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Culture Media Supplements

Further Information related to  SR0183

CAMPYLOBACTER SELECTIVE MEDIA

The revelation in the 1970's that campylobacters are important human pathogens marked the beginning of an upsurge of interest in these organisms which has continued unabated. New species are being identified and some existing species have been assigned to new genera.

In 1977, enteritis-causing campylobacters were still in the province of a few experts but with improved media and isolation procedures, clinical laboratories can isolate thermophilic campylobacters routinely. A considerable body of literature on the genus has arisen; reference to this work and an up-date on developments has been made in a review by Penner1. The table below has been adapted from this review.

Differential reaction and characteristics for species of the genera Campylobacter, Arcobacter and Helicobacter pylori. (Adapted from Penner1)

 
Growth
Susceptibility
Species
Catalase
Nitrate
H2S (TSI)
Hippurate
Urease
25°C
37°C
42°C
Nali dixic acid
Cepho-lothin
G+C content (mol%)
Campylobacter fetus subsp. fetus
+
+
-
-
-
+
+
(-)
R
S
33-34
C. fetus subsp. venerealis
+
+
-
-
-
+
+
-
R
S
33-34
C. hyointestinalis  
+
+
+
-
-
(+)
+
+
R
S
35-36
C. jejuni  
+
+
-
+
-
-
+
+
S
R
30-32
C. coli  
+
+
-
-
-
-
+
+
S
R
31-33
C. lari  
+
+
-
-
-
-
+
+
R
R
31-33
C. upsaliensis  
(-)
+
-
-
-
-
+
+
S
S
35-36
‘C. cinaedi’  
+
+
-
-
-
-
+
-
S
I
37-38
‘C. fennelliae’  
+

-

-
-
-
-
+
-

S

S
37-38
C. sputorum  
-
+
(+)
-
-
-
+
+
(S)
S
31-32
Biovar sputorum
Biovar bubulus
-
+
+
-
-
-
+
+
R
S
31-32
Biovar fecalis
+
+
+
-
-
-
+
+
R
S
32-33
C. mucosalis
-
+
+
-
-
-
+
+
R
S
38-39
C. concisus
-
+
+
-
-
-
+
+
R
R
38-39
Arcobacter cryaerophilia
+
+
-
-
-
+
+
-
d
R
29-30
A. nitrofigilis
+
+
ND
-
+
+
+
-
S
S
28-29
Helicobacter pylori
+
d
-
-
+
-
+
+
R
S
36-37

+ Positive reaction; - negative reaction; ND no test results found; (+) most strains positive but a low percentage negative: (-) most strains negative but some positive or weakly positive; d different reactions; R resistant; S suscpetible; I intermediate zones of inhibitions.
aSusceptbility to antibiotics was determined with 30m g disks.

The major step forward in recognising the importance of campylobacters in human disease was the development of isolation media which contain antibiotics. These media suppress competing faecal flora and allowed the campylobacters to grow into easily detected colonies.
The first selective supplement was developed by Skirrow1 and other workers followed with other antibiotic combinations.
Differential reactions and characteristics for species of the genera Campylobacter, Arcobacter and Helicobacter pylori. (Adapted from Penner1)Gun-Monro et al.2 carried out a laboratory and clinical evaluation of the various selective isolation media for thermophilic campylobacters. Summary tables of their findings for the above antibiotic supplements are shown.

Selective Antibiotic Supplements for the isolation of Campylobacters

Antibiotics

(mg/litre)

Butzler

SR85

Blaser-Wang

SR98

CCDA

SR155

Preston

SR117

Skirrow

SR69

Karmali

SR167

CAT

SR174

Amphotericin B

-

2

10

-

-

-

10

Bacitracin

25,000*

-

-

-

-

-

-

Cephalothin

-

15

-

-

-

-

-

Cefazolin

15

-

-

-

-

32

-

Cefoperazone

-

-

32

-

-

-

8

Colistin

10,000*

-

-

5,000*

-

-

-

Cycloheximide

50

-

-

100

-

100

-

Novobiocin

5

-

-

-

-

-

-

Polymyxin

-

2,500*

-

-

2,500

-

-

Rifampicin

-

-

-

10

-

-

-

Trimethoprim

-

5

-

10

5

-

-

Vancomycin

-

10

-

-

10

20

-

Teicoplanin

-

-

-

-

-

-

4

*IU/L
CCDA – Modified Charcoal Cefoperazone Desoxycholate Agar (Blood-free medium)

Table 1. Recovery of 70 C. jejuni strains on five selective media.

