Yersinia pseudotuberculosis produces novel superantigenic toxins designated YPMa ( Y. pseudotuberculosis -derived mitogen), YPMb, and YPMc and has a pathogenicity island termed HPI (high-pathogenicity island) and R-HPI (the right-hand part of the HPI with truncation in its left-hand part) on the chromosome. Analysis of the distribution of these virulence factors allowed for differentiation of species Y. pseudotuberculosis into six subgroups, thus reflecting the geographical spread of two main clones: the YPMa + HPI − Far Eastern systemic pathogenic type belonging to serotypes O1b, -2a, -2b, -2c, -3, -4a, -4b, -5a, -5b, -6, -10, and UT (untypeable) and the YPMs − HPI + European gastroenteric pathogenic type belonging to serotypes O1a and -1b. The YPMa + HPI + pathogenic type belonging to serotypes O1b, -3, -5a, -5b, and UT and the YPMb + HPI − nonpathogenic type belonging to non-melibiose-fermenting serotypes O1b, -5a, -5b, -6, -7, -9, -10, -11, and -12 were prevalent in the Far East. The YPMc + R-HPI + European low-pathogenicity type belonging to non-melibiose-fermenting serotype O3 and the YPMs − HPI − pathogenic type belonging to 15 serotypes were found to be prevalent all over the world. This new information is useful for a better understanding of the evolution and spread of Y. pseudotuberculosis.

A duplex real-time SYBR Green LightCycler PCR (LC-PCR) assay with DNA extraction using the QIAamp DNA Stool Mini kit was evaluated with regard to detection of 8 of 17 species of food-or waterborne pathogens in five stool specimens in 2 h or less. The protocol used the same LC-PCR with 20 pairs of specific primers. The products formed were identified based on a melting point temperature (T m ) curve analysis. The 17 species of food-or waterborne pathogens examined were enteroinvasive Escherichia coli, enteropathogenic E. coli, enterohemorrhagic E. coli, enterotoxigenic E. coli, enteroaggregative E. coli, Salmonella spp., Shigella spp., Yersinia enterocolitica, Yersinia pseudotuberculosis, Campylobacter jejuni, Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Aeromonas spp., Staphylococcus aureus, Clostridium perfringens, and Bacillus cereus. No interference with the LC-PCR assay was observed when stool specimens were artificially inoculated with each bacterial species. The detection levels were approximately 10 5 food-or waterborne pathogenic bacteria per g of stool. The protocol for processing stool specimens for less than 10 4 food-or waterborne pathogenic bacteria per g of stool requires an overnight enrichment step to achieve adequate sensitivity. However, the rapid amplification and reliable detection of specific genes of greater than 10 5 food-or waterborne pathogenic bacteria per g in samples should facilitate the diagnosis and management of food-or waterborne outbreaks.The traditional cultural techniques for the direct isolation and identification of food-or waterborne pathogens from stool specimens in food poisoning outbreaks are time-consuming and laborious. Efforts have been made in diagnostic laboratories to reduce the time required for identification of food-or waterborne pathogens. Real-time PCR is currently used for the diagnosis of infectious agents, and there are few reports of the application of real-time PCR for the direct detection of Escherichia coli strain O157:H7 (11) and Plesiomonas shigelloides (20) in stool specimens. However, traditional PCR is often used for the detection of infectious agents from stool specimens and food samples, and there are a number of previous reports of PCR use for detection of food-or waterborne pathogens, including E. coli (6,14,12,27,37,38,40), Salmonella spp. (26) (38). The published PCR protocols used for detection of food-or waterborne pathogens differ and include the use of different PCR cycler machines, annealing temperatures, and buffer systems. For routine laboratory analysis, the development of streamlined PCR methods would be ideal. In addition, not all of the published PCR methods are so sensitive or specific. Therefore, careful choice of PCR primers, modification of some existing PCR primers, and development of new PCR methods for detection and identification of a wide range of food-or waterborne pathogens are all necessary for the development of a standardized PCR protocol (38). Moreover, interference and inhibitors should be considered...

The prevalence and concentration of Shiga toxin-producing Escherichia coli (STEC) in cattle faeces (n=605) at the time of slaughter was studied in Shimane Prefecture, Japan on a monthly basis between April 2000 and March 2001. Screening with stx-PCR determined a prevalence of 37.5%. After analysis of spread faeces and enriched samples on cefixime, tellurite and sorbitol-MacConkey agar using HCl treatment, 114 STEC strains were singly or concomitantly isolated from 97 cattle (15.9%). Of the 605 cattle, 31 (5.1%) harbored O26:H11, O111:H-, O121:H19 or O157:H7, which had the stx1 and/or stx2 and eae and hlyA genes, and 7 (23%) of these 31 cattle were high level carriers that contained these typical STEC at concentrations of 10(5)-10(8) CFU/g of faeces. The predominant serotype was O26:H11 (20 strains) and the second most frequent was O157:H7 (9 strains). Of the 605 cattle, 68 (11.2%) harboured 36 other serotypes and 6 (5.9%) of the 67 cattle were high level carriers. As a comparison between the prevalence of STEC and the faecal pH, it was demonstrated that STEC can be isolated from cattle with a wide range of faecal pH values. The presence of a high-carriage animal at the abattoir increases the potential risk of meat contamination during the slaughtering process, regardless of faecal pH.

Amiodarone treatment can improve cardiac symptom, function, and sympathetic nerve activity, as evaluated by I-123 metaiodobenzylguanidine imaging in patients with dilated cardiomyopathy, which improves to a similar extent with beta-blocker treatment.

While there are several studies on the ecology of Vibrio vulnificus and Vibrio parahaemolyticus in estuarine water environments around the world, there is little information on the distribution of both organisms during the cold-weather months. Thus, we conducted a multi-year study on the ecology of both organisms in brackish environments of the Sada River, a drainage canal flowing slowly into the Japan Sea from Lake Shinji in Shimane Prefecture, Japan. Water samples were collected twice a month at five sites from August 2000 to May 2002. Both organisms were enumerated in 10 l water, 100 g sediment and 10 g shellfish by the most probable number (MPN) procedure. Isolates were confirmed as V. vulnificus using hemolysin gene PCR. During the last 7 months (including winter) of the study, water and sediment samples were also analyzed for the presence of both organisms. V. parahaemolyticus was isolated from river mouths and coastal environments of average salinity > or = 4.4+/-2.0 ppt throughout the year at cell concentrations of 10(-3) to 10(1) MPN ml(-1). Similar concentrations of V. vulnificus were isolated from coastal environments of average salinity 24.0+/-5.4 ppt, except for two times when water moved to the upper reaches due to high tide and V. vulnificus was rifted to the upper reaches. These findings suggest that both organisms are continuously distributed in the Sada estuary throughout the year regardless of water temperature.