Anja Siitonen

An international outbreak of Salmonella infections caused by alfalfa sprouts grown from contaminated seeds

An outbreak of Salmonella serotype stanley infections occurred in the United States and Finland in 1995. The outbreak was investigated through case-control studies in Arizona, Michigan, and Finland; by isolate subtyping; and by tracing and culturing of the implicated food. Alfalfa sprout consumption was the only exposure associated with S. stanley infections in Arizona (matched odds ratio [MOR] = 11.1; 95% confidence interval [CI], 1.4-513), Michigan (MOR = 5.5; CI, 1.6-23), and Finland (MOR undefined; CI, 4.9-infinity). US and Finnish patient isolates were a unique outbreak strain distinct from S. stanley isolates not linked to the outbreak. Alfalfa sprouts eaten by patients in 6 US states and Finland were traced to seed shipped by a Dutch shipper. Thus, it was concluded that alfalfa sprouts grown from contaminated seed caused an international outbreak of > or =242 S. stanley infections in > or =17 US states and Finland. This outbreak illustrates a new mechanism through which contamination of fresh produce can cause large, widely dispersed outbreaks.

An outbreak of Listeria monocytogenes serotype 3a infections from butter in Finland.

In February 1999, an outbreak of listeriosis caused by Listeria monocytogenes serotype 3a occurred in Finland. All isolates were identical. The outbreak strain was first isolated in 1997 in dairy butter. This dairy began delivery to a tertiary care hospital (TCH) in June 1998. From June 1998 to April 1999, 25 case patients were identified (20 with sepsis, 4 with meningitis, and 1 with abscess; 6 patients died). Patients with the outbreak strain were more likely to have been admitted to the TCH than were patients with other strains of L. monocytogenes (60% vs. 8%; odds ratio, 17.3; 95% confidence interval, 2.8-136.8). Case patients admitted to the TCH had been hospitalized longer before cultures tested positive than had matched controls (median, 31 vs. 10 days; P=.008). An investigation found the outbreak strain in packaged butter served at the TCH and at the source dairy. Recall of the product ended the outbreak.

Clinical Isolates of Non-O157 Shiga Toxin-Producing Escherichia coli: Serotypes, Virulence Characteristics, and Molecular Profiles of Strains of the Same Serotype

ABSTRACT All human Shiga toxin-producing Escherichia coli (STEC) non-O157 strains (n = 56) isolated in Finland from 1990 to August 2000 were characterized for the O:H serotype,stx1 and stx2 genes, production of enterohemolysin, and sensitivity to 12 antimicrobial agents. Strains of the same serotype were genotyped by pulsed-field gel electrophoresis (PFGE) after XbaI restriction of total DNA. The 56 non-O157 isolates belonged to 29 serotypes. Two of the serotypes (O102:H7 and OX181:H49) have not previously been described as being associated with STEC infections in humans or isolated from animals. Thirty-four strains (61%) within seven serotypes (O103:H2 [14 isolates], O26:H11 [6 isolates], O145:H28 [4 isolates], O145:HNM [3 isolates], O15:HNM [3 isolates], OX174:H21 [2 isolates], and O Rough:HNM [2 isolates]) were represented by more than one isolate. Of these strains, O103:H2 isolates were divided into seven, O26:H11 isolates were divided into four, and the rest within a serotype were divided into two genotypes in PFGE. In PCR, 31 (55%) of the 56 strains were positive for the stx2 gene only and 24 strains (43%) were positive for stx1 only. One strain (O43:H2) carried both stx1 andstx2. Forty-two strains (75%) produced enterohemolysin, and 39 strains (70%) possessed the eaegene. Of the latter 39 strains, 36 (92%) were enterohemolytic, whereas only 6 (35%) of the 17 isolates lacking the eae gene were enterohemolytic (P < 0.001). The majority of the strains (44 strains, 79%) were sensitive to all 12 antimicrobials tested. Of the 56 strains, 20 (36%) were associated with small family outbreaks in nine families and 14 (25%) were associated with recent travel abroad.

Clinical Escherichia coli strains carrying stx genes: stx variants and stx-positive virulence profiles.

