Dr.T.V.Rao MD

Cholera affects Millions 

CHOLERA AFFECTS millions, in endemic areas and causes thousands of deaths especially during seasonal epidemics. Robert Koch, the famous microbiologist of Germany, discovered Vibrio cholerae, the causative organism. Because of its characteristic shape he originally referred to it as comma bacilli. Nearly 7-8 epidemics 7caused by the pathogen `Vibrio cholerae affected different parts of the world.

Discovery of Cholera Organisms 

Cholera came to Florence in 1854 during the Asiatic Cholera Pandemic of 1846-63. Pacini became very 1846-63. interested in the disease. Immediately following the death of cholera patients, he performed an autopsy and with his microscope, conducted histological examinations of the intestinal mucosa. During such studies, Pacini first discovered a comma-shaped bacillus which he described commaas a Vibrio. He published a paper in 1854 entitled, "Microscopical observations and pathological deductions on cholera" in which he described the organism and its relation to the disease. His microscopic slides of the organism were clearly labeled, identifying the date and nature of his investigations.

Spread of Cholera Pandemics

many credited Robert Koch (1843(18431910) with this seminal discovery.1854  Filippo Pacini. but not until cholera. 82 years after his death. when the international committee on nomenclature in 1965 adopted Vibrio cholerae Pacini 1854 as the correct name of the choleracholera-causing organism. would gain prominence for his discovery of Vibrio cholera.Filippo Pacini . Until then. .

like most of the scientific community. was unaware of Pacini's work at the University of Florence. Since Koch's findings eventually became accepted by his scientific peers.Robert Koch Isolates V. he became the acknowledged discoverer of the cholera organism. .cholerae 1883  The German physician Robert Koch. and were widely know in the popular press. Yet both independently came to a similar conclusion.

Recent Cholerae Outbreaks .

Vibrios in Nature Vibrios are among the most common bacteria in surface water worldwide.  They appear curved aerobic rods and are motile. while other vibrios may cause sepsis or enteritis  .possesing a polar flagellum  V.cholerae serogroups O1 and O139 casue cholera in humans.

cholerae transmission in water and the development of sanitary system .Vibrio cholerae Epidemiology and spread of Cholera closely parallels the recognition of V.

vibrios may become straight rods that resemble the gramgramnegative enteric bacteria.  On prolonged cultivation.cholerae is a comma shaped curved rod 2 4 µm long¶ long¶  It is actively motile by means of polar flagellum.  .Morphology and Identification V.

Enrichment Medium   Enrichment of the fecal specimens are done on Alkaline peptone water Venktraman Ramakrishan Meidum is simple medium can be used as transport medium .

 .  Vibrios grow at a very high pH ( 8-5 ± 9-5 ) and 8are rapidly killed by acid  In resource poor laboratories MacConkey¶s MacConkey¶ agar can be used.cholerae produces convex.  Grow well at 370c on many defined media. smooth.round colonies that are opaque and granular in transmitted light.Culture V.

TCBS  V.Selective Medium . on which it produces yellow colonies that are readily visible against the dark green background of the agar. .cholerae grows well on Thiosulphate citrate bile sucrose (TCBS ) agar.

cholerae and other Vibrios Vibrio species are susceptible to compound 0/129 and differentiates from Aeromonas Vibrios usually grow on medium containing 6% NaCl Halophilic vibrios need and grow in the presence of > 6% Nacl On Blood agar Vibrios show hemodigesion .cholerae ferments sucrose and mannose but not arabinose A positve Oxidase test is key step in preliminary identification of V.Growth Characteristics       V.

cholerae strains of O group 1 and O group 139 cause classic cholera.cholerae has O lipopolysachhrade that confer serologic specificity There are at least 139 O antigen groups V. Occasionally non 01/non 0139 V.Antigenic Structure and Biological Classification      Many Vibrios share a single heatlaible H antigen.cholerae cause Cholera like disease . V. Antibodies to H antigen are probably not involved in the protection of susceptible hosts.

