In 1900, Jules Bordet along with Octave Gengou observed a small ovoid bacterium in the sputum of a 5 month old child suffering from pertussis, or whooping cough.
The bacterium was similar to Haemophilus influenza but showed distinct morphological characterstic which led Bordet and Gengou to consider it as a separate species.
The organism was unable to be isolated and cultivated on ordinary blood agar plates.
Six years later, Bordet and Gengou suceed in making a selective media called Bordet and Gengou (BG) medium, which helped in isolating this fastidous bacteria.
2. History
⢠In 1900, Jules Bordet along with Octave
Gengou observed a small ovoid bacterium in
the sputum of a 5 month old child suffering from
pertussis, or whooping cough.
Jules Bordet
Belgian Immunologist and
Microbiologist
Octave Gengou
Belgian Bacteriologist
3. History
⢠The bacterium was similar to Haemophilus
influenza but showed distinct morphological
characterstic which led Bordet and Gengou to
consider it as a separate species.
⢠The organism was unable to be isolated and
cultivated on ordinary blood agar plates.
⢠Six years later, Bordet and Gengou suceed in
making a selective media called Bordet and
Gengou (BG) medium, which helped in
isolating this fastidous bacteria.
4. Introduction
Bordetella is a genus of small gram
negative coccobacilli of the
phylum Proteobacteria.
⢠Bordetella species, with the exception of B.
petrii, are obligate aerobes, as well as highly
fastidious, or difficult to culture.
⢠Three species are human Pathogens (B.
Pertussis, B. Parapertussis,B
bronchiseptica)
⢠one of these (B. bronchiseptica) is also motile
5. Introduction contdâŚ
Species
⢠B. Pertussis causes whooping cough
⢠B. Parapertussis mild whooping cough
⢠B. bronchiseptica mild whooping cough
⢠B. avium causes respiratory disease in
turkeys
6. Bordetella pertussis
Morphology
⢠Small, ovoid,1-1.5x0.3um gm âve coccobacilli,
non motile and non sporing. It is capsulated but
loses capsule on repeated subculture
⢠Bipolar metachromatic granules may be
observed on staining with toludine blue
7. Culture
⢠It is aerobic and cannot grow anaerobically
⢠Optimum temp for growth is 35-36oC
⢠No growth on nutrient agar
⢠Requires complex media for primary
isolation
⢠Bordet-Gengou (glycerol, potato & blood
agar) is commonly used media. After 48-
72hrs small, smooth, grayish white,
refractile colonies are seen . Colonies
resemble bisected pearls or mercury drops.
A hazy zone of hemolysis surrounds the
colonies
⢠Charcoal blood agar is also used for
primary isolation of organism (commonly
used Regan-Lowe (RL) medium available
as semisolid/solid medium, also used as
transport media
9. Resistence
⢠Killed by heating at 55oc at 30 mins
⢠By drying and disinfectants
⢠Survive outside the body in droplets for
few hours
⢠Viable at low temperature
10. Pathogenesis
⢠B. Pertussis is a acute, highly contagious
pediatric disease. and is called
whooping cough
⢠Adults and adolescents are also effected
⢠95% due to B. pertussis
⢠5% by B. parapertussis
⢠Only 0.1% by B. bronchoseptica
11. Pathogenesis contdâŚ
⢠Incubation period is 1-2 weeks
⢠Source of infection- infected human
⢠Transmission â droplets
⢠Duration disease lasts for 6-8 weeks
⢠3 stages
â catarrhal,
â Paroxysmal
â convalescent each lasts for 2 weeks
12. Catarrhal stage:
⢠Stage of infectivity.
like common cold.
⢠Low grade fever, running nose, nasal
congestion, Sneezing
⢠Dry irritating cough
⢠Clinical diagnosis difficult
⢠Respond to antibiotics
13. Paroxysmal stage:
⢠Bouts of coughing: At the
end of bout a long inspratory
effort is usually accompained
by a characterstic high
pitched sound âwhoopâ
⢠Duration of the stage is 1-6
weeks
14. Paroxysmal stage:
⢠During an attack,
individual may become
Cynotic due to lack of
oxygen.
⢠Children and young
infants appears
especially ill and
distressed.
Cynotic patches on the face
15. Convalescent stage(Recover stage):
⢠3-4 weeks after an acute illness
paraxysms are decreased.
