Chapter 33: Brucella species Flashcards
Size of Brucella
Small (0.6 x 0.6 to 1.5 um)
Motility
Non-motile
Shape
Coccobacilliary
Type of bacteria
Gram-negative
Environement
Aerobic and capnophilic (high CO2)
Test for:
-Catalase
-Oxidase
-Positive
-Positive (some are negative)
Oxidase positive except
-B. ovis
-B. neotomae
Urease positive except
B. ovis
Requirement (fastidious)
-5 to 10% CO2 for primary isolation
- enriched media with blood or serum for culturing
Require 5 to 10% CO2 for primary isolation
-Brucella bovis
- some biotypes of B. abortus
-growth of other Brucella spp is enhanced in this atmosphere
Requires enriched media with blood or serum for culturing:
-B. abortus biotype 2
- B. ovis
Referred to as MZN-psotive because
They are not decolorized by 0.5% acetic acid in the modified Ziehl-Neelsen (MZN) staining technique
MZN-stained smears of body fluids or tissues, characteristically appear as
Clusters of red coccobacilli
Brucella genome is unusual because
It is composed of two circular chromosomes
(except: B. suis biovar 3- single chromosome)
Predilection for
Both female and male reproductive organs in sexually mature animals
Remain viable in
moist environment for many months
Transmission
Direct contact (infected animals/fluids and tissues associated with abortion)
Stain ____ using MZN method
Red
Some species cause ____ in humans
Undulant fever
Site of replication and dependence on host cells
They are intracellular pathogens
Usual Host: Cattle
Clinical significance: Abortion, orchitis
B. abortus
Usual Host: Goats, sheep
Clinical significance:Abortion, orchitis, arthtritis
B. melitensis
Usual Host: Pigs
Clinical significance:
-abortion
-orchitis
-spondylitis
-infertility
B. suis
Usual Host: Sheep
Clinical significance:
-Epididymitis in rams
-sporadic abortion in ewes
B. ovis
Usual Host: Dogs
Clinical significance:
-Abortion
-Epididymitis
-Discospondylitis
-Sterility in male dogs
B. canis
Usual Host: Dessert wood rat
Clinical significance:
-not isolated from domestic animals
B. neotomae
Usual Host: Cetaceans
Clinical significance:
B. ceti
Usual Host: Pinnipeds
Clinical significance:
B. pinnipedialis
Species occasionally infected:
-Sheep
-Goats
-Pigs
Clinical significance:
-Sporadic abortion
B. abortus
Species occasionally infected:
-Horses
Clinical significance:
-Bursitis
B. abortus
Species occasionally infected:
-cattle
Clinical significance:
-sporadic abortion
-brucellae in milk
B. melitensis
Species occasionally infected:
-Humans
Clinical significance:
-Malta fever
-severe systemic disease
B. melitensis
Species occasionally infected:
-Humans
Clinical significance:
-intermittent fever
-systemic disease
B. suis
Species occasionally infected:
-Humans
Clinical significance:
-Mild systemic disease
B. canis
Species occasionally infected:
-Dolphins
Clinical significance:
-May cause abortion
-Neurological disease has been described
B. ceti
Species occasionally infected:
-Humans
Clinical significance:
-little evidence of disease
B. ceti
Species occasionally infected:
-Humans
Clinical significance:
-little evidence of disease
B. pinnipedalis
B. abortus:
-Number of biotypes
-Requirement for CO2
-7
-v
B. melitensis
-Number of biotypes
-Requirement for CO2
-3
- negative
B. suis
-Number of biotypes
-Requirement for CO2
-5
- negative
B. ovis
-Number of biotypes
-Requirement for CO2
-1
- positive
B. abortus
-production of H2S
-Urease activity
- v
- positive
B. melitensis
-production of H2S
-Urease activity
-negative
-v
B. suis
-production of H2S
-Urease activity
-v
-positive
B. ovis
-production of H2S
-Urease activity
negative
B. canis
-production of H2s
-Urease activity
-negative
-positive
B. abortus:
Growth in media
-Thionin: v
-Basic fuchsin: v
B. melitensis:
Growth in media
-Thionin: positive
-Basic fuchsin: positive
B. suis:
Growth in media
-Thionin: positive
-Basic fuchsin: variable
B. ovis:
Growth in media
-Thionin: positive
-Basic fuchsin: negative
B. canis:
Growth in media
-Thionin: positive
-Basic fuchsin: negative
Conducted on bulk milk samples for monitoring infections in dairy herds. Sensitive but may not be reliable in large herds
Brucella milk ring test
Useful screening test. Antigen suspension is adjusted to pH 3.6, allowing agglutination by IgG1 antibodies. Qualitative test only, positive results require confirmation by CFT or ELISA
Rose-Bengal plate test
Widely accepted confirmatory test for individual animals
Complement-fixation test (CFT)
Reliable screening and confirmatory test
Indirect ELISA
Recently developed test with high specificity; capable of detecting all immunoglobulin classes and can be used to differentiate infected animals from S19-vaccinated cattle
Competitive ELISA (using monoclonal antibodies)
A tube agglutination test which lacks specificity and sensitivity; IgG1 antibodies may not be detected, leading to false negative results
Serum agglutination test (SAT)
Sensitive test for detecting non-agglutinating antibodies not detected by the SAT
Antiglobulin test
Differentiated by
-colonial appearance
-biochemicsl tests
-specific cultural requirements and growth inhibition by dyes
Definitive identification
-agglutination with monospecific sera
-susceptibility to bacteriophages
-Molecular methods
Occur in smooth forms, small, glistening, bluish, translucent. Colonies become opaque with age.
