H de Valk

Surveillance of listeria infections in Europe.

In addition to the economic consequences and threats associated with outbreaks, listeriosis remains of great public health concern, as it has one of the highest case fatality rates of all the foodborne infections (20%-30%), and has common source epidemic potential. Changes in the way food is produced, distributed and stored have created the potential for diffuse and widespread outbreaks involving many countries. In 2002, a survey was carried out to assess the need for and the feasibility of a European network on listeria infections in humans. Data on surveillance systems and laboratory methods were collected through two postal surveys sent to the national Centres for communicable disease surveillance and to the listeria reference laboratories. Surveillance systems for listeria infections were in operation in 16 out of the 17 countries surveyed, and 16 countries had a national reference laboratory (NRL). All countries based their case definition of listeriosis on the isolation of Listeria monocytogenes. Fourteen NRLs performed at least one typing method on human strains. At least 13 countries already carried out or expressed willingness to carry out characterisation of isolates by pulsed field gel electrophoresis (PFGE) of L. monocytogenes strains isolated from human cases following a standard protocol. The participants concluded that there was a clear added value to having a European surveillance network for listeria infections, particularly for outbreak detection and investigation, and that a surveillance network based on the existing national surveillance systems was feasible.

Local and regional spread of chikungunya fever in the Americas

Chikungunya fever (CHIKV), a viral disease transmitted by mosquitoes, is currently affecting several areas in the Caribbean. The vector is found in the Americas from southern Florida to Brazil, and the Caribbean is a highly connected region in terms of population movements. There is therefore a significant risk for the epidemic to quickly expand to a wide area in the Americas. Here, we describe the spread of CHIKV in the first three areas to report cases and between areas in the region. Local transmission of CHIKV in the Caribbean is very effective, the mean number of cases generated by a human case ranging from two to four. There is a strong spatial signature in the regional epidemic, with the risk of transmission between areas estimated to be inversely proportional to the distance rather than driven by air transportation. So far, this simple distance-based model has successfully predicted observed patterns of spread. The spatial structure allows ranking areas according to their risk of invasion. This characterisation may help national and international agencies to optimise resource allocation for monitoring and control and encourage areas with elevated risks to act.

Novel mixed-format real-time PCR assay to detect mutations conferring resistance to triazoles in Aspergillus fumigatus and prevalence of multi-triazole resistance among clinical isolates in the Netherlands

OBJECTIVES The aim of this study was: (i) to study the prevalence of triazole-resistant Aspergillus fumigatus isolates in the Netherlands; and (ii) to design rapid real-time PCR methods to identify such isolates. METHODS A novel mixed-format real-time PCR assay is described for the detection of mutations leading to triazole resistance in A. fumigatus. One set of PCR primers and a probe carrying a single fluorescent label in combination with a double-stranded DNA fluorescent dye allow simultaneous detection of (a) specific mutation(s) as well as of the amplified product that serves as an internal amplification control. The method was applied to a random collection of 209 clinical isolates from throughout the Netherlands and was compared with phenotypic susceptibility testing. RESULTS A total of four triazole-resistant isolates were identified, resulting in a prevalence of resistant isolates of <2%. All four isolates contained an identical combination of mutations leading to multi-triazole resistance, as reported before by others. Molecular testing results were 100% concordant with phenotypic susceptibility testing. CONCLUSIONS Although in specific patient populations the prevalence of resistance in A. fumigatus may be an emerging problem, in the general population it is still relatively low. The novel real-time PCR format allows rapid and reliable identification of such isolates.

An outbreak of multidrug-resistant Salmonella enterica serotype Newport infections linked to the consumption of imported horse meat in France.

In 2003, 14 cases of multidrug-resistant Salmonella Newport infections were reported. This is the first documented foodborne outbreak of multidrug-resistant S. Newport in France. The blaCMY gene was present in all isolates. All cases reported having eaten horse meat from a common wholesaler. The country of origin of the imported meat could not be identified.