Complications:
⢠Subconjunctival Hemorrhage
⢠Bronchopneumonia
⢠Lung Collapse
⢠Convulsions
⢠Coma
⢠abdominal and inguinal hernias
⢠pneumothorax,
16.
17. Epidemiology
Pediatric disease
⢠Incidence & mortality highest in first year
⢠Whooping cough is one of most infectious
of bacterial disease
⢠Immunity occurs after one attack
⢠But reinfection in adults severe
20. Laboratory Diagnosis contdâŚ
⢠Prenasal swab passed in floor of
the nasal cavity and a material
collected from pharyngeal wall
⢠Postnasal swab passed through
mouth to collect posterior
pharyngeal secretions
⢠Cough plate method Bordet-
Gengou culture plates is held 10-
15cm in front of pt mouth during
about of coughing and cough
droplets are inoculated directly
on culture plate
⢠No cotton swab used only dacron
or calcium alginate swab used for
collection
21. Laboratory Diagnosis contdâŚ
Microscopy in this bacilli are demonstrated in respiratory
secretions by fluorescent antibody technique
⢠Culture after swab collection on Bordet and Gengou
media or charcoal agar (dimidine flouride & pencillin in to
media) to make it more selective
⢠Plates incubated in high humidity at 35-36oc for 3-5
days
⢠Pearl like colonies appear in 2-3 days
⢠Microscopy & slide agglutination confirms it
⢠Immunofluroscence is useful in identifying the bacillus in
smears from culture
22. Different characterstics of Bordetella
species
Character B. pertussis B. parapertussis B. bronchiseptica
Motility - - +
Growth on
nutrient agar
- + +
Pigment
production
- + -
Oxidase + - +
Urease
production
- + +
Citrate
production
- + +
Nitrate
production
- - +
Toxins
HLT& TCT
ACT
PT
+
+
+
+
+
-
+
+
-
23. SEROLOGY
⢠Agglutination test rising titer demonstrated
in paired sera
⢠Complement fixation test
⢠Immunofluoroscent test
⢠Antibody in sera demonstrated in third week
⢠Detection of IgA antibody in
nasopharyngeal secretion by ELISA
26. Prophylaxis
⢠Immunisation âinfants & children immunised with killed B.
pertussis vaccine
⢠3 I/m inj at an interval of 4-6 weeks are given before 6 months of
age
⢠Booster at the end of first yr of life
⢠Pertussis vaccine along with diptheria & Tetanus toxoid (DPT)
⢠B. pertussis act as adjuvant for toxoids
⢠2 types of vaccine available
DwPT (whole cell pertussis vaccine )
DaPT (acellular pertussis vaccine)
Booster also given at 5yrs and every 10 yrs
DPT vaccination
27. Prophylaxis contdâŚ
⢠vaccine causes fever; injection-site pain,
erythema, and swelling; irritability
⢠Complications Encepholopathy and
convulsions
Booster dose is given to contacts of the
case along with Erythromycin for 5 days
DPT vaccination
28. Bordetella Parapertussis
⢠5% of whooping cough cases produces
mild disease
⢠Resemble B. pertussis
⢠Pertussis vaccine does not protect B.
parapertussis infection
29. Bordetella Bronchiseptica
⢠Cause 0.1% of the cases can grow on
nutrient agar and antigenically related to
B. pertussis and Brucella abortus
31. BRUCELLA
INTRODUCTION
⢠Causative organism of MEDITERRANEAN FEVER,
MALTA FEVER AND UNDULANT FEVER.
⢠1859: Marstan: Crimean war, Gastric remittent fever
⢠1862: Nocard
⢠1886: Bruce,
⢠1897: Bang,
⢠1905: Zammit
⢠1914: Traum
32. BRUCELLA
⢠Non-motile, non-sporing aerobic Gram
negative coccobacilli, may be arranged in
chains, non-capsulate
⢠Obligate intracellular parasite
⢠Infects domestic animals (goat, sheep,
cattle, pigs, camels, reindeer) from whom
man acquires disease (ZOONOTIC
DISEASE)
33. BRUCELLA
⢠Mediterranean basin, Arabian contries,
Indian subcontinent, Parts of Mexico,
Central and South America)
⢠In India:
â Brucella melitensis (71%)
â Brucella abortus (29%)
35. BRUCELLA
⢠CULTURE
⢠Strictly aerobic (only Br melitensis req 5-
10% CO2- CAPNOPHILIC)
⢠37áľC pH 6.6-7.4
⢠Ordinary media â OK but slow growth
36. BRUCELLA
⢠Enriched media with glucose / serum / liver
infusion + antibiotics (polymyxin, bacitracin,
cycloheximide) = selective medium
⢠Small translucent colonies, spheroidal shape,2-
7mm reach max size in 5-7 days.
