Outbreak of hospital-acquired gastroenteritis and invasive infection caused by Listeria monocytogenes, Finland, 2012.

During one week in July 2012, two patients from the same ward at the municipal hospital in Vaasa, Finland, were diagnosed with septicaemia caused by Listeria monocytogenes. An outbreak investigation revealed eight concomitant cases of febrile gastroenteritis caused by L. monocytogenes on the same ward. Median age of the cases was 82 years and median incubation time for listerial gastroenteritis was 21 h (range 9-107). An additional 10 cases of invasive listeriosis caused by the same outbreak strain were identified across the whole country during the summer of 2012. Environmental investigation at the affected municipal hospital ward revealed ready-sliced meat jelly as the suspected source of the infection. During inspection of the meat jelly production plant, one pooled sample taken from a floor drain and a trolley wheel in the food processing environment was positive for the outbreak strain of L. monocytogenes. After the producer stopped the production of meat jelly, no further cases of listeriosis with the outbreak strain were identified via nationwide surveillance.

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.

Investigation of increased listeriosis revealed two fishery production plants with persistent Listeria contamination in Finland in 2010.

In 2010, a marked increase in listeriosis incidence was observed in Finland. Listeria monocytogenes PFGE profile 96 was responsible for one-fifth of the reported cases and a cluster of PFGE profile 62 was also detected. Investigations revealed two fishery production plants with persistent Listeria contamination. It appears likely that the plants were at least partly responsible for the increase of listeriosis. Epidemiological investigation revealed that 57% (31/54) of cases with underlying immunosuppressive condition or medication reported eating gravad or cold-smoked fish. Two public notices were issued by THL and Evira informing which groups were most at risk from the effects of listeriosis and should therefore be cautious in consuming certain products. Systematic sampling of foods and adequate epidemiological investigation methods are required to identify the sources of Listeria infections. Continuous control measures at fishery production plants producing risk products are essential.

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.

Variations of mitochondrial DNA polymerase γ in patients with Parkinson's disease.

Parkinson's disease is associated with mitochondrial dysfunction. The POLG1 gene encodes DNA-polymerase γ, which is responsible for the replication of mitochondrial DNA. Mutations in POLG1 cause neurodegenerative diseases such as progressive external ophthalmoplegia and Alpers syndrome. In this study, we investigated if mutations in POLG1 had any correlation with Parkinson's disease. Subjects consisted of Finnish patients with early-onset Parkinson's disease (EOPD, N = 441) or late-onset Parkinson's disease (LOPD, N = 263). The POLG1 gene was screened for nine previously known mutations. Two patients were compound heterozygotes with respect to putatively pathogenic alleles. Twenty-eight patients harbored a heterozygous missense mutation, but the allele frequencies did not differ from those of the controls. Interestingly, the frequency of affected siblings was 4.6-fold higher (95 % confidence interval; 1.09, 19.5) among the patients with EOPD and with heterozygous POLG1 mutations than among patients without mutations. Clinically the patients with or without POLG1 mutations did not differ from each other. Our findings provide two lines of evidence suggesting a role for POLG1 mutations in Parkinson's disease: (1) identification of patients with compound heterozygous mutations in POLG1, and (2) higher frequency of affected siblings among the EOPD patients with heterozygous POLG1 mutations than among EOPD patients without mutations.

Multiple-locus variable-number tandem-repeat analysis in genotyping Yersinia enterocolitica strains from human and porcine origins.

Sporadic and epidemiologically linked Yersinia enterocolitica strains (n = 379) isolated from fecal samples from human patients, tonsil or fecal samples from pigs collected at slaughterhouses, and pork samples collected at meat stores were genotyped using multiple-locus variable-number tandem-repeat analysis (MLVA) with six loci, i.e., V2A, V4, V5, V6, V7, and V9. In total, 312 different MLVA types were found. Similar types were detected (i) in fecal samples collected from human patients over 2 to 3 consecutive years, (ii) in samples from humans and pigs, and (iii) in samples from pigs that originated from the same farms. Among porcine strains, we found farm-specific MLVA profiles. Variations in the numbers of tandem repeats from one to four for variable-number tandem-repeat (VNTR) loci V2A, V5, V6, and V7 were observed within a farm. MLVA was applicable for serotypes O:3, O:5,27, and O:9 and appeared to be a highly discriminating tool for distinguishing sporadic and outbreak-related strains. With long-term use, interpretation of the results became more challenging due to variations in more-discriminating loci, as was observed for strains originating from pig farms. Additionally, we encountered unexpectedly short V2A VNTR fragments and sequenced them. According to the sequencing results, updated guidelines for interpreting V2A VNTR results were prepared.

Reactive arthritis following Salmonella infection: a population-based study.

