reduced. The results indicate the need for a full representative study of unpasteurized goats' and ewes' milk on retail sale throughout the year. (+info)
Genetic subtyping of Escherichia coli O157 isolates from 41 Pacific Northwest USA cattle farms.
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Escherichia coli O157 (n = 376) from 41 cattle farms were subtyped using pulsed field gel electrophoresis of endonuclease cleaved chromosomal DNA. Cleavage with XbaI resulted in 81 subtypes. Fifty-one isolates from subtypes found in more than one herd, or in herds on multiple sample collection dates were compared using the endonuclease NotI, resulting in 23 additional subtypes. Up to 11 XbaI subtypes were found per farm with up to 7 subtypes/farm identified from a single date. Indistinguishable subtypes (both XbaI and NotI) were found to persist on 4 farms for 6-24 months. Five subtypes were found on more than one farm separated by up to 640 km. Dairy farms where cattle had moved onto the farm had a similar number of subtypes as farms with no movement of cattle, and feedlots had more subtypes than dairy farms. These data indicate that there is a mechanism for multiple herd exposure to specific subtypes, there are multiple sources of exposure for cattle on farms, and on-farm reservoirs other than cattle may exist. (+info)
Discrimination between endemic and feedborne Salmonella Infantis infection in cattle by molecular typing.
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Salmonella enterica serovar Infantis is endemic in Finnish cattle. Feed contaminated with S. Infantis was distributed to cattle farms in May 1995. Following increased sampling, S. Infantis was detected on 242 farms in 1995. Molecular typing was used to differentiate the farms that were infected by the feed-related Infantis from those infected by other endemic strains. Twenty-three isolates from feed in 1995 and 413 from cattle (72 from 19924, 324 from 1995, 17 from 1996-7) were analysed. The feed-related Infantis was clonally related to the endemic infection by the ribotype, IS200-type and XbaI-profile. The feed isolates had a distinctive plasmid that appeared in pulsed-field gel electrophoresis as a 60 kb band when cleaved with XbaI or linearized by S1-nuclease. This plasmid appeared in cattle only since the outbreak and seemed stable on the follow-up farms. In addition to contact farms, the feedborne strain was found on 19% of the farms infected with S. Infantis in 1995 but not having bought suspected feedstuffs, possibly as secondary infections. (+info)