A beef-associated outbreak of Salmonella Typhimurium DT104 in The Netherlands with implications for national and international policy.

A ten-fold increase in Salmonella Typhimurium DT104 cases in The Netherlands during September-November 2005 prompted an outbreak investigation. A population-based matched case-control study included 56 cases and 100 controls. Risk factors for infection were consumption of a pre-processed raw beef product (odds ratio 4.2, 95% confidence interval 1.5-12.0) and of food from mobile caterers (odds ratio 4.9, 95% confidence interval 1.1-22.1). Bacterial molecular typing established a link with another DT104 outbreak in Denmark caused by beef from a third European country. The incriminated beef was traced in The Netherlands and sampling yielded DT104 of the outbreak-associated molecular type. We concluded that this outbreak was caused by imported contaminated beef. Consumers should be informed about presence of raw meat in pre-processed food products. Optimal utilization of international networks and testing and traceability of foodstuffs has the potential to prevent foodborne infections.

Shiga toxin-producing Escherichia coli (STEC) O157 outbreak, The Netherlands, September-October 2005.

In September 2005, the first national food-related outbreak of Shiga toxin (Stx)-producing Escherichia coli (STEC) O157 was investigated in the Netherlands. A total of 21 laboratory-confirmed cases (including one secondary case), and another 11 probable cases (two primary and nine secondary cases) were reported in patients who became ill between 11 September and 10 October 2005. Preliminary investigation suggested consumption of a raw beef product, steak tartare (in the Netherlands also known as "filet americain"), and contact with other symptomatic persons as possible risk factors. A subsequent case-control study supported the hypothesis that steak tartare was the source of the outbreak (matched odds ratio (OR) 272, 95% confidence interval (CI) 3-23,211). Consumption of ready-to-eat vegetables was also associated with STEC O157 infection (matched OR 24, 95% CI 1.1-528), but was considered a less likely source, as only 40% of the cases were exposed. Samples of steak tartare collected from one chain of supermarkets where it is likely that most patients (67%) bought steak tartare, all tested negative for STEC O157. However, sampling was done three days after the date of symptom onset of the last reported case. Since 88% of the cases became ill within a two week period, point source contamination may explain these negative results. It is concluded that steak tartare was the most likely cause of the first national food-related outbreak of STEC O157 in the Netherlands.

Multiple cases of familial transmission of community-acquired methicillin-resistant Staphylococcus aureus.

The worldwide emergence of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) can have severe public health implications. Familial transmissions of CA-MRSA in The Netherlands were investigated. Among the families studied, two clusters of CA-MRSA could be identified. This report demonstrates that family members can serve as reservoirs of CA-MRSA which may become a serious problem in containing the spread of MRSA.

Genes for the majority of group a streptococcal virulence factors and extracellular surface proteins do not confer an increased propensity to cause invasive disease.

BACKGROUND: The factors behind the reemergence of severe, invasive group A streptococcal (GAS) diseases are unclear, but it could be caused by altered genetic endowment in these organisms. However, data from previous studies assessing the association between single genetic factors and invasive disease are often conflicting, suggesting that other, as-yet unidentified factors are necessary for the development of this class of disease. METHODS: In this study, we used a targeted GAS virulence microarray containing 226 GAS genes to determine the virulence gene repertoires of 68 GAS isolates (42 associated with invasive disease and 28 associated with noninvasive disease) collected in a defined geographic location during a contiguous time period. We then employed 3 advanced machine learning methods (genetic algorithm neural network, support vector machines, and classification trees) to identify genes with an increased association with invasive disease. RESULTS: Virulence gene profiles of individual GAS isolates varied extensively among these geographically and temporally related strains. Using genetic algorithm neural network analysis, we identified 3 genes with a marginal overrepresentation in invasive disease isolates. Significantly, 2 of these genes, ssa and mf4, encoded superantigens but were only present in a restricted set of GAS M-types. The third gene, spa, was found in variable distributions in all M-types in the study. CONCLUSIONS: Our comprehensive analysis of GAS virulence profiles provides strong evidence for the incongruent relationships among any of the 226 genes represented on the array and the overall propensity of GAS to cause invasive disease, underscoring the pathogenic complexity of these diseases, as well as the importance of multiple bacteria and/or host factors.