OBJECTIVES: To study the incidence and clinical picture of Salmonella-associated reactive arthritis (ReA), as well as other reactive musculoskeletal symptoms and the arthritogenicity of various Salmonella enterica ssp. enterica serotypes in the population. METHOD: We sent a questionnaire on enteric and extraintestinal (especially musculoskeletal) symptoms to 999 consecutive subjects with a Salmonella-positive stool culture. Analysis of self-reported musculoskeletal symptoms was supplemented with a clinical examination of subjects with recent symptoms. RESULTS: Of the 999 Salmonella-positive subjects, 496 (50%) returned the questionnaire. Of these, 4.4% (22/496) had ReA and 13.7% (68/496) had other reactive musculoskeletal symptoms [tendinitis, enthesopathy, or bursitis (ReTe)]. Among the ReA patients, all adults, Salmonella Enteritidis was the most common causative serotype. The clinical picture of patients with ReA was mostly monoarticular or oligoarticular. Human leucocyte antigen (HLA)-B27 was positive in 42% of patients with ReA. The Salmonella O antigens of the 496 subjects belonged to eight groups (B, C, D1, E, G, I, L, and O), all with different major O antigenic determinants. All 22 patients with ReA and all 68 patients with ReTe were in O antigen groups B, C, D1, or E. However, the occurrence of musculoskeletal complications showed no statistically significant difference in relation to different O antigen groups (p = 0.69). CONCLUSIONS: ReA occurred in 4.4% of patients after Salmonella infection, with an annual incidence of 1.8/100,000 in Finland. We found no differences in arthritogenicity between different Salmonella serotypes that trigger musculoskeletal complications.

Diarrhoeagenic Escherichia coli detected by 16-plex PCR in children with and without diarrhoea in Burkina Faso.

The importance of diarrhoeagenic Escherichia coli (DEC) in Africa is poorly understood, and is unknown in Burkina Faso. This study investigated the occurrence of five major DEC pathogroups in primary cultures of stool samples from 658 Burkinabe children under 5 years old using 16-plex PCR for virulence-associated genes. At least one DEC pathogroup was detected in 45% of 471 children with diarrhoea and in 29% of 187 children without diarrhoea (p <0.001). More than one DEC pathogroup was detected in 11% of children with and 1% of children without diarrhoea (p <0.001). Enteroaggregative E. coli was the most common pathogroup in both children with diarrhoea (26%) and children without diarrhoea (21%). Enteropathogenic E. coli and enterotoxigenic E. coli were detected significantly more often in children with diarrhoea (16% and 13%) than in children without diarrhoea (5% and 4%; p <0.001 for both pathogroups). Shiga toxin-producing E. coli and enteroinvasive E. coli were detected only in children with diarrhoea (2% and 1%, respectively). Diarrhoeagenic E. coli, especially enteropathogenic and enterotoxigenic, may be important, unrecognized causes of childhood diarrhoea in Burkina Faso.

A nationwide outbreak of Salmonella bovismorbificans associated with sprouted alfalfa seeds in Finland, 2009.

Salmonella enterica serotype Bovismorbificans is a rare serotype in Finland. In June 2009, a nationwide outbreak of S. Bovismorbificans infections occurred, and 42 clinical isolates were identified. We conducted a case-control study enrolling 28 cases and 48 matched controls, and found ready-to-eat alfalfa sprouts associated with the infection (odds ratio = 35.2, 95% confidence interval 2.8-435). The sprouts were traced back to a domestic producer, with the seeds originating in Italy. Although finding microbiological evidence for sprouts as a source of this outbreak was very challenging, S. Bovismorbificans was finally found in sprouts germinated in the laboratory, and in soaking and rinse waters during the germination process. The pulsed field gel electrophoresis showed that these isolates were identical to the human outbreak isolates. Before sampling, it is important to mix the seeds well and to take several large-size samples from different seed lots. Instead of analysing seeds, the analysis should be targeted to soaking or rinse water samples and to the sprouts germinated in a laboratory. Accordingly, the sprout producers should only buy seeds that have been analysed for Salmonella. The producers have to include Salmonella testing in their internal quality control to ensure that Salmonella is absent from sprouts placed on the market during their shelf-life. In order to reduce the health hazard caused by sprouts, an effective and safe decontamination procedure should also be developed for the seeds.

Cases of Salmonella Urbana in Finland, the Czech Republic and Latvia, January-February 2010.

A cluster of 14 cases of Salmonella Urbana cases in Finland, the Czech Republic and Latvia were identified in January-February, 2010. The majority of cases (11) were male and children under 16 years of age. The investigation is currently ongoing and comparison of pulsed-field gel electrophoresis (PFGE) profiles of the isolates suggests that the cases may have a common source of infection.

