The document outlines the history and characteristics of Bordetella, including its pathogenic species such as Bordetella pertussis, which causes whooping cough, and the diagnostics and treatments for infections. It details the laboratory identification methods and the epidemiology of brucellosis, a zoonotic disease caused by Brucella species, including transmission routes and clinical manifestations. Treatment protocols and preventive measures against both Bordetella and Brucella infections are discussed in depth.

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