Invasive Listeria monocytogenes infections in the Netherlands, 1995-2003.

In order to add to the limited data available about the incidence of invasive Listeria monocytogenes infection in the Netherlands, two studies were conducted. In the first study, data on hospital patients with listeriosis in the period 1995-2003 were obtained from the National Medical Registration (study 1). In the second study, hospital discharge letters for patients whose Listeria isolates were received by the Netherlands Reference Laboratory for Bacterial Meningitis (NRLBM) in the period 1999-2003 were retrieved (study 2). Serotyping and pulsed-field gel electrophoresis (PFGE) were used to subtype the various strains of Listeria. These reviews revealed 283 hospital patients and 159 patients with Listeria isolates. Discharge letters were received for 107 (67%) patients. The mean annual incidence of listeriosis in both studies was 2.0 per million inhabitants. The main clinical manifestations were meningitis (incidence: 0.9 and 1.0 per million in studies 1 and 2, respectively) and septicaemia (incidence: 0.08 and 1.0 per million, respectively). Listeriosis in pregnancy was rare (incidence: 1.3 and 2.4 per 100,000 pregnancies over 24 weeks of gestation, respectively). Predisposing conditions were present in 47 and 71% of the patients in studies 1 and 2, respectively. The mortality due to listeriosis was 18%. Serotypes 4b, 1/2a, and 1/2b were responsible for 96% of the cases of human listeriosis. Listeriosis is rare in the Netherlands, but its clinical course is severe and the resulting mortality is high. Therefore, the current recommendations for pregnant women to avoid high-risk foods should be continued. These dietary recommendations should also be given to individuals with predisposing conditions, since they, too, are at risk of Listeria infection.

Surveillance of Salmonella spp. and Campylobacter spp. in poultry production flocks in The Netherlands.

In The Netherlands, a national programme for the surveillance of zoonotic bacteria in farm animals has been operative since 1997. We describe the results of the surveillance of Salmonella spp. in flocks of laying hens and broilers and of Campylobacter spp. in broiler flocks in the period 1999-2002. The prevalence of Salmonella spp. in laying-hen flocks has significantly decreased from 21.1% in 1999 to 13.4% in 2002. This decreasing trend might indicate that the control measures taken by the poultry industry were effective. S. Enteritidis was the predominant serovar in laying hens accounting for one third of the positive flocks. Although prevalence estimates for Salmonella spp. in broiler flocks did not yield a significant decreasing trend in 1999-2002, a decrease in Salmonella prevalence to 11% was measured in 2002. During the study period, S. Paratyphi B var. Java emerged in broilers to become the predominant serovar in 2002 accounting for one third of the positive flocks. The prevalence of Campylobacter spp. in broiler flocks did not increase nor decrease continuously between 1999 and 2002, which roughly corresponds with the monitoring results from the poultry industry. In this period, the estimated flock prevalence roughly averaged around 20%, with C. jejuni being the predominant species. The approach of monitoring presented in this paper can serve as a blueprint for monitoring schemes in farm animal populations to be developed in the context of the EC Zoonoses Directive.

Risk factors for Salmonella Enteritidis and Typhimurium (DT104 and non-DT104) infections in The Netherlands: predominant roles for raw eggs in Enteritidis and sandboxes in Typhimurium infections.