Population-based surveillance study of Campylobacter infections in Finland.

The annual incidence in 14,361 campylobacteriosis cases reported in Finland in 2002-2005 varied between 61 and 76/100,000 population. The mean incidence was highest (148/100,000) in the 25-29 years age group and lowest (range 21-24/100,000) in children aged 5-14 years and patients aged ≥75 years. The number of domestic cases was low in winter and peaked in summer. A total of 622 strains isolated from domestic infections and 785 foreign travel-related strains were serotyped. Serotypes Pen 3 and Pen 37 had the strongest association with travel-related infections (96%, P<0·001), and Pen 6,7, Pen 12 and Pen 27 were significantly associated with domestic infections (>70% domestic within each serotype, P<0·001). Pen 2 and Pen 1,44 were less common in older than in younger patients. Of domestic strains, a higher proportion of Pen 2 strains was isolated in winter (18%) compared to the other serotypes (0-10%).

Phenotype MicroArray in the metabolic characterisation of Salmonella serotypes Agona, Enteritidis, Give, Hvittingfoss, Infantis, Newport and Typhimurium.

The Phenotype MicroArray (PM) technology was used to study the metabolic characteristics of 29 Salmonella strains belonging to seven serotypes of S. enterica spp. enterica. Strains of serotypes Typhimurium (six strains among definite phage types DTs 1, 40 and 104) and Agona (two strains) were tested for 949 substrates, Enteritidis (six strains of phage type PT1), Give, Hvittingfoss, Infantis and Newport strains (two of each) were tested for 190 substrates and seven other Agona strains for 95 substrates. The strains represented 18 genotypes in pulsed-field gel electrophoresis (PFGE). Among 949 substrates, 18 were identified that could be used to differentiate between the strains of those seven serotypes or within a single serotype. Unique metabolic differences between the Finnish endemic Typhimurium DT1 and Agona strains were detected, for example, in the metabolism of D-tagatose, D-galactonic acid gamma-lactone and L-proline as a carbon source. Thus, the PM technique is a useful tool for identifying potential differential markers on a metabolic basis that could be used for epidemiological surveillance.

Salmonellosis cases caused by a rare Salmonella Enteritidis PT6c associated with travel to Bulgaria, June-July 2008.

In June 2008 an outbreak of gastroenteritis was registered in Sunny Beach resort situated on the Black Sea coast in Bulgaria, affecting 14 employees of a hotel, five of whom tested positive for Salmonella Enteritidis. During June-July 2008 four sporadic S. Enteritidis cases were also reported and two of them were foreign tourists. In the same period S. Enteritidis cases connected with travel to Bulgaria were reported to the European Centre for Disease Prevention and Control (ECDC) from Finland, United Kingdom, Sweden, Germany and Norway. We describe a study performed to find out relatedness between Bulgarian and Finnish S. Enteritidis isolates using phage typing (PT) and pulse-field gel electrophoresis (PFGE). Fifteen S. Enteritidis isolates from Bulgaria and 195 from Finland (including 28 from travellers to Bulgaria) were phage typed. Within Bulgarian isolates four different PTs were found and PT6c with eight strains was predominant. Nineteen out of 28 strains isolated from the Finns visiting Bulgaria belonged also to PT6c. PFGE typing (with one enzyme) of all S. Enteritidis PT6c strains (8 Bulgarian and 19 Finnish isolates) showed indistinguishable PFGE profile. The typing results thus demonstrated a link between Bulgarian and Finnish S. Enteritidis isolates. We conclude that S. Enteritidis PT6c was the cause of salmonella outbreak in Sunny Beach and was exported to Finland, and likely to the United Kingdom, Norway, Sweden and Germany.

New 16-plex PCR method for rapid detection of diarrheagenic Escherichia coli directly from stool samples.

A rapid 16-plex polymerase chain reaction (PCR) suitable for routine diagnostics of diarrheagenic Escherichia coli (EHEC, EIEC, EAEC, ETEC, and EPEC) was developed, validated with control strains, and tested with 250 diarrhoeal stool samples. The specificity was 100% when tested with 289 control bacterial strains, and the analytical sensitivity of automated DNA extraction directly from stool samples was made by boiling the bacterial culture (10(4)-10(5) colony forming units/ml). The assay design starting directly from extraction of stool DNA allowed same day analysis without compromising sensitivity and specificity, which makes it superior compared to PCR after culturing the bacteria. The 16-plex PCR method demonstrated high prevalence of diarrheagenic E. coli in stool samples of patients returning from abroad (39.0%) in contrast to the patients with no travel history (8.7%; p < 0.001). The high prevalence of diarrheagenic E. coli suggests that their screening should be part of normal diarrhoea diagnostics, at least in the leading diagnostic laboratories.