Incidence and prevalence of Toxoplasma gondii infection in women in France, 1980-2020: model-based estimation.

Toxoplasmosis is a worldwide zoonosis due to Toxoplasma gondii, a ubiquitous protozoan parasite of warm-blooded animals including humans. In pregnant women, primary infection can cause congenital toxoplasmosis resulting in severe malformations in the newborn. Since 1978, public health authorities in France have implemented a congenital toxoplasmosis prevention programme, including monthly serological screening of all seronegative pregnant women, and treatment in case of seroconversion. However, this programme does not produce systematic surveillance data on incidence and prevalence. Our objective was to estimate the incidence and prevalence of T. gondii infection, and the incidence of seroconversion during pregnancy in women in France. We used a catalytic model to estimate incidence and prevalence of Toxoplasma infection between 1980 and 2020 in women of childbearing age. We used age- and time-specific seroprevalence data obtained from the National Perinatal Surveys (NPS) conducted in 1995, 2003 and 2010. We assumed that incidence depends both on age and calendar time, and can be expressed as the product of two unknown functions. We also estimated incidence of seroconversion during pregnancy in 2010 from the NPS and the National Surveillance of Congenital Toxoplasmosis (ToxoSurv). We combined data of 42208 women aged 15-45 years with serology available from the three NPS. For women aged 30 years the modelled incidence decreased from 7·5/1000 susceptible women in 1980 to 3·5/1000 in 2000. In 2010 the incidence was 2·4/1000. The predicted incidence and prevalence for 2020 was 1·6/1000 and 27%, respectively. The incidence of seroconversion during pregnancy in 2010 was estimated at 2·1/1000 susceptible pregnant women (95% CI 1·3-3·1) from the NPS and 1·9 (95% CI 1·8-2·1) from ToxoSurv. Incidence and prevalence of Toxoplasma infection has decreased markedly during the last 30 years. This decrease may be explained by a lower exposure to the parasite by changes in food habits and by improved hygiene practices in meat production. Modelled estimations were consistent with estimates observed in other studies conducted previously in France. The catalytic modelling provides reliable estimates of incidence and prevalence of Toxoplasma infection over time. This approach might be useful for evaluating preventive programme for toxoplasmosis.

Interlaboratory reproducibility of a microsatellite-based typing assay for Aspergillus fumigatus through the use of allelic ladders: proof of concept

An interlaboratory study was performed with the aim of investigating the reproducibility of a multiplex microbial microsatellite-based typing assay for Aspergillus fumigatus in different settings using a variety of experimental and analytical conditions and with teams having variable prior microsatellite typing experience. In order to circumvent problems with exchange of sizing data, allelic ladders are introduced as a straightforward and universally applicable concept for standardization of such typing assays. Allelic ladders consist of mixtures of well-characterized reference fragments to act as reference points for the position in an electrophoretic trace of fragments with established repeat numbers. Five laboratories independently analysed six microsatellite markers in 18 samples that were provided either as DNA or as A. fumigatus conidia. Allelic data were reported as repeat numbers and as sizes in nucleotides. Without the use of allelic ladders, size differences of up to 6.7 nucleotides were observed, resulting in interpretation errors of up to two repeat units. Difficulties in interpretation were related to non-specific amplification products (which were resolved with explanation) and bleed-through of the different fluorescent labels. In contrast, after resolution of technical or interpretive problems, standardization of sizing data by using allelic ladders enabled all participants to produce identical typing data. The use of allelic ladders as a routine part of molecular typing using microsatellite markers provides robust results suitable for interlaboratory comparisons and for deposition in a global typing database.

