Non-O157 STEC can cause severe illness that is comparable to the illness caused by STEC O157. Strains that produce Shiga toxin 2 are much more likely to cause HUS than are those that produce Shiga toxin 1 alone. Improving surveillance will more fully elucidate the incidence and pathological spectrum of these emerging agents. These efforts require increased clinical suspicion, improved clinical laboratory isolation, and continued serotyping of isolates in public health laboratories.

We examined 1,266 fecal specimens from healthy cattle during the investigations of two sporadic cases of hemolytic uremic syndrome associated with raw milk consumption and an outbreak of gastroenteritis and hemolytic uremic syndrome caused by Escherichia coli serotype 0157:117. We collected specimens from heifers, calves, and adult cows on 22 farms, in a stockyard, and in a packing house. We also collected 3 raw hamburger specimens from a restaurant and 23 raw milk samples from two farms. All specimens were examined for E. coli 0157:H7 by using sorbitol-MacConkey agar, H immobilization, 0157 agglutination, and tissue culture cytotoxicity. E. coli 0157:H7 was isolated from 16 heifers or calves and 1 adult cow on 22 farms, 1 stockyard calf, 2 beef specimens, and 1 raw milk sample. Selected fecal specimens were also examined for the presence of other Shiga-like-toxin-producing E. coli (SLTEC) by testing polymyxin B extracts of colony sweeps and then testing individual colonies for toxin production. SLTEC other than 0157 was isolated from 8 of 10 farms investigated and from the stockyard; 8% of adult cows and 19% of heifers and calves were positive for SLTEC. Several animals were positive for SLTEC by colony sweep only. This investigation demonstrates that dairy cattle are a reservoir of E. coil 0157:H7 and other SLTEC.

Vibrio, a diverse genus of aquatic bacteria, currently includes 72 species, 12 of which occur in human clinical samples. Of these 12, three species-Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus-account for the majority of Vibrio infections in humans. Rapid and accurate identification of Vibrio species has been problematic because phenotypic characteristics are variable within species and biochemical identification requires 2 or more days to complete. To facilitate the identification of human-pathogenic species, we developed a multiplex PCR that uses species-specific primers to amplify gene regions in four species (V. cholerae, V. parahaemolyticus, V. vulnificus, and V. mimicus). The assay was tested on a sample of 309 Vibrio isolates representing 26 named species (including 12 human pathogens) that had been characterized by biochemical methods. A total of 190 isolates that had been identified as one of the four target species all yielded results consistent with the previous classification. The assay identified an additional four V. parahaemolyticus isolates among the other 119 isolates. Sequence analysis based on rpoB was used to validate the multiplex results for these four isolates, and all clustered with other V. parahaemolyticus sequences. The rpoB sequences for 12 of 15 previously unidentified isolates clustered with other Vibrio species in a phylogenetic analysis, and three isolates appeared to represent unnamed Vibrio species. The PCR assay provides a simple, rapid, and reliable tool for identification of the major Vibrio pathogens in clinical samples, and rpoB sequencing provides an additional identification tool for other species in the genus Vibrio.The genus Vibrio is a highly diverse group of gram-negative bacteria that contains approximately 72 species (www.bacterio .net). The group includes symbionts and commensals that are found in or on marine animals, as well as many species that are pathogenic to animals (21). There are 12 species that are routinely isolated from human clinical samples, and the diseases in which they are implicated include diarrheal disease, septicemia, and wound infections (6).Three species account for the majority of human Vibrio infections. Toxigenic Vibrio cholerae is the causative agent of the disease cholera and is acquired through ingestion of contaminated food or water. Infection may lead to a profuse, watery diarrhea that can lead to severe dehydration and death if left untreated. Worldwide, large outbreaks are caused by toxigenic strains of serogroups O1 and O139 that produce the cholera toxin, but in the United States, nontoxigenic strains predominate among the cases reported to the Centers for Disease Control and Prevention (CDC) (4). A total of 48 cases of cholera occurred in the United States in 2004 and were reported to the CDC (4), including eight cases infected with toxigenic V. cholerae, four of which were associated with travel, and 40 cases infected with nontoxigenic cholera. Vibrio cholerae accounted for 9.8% of all Vibrio isolates reported...

Six commercially available bacterial identification products were tested with Vibrio alginolyticus (12 strains), V. cholerae (30 strains), Photobacterium (Vibrio) damselae (10 strains), V. fluvialis (10 strains), V. furnissii (4 strains), V. hollisae (10 strains), V. metschnikovii (9 strains), V. mimicus (10 strains), V. parahaemolyticus (30 strains), and V. vulnificus (10 strains) to determine the accuracy of each system for identification. The products included API 20E, Crystal E/NF, MicroScan Neg ID2 and Rapid Neg ID3, and Vitek GNI؉ and ID-GNB. Each product was tested only with those species that were listed in its database. Overall, the systems correctly identified 63.9, 80.9, 63.1, 73.6, 73.5, and 77.7% of the isolates to species level, respectively. Error rates ranged from 0.8% for the API 20E to 10.4% for the Rapid Neg ID3. The API 20E gave "no identification" for 13.1% of the isolates, while the Neg ID2, GNI؉, ID-GNB, and Crystal were unable to identify 1.8, 2.9, 5.0, and 6.9%, respectively. For V. cholerae, specifically, accuracy ranged from 50.0 to 96.7%, with the API 20E having the worst performance and Crystal having the best. V. fluvialis presented the biggest challenge for the API 20E and the GNI؉, with probabilities averaging 10%, while V. mimicus was a major problem with the Crystal E/NF, which identified none of the strains correctly. With the Neg ID2, correct answers were often obtained only after a modified inoculation of the panel with a bacterial suspension prepared with 0.85% NaCl. Additional tests required for identification often included growth in the absence of NaCl, which is not readily available in most clinical laboratories. The only product to correctly identify at least 90% of V. cholerae strains was the Crystal E/NF, and only three of the six products, the API 20E and both of the Vitek cards, correctly identified more than 90% of the V. parahaemolyticus strains. Thus, extreme care must be taken in the interpretation of answers from these six commercially available systems for the identification of Vibrio species.There are approximately 63 species of Vibrio (www.bacterio .cict.fr), of which 12 occur in human clinical specimens (11). Of particular interest to clinical microbiology laboratories is Vibrio cholerae, which has been the cause of many epidemics and deaths and which is generally acknowledged to be of serious concern to those currently involved in bioterrorism. Indeed, when the list of critical agents for bioterrorism and civil preparedness was released (21), V. cholerae was classified as a biothreat level B, which is defined as an agent having a moderate ease of transmission and morbidity with a low rate of mortality (31). Thus, correct identification of this organism and its differentiation from other species of the genus are important.Other species of Vibrio such as V. vulnificus, which has a fatality rate of approximately 50% in patients with primary septicemia, also cause significant human disease (11). Photobacterium (Vibrio) damselae, V. metschnikovii, and V. cincin...