Escherichia coli O157:H7, a toxin-producing food and waterborne bacterial pathogen, has been linked to large outbreaks of gastrointestinal illness for more than two decades. E. coli O157 causes a ...wide range of clinical illness that varies by outbreak, although factors that contribute to variation in disease severity are poorly understood. Several recent outbreaks involving O157 contamination of fresh produce (e.g., spinach) were associated with more severe disease, as defined by higher hemolytic uremic syndrome and hospitalization frequencies, suggesting that increased virulence has evolved. To test this hypothesis, we developed a system that detects SNPs in 96 loci and applied it to >500 E. coli O157 clinical strains. Phylogenetic analyses identified 39 SNP genotypes that differ at 20% of SNP loci and are separated into nine distinct clades. Differences were observed between clades in the frequency and distribution of Shiga toxin genes and in the type of clinical disease reported. Patients with hemolytic uremic syndrome were significantly more likely to be infected with clade 8 strains, which have increased in frequency over the past 5 years. Genome sequencing of a spinach outbreak strain, a member of clade 8, also revealed substantial genomic differences. These findings suggest that an emergent subpopulation of the clade 8 lineage has acquired critical factors that contribute to more severe disease. The ability to detect and rapidly genotype O157 strains belonging to such lineages is important and will have a significant impact on both disease diagnosis and treatment guidelines.
While increasing data on bacterial evolution in controlled environments are available, our understanding of bacterial genome evolution in natural environments is limited. We thus performed full ...genome analyses on four Listeria monocytogenes, including human and food isolates from both a 1988 case of sporadic listeriosis and a 2000 listeriosis outbreak, which had been linked to contaminated food from a single processing facility. All four isolates had been shown to have identical subtypes, suggesting that a specific L. monocytogenes strain persisted in this processing plant over at least 12 years. While a genome sequence for the 1988 food isolate has been reported, we sequenced the genomes of the 1988 human isolate as well as a human and a food isolate from the 2000 outbreak to allow for comparative genome analyses.
The two L. monocytogenes isolates from 1988 and the two isolates from 2000 had highly similar genome backbone sequences with very few single nucleotide (nt) polymorphisms (1 - 8 SNPs/isolate; confirmed by re-sequencing). While no genome rearrangements were identified in the backbone genome of the four isolates, a 42 kb prophage inserted in the chromosomal comK gene showed evidence for major genome rearrangements. The human-food isolate pair from each 1988 and 2000 had identical prophage sequence; however, there were significant differences in the prophage sequences between the 1988 and 2000 isolates. Diversification of this prophage appears to have been caused by multiple homologous recombination events or possibly prophage replacement. In addition, only the 2000 human isolate contained a plasmid, suggesting plasmid loss or acquisition events. Surprisingly, besides the polymorphisms found in the comK prophage, a single SNP in the tRNA Thr-4 prophage represents the only SNP that differentiates the 1988 isolates from the 2000 isolates.
Our data support the hypothesis that the 2000 human listeriosis outbreak was caused by a L. monocytogenes strain that persisted in a food processing facility over 12 years and show that genome sequencing is a valuable and feasible tool for retrospective epidemiological analyses. Short-term evolution of L. monocytogenes in non-controlled environments appears to involve limited diversification beyond plasmid gain or loss and prophage diversification, highlighting the importance of phages in bacterial evolution.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
PulseNet is a national network of pubic health and food regulatory laboratories established in the US to detect clusters of foodborne disease and respond quickly to foodborne outbreak investigations. ...PulseNet laboratories currently subtype
Escherichia coli O157:H7, non-typhoidal
Salmonella, and
Shigella isolates by a highly standardized 1-day pulsed-field gel electrophoresis (PFGE), and exchange normalized DNA “fingerprint” patterns via the Internet. We describe a standardized molecular subtyping protocol for subtyping
Listeria monocytogenes that was recently added to PulseNet. The subtyping can be completed within 30 h from the time a pure culture of the bacteria is obtained.
