EURL- Campylobacter 5th Workshop

Transcription

1 EURL- 5th Workshop Technical Report 2010 Eva Olsson Engvall The EURL- 5th workshop was held in Uppsala 4-6 October 2010 Representatives from NRLs in 26 Member States (MS), two non-mss Iceland and Norway, and Candidate country Croatia participated. Experts from DG Sanco, EFSA, and ECDC as well as other experts participated. Altogether 51 persons attended the workshop. Workshop program. The program included presentations by experts from the European Commission DG Sanco, EFSA and ECDC, by EURL staff members, NRL representatives and by other experts. The presentations are posted on the EURL- website. DG SANCO, EFSA and ECDC Klaus Kostenzer (KK) DG Sanco presented Ongoing activities on from the riskmanagement s view. The EU legal framework activities to control are regulated by EC Directives and Regulations. Different activities are carried out, i.e. collection of zoonoses data, summarized in the annual Community Summary Reports and the EFSA Scientific Opinions (one published on quantification of the risk posed by broiler meat, and one ongoing on control options and performance objectives and/or targets at different stages of the food chain). Further, the baseline survey in 2008 enabled the collection of comparable data on prevalence in broiler batches and broiler carcasses in the MSs. A next step would be setting Community targets, an issue that is currently under discussion with the MSs. KK described the process steps within Regulation (EC) 2160/2003 on the control of Salmonella and other specified food-borne zoonotic agents. A suggestion to set process hygiene criteria (PHC) for has been made, but several MSs considered it too early. Maria Teresa da Silva Felicio (MTSF), EFSA, gave two presentations. In her first presentation, results from the 2008 Community Summary Report on trends and sources of zoonoses and zoonotic agents in the EU, MTSF described the collection and analysis of zoonoses data from the MSs. is one of 8 zoonotic agents, for which it is mandatory to report findings in food, feed and animals. In addition, food-borne outbreaks have to be reported. In 2008, the number of human cases in 25 EU MSs were (to be compared with salmonellosis cases in 27 MSs). The number of samples taken from different food categories or animals categories, the sampling and analytical methods vary between countries, which means that results from different countries are not directly comparable. Nevertheless, poultry meat is still regarded the most important food-borne source of campylobacteriosis. In the second presentation, Results of the baseline survey on prevalence of in broiler batches and on broiler carcasses, in the EU 2008, MTSF summarized the work with preparing the part A and part B reports of the baseline survey results. Objectives with the baseline survey were to estimate the prevalence of in broiler batches (caecum samples at slaughter) and of in broiler meat (contamination on broiler carcasses at the end of the slaughter process), and to get information about the quantity (numbers) of in broiler meat (contamination on broiler carcasses at the end of the slaughter process). In part A, prevalence data are published and discussed. Part B aimed at studying the effects of factors, which may be associated with colonized broiler batches and contaminated carcasses at EU level. The baseline survey showed that there are great differences in prevalences between the MSs. Factors that were associated with colonized batches were previous thinning of the flock and age of birds at sampling. Factors associated with contaminated carcasses were colonization status of the slaughter