Epidemic V.cholerae .cholerae is biotyped into 1 Classic 2 El Tor  .cholerae 01 has determinants that make possible futher typing  Serotypes are Ogawa.V.typing V. Inaba and Hikojima.

Proskauer test  Resistant to Polymyxin B  Molecular techniques can also be used to type V.chlorae  Tests can be done reference laboratories and technically demanding  .El Tor Vibrios The El Tor vibrios produce a Hemolysin and positive results with VogesVoges.

Specific totals for numbers of V cholerae 0139 cases are unknown because affected countries do not report infections caused by 01 and 0139 separately. . >100.cholrae 0139  The epidemic of cholera caused by V cholerae 0139 has affected at least 1 1 countries in southern Asia. however. and appears to be closely related to V cholerae 01 biotype El Tor strains.New Epidemic of V. V cholerae 0139 produces severe watery diarrhea and dehydration that is indistinguishable from the illness caused by V cholerae 01.000 cases of cholera caused by V cholerae 0139 may have occurred.

V.cholerae 0139 donot produce 01 type lipopolysachhrade and does not have all the genes necessary to make this antigen V.cholerae 0139 make a polysachhrdie capsule like other non 01 V.cholerae strains while V.cholerae.cholerae 01 does not make a capsule .cholerae 0139 is very similar to V.choleraeV.0-139    V.cholerae 01 El Tor biotype V.

cholerae 0139 .V.

cholrae 0139 Epidemic The epidemic of cholera caused by V cholerae 0139 has affected at least 1 1 countries in southern Asia. V cholerae 0139 produces severe watery diarrhea and dehydration that is indistinguishable from the illness caused by V cholerae 01 and appears to be closely related to V cholerae 01 biotype El Tor strains. strains.V. .

and absorption of sodium and chloride is inhibited The genes for V.cholerae Enterotoxin are on the bacterial chromosome Cholerae Enterotoxin is antigenically related to LT of Escherichia . which promotes entry of subunit A into the cell Activation of subunit A1 yields increased levels of intracellular cAMP and results in prolonged hyper secretion of water and electrolytes There is increased sodium dependent chloride secretion.Vibrio cholerae .000 ) and B Ganglioside GM1 serves as a mucosal receptor for subunit B.cholerae produce heat labile enterotxoin with a Mol wt of about 84.000 consisting of sub units A ( MW 28.Enterotoxin       V.

How Enterotoxin acts .

Pathology and pathogenesis     V.cholerae are when ingested with water .cholrae But a person with normal normal gastric acidity has to consume 1010 or more V.cholerae is pathogenic to humans. When bacteria are consumed with food few organism as much as 102 104 organisms are adequate to cause an attack because of the buffering capacity of the food Any medication or conditions that decreases stomach acidity makes a person more susceptible to infection with V.

Clinical events in Cholera .

.Pathology     Cholera is not an invasive infection Organisms do not reach blood. only act locally Virulent V.cholerae organism attach to the microvillus of the brush border of epithelial cells They multiply and liberate cholera toxin and perhaps Mucinase and Endotoxin. Endotoxin.

About 60% of infections are caused with classic V.cholerae and are asymptomatic. about 75% of infections are caused by El Tor biotype .Clinical manifestations       Diarrhea occurs as much as 20 30 Liters/Day fluids are lost. Results in dehydration Shock Acidosis Can lead to death.

profuse diarrhea. depends on the size of the inoculums ingested Manifest with Nausea .Clinical features   The incubation period is 1 4 days for person who develop symptoms. . vomiting. and abdominal cramps Rice water stool characteristic of cholera loss of fluid leads to profound dehydration Circulatory collapse and anuria.

 Milder cases difficult to differentiate from other diarrheal diseases.  .Stool Examination Stool specimen appear as Rice water  On Microscopy contain Mucus.  El Tor vibrios cause milder disease than classic biotypes. epithelial cells and large number of Vibrios.

 . pans.  Specimens should not be collected from bed pans.  Vomitus not advised.Specimen Collections Stool specimens are collected in acute stage of the disease.  Simple collection of stool in a wide Mouthed container is safe and hygienic. before the antibiotics are administered.

