Previous Article | Next Article 
Journal of Clinical Microbiology, November 2001, p. 4222-4223, Vol. 39, No. 11
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.11.4222-4223.2001
LETTERS TO THE EDITOR
Efficient Isolation of Campylobacter upsaliensis
from Stools
 |
LETTER |
Byrne et al. (1) presented data on the superiority
of cefoperazone amphotericin teicoplanin (CAT) selective medium over modified cefoperazone charcoal deoxycholate selective medium for the
efficient isolation of Campylobacter upsaliensis from
stools. There are alternatives to the use of selective media for the
isolation of C. upsaliensis. Since 1977 we have routinely
isolated campylobacters from the diarrhetic stools of pediatric
patients at the Red Cross Children's Hospital, Cape Town, South
Africa. In 1990, primarily for cost containment reasons, the use of
antibiotic-containing selective media for Campylobacter
isolation was discontinued in our laboratory and the Cape Town protocol
was introduced. This isolation protocol was the first to combine both
membrane filtration onto antibiotic-free blood agar plates and
incubation in an H2-enhanced microaerobic atmosphere
(3). With the use of this protocol, the number of
stool cultures positive for campylobacteria rose to 21.8% from the
7.1% previously obtained with Skirrow's and other selective media
available at that time (3). Since the introduction of the
Cape Town protocol we have isolated over 1,200 strains of C. upsaliensis from the diarrhetic and normal stools of pediatric and
adult patients and from dogs, cats, and meercats (2). Our
laboratory could begin to isolate C. upsaliensis, Campylobacter concisus, Campylobacter curvus,
Campylobacter rectus, Campylobacter sputorum
biovar sputorum, Campylobacter
hyointestinalis, Helicobacter fennelliae,
Helicobacter cinaedi, Arcobacter butzleri, and
other campylobacteria from the stools of humans and animals only with
the introduction of the Cape Town protocol. Some strains of
campylobacteria are sensitive to antibiotics commonly used in selective
media or have an essential requirement for an H2-enhanced microaerobic atmosphere.
We have compared the efficacy of the filtration component of the Cape
Town protocol with that of CAT selective medium for C. upsaliensis isolation from 300 consecutive diarrhetic stool samples from gastroenteritis patients at the Red Cross Children's Hospital (Table 1). The antibiotic-free
filtration and CAT isolation plates were incubated under identical
conditions, in an H2-enhanced microaerobic atmosphere at
37°C. Campylobacter, Helicobacter, and
Arcobacter isolates were identified by recognized phenotypic and biochemical criteria. The data in Table 1 indicate that with filtration onto antiobiotic-free plates, 20.3% of the stools were positive for campylobacteria, while with the use of CAT selective plates only 4.7% of the same stools were positive for campylobacteria. Both methods were equally efficient for the isolation of
Campylobacter coli and A. butzleri; however,
filtration was superior to CAT selective medium for all other
campylobacteria isolated. Campylobacter jejuni subsp.
doylei, H. fennelliae, C. hyointestinalis, and C. concisus strains were isolated
with filtration but were not isolated with CAT media. Sixteen strains
of C. jejuni subsp. jejuni were isolated with
filtration, whereas nine strains were isolated with CAT medium.
Eleven C. upsaliensis strains were obtained with filtration, but only a single C. upsaliensis strain was obtained with
CAT medium. Generally, colonies of C. upsaliensis and other
campylobacteria on the antibiotic-free blood agar plates used in the
Cape Town protocol were larger, more prominent, and faster growing
(visible growth after 2 to 4 days) than those on the CAT plates.
View this table:
[in this window]
[in a new window]
|
TABLE 1.
Efficiency of filtration versus that of CAT selective
medium for isolation of C. upsaliensis and related organisms
from 300 consecutive diarrhetic stools of patients at the Red Cross
Children's Hospital
|
|
Byrne et al. (1) state that membrane filtration is costly
and labor intensive. We do not agree, as the Cape Town protocol, which
has been in continuous use over the last 11 years, has proved to be a
simple, efficient, and cost-effective alternative to the use of
antibiotic-containing selective media for the isolation of C. upsaliensis and other campylobacteria from stool. The
underdetection of C. upsaliensis and other campylobacteria
in the stools of gastroenteritis patients is an important diagnostic
problem, and application of the Cape Town protocol may help alleviate
this concern.
 |
REFERENCES |
| 1.
|
Byrne, C. D.,
A. Doherty,
M. Mooney,
D. Woodward,
W. Johnson,
F. Rodgers, and B. Bourke.
2001.