ABSTRACT Altogether, 173 Shiga toxin-producing Escherichia coli (STEC) serotype O157 (n = 111) and non-O157 (n = 62) isolates from 170 subjects were screened by PCR-restriction fragment length polymorphism for eight different stx genes. The results were compiled according to serotypes, phage types of O157, production of Stx toxin and enterohemolysin, and the presence of eae. The stx genes occurred in 11 combinations; the most common were stx2 with stx2c (42%), stx2 alone (21%), and stx1 alone (16%). Of the O157 strains, 64% carried stx2 with stx2c versus 2% of the non-O157 strains (P < 0.001). In the non-O157 strains, the prevailing gene was stx1 (99% versus 1% in O157 strains; P < 0.001). In addition, one strain (O Rough:H4:stx2c) which has not previously been described as associated with hemolytic-uremic syndrome (HUS) was found. Ten stx-positive virulence profiles were responsible for 71% of all STEC infections. Of these profiles, five accounted for 71% of the 21 strains isolated from 20 patients with HUS or thrombotic thrombocytopenic purpura (TTP). The strains having the virulence profile that caused mainly HUS or TTP or bloody diarrhea produced Stx with titers of ≥1:128 (90%) more commonly than did other strains (51%; P < 0.001). These strains were also more commonly enterohemolytic (98% versus 68% for other strains; P < 0.001) and possessed the eae gene (100%) more commonly than did other strains (74%; P < 0.001). A particular virulence profile, O157:H7:PT2:stx2:stx2c:eae:Ehly, was significantly more frequently associated with HUS and bloody diarrhea than were other profiles (P = 0.02) and also caused the deaths of two children. In this study, the risk factors for severe symptoms were an age of <5 years and infection by the strain of O157:H7:PT2 mentioned above.

International outbreak of Salmonella Oranienburg due to German chocolate

BackgroundThis report describes a large international chocolate-associated Salmonella outbreak originating from Germany.MethodsWe conducted epidemiologic investigations including a case-control study, and food safety investigations. Salmonella (S.) Oranienburg isolates were subtyped by the use of pulsed-field gel electrophoresis (PFGE).ResultsFrom 1 October 2001 through 24 March 2002, an estimated excess of 439 S. Oranienburg notifications was registered in Germany. Simultaneously, an increase in S. Oranienburg infections was noted in other European countries in the Enter-net surveillance network. In a multistate matched case-control study in Germany, daily consumption of chocolate (matched odds ratio [MOR]: 4.8; 95% confidence interval [CI]: 1.3–26.5), having shopped at a large chain of discount grocery stores (MOR: 4.2; CI: 1.2–23.0), and consumption of chocolate purchased there (MOR: 5.0; CI: 1.1–47.0) were associated with illness. Subsequently, two brands from the same company, one exclusively produced for that chain, tested positive for S. Oranienburg. In two other European countries and in Canada chocolate from company A was ascertained that also contained S. Oranienburg. Isolates from humans and from chocolates had indistinguishable PFGE profiles. No source or point of contamination was identified. Epidemiological identification of chocolate as a vehicle of infections required two months, and was facilitated by proxy measures.ConclusionsDespite the use of improved production technologies, the chocolate industry continues to carry a small risk of manufacturing Salmonella-containing products. Particularly in diffuse outbreak-settings, clear associations with surrogates of exposure should suffice to trigger public health action. Networks such as Enter-net have become invaluable for facilitating rapid and appropriate management of international outbreaks.

A Widespread Outbreak of Yersinia pseudotuberculosis O:3 Infection from Iceberg Lettuce

BACKGROUND The vehicles and sources of Yersinia pseudotuberculosis infection are unknown. In Finland, clinical microbiology laboratories routinely report Y. pseudotuberculosis isolations and submit isolates for serotype analysis. In October 1998, the number of serotype O:3 infections increased markedly. METHODS Case patients with culture-confirmed Y. pseudotuberculosis O:3 infection were identified by use of laboratory-based surveillance. We conducted a population-based case-control study. Healthy community control subjects were matched by age, sex, and postal code. Isolates were subtyped by pulsed-field gel electrophoresis (PFGE). RESULTS Nationwide, 47 case patients were identified (age range, 2-77 years; median, 19 years). One patient with bacteremia died; 5 underwent appendectomies. We enrolled 38 case patients and 76 control subjects in the case-control study. Seventy-one percent of case patients and 42% of control subjects reported having eaten iceberg lettuce (matched odds ratio, 3.8; 95% confidence interval, 1.3-9.4); a dose-response relationship was found for increasing frequency of consumption. Of the 27 isolates obtained from case patients and tested in the analysis, all had indistinguishable PFGE patterns. Four lunch cafeterias that had served iceberg lettuce were associated with clusters of case patients. The lettuce was traced back to originating farms. CONCLUSIONS Iceberg lettuce was implicated as the vehicle of a widespread foodborne Y. pseudotuberculosis outbreak. Ongoing laboratory-based surveillance and serotype analysis were essential in the rapid detection of infection. Cases of yersiniosis, which appear to be sporadic, may be part of unrecognized outbreaks caused by contaminated fresh produce.