Since 1996 Salmonella Typhimurium DT104 salmonellosis has increased in The Netherlands. This prompted a case-control study of risk factors for salmonellosis to inform transmission routes for this phage type. Cases were laboratory-confirmed patients with a Salmonella infection and controls were selected from population registries by frequency matching for age, sex, degree of urbanization and season. Cases and controls received a questionnaire on risk factors. Of the 1171 cases, 573 (49%) responded: 245 S. Enteritidis and 232 S. Typhimurium cases (both DT104 and non-DT104), of which 58 were DT104. Of the 10250 controls, 3409 (33%) responded. Use of H2 antagonists [odds ratio (OR) 4.4, 95% CI 1.6-12.2] and proton pump inhibitors (OR 4.2, 95% CI 2.2-7.9), consumption of raw eggs (OR 3.1, 95% CI 1.3-7.4) and products containing raw eggs (OR 1.8, 95% CI 1.1-3.0) were associated with endemic S. Enteritidis infection. Risk factors for endemic S. Typhimurium infection were use of proton pump inhibitors (OR 8.3, 95% CI 4.3-15.9), occupational exposure to raw meat (OR 3.0, 95% CI 1.1-7.9), playing in a sandbox (for children aged 4-12 years) (OR 2.4, 95% CI 1.6-3.7), consumption of undercooked meat (OR 2.2, 95% CI 1.1-4.1) and use of antibiotics (OR 1.9, 95% CI 1.0-3.4). Use of proton pump inhibitors (OR 11.2, 95% CI 3.9-31.9) and playing in a sandbox (OR 4.4, 95% CI 1.8-10.7) were the only risk factors for S. Typhimurium DT104 salmonellosis. This study confirms known risk factors for salmonellosis. However, playing in a sandbox was a predominant new risk factor for S. Typhimurium salmonellosis in children [population attributable risk (PAR) 14%], and especially for S. Typhimurium DT104 (PAR 32%).

Shiga toxin-producing Escherichia coli (STEC) O157 outbreak, The Netherlands, September - October 2005.

In September 2005, the first national food-related outbreak of Shiga toxin (Stx)-producing Escherichia coli (STEC) O157 was investigated in the Netherlands. A total of 21 laboratory-confirmed cases (including one secondary case), and another 11 probable cases (two primary and nine secondary cases) were reported in patients who became ill between 11 September and 10 October 2005. Preliminary investigation suggested consumption of a raw beef product, steak tartare (in the Netherlands also known as 'filet américain'), and contact with other symptomatic persons as possible risk factors. A subsequent case-control study supported the hypothesis that steak tartare was the source of the outbreak (matched odds ratio (OR) 272, 95% confidence interval (CI) 3 - 23211). Consumption of ready-to-eat vegetables was also associated with STEC O157 infection (matched OR 24, 95% CI 1.1 - 528), but was considered a less likely source, as only 40% of the cases were exposed. Samples of steak tartare collected from one chain of supermarkets where it is likely that most patients (67%) bought steak tartare, all tested negative for STEC O157. However, sampling was done three days after the date of symptom onset of the last reported case. Since 88% of the cases became ill within a two week period, point source contamination may explain these negative results. It is concluded that steak tartare was the most likely cause of the first national food-related outbreak of STEC O157 in the Netherlands.

Transmission of a Panton-Valentine leucocidin-positive, methicillin-resistant Staphylococcus aureus strain between humans and a dog.

Panton-Valentine leucocidin-positive methicillin-resistant Staphylococcus aureus (MRSA) strains with identical resistance patterns were cultured from recurrent infections of a 51-year-old patient, her healthy husband, son, and dog, and pulsed-field gel electrophoresis showed that all MRSA strains were indistinguishable.

Emergence of virulent methicillin-resistant Staphylococcus aureus strains carrying Panton-Valentine leucocidin genes in The Netherlands.

Methicillin-resistant Staphylococcus aureus (MRSA) strains carrying the Panton-Valentine leucocidin (PVL) genes have been reported worldwide and are a serious threat to public health. The PVL genes encode a highly potent toxin which is involved in severe skin infections and necrotizing pneumonia, even in previously healthy individuals. We assessed the prevalence of PVL-positive MRSA in The Netherlands for two periods of time: (i) 1987 through 1995 and (ii) 2000 and 2002, and determined their characteristics by using multilocus sequence typing and staphylococcal chromosome cassette (SCCmec) typing. It was found that up to 15% of all MRSA isolates detected in The Netherlands harbored the PVL genes. Most PVL-positive MRSA isolates were obtained from severe soft tissue infections in relatively young individuals. The first PVL-positive MRSA described in The Netherlands, isolated in 1988, was a single-locus variant of the "Berlin" epidemic MRSA clone. The 20 PVL-positive MRSA isolates studied in 2000 and 2002 consisted of five different sequence types (STs) that belonged to four clonal complexes. One of the STs, ST80, is considered to be a widespread European clone and was the most predominant ST (60%) in this study, while ST37 had never been found to be associated with PVL-positive MRSA. Most isolates harbored SCCmec type IV, a supposed marker for community-acquired MRSA. The number and type of virulence-associated genes varied among the different STs.

