Campylobacter detection in. chicken caeca

Transcription

1 Campylobacter detection in Results of Ring test number 1 March 27 chicken caeca CRL Campylobacter Ingrid Hansson

2 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.

3 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

4 Detection of Campylobacter Results from 24 NRLs participating in ringtest 1, March 27 No of NRLs 25 Correct result detection Correct result identification C.jejuni 2. C.coli 3. C.coli 4. Neg 5. C.jejuni CCUG Cb CCUG Cb

5 Detection of Campylobacter Results from 24 NRLs participating in ringtest 1, March 27 No of correct results 6 Correct result AB EBC 85B 344C 288D E6BA FC FA 754 Laboratory code 26E3 2E C C9D 3E2C F135 2B5B 5F1 2A8D 37CF 36D3

6 Species identification of Campylobacter Results from 24 NRLs participating in ringtest 1, March 27 No of correct results AB EA3F 85B 344C 288D E6BA FC FA E3 2E C C9D Laboratory code 3E2C F135 2B5B 5F1 2A8D 37CF Neg C. coli C. jejuni 36 D3

7 Campylobacter detection in Results of Ring test number 2 September 27 chicken caeca CRL Campylobacter Ingrid Hansson

8 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, E3, 2A8D, 2B5B, 2C14, 2E41, 36D3, 3E2C, F135 3 days 2 297, 37CF 7 days 2 E8C4, FC1

9 Campylobacter in chicken caeca Culture media; mccda and other Incubation time and temperatures; according to ISO 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)

10 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

11 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 -

12 Microaerobic atmosphere Number of NRL Gas generating kits 12 Incubator 7 Anoxomat 4 Zip-lock bags 1

13 Detection of Campylobacter Results from 24 NRLs Correct results from NRLs mccda (N=24) Other (N=22) Sample number in the ringtest

14 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

15 Species identification Confirmation from mccda No of NRL Sample No C. jejuni C. coli C. lari Neg

16 No of NRL 25 2 Species identification Confirmation from other selective media than mccda C. jejuni C. coli C. lari Neg Sample No

17 Species identification PCR No of NRL Sample No C. jejuni C. coli C. lari Neg

18 No of NRL 25 Species identification Inoculated with C. jejuni CCUG Sample no (mccda) C. jejuni C. coli C. lari Neg

19 Species identification Inoculated with C. coli, Cb No of NRL Sample no (mccda) C. jejuni C. coli C. lari Neg

20 Species identification Natural contaminated, from the same slaughter batch No of NRL Sample no (mccda) C. jejuni C. coli C. lari Neg

21 Species identification Negative caeca from the same slaughter batch No of NRL Sample no (mccda) C. jejuni C. coli C. lari Neg

22 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 /7* 1 + /12* 3 + /3* Sample no /7* 13 + /15* + /* Sample no /6* 12 + /16* + /* + mccda * Other plate than mccda

23 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

24 Detection of Campylobacter B 344C 288D E6BA FA 754 Correct result detection mccda 26E E C C9D 3E2C F135 2B5B 5F1 2A8D 37CF 36D3 FC1 E8C4

25 Species identification of Campylobacter Correct results phenotyping from mccda B 344C 288D E6BA FA E E41 Laboratory code C C9D 3E2C F135 2B5B 5F1 2A8D 37CF 36D3 F21 E8C4 Negative Nat contam. C. coli C. jejuni

26 Detection of Campylobacter Results from 3 NRLs ring test 2B No correct results from NRLs mccda Other Sample number in the ringtest

27 Species identification Results from 3 NRLs ring test 2B from mccda No of NRL C. jejuni C. coli C. lari Neg Sample no

28 No of NRL Species identication Results from 3 NRLs ring test 2B C. jejuni C. coli C. lari Neg Sample no, from media other than mccda

29 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)

30 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