Genus

BRUCELLA
 Other Names
Human Disease Animal Disease

 Malta Fever  Enzootic Abortion

 Undulant Fever  Epizootic Abortion
 Mediterranean Fever  Contagious Abortion
 Mediterranean remittent  Infectious abortion
Fever  Abortus fever
 Rock Fever of Gibraltar  Slinking of Calves
 Gibraltar fever  Bang’s Disease (Cattle)
 Gastric Fever  Ram Epididymitis
(Male sheep)
Overview

 Historical Background
 Biotypes
 Cultural characters
 Morphology
 Biochemical reactions
 Suceptibility
 Antigenic structures
 Bacteriophages
Overview contd…

 Epidemiology
 Transmission
 Pathogenesis
 Disease in humans
 Disease in animals
 Laboratory diagnosis
 Treatment
 Prevention & control
  Sir David Bruce (1855-1931)
– British Army physician &
microbiologist
– 1883 sent to Malta to provide
medical care to the troops
– In 1886, isolated Micrococcus
melitensis from spleens of 4
soldiers died of the disease
– Transmitted to monkey
Professor FEG Cox. The Wellcome Trust, Illustrated History of Tropical Diseases
experimentally
 Bernhard Bang
(1848-1932)
– Danish physician &
veterinarian
– In 1897, Discovered
Bacterium abortus
could infect cattle,
horses, sheep,
& goats
 Zammit (1905) proved
transmission to human by
goat’s milk

 Protected British soldiers by

prohibiting use of goat’s milk
& milk products
 In 1914, Mohler & Traum isolated B.
suis from swine

 In 1966, Jacob Tarun isolated B.

canis from aborted pups

 Later Buddle (1956) isolated B. ovis

from aborted sheep & B. neotome
from desert wood rats by Stoenner &
Lackman (1957)
Species Biovar/ Natural Host Human
Serovar Pathogen

B. abortus 1-6, 9 cattle, bison, buffalo yes

B. melitensis 1-3 goats, sheep yes

B. suis 1, 2, 3 swine yes

2 European hares yes
4* reindeer, caribou (deer) yes

5 rodents yes
B. canis none dogs, other canids yes

B. ovis none sheep no

B. neotomae none rodents yes

B. maris marine mammals yes

B. pinnipedialis, Sea lions
B. cetaceae
Whales, dolphins
Serum dextrose agar, serum potato agar, trypticase
soya agar, tryptose agar, Farrell’s agar medium
 Addition of Bacitracin, Polymyxin &
Cycloheximide to above media makes
selective (Farrell’s medium)
 Serum or liver extract improves
growth
 Erythritol has stimulating effect on
growth
 Don’t grow anaerobically
 Optimum temperature for growth 37O C (20 -40OC)

 Optimum pH 6.6 - 7.4

 Environmental persistence
 Withstands drying
 Temperature, pH, humidity
 Frozen & aborted materials, urine (6 D),dust (6Wk),
water (10Wk) & soil
 MORPHOLOGY

 Gram negative, Aerobic, Non-motile,

Non-capsulated, Non spore forming,
Coccobacillus or Short rods arranged
singly or short chains
(0.5-0.7 x 0.6-1.5 µm size)

 Many strains require 5-10% CO2

(capnophilic)
 Facultative intracellular organisms

 Bipolar staining not uncommon

 Biochemical
reactions

 Catalase & oxidase positive,

 Reduces nitrate

 Rapidly positive for urease

 Negative for IMViC reactions

 Susceptibility

 Killed at 600C in 10 minutes & 1%

phenol in 15 minutes

 Sensitive to pasteurization temperature

 Survival is long in refrigerated milk

(10D), ice creams (1M), butter (4M) &
cheese; meat; soil & manure several
weeks
Antigenic structure

 Cross react with V. cholerae, E. coli O116

& E. coli O157, Salmonella serotypes
group N, P. multophila, Y. enterocolitica &
F. tularensis
 Epidemiology
Distribution

 B. abortus most common worldwide

 Order of pathogenicity to man considered as
B. melitensis, B. suis, B. abortus & B. canis
 Affects wide range of hosts including Cattle,
buffalo, swine, sheep, goat, camel, horse &
dog

 Low prevalence in developed countries, 17

countries declared brucellosis free including
Norway, Finland, Sweden & Denmark
 Brucellosis in India
IN Animals

 Prevalence rate varies from country to

country, more in organized dairy farms
than village & reported from all states of
India

 More common in areas with heavy rainfall

& humidity

 Increasing prevalence in Tarai Region

 No isolation of Brucella spp in camel &
poultry except their serological evidence
 In Man

 Mostly contracted from goat & sheep,

more common in shepherds & villagers
 In urban areas, contracted through milk
of sheep & goat, happens in summer
months in scarcity of bovine milk

 More common between 20 to 50 years

age
Who is at Risk?