[Sharp increase of infections with Salmonella serotype Typhimurium DT104 in The Netherlands]. / Sterke toename van infecties met Salmonella serotype Typhimurium DT104 in Nederland.

Salmonella remains an important source of food-related outbreaks of gastro-enteritis. In The Netherlands, regional laboratories send cultured Salmonella isolates to the National Institute of Public Health and the Environment (RIVM) for sero- and phage typing. Abnormal increases in the incidence of Salmonella infections are monitored by means of surveillance. Since the middle of September 2005, there has been a sharp increase in Salmonella Typhimurium DT104 (Dutch phage type 506 and 401) isolates, from all parts of the country and mainly from 6- to 20-year-olds. Since a first series of telephone interviews did not yield a clear source of the outbreak, a more extensive, written study has been started in a larger group of patients and corresponding controls.

Panton-Valentine leukocidin positive MRSA in 2003: the Dutch situation.

Analysis of methicillin-resistant Staphylococcus aureus (MRSA) isolates in the Netherlands in 2003 revealed that 8% of the hospital isolates carried the loci for Panton-Valentine leukocidin (PVL). Molecular subtyping showed that most Dutch PVL-MRSA genotypes corresponded to well-documented global epidemic types. The most common PVL-MRSA genotypes were sequence type ST8, ST22, ST30, ST59 and ST80. MRSA with ST8 increased in the Netherlands from 1% in 2002 to 17% in 2003. It is emphasised that PVL-MRSA might not only emerge in the community, but also in the hospital environment.

Methicillin-resistant staphylococci isolated from animals.

Staphylococci isolated from animals (n=311) were screened for methicillin resistance by oxacillin agar screening. Oxacillin-resistant strains were tested for the presence of the mecA gene by PCR. Isolates were identified by standard techniques and 16S rDNA analysis, and their antimicrobial susceptibilities were tested using an agar diffusion method. MecA-positive strains were further analyzed using pulsed-field gel electrophoresis (PFGE). From 11 multidrug-resistant staphylococci, 6 were mecA-positive: 2 methicillin-resistant Staphylococcus aureus (MRSA) and 4 Staphylococcus haemolyticus. Screening of 300 staphylococci (100 S. aureus, 100 S. intermedius and 100 coagulase-negative staphylococci (CNS)) randomly chosen from the strain collection of the Veterinary Microbiological Diagnostic Center yielded five oxacillin-resistant coagulase-negative staphylococci, four of which were mecA-positive. PFGE showed that all mecA-positive staphylococci isolated from animals had distinct patterns. However, one MRSA isolated from a flank fistula of a dog showed homology to a human epidemic MRSA cluster, suggesting that transfer of MRSA between humans and dogs might occur.

Widespread dissemination in The Netherlands of the epidemic berlin methicillin-resistant Staphylococcus aureus clone with low-level resistance to oxacillin.

Methicillin-resistant Staphylococcus aureus (MRSA) is an important human pathogen and represents a growing public health burden due to the emergence and spread of epidemic strains, particularly within the hospital environment. An epidemic MRSA clone, with characteristic low-level resistance to oxacillin, emerged in the year 2000 and became endemic in the Netherlands. Multilocus sequence typing characterized the strain as sequence type 45, which was previously designated the Berlin epidemic MRSA clone. In 2 years, this strain has become the predominant MRSA clone in the Netherlands.

A large increase of Salmonella infections in 2003 in the Netherlands: hot summer or side effect of the avian influenza outbreak?

In June 2003, the Dutch national Salmonella centre reported a significant excess isolation rate of Salmonella Enteritidis when compared with earlier years in most regional public health laboratories. By the end of 2003, this amounted to an extra 540 laboratory confirmed cases for the whole of the Netherlands, which implies an estimated 7500 extra cases of gastroenteritis caused by S. Enteritidis in the general population, an increase of 50% on previous years. The hot summer could not explain the findings. Strong evidence has been found to suggest that the increase in importation of salmonella contaminated eggs, as a side effect of a concurrent avian influenza outbreak, was the most probable reason for this excess.

