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Material Safety Data Sheet

Further Information

Organisms

Organisms in the Clinical sector this product works with:

Dehydrated Culture Media

Sector: Clinical

CAMPYLOBACTER SELECTIVE AGAR (PRESTON)

a selective medium which when prepared from Campylobacter Agar Base CM0689, Preston Campylobacter Selective Supplement SR0117 and Lysed Horse Blood SR0048, can be used for the selective isolation of Campylobacter jejuni and C. coli from human, animal, avian and environmental specimens

CAMPYLOBACTER AGAR BASE

Code: CM0689

Typical Formula*

gm/litre

`Lab-Lemco’ powder

10.0

Peptone

10.0

Sodium chloride

5.0

Agar

12.0

pH 7.5 ± 0.2 @ 25°C

 
* Adjusted as required to meet performance standards

CAMPYLOBACTER SELECTIVE SUPPLEMENT (PRESTON)

Code: SR0117

Vial contents (each vial is sufficient for 500ml of medium)

per vial
per litre

Polymyxin B

2,500IU

5,000IU

Rifampicin

5.0mg

10.0mg

Trimethoprim

5.0mg

10.0mg

Cycloheximide

50.0mg

100.0mg

MODIFIED PRESTON CAMPYLOBACTER SELECTIVE SUPPLEMENT

Code: SR0204

Vial contents (each vial is sufficient for 500ml of medium)

per vial
per litre

Polymyxin B

2,500IU

5,000IU

Rifampicin

5.0mg

10.0mg

Trimethoprim

5.0mg

10.0mg

Amphoteracin B

5.0mg

10.0mg

Directions (to prepare Preston Campylobacter Selective Agar)
Suspend 18.5g of Campylobacter Agar Base in 475ml of distilled water and bring to the boil to dissolve completely. Sterilise by autoclaving at 121°C for 15 minutes. Cool to 50°C. Aseptically add 25ml of Lysed Horse Blood (SR0048), and 1 vial of Preston Campylobacter Selective Supplement (SR0117 or SR0204) reconstituted as directed and one vial of Campylobacter Growth Supplement (SR0232). Mix well and pour into sterile Petri dishes.

Directions (to prepare Preston Campylobacter Selective Enrichment Broth)
Dissolve 12.5g of Nutrient Broth No.2 (CM0067) in 475ml of distilled water and sterilise by autoclaving at 121°C for 15 minutes. Cool to 50°C or below. Aseptically add 25ml of Lysed Horse Blood (SR0048), 1 vial of Preston Campylobacter Selective Supplement (SR0117 or SR0204) and 1 vial of Campylobacter Growth Supplement (SR0232). Aseptically dispense 5ml volumes in sterile small screw-capped bottles. The Selective Enrichment Broth may be stored for up to 7 days at 2-8°C.

It is essential that the head space above the liquid should be as small as possible to ensure microaerobic conditions.

Description
The Preston Campylobacter Selective Agar is based on the formulation described by Bolton and Robertson1. This medium was specifically formulated to be suitable for isolation of Campylobacter species from all types of specimens (human, animal, avian and environmental).
In comparative studies2 of the selective media of Skirrow, Butzler, Blaser, Campy-BAP and Preston, the Preston medium was found to give the maximum isolation rate of Campylobacter species from all types of specimens tested and also to be the most selective.

Oxoid Campylobacter Agar Base has been prepared from materials described by Bolton and Robertson1. It is suitable as a basal medium for the selective supplements of Blaser-Wang, Skirrow and Butzler.

Preston Campylobacter Selective Supplement (SR0117) can also be used to prepare Preston Campylobacter Selective Enrichment Broth2.
The selective enrichment technique is recommended for specimens and food samples that are expected to be heavily contaminated and/or carry small numbers of viable colony forming units. The Preston Campylobacter Selective Enrichment Broth which is supplemented with the growth supplement SR0232, made to the formulation of George et al.3 effectively quenches toxic compounds which may form on exposure of the medium to light and air4.

Technique
Direct Selective Plating Method

  1. Prepare the Preston Campylobacter Selective Agar as directed.
  2. Emulsify the specimen under test in 2ml of 0.1% peptone water.
  3. Inoculate onto the selective medium with cotton tipped swabs so that single isolated colonies are formed.
  4. Incubate the plates in an atmosphere consisting of approximately 5-6% oxygen, 10% carbon dioxide and 84-85% nitrogen for 24-48 hours at 42°C. This can best be achieved by using the Oxoid Gas Generating Kit for Campylobacters (BR0056) in conjunction with the Oxoid Anaerobic Jar and an active catalyst. For jars of smaller capacity (2.5 litres) use the Oxoid Gas Generating Kit for Campylobacters BR0060. Alternatively use CampyGen (CN0025 or CN0035). CampyGen does not require the addition of water or a catalyst.
  5. Examine the plates and confirm the typical colonies as Campylobacter jejuni or Campylobacter coli by the standard methods.

When few Campylobacter colony forming units are present 48 hours incubation is necessary.

Selective Enrichment Broth Technique

  1. Prepare the Preston Selective Enrichment Broth as directed.
  2. Emulsify the specimen under test in the selective enrichment broth.
  3. Incubate the broth aerobically at 42°C for 24 hours.
  4. Subculture on to Preston Campylobacter Selective Agar or Campylobacter Blood-Free Selective Agar.

CAMPYLOBACTER TRANSPORT MEDIUM
Campylobacter Selective Supplement (Preston) is incorporated in an improved medium for storage and transportation of thermophilic Campylobacters 5.

Storage conditions and Shelf life
Store the dehydrated medium at 10-30°C and use before the expiry date on the label.
Store the prepared plates at 2-8°C.

Appearance
Dehydrated medium: Straw coloured, free-flowing powder
Prepared medium: Agar - Straw coloured gel
Broth - Straw coloured solution

Quality control

Positive control:

Expected results

Campylobacter jejuni ATCC® 29428 *

Good growth; grey-brown coloured colonies

Negative control:

 

Escherichia coli ATCC® 25922 *

Inhibited

* This organism is available as a Culti-Loop®

References
1. Bolton F.J. and Robertson L. (1982) J. Clin. Pathol. 35. 462-467.
2. Bolton F.J., Coates D., Hinchliffe P.M. and Robertson L. (1983) J. Clin. Pathol. 36. 78-83.
3. George H.A., Hoffman P.S., Kreig N.R. and Smibert R.M. (1979) Can. J. Microbiol. 25. 8-16.
4. Bolton F.J., Coates D. and Hutchinson D.N. (1984) J. Appl. Bact. 56. 151-157.
5. Rogol M., Schnaidman B., Katzenelso E. and Sechter I. (1990) Eur. J. Clin. Microbiol. Inf. Dis. 9. 760-762.

 
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