-B. abortus
-B. melitensis
-Brucella suis
Primary isolates always occur in rough forms. They are dull, yellowish, opaque and friable.
-B. ovis
-B. canis
Brucella on blood agar
Non-haemolytic
Used to detect presence of important surface antigens
Slide agglutination with monospecific antisera
Important surface antigens
-abortus antigen A
-melitensis antigen M
Feature of the rough brucellae B. ovis and B. canis, can be detected by anti-R serum
R antigen
Isolates of B. abortus is lysed by a specific bacteriophage at routine test dilution
Tbilisi phage
Could differentiate between:
-B. abortus biovars 1,2 and 4
-B. melitensis
-B. ovis
-B. suis
AMOS-PCR
A new multiplex PCR assay described in the OIE Terrestrial Manual identifies most Brucella species in a single step, including newly describe species from marine mammals and B. abortus and B. melitensis vaccine strains
Bruce-ladder
Less virulent Brucella
-lacks outer membrane
-produce rough colonies
-rough forms are usually eliminated unlike smooth which persist and multiply
Brucella persist within ____ but not within____
-macrophage
-neutrophils
Non-opsonized brucellae are taken up through interaction of the O side- chains of LPS with cholesterol-rich regions of the phagocyte plasma membrane, termed as
Lipid rafts
Once engulfed, brucellae persist within the acidified phagosome or
‘Brucella-containing vacuole’
Is important as it induces changes in gene expression of the Brucella organism, which favor intracellular survival
Acidification of phagosome
Constituents of the outer membrane. Helps in the prevention of phagolysosome fusion
Cyclic beta-1,2-glucans
Major mechanism for intracellular survival. Important determinant of bacterial virulence.
Inhibition of phagosome-lysosome function
Effectors secreted by a type ____ secretion system encoded by the ____. Important in maturation of the vacuole and it’s transport to, and fusion with, the rough endoplasmic reticulum
-type IV system
-virB operon
Virulent brucellae are transported to
Regional lymph nodes
Spread and localization in the reproductive organs and associated glands in sexually mature animals
Intermittent bacteremia
-Acts as a growth factor for brucellae.
-Present in high concentrations in the placentae of cattle, sheep, goats and pigs
-Growth factor is also found in other organs such as mammary gland and epididymis (targets of brucellae)
Erythritol (polyhydric alcohol)
-Strongly influenced by the stage gestation.
- Increases in late gestation, when the cells actively secrete steroid hormones
Intracellular replication in trophoblastic cells
Organisms may localize in joints or intervertebral discs
Chronic brucellosis
Responsible in part for the persistent infections seen with this pathogen.
brucellae may inhibit or delay the host immune repsonse
Although each Brucella spp has its own natural host,______, ______, and ______ can infect animals other than their preferred host.
-B. abortus
-B. melitensis
-biotypes og B. suis
Bovine brucellosis
B. abortus
Caprine and Ovine brucellosis
B. melitensis
Ovine epididymitis
B. ovis
Ovine epididymitis (B. ovis):
-Rams
-Ewes
-epididymitis
-placentitis
Porcine brucellosis
B. suis
B. suis:
Host: Infects pigs
Brucella suis biovar 1 to 3
Brucella suis:
-wild boat (host)
-European hare (reservoir host)
-produces military lesions particularly in reproductive tract
B. suis biovar 2
B. suis:
-Infects reindeer and caribou in Northern Canada, Alaska, Siberia
B. suis biovar 4
B. suis:
-Infects wild rodents
B. suis biovar 5
-Canine brucellosis
- Rough form (low virulence)
B. canis
Brucellosis in humans known as undulant fever
Severe:
-B. melitensis (Malta fever)
-B. suis biovars 1 and 2
Moderately severe:
-B. abortus
Mild:
-B. canis