Molecular typing of Aspergillus species

Aspergillus species are widely distributed fungi that release large amounts of airborne conidia, which are dispersed in the environment. Several Aspergillus species have been described as human pathogens. Molecular techniques have been developed to investigate the epidemiological relation between environmental and clinical isolates. Several typing methods have been described for Aspergillus species, most of them with reference to Aspergillus fumigatus. Here, we summarise all the different available molecular typing techniques for Aspergillus. The performance of these techniques is evaluated with respect to their practical feasibility, and their interpretation and discriminatory power assessed. For A. fumigatus isolates, a large extent of genetic variability is demonstrated and therefore fingerprinting techniques with high discriminatory power and high reproducibility are required for this species. Afut1‐restriction fragment length polymorphism and microsatellite typing showed the highest discriminatory power. In addition, the microsatellites show excellent reproducibility. Other typing techniques are still useful for smaller epidemiological problems and for less well‐equipped laboratories.

Evaluation of the impact on human salmonellosis of control measures targeted to Salmonella Enteritidis and Typhimurium in poultry breeding using time-series analysis and intervention models in France.

In France, salmonellosis is the main cause of foodborne bacterial infection with serotypes Enteritis (SE) and Typhimurium (ST) accounting for 70% of all cases. French authorities implemented a national control programme targeting SE and ST in poultry and eggs from October 1998 onwards. A 33% decrease in salmonellosis has been observed since implementation. We designed an evaluation of the impact of this control programme on SE and ST human infections in France. Using monthly Salmonella human isolate reports to the National Reference Centre we defined two intervention series (SE and ST) and one control series comprising serotypes not know to be associated with poultry or eggs. The series, from 1992 to 2003, were analysed using autoregressive moving average models (ARMA). To test the hypothesis of a reduction of SE and ST human cases >0 after the programme started and to estimate its size, we introduced an intervention model to the ARMA modelling. In contrast to the control series, we found an annual reduction of 555 (95% CI 148-964) SE and of 492 (95% CI 0-1092) ST human infections, representing respectively a 21% and 18% decrease. For SE, the decrease occurred sharply after implementation while for ST, it followed a progressive decrease that started early in 1998. Our study, suggests a true relation between the Salmonella control programme and the subsequent decrease observed for the two targeted serotypes. For ST, however, the decrease prior to the intervention may also reflect control measures implemented earlier by the cattle and milk industry.

Burden of acute gastroenteritis and healthcare-seeking behaviour in France: a population-based study.

In France surveillance underestimates the true burden of acute gastroenteritis (AG). We conducted a population-based, retrospective cross-sectional telephone survey between May 2009 and April 2010 in order to obtain more accurate estimates of the incidence and the burden of AG and to describe healthcare-seeking behaviour for AG. Of the 10 080 persons included in the survey, 260 respondents reported 263 episodes of AG. The incidence rate of AG was estimated at 0·33 cases/person-year (95% CI 0·28-0·37). It was highest in children aged <5 years and declined with age. Thirty-three percent (95% CI 27-40) of the AG cases consulted a physician and 76% (95% CI 70-82) used medication. Our results indicate that there are more than 21 million episodes of AG each year in France. These results allow a more accurate interpretation of the data derived from existing AG surveillance systems.

A community--wide outbreak of Salmonella enterica serotype Typhimurium infection associated with eating a raw milk soft cheese in France.

In 1997, a community-wide outbreak of Salmonella enterica serotype Typhimurium (S. typhimurium) infection occurred in France. The investigation included case searching and a case-control study. A case was defined as a resident of the Jura district with fever or diarrhoea between 12 May and 8 July 1997, from whom S. typhimurium was isolated in stool or blood. One hundred and thirteen cases were identified. Thirty-three (83 %) of 40 cases but only 23 (55 %) of 42 community controls, matched for age and area of residence, reported eating Morbier cheese (Odds ratio: 6.5; 95 % Confidence Interval: 1.4-28.8). Morbier cheese samples taken from the refrigerators of two case-patients and one symptom-free neighbour cultured positive for S. typhimurium of the same phage type as the human isolates. The analysis of distribution channels incriminated one batch from a single processing plant. These findings show that an unpasteurized soft cheese is an effective vehicle of S. typhimurium transmission.