CHANGE
IN BIOTYPE TREND OF Vibrio cholerae IN NEPAL
Rojina Maharjan1, Dina Shrestha1,
Jyoti Acharya2, Nabaraj Adhikari1 and Upendra Thapa
Shrestha1*
1Department of
Microbiology, Kantipur College of Medical Science, Sitapaila, Kathmandu
2Sukraraj
Tropical and Infectious Disease Hospital, Teku, Kathmandu
*Corresponding
email: upendrats@gmail.com
ABSTRACT
Background:
Cholera is an acute diarrheal illness caused by the
toxigenic bacteria Vibrio cholerae
serogroup O1 and O139 and is associated with rapid loss of body fluids leading
to dehydration, electrolyte disturbances and hypovolemic shock; without
treatment, death can occur within hours. V.
cholerae is classified into more than 200 serogroups based on the O antigen
of the lipopolysaccharide; of these, only O1 and O139 serogroups cause epidemic
cholera. Again, V. cholerae O1 is
further classified into two biotypes: classical and El Tor and contain two
major serotypes: Ogawa, Inaba and additional serotype Hikojima contains both
specific antigens, is rare.
Method: The
study was carried out in Sukraraj Tropical and Infectious Disease Hospital,
Teku, Kathmandu, Nepal from June to November 2012. This research was ethically
approved by Nepal Health Research Council, Kathmandu, Nepal. A total of 450
stool samples from diarrheal patients were collected and transported to the
laboratory as soon as possible. Firstly, inoculating alkaline (pH 8.5) peptone
broth with the specimen and then streaking for isolation after an approximate
6-hours incubation period; this process both enables the rapidly growing
vibrios to multiply and suppresses much of the commensal microflora. For
cultural diagnosis, culture was done on both nonselective media MacConkey agar (MA)
and selective media thiosulfate-citrate-bile salts-sucrose (TCBS)agar on which
the sucrose-fermenting cholera vibrios produce a distinctive yellow colony. The
colony characteristics were observed and subcultured on Nutrient Agar (NA),
futher identification done by gram staining and appropriate biochemical tests.
Isolated V. cholerae were subjected
for serotyping using kit Mast Group as per kit's instruction. All the isolated
strains of V. cholerae were subjected to biotyping by Polymyxin B (50U)
sensitivity test, Voges Proskauer reaction and chicken RBC agglutination tests.
Result:
Out of 450 samples, 22 (4.9%) were
positive for V. cholerae. All isolated strains of V. cholerae strains belonged to serogroup O1, serotype Ogawa and biotype
classical. Children and the elderly people are mostly affected by
cholera. Cholera outbreaks
occurred more in moonsoon season (July and August) . Patients consuming
untreated water for drinking purpose were having more cholera cases 10 (10.9%);
whereas patients drinking boiled water did not have any cholera case. Vomiting,
dehydration, abdominal pain, anorexia and nausea and stool passage frequency ≥
10 were the clinical features and Patients suffering from diarrhea for 2 or
more days yet unrecovered were having more cholera cases. However, all V. cholerae isolates
were sensitive to tetracycline and found to be multi-drug resistant.
Conclusion:
It is suggested
that V. cholerae infection
remain a serious problem like in our developing countries due to the poor
hygienic condition. It can be concluded that proper identification of V. cholerae can be done
by phenotyping method which helps in manangement for diarrhoeal population.
Key
words: Diarrhoea, V.
cholerae, serotyping, Nepal
Note: The full version of this manuscript is coming soon.