2 batch, age of the birds and time of sampling during the day. The colonized batches were also strongly associated with higher numbers of on the carcasses. The analyses of results also showed that there was an unexplained variance attributed to slaughter house specific factors but for which no data were gathered during the survey. Some of the results of contaminated carcasses should be interpreted with caution, since sensitivity of the detection test may have varied between the MSs. Johanna Takkinen (JT), ECDC (European Centre for Disease Prevention and Control), presented Surveillance of food- and water borne diseases and zoonoses in the EU. Focus on. JT presented the ECDC and its mission to identify, assess and communicate current and emerging health threats to human health from communicable diseases. A long- term surveillance strategy for includes the enhanced surveillance of, among other diseases, food and waterborne diseases (FWD). General FWD surveillance objectives have been set up and a FWD surveillance network. The network builds on the former Enter-net network which has nominated six priority diseases: salmonellosis, campylobacteriosis, STEC/VTEC infection, listeriosis, shigellosis, and yersiniosis. Currently there is a special focus on salmonellosis and VTEC. For strengthening capacity the need for training and quality issues, especially as regards, have been raised. New EU project Merete Hofshagen, National Veterinary Institute, NO, presented a new EU project CamCon control novel approaches in primary poultry production. The project, with 10 partners from 7 countries (Norway, Denmark, UK, The Netherlands, Portugal, Spain and Poland), started in May 2010 and will continue for 4 years. There are 6 work packages (WPs): 1. Epidemiology, 2. Pre-harvest interventions, 3. Diagnostic tools for monitoring and control, 4. Risk assessment and economics, 5. From Science to industry, and 6. Management. A webpage has been set up: Analytical methods, revision of standards, validation of methods Enne de Boer (EdB), VWA, NL, presented Revision of ISO parts 1 and 2 including proposals from the Ad hoc group ISO/CEN that had a meeting on the 4 th of October (before the workshop started). The group working on the revision of the ISO standards EN ISO :2006 and ISO/TS :2006 has decided (i) to launch a revision of part 1, to improve the enrichment and confirmation steps and microaerobic conditions, and (ii) to transform part 2 into an EN ISO standard including a modification of confirmation steps. EdB went through a number of proposals made by the ad hoc group in order to improve the standards. For revision of the standards, further studies are necessary to form a solid basis for making decisions. A call for participation in a comparative trial for detection of in different matrices including samples from the primary production was made last fall (2009). Eight laboratories participated, however different matrices had been used and the results were rather diverse. More comparative trials are therefore welcome. The ad hoc group will prepare a first draft of the revised standard (parts 1 and 2) based on the current proposals and the draft will be sent to ISO and CEN and a voting procedure will follow. Wilma Jacobs- Reitsma (WJR) RIVM, NL, presented MicroVal validation of a enumeration medium. The standard EN ISO 16140: 2003 Microbiology of food and animal feeding stuffs Protocol for the validation of alternative methods, is under revision, a process that may take some time. WJR described the different steps in the validation process including both qualitative and quantitative method validation, according to this standard. WJR then described the validation of a new medium (selective agar) for enumeration of thermotolerant species, in particular C. jejuni and C. coli in poultry products.

