Campylobacter detection in Results of Ring test number 1 March 27 chicken caeca CRL Campylobacter Ingrid Hansson
Ring test 1, Swab samples Detection of Campylobacter in chicken caecal contents. Ring test number 1, swab samples Regarded as pilot study. Test the ability to detect Campylobacter in chicken faeces. Give information about how well the transportation of samples and communication works between the CRL and NRLs.
Ring test number 1, swab samples Five swab samples, transported in Amies transport medium containing charcoal. Overnight culture of Campylobacter serum broth has been mixed with chicken caecal contents. Four out of five swabs contain Campylobacter, but all swabs have been in contact with the chicken caecal contents. Direct culture were carried out on a selective medium suitable for Campylobacter
Detection of Campylobacter Results from 24 NRLs participating in ringtest 1, March 27 No of NRLs 25 Correct result detection Correct result identification 2 15 1 5 1. C.jejuni 2. C.coli 3. C.coli 4. Neg 5. C.jejuni CCUG 11284 Cb 671-3 CCUG 11283 Cb 15398-3
Detection of Campylobacter Results from 24 NRLs participating in ringtest 1, March 27 No of correct results 6 Correct result 5 4 3 2 1 135 7AB EBC 85B 344C 288D E6BA FC1 332 31FA 754 Laboratory code 26E3 2E41 2657 2C14 82 3C9D 3E2C F135 2B5B 5F1 2A8D 37CF 36D3
Species identification of Campylobacter Results from 24 NRLs participating in ringtest 1, March 27 No of correct results 6 5 4 3 2 1 135 7AB EA3F 85B 344C 288D E6BA FC1 332 31FA 754 26E3 2E41 2657 2C14 82 3C9D Laboratory code 3E2C F135 2B5B 5F1 2A8D 37CF Neg C. coli C. jejuni 36 D3
Campylobacter detection in Results of Ring test number 2 September 27 chicken caeca CRL Campylobacter Ingrid Hansson
Days of transport of packages Time No of labs NRL Lab codes 1 day 1 5F1, 754, 82, 135, 288D, 31FA, 332, 344C, 3C9D, E6BA 2 days 11 7AB, 85B, 2657. 26E3, 2A8D, 2B5B, 2C14, 2E41, 36D3, 3E2C, F135 3 days 2 297, 37CF 7 days 2 E8C4, FC1
Campylobacter in chicken caeca Culture media; mccda and other Incubation time and temperatures; according to ISO 1272-1 26 Confirmation tests; morphology, motility, growth at 25ºC (microaerobic) and 41.5 ºC aerobic, oxidase Species identification; catalase, Hippurate hydrolysis, Indoxyl acetate, Nalidixic acid (optional), Cephalotin (optional), PCR (optional)
Start of the analyses Days between samples left CRL and start of analysis Correct results, detection Start No of labs mccda Other selective media 1 day 6 4/6 4/5 2 days 11 1/11 8/1 3 days 4 4/4 4/4 7 days 3 1/3 1/3
Detection of Campylobacter Second media No of NRL Correct result detection Karmali 11 8/11 Skirrow 6 4/6 Abeyta-Hunt-Bark (AHB) 1 2/2 Butzler 1 1/1 Campy-cefex 1 1/1 CAT 1 1/1 Preston 1 1/1 No second media 2 -
Microaerobic atmosphere Number of NRL Gas generating kits 12 Incubator 7 Anoxomat 4 Zip-lock bags 1
Detection of Campylobacter Results from 24 NRLs Correct results from NRLs 3 25 2 15 1 5 mccda (N=24) Other (N=22) 1 2 3 4 5 6 7 8 9 1 11 12 Sample number in the ringtest
