Severe Outbreak of Sorbitol-Fermenting Escherichia coli O157 via Unpasteurized Milk and Farm Visits, Finland 2012.

Shiga toxin-producing, sorbitol-fermenting Escherichia coli O157 (SF O157) has emerged as a cause of severe human illness. Despite frequent human findings, its transmission routes and reservoirs remain largely unknown. Foodborne transmission and reservoir in cattle have been suspected, but with limited supporting evidence. This study describes the outbreak of SF O157 that occurred in Finland in 2012. The outbreak originated from a recreational farm selling unpasteurized milk, as revealed by epidemiologic and microbiological investigations, and involved six hospitalized children and two asymptomatic adults with culture-confirmed infection. An identical strain of SF O157 was isolated from patients, cattle and the farm environment, and epidemiologic analysis suggested unpasteurized milk as the vehicle of transmission. This study reports the first milkborne outbreak of SF O157, provides supporting evidence of cattle as a reservoir and highlights the health risks related to the consumption of unpasteurized milk.

Hybrids of Shigatoxigenic and Enterotoxigenic Escherichia coli (STEC/ETEC) Among Human and Animal Isolates in Finland.

Diarrhoeagenic Escherichia coli (DEC) cause serious foodborne infections in humans. Total of 450 Shigatoxigenic E. coli (STEC) strains isolated from humans, animals and environment in Finland were examined by multiplex PCR targeting the virulence genes of various DEC pathogroups simultaneously. One per cent (3/291) of the human STEC and 14% (22/159) of the animal and environmental STEC had genes typically present in enterotoxigenic E. coli (ETEC). The strains possessed genes encoding both Shiga toxin 1 and/or 2 (stx1 and/or stx2 ) and ETEC-specific heat-stable (ST) enterotoxin Ia (estIa). The identified stx subtypes were stx1a, stx1c, stx2a, stx2d and stx2g. The three human STEC/ETEC strains were isolated from the patients with haemolytic uraemic syndrome and diarrhoea and from an asymptomatic carrier. The animal STEC/ETEC strains were isolated from cattle and moose. The human and animal STEC/ETEC strains belonged to 11 serotypes, of which O2:H27, O15:H16, O101:H-, O128:H8 and O141:H8 have previously been described to be associated with human disease. Identification of multiple virulence genes offers further information for assessing the virulence potential of STEC and other DEC. The emergence of novel hybrid pathogens should be taken into account in the patient care and epidemiological surveillance.

High frequency of reactive arthritis in adults after Yersinia pseudotuberculosis O:1 outbreak caused by contaminated grated carrots.

OBJECTIVE: We describe the epidemiological and microbiological process in the clearing of a foodborne outbreak of Yersinia pseudotuberculosis O:1 linked to raw carrots and frequency of the associated reactive extra-gastrointestinal manifestations. METHODS: The patient samples were investigated by routine culture or antibody testing methods. The real-time bacterial PCR was used to detect Y pseudotuberculosis in samples from the grated carrots and in those taken from the carrot storage. Genotype of bacterial isolates was determined by pulsed-field gel electrophoresis. For case identification, we retrospectively looked over the laboratory files of the central hospital focusing on the time period of the outbreak. RESULTS: Altogether 49 case patients were identified. Y pseudotuberculosis was detected by real-time PCR analysis in samples taken from grated carrots and from the carrot distributor. Bacterial isolates originating from the farm environment showed identical serotype (O:1) and genotype (S12) with the patients' isolates. Among 37 adults, reactive arthritis (ReA) was found in 8 (22%) and three adults had probable ReA. Six (67%) out of nine human leucocyte antigen (HLA) typed patients with ReA were HLA-B27 positive. Erythema nodosum was found in 42% of the 12 children, whereas none of them had definite ReA. CONCLUSIONS: In this outbreak, Y pseudotuberculosis was for the first time detected in both patient and food samples. ReA was more common than earlier reported in the outbreaks associated with this pathogen; the reason may be that the previous outbreaks have occurred among children. HLA-B27 frequency was higher than usually reported in single-source outbreaks of ReA.

Yersinia pseudotuberculosis causing a large outbreak associated with carrots in Finland, 2006.

A large outbreak of Yersinia pseudotuberculosis O:1 infection affected over 400 children from 23 schools and 5 day-care centres in two municipalities in southern Finland in August-September, 2006. A retrospective cohort study conducted in a large school centre showed that the outbreak was strongly associated with the consumption of grated carrots served at a school lunch. The risk of illness increased with the amount of carrots eaten. Poor quality carrots grown the previous year had been delivered to the school kitchens in the two municipalities affected. In the patients' samples and in the environmental samples collected from the carrot distributor's storage facility, identical serotypes and genotypes of Y. pseudotuberculosis were found, but the original source and the mechanism of the contamination of the carrots remained unclear. Outbreaks of Y. pseudotuberculosis linked to fresh produce have been detected repeatedly in Finland. To prevent future outbreaks, instructions in improved hygiene practices on the handling of raw carrots have been issued to farmers, vegetable-processing plants and institutional kitchens.

TaqMan-based real-time PCR method for detection of Yersinia pseudotuberculosis in food.

A sensitive and specific assay for detection of food-borne pathogenic Yersinia pseudotuberculosis was developed. The primer-probe set was designed to target a 157-bp sequence of the chromosomally located gene ail. The complete method, including an internal amplification control, was evaluated for several different food items.

