CHOLERA Vibrio cholerae Brenda Anna Kwambana Wangeci Kagucia.
Vibrio cholera - University of Mosulmedicinemosul.uomosul.edu.iq/files/pages/page_2106458.pdf ·...
Transcript of Vibrio cholera - University of Mosulmedicinemosul.uomosul.edu.iq/files/pages/page_2106458.pdf ·...
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Vibrio cholera
احمد معن سعدهللا •
بالل مؤيد جاسم •
بالل محمد حازم •
فراس الطائي: بأشراف الدكتور •
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Genus : Vibrio
the main species:
.choleraCauses epidemic and pandemic : Vibrio cholerae .1
2. Non-cholera Vibrios: Cause ear, wound, soft tissue, and extra-intestinal infections.
3. Vibrio parahaemolyticus: Causes gastroenteritis, and
possibly extra-intestinal infections.
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Vibrionacae
• Catalase +
• Oxidase +
• They ferment glucose, maltose, mannite, and sucrose with acid production only
• The TSI shows A/A profile (yellow colour)
Pseudomonas
• Catalase +
• Oxidase +
• Only glucose fermented without gas
• The TSI shows K/A , pink /yellow
Enterobacteae
• Catalase +
• Oxidase –
• LF : E.coli , klebseilla , citrobacter , enterobacter .
• NLF : shigella , salmonella , proteus .
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Pseudomonas Vibrionacae Enterobacteae
Catalase +ve Catalase +ve Catalase +ve
Oxidase +ve Oxidase +ve Oxidase –ve
NLF Ferment
maltose,lactose,manitol
Ferment glucose
motile motile Motile / shigella ,
klebsiella ,yersinia
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• Gram negative
• Short curved rods like commas with marked pleomorphism size
• These m.o. are : highly aerobic
• very actively motile (described as shooting stars or darting motility)
• The best way to detect motility is by dark-field or contrast microscopy
• These m.o. can tolerate temprature of range 18-37.
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Biochemical characteristics of V. cholerae:
•They ferment glucose, maltose, mannite, and sucrose with acid production only.
• These m.o. are:
respiratory (oxidase positive)
fermentative (catalase positive).
• They give positive cholera-red reaction on nitrate peptone medium by reducing nitrate to nitrite, and produce indol (+ve) on tryptophan medium.
•The TSI shows A/A profile (yellow colour).
• The optimum pH for the growth of these m.o. is 7.0 , but they can tolerate up to 8.5 - 9.0.
• They are very sensitive to acidic pH (less than 6), therefore, they are quite susceptible to gastric juice (a major barrier against V. cholerae).
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بالل محمد حازم
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Culture of V. cholerae
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This m.o. can be cultivated on different media:
Alkaline Peptone Water (APW;
Sea Water): This medium is of pH 8.5 – 9.0 and used for the
primary (first) isolation from clinical specimens. The
m.o. grow forming a surface pellicle within 8 hours.
Then, from this medium the m.o. can be sub-
cultured on solid media
Many m.o. can not tolerate such pH, but
Pseudomonas does.
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2. Thiosulfate Citrate Bile Salt Sucrose Agar (TCBS):
The non-inoculated medium is olive green in colour, and contains an indicator called bromo-thymol blue.
V.Cholerae is sucrose- fermenter, and due to acid production, the pH drops and colour of the medium and colonies become yellow. Aeromonas can produce similar chages in this medium.
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TCBS
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3. Tellurite Taurocholate Gelatin Agar (TTGA)
On this medium the colonies show
iridescence and are surrounded by cloudy
zone of gelatin hydrolysis.
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4. Meat Extract Agar: On this medium the colonies appear
translucent green to red bronzy in colour and
in old cultures the colonies become opaque
and rough
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5. Taurocholate Pepton Broth:
This medium can be used primary
cultivation of the m.o., then sub-
cultured on other media
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6. MacConkey’s Agar:
V.Cholerae colonies are of non-lactose
fermenters
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7. Blood Agar:
El-Tor growth shows beta-haemolysis ( versus the
classical).
In general, the colonies of V. cholerae are convex,
smooth, granular.
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احمد معن سعدهللا
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Practical procedure of cultivation of V. cholera :
1. Direct stool examination:
a. Motility: Dark field or contrast microscopy.
b. Microscopical exam.: mucous,epith.cells,and large
number of m.o. , but no pus cells are seen.
c. Fluorescent antibody staining.
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2. From the APW, TCBS and
another APW are inoculated and
left either for 5 hours or
overnight.
3.The colonies are subjected to
further identification.
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Further Identification of V. cholerae:
1. String test: A drop of 0.5% Na-deoxycholate is mixed with a colony ;
after 60 seconds, with a loop there will a thread of
tenacious nature. If positive, it means V. cholerae.
2. Polyvalent anti-sera (anti-O1,anti- O139): To confirm that the V. cholerae belongs to O-1 or
O-139, agglutination test with specific polyvalent sera and
emulsified colonies is carried out.
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3. Biochemical tests including cholera
red reaction.
4. Differentiation between classical & El-
Tor Biotypes.
5. Serotyping: by specific antisera
against A, B and A antigenic factors to
determine Ogawa, Inaba & Hikojima.
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