CCDA Supplement (Campylobacter Blood free)

Technique

Collect, dilute and prepare samples and volumes as required according to specifications, directives, official standard regulations and/or expected results.

Reconstitute the vial with 5 mL of sterile diluent, pre-warmed to approx. 37 °C, and add it to 500 mL of Campylobacter Agar Base Blood Free (CCDA) ISO (LC-1129) cooled to 50 °C before pouring into Petri dishes.

Once solidified on a flat surface, spread the plates by streaking methodology or by spiral method.

Incubate the plates in microaerophilic atmosphere at 40-42 °C for 24-48 h.

(Incubation times longer than those mentioned above or different incubation temperatures may be required depending on the sample or the specifications.)

After incubation, count all the colonies that have appeared on the surface of the agar.

Presumptive isolation of Campylobacter spp. must be confirmed by further microbiological and biochemical tests.

Reconstitute the original freeze-dried vial by adding 5 mL of sterile distilled water.

Note: each vial is sufficient to supplement 500 mL of Blood Free Campylobacter Agar Base.

Physical/chemical control

Microbiological control

Reconstitute 1 vial as indicated in Composition; shake and dissolve completely.

Add 1 vial to 500 mL of medium base. DO NOT HEAT once supplemented.

Microaerophilia. Incubation at 35 ± 2 °C or 42 ± 2 °C for 24-48 hours.

Distribute the complete medium, cooled to 50 °C, into 90 mm plates.

Incubate according to the instructions for the complete medium indicated in Composition.

Microaerophilia. Incubation at 41.5 ± 1 °C; reading at 44 ± 4 h.

Sterility control

Add 5 mL of the sample to 100 mL of TSB and to 100 mL of Thioglycollate.

Incubation 48 hours at 30-35 °C and 48 hours at 20-25 °C: NO GROWTH.

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• BAYLIS, C.L., (editor) (2007) Manual of Microbiological Methods for the Food and Drinks Industry. 5th Edition Method 3.3.1:2007. CCFRA. Chipping Campden. U.K.
• BOLTON, F.J. (2000) Methods for isolation of campylobacters from humans, animals, food and water. In "The increasing incidence of human campylobacteriosis" Report and Proceedings of a WHO Consultation of Experts. Copenhagen Denmark 21-25 November 2000, WHO/CDS/CSRAPH 2001. p. 87-93.
• BOLTON, F.J. & D. COATES (1983) Development of a blood-free campylobacter medium: screening tests on basal media and supplements, and the ability of selected supplements to facilitate aerotolerance. J. Appl. Bacteriol. 54:115-125.
• BOLTON, F.J., D. COATES & D.N. HUTCHINSON (1984) The ability of Campylobacter media supplements to neutralize photochemically induced toxicity and hydrogen peroxide. J. Appl. Bacteriol. 56:151-157.
• CORRY, J.E.L., H. IBRAHIM ATABAY, S.J. FORSYTHE & L.P. MANSFIELD (2003) Culture Media for the isolation of campylobacters, helicobacters and arcobacters. In Handbook of Culture Media for Food Microbiologists. J.E.L. Corry et al. (Eds.) Elsevier Science B.V. Amsterdam.
• CORRY, J.E.L., G.D.W. CURTIS & R.M. BAIRD (2003) Handbook of culture media for food Microbiology. Elsevier Sci. B.V. Amsterdam.
• FDA (Food and Drug Administration) (1998) Bacteriological Analytical Manual. 8th Edition. Rev. A. AOAC International. Gaithersburg, Maryland, USA.
• HUNT, J.M., C. ABEYTA & T. TRAN (1998) Campylobacter. In FDA BAM 8th Edition (rev. A) 7.01-7.027 AOAC International. Gaithersburg, Md, USA.
• HUTCHINSON, D.N. & F.J. BOLTON (1984) Improved blood-free selective medium for the isolation of Campylobacter jejuni from faecal specimens. J. Clin. Pathol. 37:956-957.
• ISO 10272-1 Standard (2017) Microbiology of the food chain - Horizontal Method for detection and enumeration of Campylobacter spp. - Part 1: Detection method.
• ISO 10272-2 Standard (2017) Microbiology of the food chain - Horizontal Method for detection and enumeration of Campylobacter spp. - Part 2: Colony count technique.
• ISO 11133:2014/Amd 1:2018. Microbiology of food, animal feed and water. Preparation, production, storage and performance testing of culture media.
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