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.

1. Bordetella

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 contd…
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 –ve 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

8. Biochemical reactions
• Doesn’t ferment sugars
• Indole +ve
• Reduce nitrates to nitrites
• Split urea
• Citrate positive
• Oxidase & catalase +ve

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 contd…
• Incubation period is 1-2 weeks
• Source of infection- infected human
• Transmission – droplets
• Duration disease lasts for 6-8 weeks
• 3 stages
– catarrhal,
– Paroxysmal
– 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 “whoop”
• 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,

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

18. Laboratory Diagnosis
Specimen
• prenasal swab
• postnasal swab
• Cough plate method

19. Pre nasal swab

20. Laboratory Diagnosis contd…
• 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 contd…
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

24. Treatment
• Tetracycline, Chlorophenicol,
Erythromycin (DOC) and ampicillin

25. Early Immunization is best
solution to prevent the Pertusis

26. Prophylaxis
• Immunisation –infants & 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 contd…
• 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

30. Brucella

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:
– Brucella melitensis (71%)
– Brucella abortus (29%)

34. BRUCELLA
• CLASSIFICATION
• Br melitensis - goats and sheep-MALTA
FEVER
• Br abortus – cattle – UNDULANT FEVER
• Br suis – pigs
• Br canis – dogs
• Br ovis – sheep
• Br neotomae – desert rodents

35. BRUCELLA
• CULTURE
• Strictly aerobic (only Br melitensis req 5-
10% CO2- CAPNOPHILIC)
• 37ᵒC pH 6.6-7.4
• Ordinary media – 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:
– CO2 REQUIREMENT(Br abortus needs CO2)
– H2S production (Br abortus and American
strains of suis produce H2S)

39. BRUCELLA
• RESISTANCE
• 60 ᵒ C 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
– INGESTION
– INHALATION
– INOCULATION
– DIRECT CONTACT

42. Transmission of Brucella

43. Transmission of Brucella to Human

44. Pathogenesis
• It is a facultative intracellular parasite
• Organism is opsonized by human serum.

45. BRUCELLA
• PATHOGENESIS
Alimentary or Respiratory tract

Lymphatics

Lymph nodes

Blood (bacterimia IP 2-3 weeks to months)

Colonise in organs especially in lymphoreticular system

Proliferation of macrophages, endothelial cells, lipocytes

Non-caseating granulomata (lymph nodes, liver, spleen and bone
marrow)

46. BRUCELLA
• TYPES OF INFECTION
• SUBCLINICAL / LATENT – Diagnosis only by
serological tests
• ACUTE INFECTION – Fever with chills
(undulant), headache, bone and joint pains,
lymphadenopathy, hepatosplenomagaly
• Chronic – low grade fever with periodic
exacerbations

47. BRUCELLA
• Skeletal system: Bony pain, synovitis,
synovial effusion
• Neurologic: Meningitis, encephalitis,
peripheral neuropathy, intracerebral
abscess, psychosis
• CVS: Endocarditis, myocarditis,
pericarditis

48. BRUCELLA
• Pulmonary: Bronchopneumonia, lung
abscess, military lesions, pleural effusion
• Hematology: anemia, leucopenia,
thrombocytopenia
• Systemic: Can cause features of acute
cholecystitis and pancreatitis

49. Clinical manifestation of Brucella

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

51. BRUCELLA
• CFT
• ELISA
• MOLECULAR TECHNIQUES
• SKIN TEST – BRUCELLIN

52. BRUCELLA
• DETECTION OF ANIMAL INFECTION
• ANTIBODIES IN MILK OF ANIMALS-
MILK-RING TEST

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

55. • Thank you