Detection of Campylobacter in chicken caeca Four different kind of caecum samples Negative caeca; sample no 3, 6 and 1 Negative caeca inoculated with Campylobacter jejuni, sample no 1, 7 and 11 Negative caeca inoculated with Campylobacter coli, sample no 2, 5 and 9 Natural contaminated caeca with Campylobacter jejuni, sample no 4, 8 and 12
Species identification Confirmation from mccda No of NRL 25 2 15 1 5 1 2 3 4 5 6 7 8 9 1 11 12 Sample No C. jejuni C. coli C. lari Neg
No of NRL 25 2 Species identification Confirmation from other selective media than mccda 15 1 5 C. jejuni C. coli C. lari Neg 1 2 3 4 5 6 7 8 9 1 11 12 Sample No
Species identification PCR No of NRL 16 14 12 1 8 6 4 2 1 2 3 4 5 6 7 8 9 1 11 12 Sample No C. jejuni C. coli C. lari Neg
No of NRL 25 Species identification Inoculated with C. jejuni CCUG 11284 2 15 1 5 1 7 11 Sample no (mccda) C. jejuni C. coli C. lari Neg
Species identification Inoculated with C. coli, Cb 671-3 No of NRL 3 25 2 15 1 5 2 5 9 Sample no (mccda) C. jejuni C. coli C. lari Neg
Species identification Natural contaminated, from the same slaughter batch No of NRL 3 25 2 15 1 5 4 8 12 Sample no (mccda) C. jejuni C. coli C. lari Neg
Species identification Negative caeca from the same slaughter batch No of NRL 3 25 2 15 1 5 3 6 1 Sample no (mccda) C. jejuni C. coli C. lari Neg
Differences between selective media Sample No 3, 6 and 1 from the same slaughter batch Neg sample no No growth +/ * Other +/ * C. coli +/ * Sample no 3 11 + /7* 1 + /12* 3 + /3* Sample no 6 11 + /7* 13 + /15* + /* Sample no 1 12 + /6* 12 + /16* + /* + mccda * Other plate than mccda
Time to start the analyses Ring test number 2B October 27 Correct result Correct result Time No of labs mccda Other 2 days 2 1/2 1/2 3 days 1 1/1 1/1
Detection of Campylobacter 14 12 1 8 6 4 2 135 85B 344C 288D E6BA 332 31FA 754 Correct result detection mccda 26E3 297 2E41 2657 2C14 82 3C9D 3E2C F135 2B5B 5F1 2A8D 37CF 36D3 FC1 E8C4
Species identification of Campylobacter Correct results phenotyping from mccda 14 12 1 8 6 4 2 135 85B 344C 288D E6BA 332 31FA 754 26E3 297 2E41 Laboratory code 2657 2C14 82 3C9D 3E2C F135 2B5B 5F1 2A8D 37CF 36D3 F21 E8C4 Negative Nat contam. C. coli C. jejuni
Detection of Campylobacter Results from 3 NRLs ring test 2B No correct results from NRLs 5 4 3 2 1 mccda Other 1 2 3 4 5 6 7 8 9 1 11 12 Sample number in the ringtest
Species identification Results from 3 NRLs ring test 2B from mccda No of NRL 5 4 3 2 1 1 2 3 4 5 6 7 8 9 1 11 12 C. jejuni C. coli C. lari Neg Sample no
No of NRL 5 4 3 Species identication Results from 3 NRLs ring test 2B 2 1 1 2 3 4 5 6 7 8 9 1 11 12 C. jejuni C. coli C. lari Neg Sample no, from media other than mccda
Conclusions Campylobacter in chicken caeca Campylobacter spp. could survive in chicken caeca for at least 4 days mccda seems to be one of the most selective agar plate for Campylobacter spp. 19 (79%) out of 24 NRLs could detect Campylobacter in the caeca (mccda) 16 (67%) out of 24 NRLs could identify the correct Campylobacter species in the caeca (mccda)
Acknowledgement To all Member States, Norway and Island participating in the ring test To the staff of the CRL- Campylobacter To the Commission for funding the work of the CRL