Application of molecular genetic methods in diagnostics and epidemiology of food-borne bacterial pathogens†

Salmonella enterica, Campylobacter and Yersinia species, Shiga toxin‐producing Escherichia coli (STEC), Listeria monocytogenes and Clostridium perfringens are the bacterial pathogens constituting the greatest burden of food‐borne disease in Finland. Several molecular genetic methods have been applied to diagnose, discriminate and survey these bacteria. PCR, PCR‐RFLP and PFGE are the most widely and successfully used. However, these methods are unable to replace conventional and internationally standardised phenotyping. Electronic database libraries of the different genomic profiles will enable continuous surveillance of infections and detection of possible infection clusters at an early stage. Furthermore, whole‐genome sequence data have opened up new insights into epidemiological surveillance. Laboratory‐based surveillance performed in a timely manner and exploiting adequate methods, and co‐operation at local, national and international levels are among the key elements in preventing food‐borne diseases. This paper reviews different applications of molecular genetic methods for investigating enteric bacterial pathogens and gives examples of the methods successfully used in diagnostics and epidemiological studies in Finland.

An Outbreak of Gastrointestinal Illness and Erythema Nodosum from Grated Carrots Contaminated with Yersinia pseudotuberculosis

BACKGROUND Outbreaks of Yersinia pseudotuberculosis infection have been epidemiologically linked to fresh produce, but the bacterium has not been recovered from the food items implicated. In May 2003, a cluster of gastrointestinal illness and erythema nodosum was detected among schoolchildren who had eaten lunches prepared by the same institutional kitchen. METHODS We conducted a case-control study and trace-back, environmental, and laboratory investigations. Case patients had culture-confirmed Y. pseudotuberculosis O:1 infection, erythema nodosum, or reactive arthritis. Bacterial isolates from clinical and environmental samples were compared using pulsed-field gel electrophoresis (PFGE). RESULTS Of 7392 persons at risk, 111 (1.5%) met the case definition; 76 case patients and 172 healthy control subjects were enrolled in the case-control study. Only raw grated carrots were significantly associated with illness in a logistic-regression model (multivariable odds ratio, 5.7 [95% confidence interval, 1.7-19.5]); a dose response was found for increasing amount of consumption. Y. pseudotuberculosis O:1 isolates from 39 stool specimens and from 5 (42%) of 12 soil samples that contained carrot residue and were obtained from peeling and washing equipment at the production farm were indistinguishable by PFGE. CONCLUSIONS Carrots contaminated early in the production process caused a large point-source outbreak. Our findings enable the development of evidence-based strategies to prevent outbreaks of this emerging foodborne pathogen.

Molecular Epidemiology of 3 Putative Virulence Genes for Escherichia coli Urinary Tract Infection—usp, iha, and iroNE. coli

This study describes the epidemiological association of 3 putative genes for virulence of uropathogenic Escherichia coli; uropathogenic specific protein (usp), a Vibrio cholerae zot gene homologue; IrgA homologue adhesin (iha), a nonhemagglutinating adhesin; and iroN(E. coli), a catechole siderophore receptor homologue. We compared the relative frequency in urinary tract infection (UTI) isolates (n=508), compared with non-UTI isolates (n=416). iroN(E. coli) occurred 2.1-3.6 times more frequently in UTI isolates than in rectal isolates (P=1.1x10-18 to P=2.7x10-5) and was associated with several uropathogenic virulence genes found on pathogenicity islands. usp occurred more frequently in isolates from patients with pyelonephritis (P=3.6x10-9), in periurethral isolates (P=.001), and in isolates from patients with UTI who were aged 40-65 years (P=.004), when compared with the rectal isolates; iha was not associated with UTI in this study.

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.