Genus BRUCELLA

Dr.P.Sreenivasulu Reddy MD.,

Professor of Microbiology
NARAYANA MEDICAL COLLEGE
Nellore-1
Andhra Pradesh INDIA
 Many Names of Brucellosis

Human Disease Animal Disease

 Malta Fever  Bang’s Disease

 Undulant Fever  Enzootic Abortion

 Mediterranean Fever  Epizootic Abortion

 Rock Fever of Gibraltar  Slinking of Calves

 Gastric Fever  Ram Epididymitis

 Contagious Abortion
 Sir William Burnett
(1779-1861)

•Physician General to
the British Navy
•Differentiated the
various fevers
affecting soldiers
 Jeffery Allen
Marston

•Contracted Malta
fever.
•Described his own
case in great detail.
 Sir David Bruce
(1855-1931)

•British
Army physician
and microbiologist
•Discovered Micrococcus
melitensis (July 9, 1887)
 Bernhard Bang
(1848-1932)

•Danish physician and

veterinarian
•Discovered Bacterium
abortus (1897) could
infect cattle, horses,
sheep and goats.
 Alice Evans, American bacteriologist
 Credited with linking the organisms
 Similar morphology and pathology between:
 Bang’s Bacterium abortus
 Bruce’s Micrococcus melitensis
 Nomenclature today credited to
Sir David Bruce
 Brucella abortus and Brucella melitensis.
 Morphological Profile

Brucella spp. are small,

Gram-negative, non-motile,
non-spore-forming,
coccobacilli .
Intracellular microbes
causing chronic disease.
 Cultural characters

Aerobe.
Best medium is Trypticase
Soya Agar with selective
agents
(Polymyxin,Cyclohexamide).

Small,moist,translucent and
glistening colonies after 3 or
more days of incubation.
Species Biovar/ Natural Host Human
Serovar Pathogen
B. abortus 1-6, 9 cattle yes
B.melitensis 1-3 goats, sheep yes

B. suis 1, 3 swine yes

2 hares yes
4 reindeer, caribou yes

5 rodents yes
B. canis none Dogs yes

B. ovis none Sheep no

B. neotomae none Desert wood rat no

B. maris (?) marine mammals ?

 Biochemical reactions

Catalase and Oxidase positive.

Nitrates are reduced.
Rapidly positive for urease.
Negative for IMViC reactions.

Susceptibility
 Killed at 600 C in 10 minutes
 Pasteurization of milk .
 Survival is long in refrigerated milk,
ice creams and cheese.
 Transmission to Humans

 Conjunctiva or broken skin on contact with

infected tissues
 Blood, urine, vaginal discharges, aborted
fetuses, placentas
 Ingestion
 Raw milk & unpasteurized dairy products
(Cheese)
 Rarely through undercooked meat
 Transmission to Humans
 Inhalation of infectious aerosols
 Pens, stables, slaughter houses
 Inoculation with vaccines
 B. abortus strain 19, RB-51
 B. melitensis Rev-1
 Conjunctival splashes, injection
 Person-to-person transmission is rare
 Incubation varies
 5 days to three months
 Transmission in Animals

 Ingestion / contact with infected

tissues or body fluids
 Mucous membranes, injections
 Venereal
 Swine, sheep, goats, dogs
 Fomites
 Who is at Risk?
 Occupational Disease
 Cattle ranchers/dairy farmers
 Veterinarians
 Abattoir workers
 Meat inspectors
 Lab workers
 Hunters
 Travelers
 Consumers of unpasteurized dairy products
 Pathogenecity and Immunity
Followed by Entry
Ingested by PMN and not killed (super oxide dismutase ,LPS and
nucleotide substance).
Spread to local LN, multiply and spread to blood.
Reaches to RES (Spleen,Liver,BM,LN).
Resist phagocytosis (production of AMP/GMP and TNF Inhibits the
phagolysosome fusion).
Spread (CNS,Heart,Joints,Respiratory,Skin,GU system)
Host response leads to granuloma formation and necrosis.
 B. abortus
(Contagious/ Infectious abortion)

 Worldwide
 Some countries have
eradicated it
 Notifiable disease
in many countries
 abortions, arthritic joints.
 Fever of Unknown Origin (FUO)

Main causes for spontaneous abortion in animals are erythritol, which can promote infections in the fetus and
placenta and lack of anti-Brucella antibody in amniotic fluid.
 B. canis

 Poorly understood
 1-19% prevalence in United
States
 contact with aborted fetuses
and semen.
 Rarely causes disease in
humans.
 B. suis
 Biovars 1 and 3
 Worldwide problem.
 Free
 United Kingdom, Canada
 Eradicated
 Holland, Denmark
 Low Incidence
 Middle East, North Africa
 Human Disease