3 Proficiency Tests in 2010 Ingrid Hansson, EURL-, presented results from Proficiency Test (PT) (previously named Ring test) number 7 Detection and enumeration of spp in broiler meat. The PT was distributed in April 2010 by courier to 34 NRLs. The PT included detection and enumeration of using ISO part 1 and 2. The samples consisted of broiler meat, which should be used together with 10 vials with or without spp. The objectives were to: (i) assess the performance of the NRLs to detect and enumerate in broiler meat samples, and (ii) to evaluate the standard operating procedures (SOPs) prepared for the analyses. Out of the 34 NRLs, 19 reported correct results of detection of in all ten samples after enrichment in Bolton broth followed by culture on mccda and 22 NRL after enrichment in Bolton broth followed by culture on second selective agar. The isolated spp. were speciated by the use of phenotypic methods and/or molecular methods such as Polymerase Chain Reaction (PCR) techniques. In general, identification by PCR gave more correct results compared to the phenotyping in this proficiency test. Thirty-one of the 34 NRLs reported results from enumeration. 20 NRLs correctly reported negative or spp. found in the 10 samples by direct plating on mccda for enumeration. Most of the NRLs found two log differences between the samples with different concentrations of C. jejuni, C. lari and C. coli. Eva Olsson Engvall, EURL-, presented the results of PT no 6, 2010, Swab ring test. It had been agreed at the workshop in 2009 that a voluntary ring test on detection and identification of species should be organized by the EURL. The test consisted of 10 bacterial cultures of spp (n = 8) and related organisms (n = 2), which were distributed as live cultures on swabs in Amies transport medium. The swabs were sent by regular airmail on 8 th of March The NRLs were free to use any method for identification. Thirty-three NRLs participated in the test, 30 NRLs in the EU MSs and three in non-eu countries (CH, MK and NO). Eighteen NRLs used a combination of phenotypic tests and PCR for identification, nine used only phenotypic tests and six NRLs used only PCRs. In addition, 16S rdna sequencing, latex agglutination, API- Campy and mass spectrometry by Maldi-Tof MS were applied by some NRLs. The overall results showed that the test had been relatively difficult to perform and some strains were not culturable when they arrived at the NRLs. However, the majority of NRLs correctly identified C. jejuni, C. coli and C. lari strains as spp. One sample, containing C. avium, was excluded in the evaluation of PT results since it was reported as no growth at 24 NRLs. Two NRLs could identify it as C. avium (by sequencing), 4 NRLs reported it as spp and one each as C. jejuni, C. coli or not spp. The sample with C. upsaliensis also caused problems; only 17 NRLs correctly identified the species, and 3 NRLs reported it as spp. From 12 NRLs it was reported as no growth. It was concluded that: the PT should have been sent by courier (door-to-door delivery) to secure short time for transport, most NRLs could correctly identify the thermophilic spp C. jejuni, C. coli and C. lari as such or at least as spp, identification by PCR, either alone or in combination with phenotypic tests was more reliable than using only phenotypic tests for identification of species in this PT. Ann Nyman, EURL-, presented What is good performance in a proficiency test? In previous proficiency tests (PTs) of no limits have been set for defining good performance. However, since the PTs have developed and the bacterial reference materials used in two PTs since 2009 have been freeze dried bacterial cultures, which are more stable and comparable, there is also a need for a more standardized assessment of the performance in the PTs. Other EU-RLs have already defined their limits or cut-offs for what is considered good performance in their PTs, and the EU- RL believes that applying a standardized assessment will give a useful tool for both the EU-RL and the NRLs to evaluate the performance in the tests. A proposal on what could be considered

4 reasonable limits and cut-offs for classification of good performance in the different parts of the PT ( Detection and enumeration ) was presented. The proposal was received positively by the participants and it was agreed that the reports of PT 6 and PT 7 should include a first outline of criteria for good performance in the PTs. Group discussions Elina Lahti, EURL-. Summary of discussions. The participants were given a couple of questions to be discussed in groups of about 8 persons. First, the delegates discussed the baseline study. The results of the study were mainly consistent with previous findings in the respective countries. For economical reasons no major actions have been taken after the study. However, in Belgium an upper limit of 100 cfu/g has been set for in chicken meat. National monitoring projects are conducted in some countries every 3 rd or 4 th year according to a previous plan. In some countries, research projects are in progress. The new proposal for a revised ISO method for detection and enumeration of was discussed. Fewer steps in confirmation were supported by most participants. However, the suggestion to allow two options for enrichment based on whether a low or high background flora is expected was considered more problematic. Difficulties to estimate the background flora and the need for thorough testing before taking the standard into use were mentioned. National presentations Karl Pedersen, DK: activities in Denmark. Surveillance data on in poultry and on antimicrobial resistance were presented. The data are also available from: and respectively. DTU Food will organize an EU course in microbiology for 3 rd countries. The course will be held in November-December 2010 and in May The course is part of the EU programme Better Training for Safer Food. Marina Mikulic, HR: Baseline study on the prevalence of preliminary results. A baseline study on the prevalence and antimicrobial resistance to spp in broiler batches and of spp and Salmonella spp in broiler carcasses, is carried out in Croatia in The study is performed according to technical specifications for the EU baseline survey in Preliminary results were presented. Alexander Maramski, BG: NRL Microbial safety of food products. The activities at the Bulgarian NRL in 2010 were presented including results of a research study on prevalence of spp in pig and lamb carcasses. Elisabetta Di Giannatale, IT: Italian NRL for spp. The activities at the Italian NRL were presented. Projects include molecular typing, genome sequencing, serotyping, antimicrobial sensitivity testing, and a study of the prevalence and role of risk factors for in the broiler and pig production chains. Ruta Bubuliene, LT: National Food and Veterinary Risk Assessment Institute, NRL for in Lithuania. Epidemiological data of prevalences of in humans, poultry, and other animals and in food in Lithuania were presented.

