BORDETELLA PERTUSSIS

WHOOPING COUGH
Dr.T.V.Rao MD

DR.T.V.RAO MD

A TRIBUTE TO BORDET - GENGOU

DR.T.V.RAO MD

WHAT IS WHOOPING COUGH

Whooping Cough (Pertussis) is a bacterial infection of the lungs which is caused by a bacterium Bodetella pertussis. It is a very contagious disease which causes coughing with little or no fever. The coughing may be so severe that it leads to vomiting and aspiration.
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HOW THE NAME WHOOPING DERIVED Whooping cough is an infectious bacterial disease that causes uncontrollable coughing. The name comes from the noise you make when you take a breath after you cough. You may have choking spells or may cough so hard that you vomit.
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IDENTIFICATION BODETELLA
Jules Bordet and Gengou contributed for discovery 1900 Identified as small bacilli in children with Whooping cough. Bodetella pertussis ( Intense cough ) Other related Bacteria B.parapertussis B.brochoseptica B.avium
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BORDETELLA PERTUSSIS ( B G BACILLUS )

Gram negative organism Small, ovoid,cocobacillus. Length is 0.5 microns Have bipolar metachromatic granules when stained with Toluidine blue
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BODETELLA PERTUSSIS ( B G BACILLUS)

Small ovoid coccobacillus 0.5 microns On repeated cultures becomes become larger thread like bacilli. Non motile, Non sporing Capsulated loose on repeated culturing

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OTHER CHARACTERS
Do not swell in the presence of antigen. Loose clumps of bacilli appear as thumb print appearance with clear space between the organisms. Freshly isolated strains have fimbria.

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CULTURE CHARACTERS
Aerobic Not anaerobic Grows optimally at 35 0 to 370 c Preferred medium Bordet Gengou glycerin potato blood agar

Blood for neutralizing inhibitory substances formed during bacterial growth.

Charcoal also serves the same purpose.
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MERCURY DROP COLONIES ON BORDET-GENGOU MEDIUM

Growth takes longer up to 48 72 hours On blood agar appear as small dome shaped opaque viscid grayish white retractile Resembles bisected pearly or mercury drops
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ALUMINUM PAINT APPEARANCE

Colonies surrounded by hazy zone of hemolysis Confluent growth presents as aluminum paint.
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BIOCHEMICAL REACTIONS
In active do not ferment sugars Indole test + Reduce Nitrates Utilize citrates Splits urea

Catalase +
Oxidase +
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RESISTANCE
Killed by heat at 550c for 30 mt Drying and disinfectants kill the organism Survive outside for 5 days 3 days on cloths Few hours on paper

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ANTIGENIC CHARACTERS AND VIRULENCE

Agglutinogens - Species specific surface agglutinogens with capsule K antigens or fimbria 14 agglutinin factors are identified Factors 7 is common in all species Factor 1- 6 in only B pertussis Factor 12 in B.brochoseptica

Factor 14 in B Para pertussis

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VIRULENCE FACTORS
These virulence factors include adhesions such as filamentous hem agglutinin, agglutinogens, peractin, and fimbriae as well as a number of toxins including pertussis toxin, acetylate cyclase toxin, trachael cytotoxins, Dermonecrtoic toxin and heat-labile toxin (CDC, 2005).
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PATHOGENESIS OF B.PERTUSSIS
Like most Gram negative pathogens, B. pertussis also contains a Lipopolysaccharide coat that acts as an Endotoxin and can aid colonization by agglutinating human cells (Steele, 2004).
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VIRULENT MOLECULES

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TOXIN CELLULAR ACTION.

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MECHANISM OF INFECTION
1,2,3 are common infective strains vaccines contain all the three Agglutinogens promoting virulence by helping bacteria to attach to respiratory epithelial cells
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PERTUSSIS TOXIN
Pertussis toxin MW 1,17,000 Hexamer protein composed of 6 subunits with A B structure A has enzymatic activity it can be toxoided Pertussis toxin is the major component of Acellular Pertussis vaccine.

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NATURE OF TOXIN
It produces a highly lethal toxin (formerly called Dermonecrtoic toxin) which causes inflammation and local necrosis adjacent to sites where B. pertussis is located. The lethal toxin is a 102 kDa protein composed of four subunits, two with a mw of 24kDa and two with mw of 30 kDa.
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PERTUSSIS TOXIN
Causes pathogenesis

Present only in B.pertussis

Pertussis toxin is expressed on the surface, secreted into the surrounding medium

Posses Biochemical and Biological activity of producing lymphocytosis producing factor causes Lymphocytosis
Acts as Histamine sensitizing factor Islet activating function causes excessive Insulin secretion.
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FILAMENTOUS HEMAGGLUTININ
One of the Hemagglutinins produced by B.pertussis Filamentous Haemagglutinnins adheres to cilia of the respiratory epithelium and to erythrocytes

Helps in binding to respiratory epithelium

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OTHER TOXINS
Adenylate cyclase
Enters the target cells and acts as toxin

It acts by catalyzing the production of cAmp by various types of cells. Heat labile Toxin
Cytoplasmic protein present in Bordetella

Dermonecrtoic and lethal in Mice

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TRACHEAL TOXIN
L M W peptidoglycan Causes ciliary damage, produced by all Bodetella It induces ciliary damage in hamster tracheal ring Lipolysacchardie acts as in Gram ve bacilli

Pertactin OMP produces immunity in mice.