 Can affect any organ or system

 All patients have a cyclical fever
 Headache, weakness,
arthralgia, depression,
weight loss, fatigue,
liver dysfunction
 Human Disease

 20-60% of cases
 Osteoarticular complications
 Arthritis, spondylitis, osteomyelitis

 Hepatomegaly may occur

 Gastrointestinal complications
 2-20% of cases
 Genitourinary involvement
 Orchitis and epididymitis most common
 Human Disease

 Neurological
 Depression, mental fatigue
 Cardiovascular
 Endocarditis resulting in death
 Chronic brucellosis is hard to define
 Length, type and response to treatment variable
 Localized infection
 Blood donations of infected persons should not
be accepted
 Human Disease
 Congenitally infected infants
 Low birth weight
 Failure to thrive
 Jaundice
 Hepatomegaly
 Splenomegaly
 Respiratory difficulty
 General signs of sepsis (fever, vomiting)
 Asymptomatic
Chronic brucellosis:
It is hard to define
- Length, type and response to

treatment variable

The localized form:

-Bones and joints.
-Lumbar spondylodiscitis,
sacroiliitis is typical.
-Orchitis also frequent.
 Sequelae are highly variable

- Granulomatous hepatitis,
- Arthritis, spondylitis,
- Anaemia, leukopenia,thrombocytopenia,
- Meningitis, uveitis, optic neuritis,
- Endocarditis.

Granuloma of Liver
 Diagnosis in Humans
 Isolation of organism
 Blood, bone marrow, other tissues
 Serum agglutination test
 Four-fold or greater rise in titer
 Samples 2 weeks apart

 Immunofluorescence Method
 Organisms in clinical specimens
 PCR
Blood cultures in trypticase
soya broth (Castaneda’s
method): Biphasic medium.
Sample inoculated bottle has to
be tilted to flow the broth over
solid medium and keep it upright
position at 370C.
The growth of brucellae is
extremely slow.
Bone marrow cultures are more
sensitive than blood.
Demonstration of IgM , IgG and Ig A
ELISA is the best method.

Standard tube agglutination test:

Anti -O-polysaccharide of LPS. Killed strains
of B. abortus as antigen. Not useful for
B.canis.

Modified Tube agglutination test: 2-

mercaptoethanol is added to serum before
testing which causes disruption of IgM and
only IgG are detected.

.
Problems with TA test: Prozone phenomenon, Presence of blocking or non-agglutinating antibodies. False positives with cholera, tularemia,yersinia or vaccination

Contd…..
Castaneda strip test: Strip with colored brucella antigen. On
addition of patients serum, if antibodies are present, prevents
the flow of serum.
Brucellin Skin test: Delayed hypersensitivity.
Molecular techniques: PCR
Radiology: Alterations in infected vertebrae; Pedro Pons sign
( erosion of antero-superior corner of lumbar vertebrae) and marked osteophytosis are suspecious of brucellic
spondylitis.
 Diagnosis in Animals
Culture of urine and Milk.
Rapid latex agglutination test,
Rose Bengal card test. (Rose Bengal + B.abortus +
Sodium azide)

Contd….
Milk Ring Test: Frequently used test.
A drop of colored brucella antigen
(B.abortus/B.melitensis with hematoxylin) is added to
milk in a test tube, incubated in a water bath at
700C for 40-50 minutes.
Positive: Blue ring at the top leaving the milk
unstained.
Negative: No ring. Milk remains uniformly blue.
 Treatment of Choice

 Combination therapy has the best efficacy

 Doxycycline for 6 weeks in combination with
Streptomycin for 2-3 weeks or Rifampin for 6
weeks

 CNS cases to treat for 6-9 months.

 Same for endocarditis cases plus surgical
replacement of valves.
 Prognosis
 May last days, months, or years
 Recovery is common
 Disability is often pronounced
 About 5% of treated cases relapse
 Failure to complete the treatment regimen
 Sequestered infection requiring surgical drainage

 Case-fatality rate: <2% ( untreated)

 Endocarditis caused by B. melitensis
 Prevention and Control
 Education about risk of transmission
 Farmer, veterinarian, abattoir worker,
butcher, consumer, hunter, public
 Wear proper attire if dealing with
infected animals/ tissues
 Gloves, masks, goggles
 Avoid consumption of raw dairy
products
 Prevention and Control

 Immunize in areas of high prevalence

 Young goats and sheep with Rev-1
 Calves with RB51
 No human vaccine

 Eradicate reservoir
 Identify, segregate, and / or cull
infected animals
 References:

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