The epidemiology of human listeriosis Swaminathan, Bala; Gerner-Smidt, Peter
Microbes and infection,
08/2007, Letnik:
9, Številka:
10
Journal Article
Recenzirano
Odprti dostop
Listeriosis is a serious invasive disease that primarily afflicts pregnant women, neonates and immunocompromised adults. The causative organism,
Listeria monocytogenes, is primarily transmitted to ...humans through contaminated foods. Outbreaks of listeriosis have been reported in North America, Europe and Japan. Soft cheeses made from raw milk and ready-to-eat meats are high risk foods for susceptible individuals. Efforts by food processors and food regulatory agencies to aggressively control
L. monocytogenes in the high risk foods have resulted in significant decreases in the incidence of sporadic listeriosis.
Since the establishment of the well-known Tenover criteria in 1995 (Tenover et al., 1995), relatively few papers have been published about the interpretation of subtyping data generated by ...pulsed-field gel electrophoresis (PFGE). This paper describes the approach that has been used in the PulseNet network during the past 10 years. PFGE data must always be interpreted in the proper epidemiological context and PFGE data can not alone prove an epidemiological connection. The Tenover criteria are not generally applicable to the interpretation of PFGE subtyping data of foodborne pathogens. The reproducibility of the method with a particular organism, the quality of the PFGE gel, the variability of the organism being subtyped, and the prevalence of the pattern in question must always be considered. Only isolates displaying indistinguishable patterns should be included in the detection of clusters of infections or the initial case definition in a point-source outbreak. More variability (patterns differing from each other in two to three band positions) may be accepted if the outbreak has been going on for some time or if person-person spread is a prominent feature. If epidemiological information is sufficiently strong, isolates with markedly different PFGE patterns may be included in an outbreak.
PulseNet USA, the national molecular subtyping network for foodborne disease surveillance, began functioning in the United States in 1996 and soon established itself as a critical early warning ...system for foodborne disease outbreaks, particularly those in which cases may be geographically dispersed. The PulseNet network is now being replicated in different ways in Canada, Europe, the Asia Pacific region, and Latin America. These independent networks work together in PulseNet International allowing public health officials and laboratorians to share molecular epidemiologic information in real-time and enabling rapid recognition and investigation of multi-national foodborne disease outbreaks. Routine communication between the various international PulseNet networks will provide early warning on foodborne disease outbreaks to participating public health institutions and countries.
During May and June of 2005, 26 persons in several states were infected by a single strain (isolates indistinguishable by pulsed-field gel electrophoresis) of Salmonella enterica serotype Typhimurium ...after eating cake batter ice cream. The cake mix used to prepare the cake batter in the ice cream was implicated by epidemiologic investigation as the source of Salmonella contamination. Initial tests did not detect Salmonella in cake mix collected during the outbreak investigation. The objective of this study was to evaluate different procedures to isolate Salmonella from the implicated cake mix, cake, and ice cream. All outbreak-associated food samples (14 samples) were collected during the outbreak investigation by health departments of several of the states involved. Different combinations of Salmonella isolation procedures, including sample size, preenrichment broth, enrichment broth, enrichment temperature, and isolation medium, were used. Salmonella Typhimurium was isolated from two cake mix samples; the food isolates were indistinguishable from the outbreak pattern by pulsed-field gel electrophoresis subtyping. Universal preenrichment broth was substantially better than was lactose broth for preenrichment, and tetrathionate broth was better than was Rappaport-Vassiliadis broth for isolating Salmonella from the two positive cake mix samples. Although more typical Salmonella colonies were observed on plates from enrichment cultures grown at 35°C, more confirmed Salmonella isolates were obtained from plates of enrichment cultures grown at 42°C. Brilliant green agar, xylose lysine tergitol 4 agar, xylose lysine desoxycholate agar, Hektoen enteric agar, and bismuth sulfite agar plates were equally effective in isolating Salmonella from cake mix. The best combination of preenrichment-enrichment conditions for isolating the outbreak strain of Salmonella was preenrichment of cake mix samples in universal preenrichment broth at 35°C for 24 h, followed by enrichment in tetrathionate broth at 42°C for 24 h.
Recent advancements in biotechnology are rapidly altering the diagnostic procedures used in microbiologic analysis of foods. Biochemical identification tests have been miniaturized and automated, ...making them faster and more economical. Pathogenic bacteria that were previously isolated and identified after labor- and time-intensive enrichment and plating procedures can now be detected by measuring specific physicochemical changes resulting from their growth or metabolic activity. Nucleic acid and antibody-based assays are now used to rapidly and reliably detect pathogenic bacteria in foods. Nevertheless, foods offer unique challenges to the application of these techniques because of their complexity and variety, their interference with the rapid detection methods, and the need to detect pathogenic bacteria when they are present in foods at very low levels. Methods to sequester target pathogenic bacteria from interfering food components and to concentrate them in small volumes are needed to enable the efficient application of rapid detection and identification methods.
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