A 70-year-old woman fell seriously ill overnight with meningitis and was admitted to hospital. Cerebrospinal fluid culture yielded Listeria monocytogenes. One of the first problems in solving a human case of listeriosis suspected to be foodborne is to find the foods likely to have been transmitting L. monocytogenes. Two enrichment procedures and a direct plating procedure were used for isolation of the bacteria from different food items collected from the patient's refrigerator, local retail store and producer. Samples of vacuum-packed products of sliced pork brawn, sliced cooked medwurst and berliner wurst of the same brand harboured L. monocytogenes. Serotyping and restriction enzyme analysis (REA) with pulsed-field gel electrophoresis (PFGE) were used to characterize and compare 41 isolates, including the human strain. At least three clones were present in the foods investigated, and one of these was identical to the human clone. This clone was present in samples of medwurst from the patient's refrigerator and the local retail store. This is, to our knowledge, the first proven foodborne case of listeriosis reported in Sweden.
Since 1986, 68% of the Listeria monocytogenes isolates from human cases of invasive listeriosis in Sweden are available for retrospective studies. The aim of the present study was to characterize 601 human invasive isolates of L. monocytogenes in Sweden from 1986 to 2007 by using serotyping and pulsed-field gel electrophoresis. Since 1996, serovar 4b was permanently reduced to the second or third most common serovar in human cases in Sweden. During the latter period, 2000-2007, only 13% belonged to serovar 4b and 71% to 1/2a. The dendrogram, based on pulsovars, reveals two clusters with different serovars. Cluster 1 exhibits serovars 4b and 1/2b, whereas cluster 2 consists of serovar 1/2a. Serovar 1/2a seems to be more heterogeneous than serovar 4b.
In 1958-74 altogether 64 cases of bacteriologically verified infections of Listeria monocytogenes were diagnosed in Sweden in children, aged more than 27 days, and in adults. Immunosuppression predisposed to the disease. Thus, many patients had co-existing disorders, such as leukemia and alcoholism. Sixteen patients had been treated with corticosteroids, which were combined with cytostatic drugs in nine. Meningoencephalitis was diagnosed in 52 patients and was fatal in 16. The clinical symptoms did not differ from those in purulent meningitis caused by other bacteria. In the cerebrospinal fluid the cellular response was dominated by polymorphonuclear cells in 29 patients and by mononuclear cells in 20. Ten patients had septicemia, which was fatal in four. Clinical symptoms were dominated by chills, high fever and general prostration. One patient had pleurisy and one an abscess of the neck; both recovered. Serotypes 1 and 4b prevailed and were equally common. Many patients developed raised antibody titers in both the O-agglutination test and the complement fixation test. The titers were often not positive until after a month. Moderate granulocytosis was the rule and monocytosis was rarely seen. Ampicillin alone or combined with an aminoglycoside seemed to be the drug of choice in the treatment of listeriosis. An alternative drug was tetracycline. Most deaths occurred within six days of onset of the illness. Early diagnosis and treatment were imperative. Most patients recovered and serious sequelae were rare.
In this study, we investigated the fate of Listeria monocytogenes , pathogenic Yersinia enterocolitica , and Escherichia coli O157:H7 gfp(+) inoculated in low numbers into ready-to-eat baby spinach and mixed-ingredient salad (baby spinach with chicken meat). Samples were stored at recommended maximum refrigerator temperature (8°C in Sweden) or at an abuse temperature (15°C) for up to 7 days. Mixed-ingredient salad supported considerable growth when stored at 15°C during shelf life (3 days), with populations of L. monocytogenes , pathogenic Y. enterocolitica , and E. coli O157:H7 gfp(+) increasing from less than 2.0 log CFU/g on day 0 to 7.0, 4.0, and 5.6 log CFU/g, respectively. However, when mixed-ingredient salad was stored at 8°C during shelf life, only L. monocytogenes increased significantly, reaching 3.0 log CFU/g within 3 days. In plain baby spinach, only pathogenic Y. enterocolitica populations increased significantly during storage for 7 days, and this was exclusively at an abuse temperature (15°C). Thus, mixing ready-to-eat leafy vegetables with chicken meat strongly influenced levels of inoculated strains during storage. To explore the food safety implications of these findings, bacterial numbers were translated into risks of infection by modeling. The risk of listeriosis (measured as probability of infection) was 16 times higher when consuming a mixed-ingredient salad stored at 8°C at the end of shelf life, or 200,000 times higher when stored at 15°C, compared with when consuming it on the day of inoculation. This indicates that efforts should focus on preventing temperature abuse during storage to mitigate the risk of listeriosis. The storage conditions recommended for mixed-ingredient salads in Sweden (maximum 8°C for 3 days) did not prevent growth of L. monocytogenes in baby spinach mixed with chicken meat. Manufacturers preparing these salads should be aware of this, and recommended storage temperature should be revised downwards to reduce the risk of foodborne disease.