 Occupational disease
– Cattle ranchers/dairy farmers
– Veterinarians
– Abattoir workers
– Meat inspectors
– Lab workers
 Hunters
 Travelers
 Consumers
– Unpasteurized dairy products
Transmission

In animals

 Excrete in milk, urine, faeces & uterine

discharges
 Recovered may become carrier &
excretes intermittently
 More chances through AI than natural
service
 Environment not a good source
 In cattle, oral route through licking aborted
fetuses/genital discharge, fomites
 Also occurs in-utero or through
colostrum/milk
 Ingestion/ contact with infected tissue or
body fluid

 Through placental tissue & venereal

discharge in swine, sheep, goat & dogs
In Humans

 Conjunctiva or broken skin on

contact with infected tissues
- Blood, urine, vaginal discharge,
aborted fetus & placentas

 Ingestion of Contaminated animal

products
– Raw milk, unpasteurized dairy
products (cheese)
– Rarely through undercooked
meat
 Incubation varies from 5 days to 3
months
Transmission in
Humans Contd..

 Aerosol
– Laboratory, abattoirs
– Pens, stables, slaughter houses

 Inoculation with vaccines

– B. abortus strain 19, RB-51
– B. melitensis Rev-1

 Person-to-person transmission through

congenital or breast milk rare
Pathogenesis

 Primarily an intracellular pathogen

 Through oral regional lymph nodes reaches

to blood causing bacteremia

 Survive & replicate within Phagocytes &

Monocytes

 Infected macrophages localized within

reticuloendothelial system (Granuloma
formation in spleen, liver, bone marrow
including uterus)
 Infiltration of epithelial cells, giant
cells, lymphocytes & plasma cells seen
with granulomas

 In pregnant uterus causes necrotic

inflammation of uterine wall & fetal
membrane

 Causes detachment of placenta from

maternal cotyledons

 Resulting expulsion of fetus

DISEASE IN HUMANS
Disease in Humans

 Asymptomatic brucellosis

 Acute brucellosis

 Chronic brucellosis

 Relapsing brucellosis
Asymptomatic
brucellosis

 May be mild & self limiting

 Veterinarians, farmers,
abattoir workers , meat
processors more involved
Acute brucellosis

 IP 7-21 days
 Lassitude, body ache, head ache, insomnia,
anorexia, rapid wt. loss, weakness,
depression & fatigue
 Polyarthritis, pharyngitis & abdominal pain
 Profuse nocturnal sweating with peculiar
odour & shaking chill
 Biphasic fever (101 – 1040F) usually high in
evening
 Death due to toxaemia, thrombopenia &
endocarditis
 B. abortus milder than B. melitensis
 Chronic brucellosis

 When persist over 6 months

 Loss of body weight, lassitude, headache,
weakness, pain & sweats

 Enlargement of Cervical & Axillary lymph node

 Spleenomegaly, hepatomegaly & polyarthritis

 About 6% male have

orchitis but less
frequent than animals
 Relapsing brucellosis

 Can occur within first three months

after treatment

 Shows high fever, fatigue & chills

 More frequent nocturnal sweating

Dermal Brucella
Allergy
 Due to hypersensitivity to placenta protein
(erythema brucellumin)
 Shows necrotic papules or pustules on hands
of veterinarian
DISEASE IN ANIMALS
In bovines,
sheep & goat

 Abortions occur in Third trimester &

only once
 Although reinvasion of uterus may
take place
 Still birth & sterility may also
occur
 Erythritol in placenta favours
growth which act as predisposing
factor for abortion
Clinical Signs:
Cattle