Yersinia enterocolitica and Y. enterocolitica-like species in clinical stool specimens of humans: identification and prevalence of bio/serotypes in Finland.

This study investigated the prevalence of Yersinia enterocolitica (YE) bio/serotypes and YE-like species in clinical stool specimens. The special aim was to find the best methods for accurate identification of YE species and, further, pathogenic strains among YE isolates. Of the 41,848 specimens cultured in ten laboratories during a 12-month period, 473 Yersinia strains were isolated from 462 patients. The strains were identified by 21 biochemical tests, serotyping, colony morphology, as well as by 16S rRNA and gyrB gene sequencing. The most prevalent Yersinia findings were YE biotype 1A (64% of the strains) and pathogenic bio/serotype 4/O:3 (16%). The cold-enrichment increased the number of all isolates, and 25% of the bio/serotype 4/O:3 and 2/O:9 strains were only found by cold-enrichment. In routine diagnostic laboratories, 50% of the YE-like species were identified as YE and in 26% the identification differed from that of the reference laboratory. The microscopic colony identification on CIN agar with positive CR-MOX test, combined with several biochemical tests, identified reliably the pathogenic YE bioserotypes and most YE BT 1A strains, but some strains of the YE-like species were so heterogenic that gene sequencing was the only way to identify them.

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.

Point-of-care testing of group A streptococcal antigen: performance evaluated by external quality assessment.

The purpose of this study was to evaluate the overall performance of rapid antigen detection (RAD) in group A streptococcus (GAS) in Finland by using the results of external quality assurance (EQA) samples. We also compared the performance of laboratory professionals to that of nursing professionals. Around 22,800 EQA results among a total of 383 laboratories and physician's offices were analysed. Vocational data on the personnel who carried out the tests were available for 10,088 EQA samples, 7,428 of which were tested by laboratory technicians and 2,531 by nursing staff. The best overall performance was found with GAS-negative samples: 99% of the reports were correct. In contrast, the overall performance was only 76% when the samples were weakly positive for GAS antigen. The laboratory technicians performed statistically significantly better than the nursing staff, with both strongly positive (correct results 98.9% vs. 95.1%, respectively; p<0.001) and weakly positive (79.3% vs. 65.3%, respectively; p<0.001) samples. With negative samples, no difference in performance between the laboratory and nursing staff was found (99.5% vs. 99.0%, respectively). The professional skills of the person performing the RAD test for GAS have a major impact on the sensitivity of the test. Based on the results of this study, we suggest that EQA-like artificial specimens could be used as a tool to improve and validate the quality of RAD testing in individual testing sites.

Correct identification and discrimination between Campylobacter jejuni and C. coli by a standardized hippurate test and species-specific polymerase chain reaction.

Hippurate hydrolysis test results of 240 Campylobacter strains were compared with those of two multiplex polymerase chain reaction (PCR) assays. Of the 152 strains identified in Finnish clinical microbiology routine laboratories as C. coli (hippurate-negative), 11% were C. jejuni (hippurate-positive) by standardized hippurate test and 39% by PCR in the reference laboratory. Two of the 81 hippurate-positive strains were identified as C. coli. Standardizing the hippurate test by determining minimum and maximum turbidity limits (McFarland 6 and McFarland 10, OD(450) values 0.8 and 1.4, respectively) for the bacterial cell suspension eliminated the false-positive results, but 32% of the 145 hippurate-negative strains were still identified as C. jejuni by PCR. The species identification of Campylobacter isolates in Finland could be improved by using a standardized hippurate hydrolysis test to identify hippurate-positive C. jejuni and testing hippurate-negative strains by molecular methods. This would also improve the epidemiological data on this important zoonotic pathogen.

Emerging resistance to newer antimicrobial agents among Shigella isolated from Finnish foreign travellers.

In Finland, most cases of shigellosis are related to travel abroad. Antimicrobial drug resistance of 1814 Shigella strains isolated from Finnish patients during 1990-2005 was studied using discs of 12 antimicrobial agents. Since 2000, the E-test has been performed to determine ciprofloxacin minimum inhibitory concentrations of nalidixic acid-resistant isolates. The proportion of multi-resistant strains (resistant to >or =4 antimicrobials) was highest among isolates from China and India, but is increasing significantly in other parts of Asia. Resistance to nalidixic acid has become common among the strains from the Far East, and the first isolates also resistant to ciprofloxacin were detected during 2004-2005. All the ciprofloxacin-resistant isolates belonged to the S. flexneri 2a serotype. All the nalidixic acid-resistant S. flexneri strains had reduced susceptibility to ciprofloxacin, whereas 23% of the nalidixic acid-resistant S. sonnei strains were still completely susceptible to ciprofloxacin.