Panton-Valentine leukocidin positive MRSA in 2003: the Dutch situation.

Analysis of methicillin-resistant Staphylococcus aureus (MRSA) isolates in the Netherlands in 2003 revealed that 8% of the hospital isolates carried the loci for Panton-Valentine leukocidin (PVL). Molecular subtyping showed that most Dutch PVL-MRSA genotypes corresponded to well-documented global epidemic types. The most common PVL-MRSA genotypes were sequence type ST8, ST22, ST30, ST59 and ST80. MRSA with ST8 increased in the Netherlands from 1% in 2002 to 17% in 2003. It is emphasised that PVL-MRSA might not only emerge in the community, but also in the hospital environment.

Antimicrobial susceptibilities of salmonella strains isolated from humans, cattle, pigs, and chickens in the Netherlands from 1984 to 2001.

We monitored antimicrobial susceptibility data for Salmonella strains isolated from humans, cattle, pigs, and chickens in The Netherlands from 1984 to 2001 in order to provide insight into the dynamics of resistance over time. The strains were tested for their susceptibilities to seven antimicrobial agents by the agar diffusion method. Resistance was most common in Salmonella enterica subsp. enterica serovar Typhimurium. Among the strains from humans, pigs, and chickens, it was found that the level of resistance of serovar Typhimurium strains to tetracycline, ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole increased from 1984 to 2001. This increase could be attributed to the emergence of multidrug-resistant serovar Typhimurium DT 104. Among the strains from cattle, it was found that the level of resistance of serovar Typhimurium strains, which was already very high in the 1980s, declined during the study period to the same levels as those for the strains from the other species from 1996 to 2001. Serovar Enteritidis isolates remained susceptible during the entire survey period. Among serovar Paratyphi B variation Java strains isolated from chickens, resistance to furazolidone, flumequine, trimethoprim-sulfamethoxazole, and ampicillin emerged, although furazolidone was not used after 1990. Together, the data indicate that the levels and patterns of resistance differed considerably between Salmonella serovars isolated from one host species.

Laboratory surveillance of bacterial gastroenteric pathogens in The Netherlands, 1991-2001.

Results of the Dutch laboratory surveillance of bacterial gastroenteritis between 1991 and 2001 are presented and compared with recent findings in general practices and in the community. Between 1996 and 2000 the mean annual number of stools screened by sentinel laboratories was about 1000 samples/100,000 inhabitants, which is 4% of the estimated annual incidence of gastroenteritis in the Dutch population. Campylobacter (36/100,000 inhabitants) and salmonella (24/100,000 inhabitants) were the main pathogens isolated. Since 1996, the incidence of laboratory confirmed salmonellosis decreased by 30%, predominantly among young children. The incidence of campylobacter was highest in urban areas and Salmonella Enteritidis emerged as the predominant serotype in urban areas. Between 1991 and 2001, multi-resistant Salmonella Typhimurium DT104 emerged to comprise up to 15% of all salmonella isolates in 2001. Reported rates of Shigella spp. and Yersinia spp. varied little, with average annual incidences of 3.2 and 1.2 cases/100,000 inhabitants, respectively. Escherichia coli O157 (90% STEC) was scarcely found (0.26/100,000).

[Methicillin resistant Staphylococcus aureus (MRSA) infection in a dog in the Netherlands]. / Een meticilline-resistente Staphylococcus aureus (MRSA)-infectie bij een hond in Nederland.

In the Netherlands, methicillin resistant Staphylococcus aureus (MRSA) are regularly isolated from humans. We present the first isolation of MRSA from animal origin in the Netherlands. A coagulase positive staphylococcus was cultured from an infected wound in a Dutch dog that recently underwent surgery abroad. The staphylococcus was resistant to methicillin, ampicillin, amoxycillin + clavulanic acid, cephalexin, erythromycin, lincomycin, tetracycline, gentamicin and enrofloxacin. It was identified as S. aureus by fermentation of mannitol and Martineau-PCR. The presence of mecA was confirmed by PCR.