Smooth
Rough
⢠CAM: intracellular growth
37. BRUCELLA
⢠BIOCHEM REACTIONS
⢠Ferment sugars but negligible A + G
(Ornithine, glutamic acid, lysine, ribose)
⢠Catalase +
⢠Oxidase +
⢠H2S=some +
38. BRUCELLA
⢠DIFFERENTIAL CHARACTERS OF
BRUCELLA SPECIES BASED ON:
â CO2 REQUIREMENT(Br abortus needs CO2)
â H2S production (Br abortus and American
strains of suis produce H2S)
39. BRUCELLA
⢠RESISTANCE
⢠60 ᾠC X 10 min
⢠4 months in butter
⢠1 month in ice-cream
⢠10 days in fridge milk
⢠Weeks in soil and manure
⢠Sensitive to Streptomycin, Tetracycline,
Chloromycetin, Ampicillin
⢠Resistant to Penicillin
40. Transmission
⢠Human brucellosis is usually
not transmitted from human to human;
⢠people become infected by contact with fluids
from infected animals (sheep, cattle or pigs)
⢠or derived food products like unpasteurized milk
and cheese.
⢠Brucellosis is also considered an occupational
disease because of a higher incidence in people
working with animals (slaughter house cases).
⢠People may also be infected by inhalation of
contaminated dust or aerosols.
⢠Globally, there are an estimated 500,000 cases
of brucellosis each year
41. BRUCELLA
⢠MODES OF INFECTION
â INGESTION
â INHALATION
â INOCULATION
â DIRECT CONTACT
44. Pathogenesis
⢠It is a facultative intracellular parasite
⢠Organism is opsonized by human serum.
45. BRUCELLA
⢠PATHOGENESIS
Alimentary or Respiratory tract
ďŻ
Lymphatics
ďŻ
Lymph nodes
ďŻ
Blood (bacterimia IP 2-3 weeks to months)
ďŻ
Colonise in organs especially in lymphoreticular system
ďŻ
Proliferation of macrophages, endothelial cells, lipocytes
ďŻ
Non-caseating granulomata (lymph nodes, liver, spleen and bone
marrow)
46. BRUCELLA
⢠TYPES OF INFECTION
⢠SUBCLINICAL / LATENT â Diagnosis only by
serological tests
⢠ACUTE INFECTION â Fever with chills
(undulant), headache, bone and joint pains,
lymphadenopathy, hepatosplenomagaly
⢠Chronic â low grade fever with periodic
exacerbations
48. BRUCELLA
⢠Pulmonary: Bronchopneumonia, lung
abscess, military lesions, pleural effusion
⢠Hematology: anemia, leucopenia,
thrombocytopenia
⢠Systemic: Can cause features of acute
cholecystitis and pancreatitis
50. BRUCELLA
⢠LABORATORY DIAGNOSIS
⢠DETAILED HISTORY IS IMPORTANT
⢠BLOOD CULTURE- 10 cc blood in Liver infusion
broth and 3% NA slope (CASTANEDA medium)
⢠8 WEEKS incubation before reporting negative
⢠ONLY 50% are blood culture positive; hence
SEROLOGY is important
53. Treatment
⢠There are no clinical trials for optimal
treatment, but a 3-6 week course
of rifampicin and doxycycline twice daily is the
combination most often used, and appears to be
efficacious;
⢠the advantage of this regimen is that it is oral
medication and there are no injections; however,
a high rate of side effects (nausea, vomiting,
loss of appetite) has also been reported.
54. Prophylaxis
⢠Person handling animal use protective
clothing and gloves
⢠Pasteurisation of milk or boiling of milk
⢠Vaccination of cattle's
⢠Unimmunised animals should be slaughtered
⢠Human vaccine trial in Russia given
intradermally