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VARIATION SMOOTH TO ROUGH

B pertussis may alter from smooth to rough variation

Phase I to Phase II Phase III Phase IV( rough stage ) which is rough and avirulent form
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PATHOGENICITY
An obligate parasite Intranasal inoculation in mice induces a characteristic patches and intensive pneumonia like In humans Incubation is 1 to 2 weeks
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INCUBATION IN WHOOPING COUGH

The incubation period (the time between infection and the onset of symptoms) for whooping cough is usually 7 to 10 days, but can be as long as 21 days.
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STAGES OF INFECTION
1 Catarrhal 2 Paroxysmal 3 Convalescent Each stage lasts 2 weeks Catarrhal stage is Maximal infective Antibiotics are useful.
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PAROXYSMAL STAGE
Cough increases distinctive bouts

Violent spasms of continuous coughing

With violent act of cough, air enters into empty lung with characteristic whoop

Enters into next stage

Leads to convalescence And severity of cough decreases Total disease lasts for 6- 8 weeks.
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VIOLENT PAROXYSMS OF COUGH

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COMPLICATIONS
The violent bouts of cough leads to Subconjuctival hemorrhage Subcutaneous emphysema Bronchopneumonia Lung collapse Neurological complications Epilepsy, paralysis, mental retardation, blindness, deafness.
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EPIDEMIOLOGY
Predominately a pediatric disease Highest in the 1st year of life Maternal antibodies are not protective. Females suffers more than males. World wide in distribution Epidemics occurs periodically. In early stage of infection droplets and fomites contaminated by oropharengeal secretion are infective. Non immune rarely escape infection
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EPIDEMIOLOGY
House hold contacts at risk Chronic carriers are not known B.pertussis - 95 %

B.parapertussis 5%
B.brochoseptica occasionally occur Some times Adenovirus, Mycoplasma pneumonia may mimics whooping cough.

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HOW WHOOPING COUGH DIAGNOSED

Since the early symptoms are so non-specific, pertussis is usually not diagnosed until the appearance of the characteristic cough. Pertussis can be confirmed by taking cultures of respiratory fluids for examination in the laboratory. This involves taking a sample of secretions from the nose or throat and identifying the pertussis bacteria in the secretion

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DIAGNOSIS
Isolation by culture PCR Direct fluorescent antibody Serological testing

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http://medinfo.ufl.edu/year2/mmid/bms5300/images/d7053.jpg

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LABORATORY DIAGNOSIS
Microscopy Culture. Microscopy Demonstration of Bacilli in respiratory secretions. Florescent Antibody methods

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COUGH PLATE METHOD

Culture plate held at 1015 cm infront of the mouth when the patient is coughing spontaneously or induced cough Droplets of respiratory exhaled impinge on the media. Helpful as bed side investigation
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COUGH PLATE METHOD

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NASOPHARYNGEAL SWAB
Secretion from the posterior pharyngeal wall are collected with cotton swab on a bent wire passed from the oral cavity A Wests post nasal swab is used for collection of specimen.
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PER NASAL SWAB

Swab on a flexible wire is passed along the floor of the nasal cavity and material collected from Pharyngeal wall Dacron or Calcium alginate swabs are better
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TRANSPORT MEDIUM
Transferred into Casamio acid solution at pH 7.2 in modified Stuarts medium Glycerin potato blood agar of Bordet Gengou Adding Pencillin becomes more selective
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IDENTIFICATION OF BACTERIA
The culture plates are incubated at 360c The bacteria are identified by Microscopy and slide agglutination

Immunofluorescence methods
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SEROLOGY
Paired serum sample for detection of antibodies Gel precipitation testing Complement fixation test Detection of Ig A by ELISA from nasopharyngeal secretions.
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EARLY IMMUNIZATION IS BEST SOLUTION TO PREVENT THE PERTUSSIS

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HOW WHOOPING PREVENTED

Pertussis can be prevented by the pertussis vaccine, which is part of the DTaP (diphtheria, tetanus, a cellular pertussis) vaccine. These important immunizations are routinely given in five doses before a child's sixth birthday.
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PROPHYLAXIS
Alum absorbed vaccine is better Administered in combination with Diphtheria, and tetanus toxoid B pertussis acts as an adjuvant Early immunization, is essential in prevention of infection. Later doses are given at the interval of 4 6 weeks intervals, before 6 moths 3 doses are completed.

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BOOSTER DOSES
A booster at the end of the 1 st year Another dose at 4th year Chemoprophylaxis with Erythromycin when exposed to contacts in the vicinity Complications with vaccination Post vaccinial encephalopathy 5 10 million doses Neurotic complications Stop further vaccination Do not vaccinate after 7 years
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ADVANTAGES OF ACELLULAR VACCINE

An acellular vaccine containing whole antigen has been developed and found to elicit good antibody response with fewer side effects. It has replaced the classical vaccine in Japan since 1981 with success, with fewer out breaks and less side effects. whooping cough vaccine can be made from various components of the Bodetella pertussis bacterium, rather than the whole organism. This "acellular" vaccine works well but has fewer side effects than the traditional "whole cell" version.
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ACELLULAR VACCINES
Contain the Pertussis bacilli Contain PT FHA Agglutinogens 1, 2, 3 Produces immunity in 90 % of individuals Immunity in only 50 % by 12th year

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TREATMENT
Penicillin is not useful 10 days of Erythromycin is useful in early infection

Chloramphenicol and Cotromoxazole are effective.

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 Programme Created by Dr.T.V.Rao MD for Medical and Paramedical Students

Email doctortvrao@gmail.com

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