Medium

Colony Count

P valueb

Blood agar control

7.95 ± 0.36

 

Skirrow

7.82 ± 0.48

Nsc

Butzler

7.77 ± 0.51

<0.05

Blaser-Wang

7.70 ± 0.56

<0.05

Preston

7.76 ± 0.52

<0.05

Modified CCDA

7.91 ± 0.36

NS

Table 2. Isolation of C. jejuni from 70 simulated positive faeces samples.

Medium

24 Hours

48 Hours

Blood agar control

69 (99)

70 (100)

Skirrow

39 (56)

67 (96)

Butzler

38 (54)

60 (86)

Blaser-Wang

17 (24)

31 (41)

Preston

32 (46)

64 (91

Modified CCDA

61 (87)

69 (99)

aLog10 mean colony counts ± standard deviation.
bsignificance determined by Student's t test for unpaired samples.
cNS, Not significant.

Table 3. Suppression of faecal flora from 70 simulated positive faecal samples.

No. (%) of plates with 75% reduction of faecal flora compared with control

 

Medium number (%) of strains isolated after incubation for:

Medium

24 Hours

48 Hours

Skirrow

46 (66)

38 (54)

Butzler

56 (80)

47 (67)

Blaser-Wang

28 (40)

15 (21)

Preston

58 (83)

50 (71)

Modified CCDA

64 (91)

59 (84)

The general conclusions reached by Gun-Munro et al. were confirmed by Griffiths and Ribeiro3.

Enrichment broth cultures - the value of enrichment media for campylobacters is controversial1 but in food and environmental studies enrichment may be essential4. Enrichment at 42° C4,5 and cold enrichment at 4° C6 have been reported. Where enrichment increases competitive flora, the use of membrane filters on the surface of the agar can help select campylobacters7.
A comprehensive review of selective media for Campylobacter and Arcobacter species has been published in a special issue of International Journal of Food Microbiology.

Reference
Corry J.E.L., Post D.E., Colin P. and Laisney M.J. (1995). Int. J. Food Microbiol. 26. 43-76.

Laboratory growth environment
Atmosphere
- members of the Campylobacter genus require a wide spectrum of atmospheres for optimum growth, ranging from complete anaerobiosis to ambient air tolerance8. Most species, however, lie between these extremes and are micro-aerophilic. The Oxoid Campylobacter Gas Generating Kits (BR56 and BR60) ensure that the correct oxygen and carbon dioxide levels are produced for the optimum growth of these micro-aerophilic organisms. Alternatively use CampyGen CN025A or CN035A. CampyGen does not require the addition of water or a catalyst.
Temperature - the temperature range for incubation of Campylobacter species and related organisms varies from 15° C for Arcobacter cryaerophilea to 42° C for the thermophilic species. However, most strains have a considerable tolerance of growth temperatures around those required for optimum growth.
Helicobacter pylori - this organism is implicated as a cause of gastritis and peptic ulceration9,10,11.
A specific selective culture medium, prepared from Helicobacter pylori Selective Supplement (Dent) SR147 and Columbia Blood Agar Base CM331 is required to isolate this organism from gastric biopsy specimens.

References
1 Penner J. L. (1988) Clin. Microbiol. Reviews. 1. 157-172.
2 Gun-Munro J., Rennie R. P. , Thornley J. H., Richardson H.L., Hodge D. and Lynch J. (1987) J. Clin. Microbiol. 25. 2274-2277.
3 Griffiths A. and Ribeiro C. D. (1988) J. Clin. Path. 41. 704-705.
4 Marinescu M., Festy B., Derimay R. and Megraud F. (1987) Eur. J. Clin. Microbiol. 6. 693-694.
5 Bolton F. J., Coates D., Hinchliffe P. M. and Robertson L. (1983) J. Clin. Path. 36. 78-83.
6 Rubin S. J. and Woodward N. (1983) J. Clin. Microbiol. 18. 1008-1010.
7 Steele T. W. and McDermott S. N. (1984) Pathology 16. 263-265.
8 Neill S. D., Campbell J. N., O'Brien J. J., Weatherup S. T. and Ellis W. A. (1985) Int. J. Sys. Bacteriol. 35. 342-356.
9 Marshall B. K., Warren J. R., Blincow E. D., Phillips M., Goodwin C. S., Murray R., Blackbourne S. J. and Waters T. E. (1988) Lancet ii. 626-627.
10 Dent J. C. and McNulty C. A. M. (1988) Eur. J. Clin. Microbiol. Infect. Dis. 7. 555-568.
11 Buck G. E. (1988) Lab. Managemt. 26. 9.

 
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