Real-time PCR method for detection of pathogenic Yersinia enterocolitica in food.

The current methods for the detection of pathogenic Yersinia enterocolitica bacteria in food are time consuming and inefficient. Therefore, we have developed and evaluated in-house a TaqMan probe-based real-time PCR method for the detection of this pathogen. The complete method comprises overnight enrichment, DNA extraction, and real-time PCR amplification. Also included in the method is an internal amplification control. The selected primer-probe set was designed to use a 163-bp amplicon from the chromosomally located gene ail (attachment and invasion locus). The selectivity of the PCR method was tested with a diverse range (n = 152) of related and unrelated strains, and no false-negative or false-positive PCR results were obtained. The sensitivity of the PCR amplification was 85 fg purified genomic DNA, equivalent to 10 cells per PCR tube. Following the enrichment of 10 g of various food samples (milk, minced beef, cold-smoked sausage, fish, and carrots), the sensitivity ranged from 0.5 to 55 CFU Y. enterocolitica. Good precision, robustness, and efficiency of the PCR amplification were also established. In addition, the method was tested on naturally contaminated food; in all, 18 out of 125 samples were positive for the ail gene. Since no conventional culture method could be used as a reference method, the PCR products amplified from these samples were positively verified by using conventional PCR and sequencing of the amplicons. A rapid and specific real-time PCR method for the detection of pathogenic Y. enterocolitica bacteria in food, as presented here, provides a superior alternative to the currently available detection methods and makes it possible to identify the foods at risk for Y. enterocolitica contamination.

Multiple outbreaks of Yersinia pseudotuberculosis infections in Finland.

During 2001, 89 culture-confirmed cases of Yersinia pseudotuberculosis were reported in Finland; 55 (62%) were serotype O:1, and 34 (38%) were serotype O:3. Four major pulsed-field gel electrophoresis profiles were identified. A case-control study of 25 case patients and 71 healthy controls identified eating outside the home as a risk factor for infection.

Correspondence of genotypes of sporadic Yersinia enterocolitica bioserotype 4/O:3 strains from human and porcine sources.

The sources and transmission routes of sporadic Yersinia enterocolitica bioserotype 4/O:3 infections in Finland were studied. A total of 212 human strains were compared with 334 non-human strains, including 163 strains from pig slaughterhouses, 164 strains from retail outlets and 7 strains from pet animals. All strains were characterized using pulsed-field gel electrophoresis (PFGE) with NotI enzyme. When the 194 human and 287 non-human strains of 22 identical NotI profiles were further characterized with ApaI and XhoI enzymes, 126 genotypes (DI = 094) were distinguished. Of all 212 human strains, 80% were genetically indistinguishable from the strains found in samples of pig origin when characterized with the three enzymes. A major contamination source of sporadic Y. enterocolitica 4/O:3 infections was revealed to be edible pig offal: 71% of the human strains were indistinguishable from the strains isolated from tongues, livers, kidneys and hearts of pigs. These results reveal that in Finland contaminated pig offal is an important vehiclein the transmission of Y. enterocolitica bioserotype 4/O:3 from slaughterhouses to humans.

Bacterial adherence to silver nitrate coated poly-L-lactic acid urological stents in vitro.

The purpose of this study was to see whether it is possible to prevent bacterial adherence to bioabsorbable self-reinforced L-lactic acid polymer (SR-PLLA) urological stents. The SR-PLLA stents were coated with silver nitrate blended epsilon-caprolactone/L-lactide copolymer. The adherence of five bacterial strains (Pseudomonas aeruginosa, Enterococcus faecalis, Proteus mirabilis and two strains of Escherichia coli) to coated and non-coated SR-PLLA wires were tested. It was found that silver nitrate coating prevented the adherence of bacteria (except E. faecalis) to SR-PLLA stents. The preventive effect correlated with the silver nitrate concentration. It was also found that silver nitrate coating reduced the amount of bacteria in ambient urine. In conclusion, silver nitrate coating may reduce stent-associated bacterial infections by preventing the adherence of bacteria. Further studies are needed to confirm its efficacy and safety in clinical practice.

Bacterial adherence to ofloxacin-blended polylactone-coated self-reinforced L-lactic acid polymer urological stents.

OBJECTIVE: To determine whether ofloxacin coating has any effect on bacterial adherence to bioresorbable self-reinforced L-lactic acid polymer (SR-PLLA) urological stents. MATERIALS AND METHODS: SR-PLLA stents were coated with epsilon-caprolactone/L-lactide copolymer blended with ofloxacin at three different concentrations of ofloxacin (0.5, 2 and 5% w/w). The adherence of five bacterial strains (Pseudomonas aeruginosa, Enterococcus faecalis, Proteus mirabilis and two strains of Escherichia coli) to the coated SR-PLLA stents was analysed. Uncoated stent pieces were used as controls. The effect of ofloxacin coating on bacterial growth in the microenvironment of the stent pieces was also analysed. RESULTS: Ofloxacin coating prevented bacterial adherence to SR-PLLA stent material; this effect correlated significantly with the ofloxacin concentration of the caprolactone coating. Ofloxacin coating reduced the amount of bacteria in the microenvironment of the stent, but because of natural resistance, ofloxacin coating had little effect on E. faecalis. CONCLUSION: Except for E. faecalis, ofloxacin coating may reduce stent-associated infections. However, further studies are needed to confirm its biocompatibility and efficacy in clinical use.