Prepacked ready-to-eat mixed ingredient salads (RTE salads) are readily available whole meals that include a variety of ingredients such as raw vegetables, cooked meat, and pasta. As part of a trend toward healthy convenience foods, RTE salads have become an increasingly popular product among consumers. However, data on the incidence of foodborne pathogens in RTE salads are scarce. In this study, the microbiological safety of 141 RTE salads containing chicken, ham, or smoked salmon was investigated. Salad samples were collected at retail and analyzed using standard methods for Listeria monocytogenes, Shiga toxin-producing Escherichia coli (STEC), pathogenic Yersinia enterocolitica, Salmonella, and Campylobacter spp.L. monocytogenes was isolated from two (1.4%) of the RTE salad samples. Seven (5.0%) of the samples were positive for the ail gene (present in all human pathogenic Y. enterocolitica isolates) and three (2.1%) of the samples were positive for the Shiga toxin genes stx1 and/or stx2. However, no strains of pathogenic Y.enterocolitica or STEC were isolated. Thus, pathogens were found or suspected in almost 1 of 10 RTE salads investigated, and pathogenic bacteria probably are present in various RTE salads from retail premises in Sweden. Because RTE salads are intended to be consumed without heat treatment, control of the ingredients and production hygiene is essential to maintain consumer safety. The recommended maximum storage temperature for RTE salads varies among countries but can be up to 8°C (e.g., in Sweden). Even during a short shelf life (3 to 5 days), storage at 8°C can enable growth of psychrotrophs such as L. monocytogenes and Y. enterocolitica. The maximum storage temperature should therefore be reduced.
Among 504 clinical lineage II isolates of Listeria monocytogenes isolated during 1958-2010 in Sweden, 119 pulsed-field gel electrophoresis (PFGE) types (AscI) have been identified based on the number and distribution of all banding patterns in each DNA profile. In this study, these types were further divided into PFGE groups based on the configuration of small bands with sizes 145.5?kb.
'Gravad' rainbow trout artificially contaminated with Listeria monocytogenes was analyzed by use of a 4 h enrichment period followed by extraction of DNA and PCR amplification. This procedure made it possible to detect 10-100 cfu L. monocytogenes per gram 'gravad' rainbow trout, within 12 h. After a prolonged enrichment period of 24 h, numbers as low as 1-10 cfu L. monocytogenes per gram could be detected. The method described may be a useful tool for screening samples of 'gravad' rainbow trout for the presence of L. monocytogenes, since it is sensitive, rapid and simple.
In Sweden, many Listeria monocytogenes strains belonging to serovar 4b and isolated during the last five years from different sources share the same phagovar--2389:2425:3274:2671:47:108:340. The object of the present study was to investigate if 31 L. monocytogenes serovar 4b strains belonging to this particular phagovar could be differentiated by use of a simple restriction endonuclease analysis (REA). Among the enzymes tested, Xho I was found to be the most useful, since this enzyme could divide the 31 strains into five groups. The profiles of all human clinical isolates were indistinguishable from each other, which indicates that these strains may represent a single clone. The food isolates and the strains of human origin did not share the same profile. This further characterization may be of epidemiological importance as this phagovar of L. monocytogenes has been associated with at least two outbreaks of human listeriosis in Europe.