5 Evaluation of the workshop. A paper form was handed out to the workshop participants for evaluation of the workshop. Thirty-five participants returned a completed form at the end of the workshop. The majority of participants was satisfied with the organization and contents of the workshop program and gave high scores, Excellent or Good on all questions. A summary of answers and comments will be posted on the website. Plans for EURL- NRL activities in 2011 Eva Olsson Engvall & Ingrid Hansson, EURL-, summarized the planned activities. The workprogram and budget for the EURL- for 2011 was submitted to the Commission in August The following key activities are planned: Proficiency test(s) Detection and enumeration of in meat matrix using different enrichment broths and different selective agars. Date for submission: 4 April A second PT, like the swab ring test in 2010 was discussed, but the possibility to organize such a PT in 2011 is uncertain. Annual workshop, suggested date: 3-5 October 2011 Training course(s): basic training course in PFGE in January The participation in the course is free, but the EURL will not be able to pay costs for travel and accommodation (those costs have to be covered by the NRLs). Short PCR training courses can be arranged on an ad hoc basis. Research molecular typing, assessment/validation of analytical methods. Testing of media and matrices for the PT Scientific and technical assistance to the Commission and NRLs ISO/CEN activities, participation in work with the revision of ISO Missions, participation in international networks and meetings Communication, much of the communication is done by s. More information will be posted on the website Uppsala 29 November 2010 Eva Olsson Engvall EURL- National Veterinary Institute SE Uppsala Sweden

6 EU-RL (CRL) - workshop 4-6 October 2010, Uppsala, Sweden Programme MONDAY 4 OCTOBER Registration at First Hotel Linné Get-together dinner at First Hotel Linné. TUESDAY 5 OCTOBER Lecture room Linnésalen First Hotel Linné Welcome and introduction. Ongoing activities on - from the risk-management s view. results from the 2008 Community Summary Report on trends and sources of zoonoses and zoonotic agents in the EU. Surveillance of food- and water borne diseases and zoonoses in the EU- focus on. Coffee/tea break. Results of the baseline survey on prevalence of in broiler batches and on broiler carcasses. Group discussion of results. CamCon- control- novel approaches in primary poultry production. Staffan Ros, Dep Director General, SVA Eva Olsson Engvall, EU-RL- Klaus Kostenzer, DG- SANCO Maria Teresa Da Silva Felicio, EFSA Johanna Takkinen, ECDC Maria Teresa Da Silva Felicio, EFSA Merete Hofshagen, National Veterinary Institute, NO Lunch Revision of ISO standards. Results of Ring Test No 7, Detection and Enumeration of in Poultry Meat. Group discussions of results and Enne de Boer, VWA, NL Ingrid Hansson, EU-RL

7 methodology Coffee/tea break. Interlaboratory study for MicroVal validation of method for enumeration of in poultry meat products. Summary and end of day. Wilma Jacobs-Reitsma, RIVM, NL Karl Pedersen, DK Marina Mikulic, HR Alexander Maramski, BG Eva Olsson Engvall, EU-RL Dinner WEDNESDAY 6 OCTOBER Lecture room Linnésalen First Hotel Linné Results of Ring Test No 6, Swab samples. Good performance of proficiency tests. Discussion of limits for passing/failing the tests. Coffee/tea break. Summary and conclusions made from Tuesday s Discussions. Plans for future activities. Eva Olsson Engvall, EU-RL- Ann Nyman, EU-RL- Elisabetta Di Giannatale, IT Ruta Bubuliene, LT Elina Lahti, EU-RL- Eva Olsson Engvall and Ingrid Hansson, EU- RL Lunch Bus from Hotel Linné to Arlanda Airport