Basis of the superiority of cefoperazone amphotericin teicoplanin for isolating Campylobacter upsaliensis from stools.
J. Clin. Microbiol.
39:2713-2716[Abstract/Free Full Text].
|
| 2.
|
Lastovica, A. J., and E. Le Roux.
2000.
Efficient isolation of campylobacteria from stools.
J. Clin. Microbiol.
38:2798-2799[Free Full Text].
|
| 3.
|
Le Roux, E., and A. J. Lastovica.
1998.
The Cape Town protocol: how to isolate the most campylobacters for your dollar, pound, Franc, yen, etc., p. 31-33.
In
A. J. Lastovica, D. Newell, and E. E. Lastovica (ed.), Proceedings of the 9th International Workshop on Campylobacter, Helicobacter and related organisms. Institute of Child Health, Cape Town, South Africa.
|
| | | | |
Albert Joseph Lastovica
Elza Le Roux
Department of Medical Microbiology University of Cape Town and Red Cross Children's Hospital Cape Town, South Africa
|
 |
AUTHORS' REPLY |
We are most grateful to Drs. Lastovica and Le Roux for their interest
in our recent paper on Campylobacter upsaliensis isolation. During the course of experiments aimed at identifying the basis of the
differences in productivity between two widely available Campylobacter selective media for isolating C. upsaliensis (2), we also examined the effect of the
Cape Town protocol conditions on the growth of 15 of our isolates
(unpublished data). When the growth of the 15 isolates was compared to
that observed with conventional incubation using the CampyGen system,
we found that 9 C. upsaliensis isolates showed reduced
growth (using the ecometric plating system) and 3 isolates failed to
grow at all under the Cape Town protocol conditions.
Nevertheless, the Cape Town protocol undoubtedly has contributed
greatly to the rate of isolation of campylobacters and related organisms at the Red Cross Children's Hospital, Cape Town, South Africa. The findings presented by Lastovica and Le Roux concerning the
superiority of this protocol compared with the use of cefoperazone amphotericin teicoplanin selective medium for isolation of C. upsaliensis in their hands indicates the potential for application of their isolation methodology in investigating the epidemiology of
enteric Campylobacter infection. Studies by other
investigators comparing the productivities of the Cape Town protocol
and of selective media among populations with lower prevalences of
campylobacters clearly are warranted.
However, for the present, we feel that our own findings, together with
concerns regarding the sensitivity of filtration methods for low
numbers of organisms (1, 3), the possible biohazard of
high hydrogen levels (4), and the perceived awkwardness of
filtration methodology, pose a substantial barrier to the
attractiveness of filtration-based techniques in the clinical
laboratory setting.
 |
REFERENCES |
| 1.
|
Bourke, B.,
V. L. Chan, and P. Sherman.
1998.
Campylobacter upsaliensis: Waiting in the wings.
Clin. Microbiol. Rev.
11:440-449[Abstract/Free Full Text].
|
| 2.
|
Byrne, C.,
D. Doherty,
A. Mooney,
M. Byrne,
D. Woodward,
W. Johnson,
F. Rodgers, and B. Bourke.
2001.
Basis of the superiority of cefoperazone amphotericin teicoplainin for isolating Campylobacter upsaliensis from stools.
J. Clin. Microbiol.
39:2713-2716.
|
| 3.
|
Corry, J. E. L.,
D. E. Post,
P. Colin, and M. J. Laisney.
1995.
Culture media for the isolation of campylobacters.
Int. J. Food. Microbiol.
26:43-76[CrossRef][Medline].
|
| 4.
| Engberg, J., P. Gerner-Smidt, S. W. L. On, and
C. S. Harrington. 2000. Efficient isolation of
Campylobacteria from stools. 38:2798-2799.
|
| | | | |
Catherine Byrne
Billy Bourke
Department of Paediatrics The Conway Institute University College Dublin, and Childrens Research Centre Our Lady's Hospital For Sick Children Crumlin, Dublin 12, Ireland
|
Journal of Clinical Microbiology, November 2001, p. 4222-4223, Vol. 39, No. 11
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.11.4222-4223.2001
This article has been cited by other articles:
-
Lastovica, A J, Le Roux, E
(2003). Optimal detection of Campylobacter spp in stools. J. Clin. Pathol.
56: 480-480
[Full Text]