 I P 2 Wks- 6 Months
 Late abortions with B. abortus
 Abortion occurs in storm or wave
 Retained placenta
–Once expelled will have a
leathery appearance
 Birth of dead or weak calves
 Endometritis,
 Low milk yield, mastitis
 Orchitis, epididymitis & arthritis
 Associated with Hygroma of fore knee of cattle
Clinical Signs:
Sheep and Goats

 B. melitensis
– Late term abortions
» Retained placenta
» Birth of dead or
weak lambs/kids
 Goats
– Articular, periarticular hygroma localizations
 B. ovis
– Mainly ram epididymitis
– Fertility problems in female
Clinical Signs: Swine

 B. suis
–Prolonged bacteremia
–Abortion, early or
late gestation
–Fertility problems
–Lameness, posterior
paralysis, spondylitis,
metritis, abscesses
Clinical Signs: Horses

 B. abortus most common

–Susceptible to B. suis
 Fistulous Withers or Poll Evil
–Inflammation of the
supraspinous bursa
Clinical Signs: Dogs

B. canis
–Abortions
» Last trimester
» Prolonged
vaginal discharge
–Failure to conceive, stillbirths
– Prostatitis & unilateral or bilateral
testicular hypertrophy in males
 Also susceptible to
–B. melitensis, B. abortus & B. suis
Laboratory
Diagnosis

 Cultural examination

 Serological examination

 Molecular detection

 Delayed hypersensitivity skin

tests
Culture Method
Bio safety

 Bio safety level 2 precautions for handling

specimens

 Bio safety level 3 precautions needed for

culture manipulations & Experimental
animal studies
 Specimens
for culture

 Blood culture is most definitive, however,

positive only in 30-50% cases
 As bacteria in blood are scanty, 5 ml blood
inoculums needed with sub culturing on
every fourth day up to 6-8 weeks
 Bone marrow, lymph node aspirates, urine,
synovial fluid, abscess aspirates, liver &
spleen biopsy most successful
Serological
Examination

 Standard Tube Agglutination Test (SAT)

 Complement Fixation Test (CFT)

 ELISA, Dot-ELISA

 Fluorescence Polarization Assay (FPA)

Standard tube
Agglutination Test (SAT)

Detects antibodies in
serum, uterine discharge,
vaginal mucus & milk or
semen plasma

» which level is diagnostic ??

» 1 : 80 IU - positive in cattle
» 1 : 40 IU positive in sheep & goat
• Four fold increase at 2 week
interval
 Rose Bengal Plate
Agglutination Test

 Milk ring test (ABR

test) – Herd test
Antiglobulin (Coombs)
Test

 Mainly detecting antibodies to Br. abortus

in man & experimentally in animals
 Detects the presence of Ig G antibodies
 Applied in diagnosis of chronic human
brucellosis
Treatment

“Treatment of brucellosis in
animals neither advisable nor
practicable”
All monotherapies have
unacceptably high relapse rate,
so combination regimens of
antibiotics that can penetrate
macrophages & act in the
acidic intracellular environment
are recommended
1986 WHO guidelines for the treatment
of brucellosis

 Regimen A:  Regimen B:
Doxycycline 100 Doxycycline 200
mg PO bid for 6 mg PO +
weeks + Rifampicin 600 to
streptomycin 1 g 900 mg (15 mg/kg)
IM daily for first PO daily for 6
14-21 days weeks
 Streptomycin & tetracycline contraindicated
in pregnancy, however rifampicin is safe
Prevention & Control

In Human
 Control of animal brucellosis

 Routine pasteurization of milk &

milk products

 Depends on good personal

hygiene & environmental
sanitation

 No human vaccine
Hygienic measures

 Proper disposal of placenta & aborted

fetus

 Disinfection of farm premises & animal

sheds

 Proper ventilation system in slaughter

houses

 In labs strict biosafety precautions

 Education programmed to high risk

groups
In Cattle

 Test & slaughter / segregation of

positive reactors
- RBPT for screening &
- CFT for confirmation

 Regular testing with MBRT

 Restricted movement & proper disposal

Immunization

 Immunization of female calves between 4 - 8

months age with live attenuated B. abortus S-
19 & B. abortus RB-51 (lack of LPS) (5ml , S/C)
if herd have 5% or more infected animals
Thank You