CAMPY-TRACE – analytical - PowerPoint Presentation

1. CAMPY-TRACE – analytical innovation for improved food safetyKerstin Stingl1|Ingrid Huber2|Azuka Iwobi2 |Ewa Pacholewicz1|Janine Heise1|Nguyen Hung Long3Hoang Thi Thu Ha4|Dang Duc Anh4|Nguyen Dong Tu4|Pham Thi Ngoc5|Luu Quynh Huong51German federal Institute for Risk Assessment (BfR), National Reference Laboratory for Campylobacter, Germany|2Bavarian Health and Food Safety Authority (LGL), Germany| 3Vietnam Food Aministration (VFA)|4National Institute of Hygiene and Epidemiology, Vietnam (NIHE)|5National Institute of Veterinary Research, Vietnam (NIVR)|Campylobacter is the most frequent foodborne bacterium. The acute symptoms caused in humans are diarrhea, abdominal cramps, fever and vomiting. However, in rare cases Campylobacteriosis might also lead to autoimmune sequelae, e.g. Guillain-Barré syndrome or reactive arthritis in Europe and North America. The impact of Campylobacter is likely to be underestimated in Asian countries due to a lack of data. CAMPY-TRACE: The project aims at collecting comprehensive prevalence data and first quantitative data of this fastidious pathogen along the chicken food chain in Vietnam. Furthermore, the dogma of CFU was challenged by developing a culture-independent detection method for the quantification of live Campylobacter by qPCR. The project is funded by both the German Federal Ministry of Education and Research (BMBF) and by the Vietnamese Ministry of Science and Technology (MOST).In the framework of CAMPY-TRACE also 200 stool samples from diarrhea patients were analysed for Campylobacter. However, only few samples were positive, probably due to prior self-treatment of the patients with antibiotics before contact to the hospital.locationnpos% posHanoi22918078,6Hai Phong21816475,2total44734477Fig. 1. Prevalence of Campylobacter in farm chickens. Pool samples of 3-5 fresh feces from farm chickens per flock in two regions (Ha Noi and Hai Phong) were analysed according to ISO 10272-1C:2017. The sampling period was from Oct 2016 to Dec 2017.The average age of chicken on farms was 4.1 ± 0.8 months and the average weight of chickens was 1.9 ± 0.5 kg. A high prevalence of Campylobacter in chickens of 77 % was observed in North Vietnam, independent of the sampling location (Ha Noi or Hai Phong) (Fig. 1). As expected and in contrast to Germany, no saisonality was observed for the Campylobacter prevalance in chicken in North Vietnam throughout the sampling period.Subsequently, cecal and neck skin samples were analysed at the level of the slaughterhouse. The median quantitative level of CFU in cecal samples was log 6.75 CFU/g, which is lower than observed in European samples, ranging between log 8 and log 10 CFU/g and might be attributed to the higher age of the chicken and different chicken breeds in Vietnam. Microbial enumeration on neck skin samples from Vietnam showed that the majority of carcasses were quantitatively positive, with a peak of CFU between 100 and 1000 CFU per g. In comparison to zoonosis monitoring data from Germany, the distribution of quantitative contamination was different (Fig. 2).An alternative method for the quantification of live Campylobacter by qPCR was developed (Fig. 3), including an internal sample process control (ISPC) (Pacholewicz et al., 2019). This method is currently validated according to ISO 16140-2 against the reference method ISO 10272-2:2017. An international ring trial is conducted in autumn 2019 and the method should be applicable for chicken skin, chicken meat and pork/beef minced meat rinses at an approximate LOQ of 300 CFU/ml.Fig. 2. Comparison of quantitative levels of Campylobacter spp. at slaughtherhouses in Vietnam and Germany. The pro-cess hygiene criterion (1000 CFU/g) was exceeded in 22 % of the German and in 14 % of the Vietnamese samples. However, a large fraction of Viet-namese neck skin samples was con-taminated between 100 and 1000 CFU/g.Fig. 3. Novel method for quantification of live Campylobacter by qPCR+ISPC+PMADNA extraction> 102-103> 103-104> 10440-10010-39≤10% positive