Dark field microscopy shows rapidly motile vibrios.Laboratory Diagnosis    Mucus flecks from stool are cultured. Smears are not useful for diagnosis. .

 The stool specimens can be transported in Venkatraman Ramakrishnan medium  Alkaline peptone water is ideal enrichment medium  .  On TCBS medium typical colonies can be picked in 18 hours.Laboratory Diagnosis Culture Grwoth is rapid on Blood agar.

cholerae( Classical ) Hemolysis -ve VogesVoges-proskauer test -ve Polymyxin sensitivity +ve Group IV phage Susceptibility +ve Chick erythrocyte Agglutination -ve V.Bio Chemical Reactions V.cholerae (El Tor) Tor) +ve +ve -ve -ve +ve .

chlorae organisms are further identified by slide agglutination tests using anti -0 group 1 or group 139 Antisera and by Biochemical reactions .Confirmatory Tests for V.cholerae  V.

In experimental animals specific IgA antibodies occur in the lumen of intestine Vibriocidal antibodies in the serum titer > 1:20 have been associated with protection against colonization and disease The presence of antitoxin antibodies have not been asociated with protection . An attack of cholera if followed by immunity to reinfection but the duration and degree of immunity are not known.Immunity in Cholerae      Gastric acid produces some protection against cholera vibrios.

Patients with suspected cholera should be reported immediately to local and state health departments.When you suspect Cholera  The diagnosis of cholera should be considered in patients with watery diarrhea who have recently (i.e.. . within 7 days) returned from cholera-affected choleracountries.

Treatment    The most important part of therapy consists of correction water and electrolyte imbalance to correct severe dehydration and salt depletion. Oral Tetracycline tends to reduce stool output in cholera and shortens the period of excretion of vibrios In some endemic areas tetracycline resistance has emerged the genes are carried by transmissible plasmids .

Vaccines for Cholera  The licensed parenteral cholera vaccine provides only limited and brief protection against V cholerae 01. . and has a high cost-benefit ratio. New oral cholera vaccines are being developed. the vaccine is not recommended for travelers. may not provide any protection against V cholerae 0139. costtherefore.

None of these vaccines have attained the combination of high efficacy. although still at a high cost per case averted.Newer Vaccines  New oral cholera vaccines are being developed and provide more reliable protection. and low cost necessary to make mass vaccination feasible in choleracholeraaffected countries. . long duration of protection. simplicity of administration.

nor recommended for the vast majority of travelers by the Centers for Disease Control and Prevention (CDC). .CDC j On Vaccination for Cholerae Cholera vaccine is no longer required.

cholerae 01 of Classical type contributed to pandemics.Epidemiology of Cholerae Six Pandemics of Cholera occurred between 1817 1923.  All pandemics originated in Indian continent.  The seventh pandemic originated in Celebes Islands in Indonesia on 1961.  Most likely V.  Spread far and wide  .

8th Pandemic ? Spread of 0-139 0     Several identify that onset of 0-139 is considered 0as 8th pandmeic started in India. 3- . flies Only 1 -5% of exposed will get effected Carrier stage seldom exceeds 3.4 weeks. food. Cholerae is spread by contact with persons in early or even mild illness. By contaminated water.

Excreta disinfected All contacts to be followed up . Patients infected preferably isolated .Control of Cholerae     Needs improvement of Sanitation associated with water treatment and food.

 Vaccines not useful in Epidemic controls  .  Repated injection of vaccine containing either Lipopolysaccharides extracted from Vibrios or dense Vibrios suspension can offer limited prevention to heavely exposed persons.Use of Vaccine and Chemoprophylaxis Chemoprophylaxis with antibiotics is effective.

however. everyone.Prevention of Cholera   Although cholera can be life-threatening. because of advanced water and sanitation systems. it is lifeeasily prevented and treated. Several regions in the Devloping countries continue to be endemic locations. should be aware of how the disease is transmitted and what can be done to prevent it. cholera is not a major threat. especially travelers. . In the United States.

com .T.V.Rao MD Email doctortvrao@gmail.Created for Health awarness in Developing countries Dr.

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