RESEARCH NOTE
Laboratory-acquired Vibrio cholerae O1
infection in Austria, 2008
Steliana Huhulescu, Eva Leitner, Gebhard Feierl and Franz
Allerberger
Austrian Agency for Health and Food Safety (AGES), National Reference
Centre for Vibrio cholerae, Vienna, Austria
Abstract
Vibrio cholerae infection is a rare but well-documented cause of
laboratory-associated illness. We report on the first case of
indigenous cholera documented in Austria after more than
fifty years. In April 2008, the National Reference Centre for
V. cholerae received an isolate of V. cholerae O1, serotype
Ogawa, cultured from the stool specimen of a patient consulting
a general practitioner because of watery diarrhea. The 23 year
old microbiology student had been working with viable V. cho-
lerae for 4 weeks in a practical laboratory course. Two days
before onset of symptoms an open 300 mL Erlenmeyer flask
with approx. 30 mL of overnight V. cholerae culture tipped over
and spilled into a laboratory shaker near the student’s working
place. Wearing gloves and protective gowns, the student and
her supervisor immediately cleaned and decontaminated the
shaker. As a consequence of this laboratory incident, the insti-
tution in question replaced the clamp-less shaker plate by a
traditional shaker plate with mechanical clamps.
Keywords: Cholera, clamp-less shaker, laboratory-acquired
infection, laboratory shaker, Vibrio cholerae
Original Submission: 14 July 2009; Revised Submission:
19 August 2009; Accepted: 27 August 2009
Editor: D. Raoult
Article published online: 3 September 2009
Clin Microbiol Infect 2010; 16: 1303–1304
10.1111/j.1469-0691.2009.03051.x
Corresponding author and reprint requests: F. Allerberger,
Austrian Agency for Health and Food Safety (AGES), National
Reference Centre for Vibrio cholerae, Währingerstr. 25a, 1090 Vienna,
Austria
E-mail: Franz.Allerberger@ages.at
In Austria, epidemic cholera has not occurred since 1950
(start of statutory reporting) [1]. During 1990–2009, a total
Journal Compilation ª2010 European Society of Clinical Microbiology and Infectious Diseases
EPIDEMIOLOGY
of 11 cases of cholera were reported to the public health
authorities. In all but one case, cholera infection was
acquired during foreign travel. This report concerns the first
case of indigenous cholera reported in Austria for more than
50 years.
In April 2008, the National Reference Centre for Vibrio
cholerae received an isolate cultured from the stool specimen
of a patient consulting a general practitioner because of
watery diarrhoea. The 23-year-old microbiology student,
who had been working with viable V. cholerae for 4 weeks in
a practical laboratory course, asked her general practitioner
to consider V. cholerae as a possible causative agent.
Three days later, the isolate was reported to be V. chole-
rae O1, serotype Ogawa, sensitive to ampicillin, ciprofloxa-
cin, tetracycline, gentamicin and cefotaxime, and resistant to
trimethoprim. The diarrhoea ceased spontaneously after
3 days, hours before ciprofloxacin therapy was initiated (oral
ciprofloxacin, 250 mg every day for 5 days). The patient had
not been abroad for more than 6 months prior to the onset
of illness. She had no concomitant comedication and had
never taken proton-pump inhibitors. (The risk of infection
following oral exposure may be increased in achlorhydric
individuals [2,3].)
The incubation period of cholera is estimated to range
from a few hours to 5 days, and is usually 2–3 days [4]. No
specific event explaining this infection in Austria is known.
Pipetting by mouth can be excluded as the cause of infection.
Pipetting by mouth of any infectious material should be
strictly prohibited.
Contact with non-intact skin or mucosa and ingestion
have previously been described as routes of laboratory-asso-
ciated infection with V. cholerae [5]. Two days before onset
of the symptoms, an open 300-mL Erlenmeyer flask contain-
ing c. 30 mL of an overnight V. cholerae culture tipped over
and spilled into a laboratory shaker near the student’s work-
place. Wearing gloves and protective gowns, the student and
her supervisor immediately cleaned and decontaminated the
shaker. Whether the flask tipped over spontaneously or was
inadvertently pushed over (perhaps when another flask was
removed) could not be ascertained in retrospect. In the
reported event, a completely new laboratory shaker, with
adhesive mats instead of clamps for holding the flasks, was
used. In order to limit the possibility of mishandling of bio-
safety level 2 (BSL2) organisms, any new laboratory machine
should be assessed for maximally secure handling.
As a consequence of this laboratory incident, the institu-
tion in question replaced the clampless shaker plate with a
traditional shaker plate with mechanical clamps.
Laboratory-acquired infections are important for evaluat-
ing the effectiveness of biosafety measures.
ª2010 The Authors
1304 Clinical Microbiology and Infection, Volume 16 Number 8, August 2010
BSL2 practices, BSL2 containment equipment and BSL2
facilities are recommended for activities involving cultures of
V. cholerae [6]. V. cholerae infection is a rare but well-docu-
mented cause of laboratory-associated illness [7]. This is
rather surprising, as the infectious dose is fairly high. In stud-
ies with human volunteers, the infectious dose was deter-
mined to range from 106 to 1011 CFUs [5,6,8,9].
In many reports of laboratory-acquired infections, there
has been mention of lack of compliance with biosafety prac-
tices. This observation may be reassuring regarding the effec-
tiveness of such practices, at least if they are followed [10].
It illustrates that educating laboratory personnel to comply
with the rules remains the top priority [10]. According to
BSL2 criteria, all work that may create aerosols or splatter
must be performed inside a biological safety cabinet [6].
These precautions were apparently not taken in the present
case, where an open 300-mL Erlenmeyer flask containing
c. 30 mL of overnight culture of V. cholerae was placed in a
clampless shaker in the laboratory.
Transparency Declaration
All authors declare no conflicts of interest.
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ª2010 The Authors
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