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RUSVM Bacteriology Gram Negatives > Brucella > Flashcards

Brucella Flashcards

1
Q

Brucella Gram____? Shape?

A

Gram negative, small, coccobacilli (more coccoid) Pairs or clusters

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2
Q

Brucella Motile/Non-motile?

A

Non-Motile

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3
Q

Brucella Capsulated?

A

Non-encapsulated

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4
Q

Brucella Flagellated?

A

No flagella

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5
Q

Brucella Spore-forming?

A

Non-spore forming

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6
Q

Brucella Name the dominant surface antigens in LPS in rough versus smooth colonies

A

Rough: “R” antigen Smooth: “A” abortus or “M” melitensis antigens

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7
Q

Brucella What is significant about the cell wall?

A

Peptidoglycan layer = thicker than other Gram negative (modified acid fast stains for diagnosis)

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8
Q

Brucella Distribution?

A

Worlwide

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9
Q

Brucella Stable in environment?

A

No, obligate pathogen Survives 4 months in milk, urine, water, damp soil but NEEDS MAMMALIAN HOST

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10
Q

Brucella Often chronic or acute infections?

A

Mainly chronic infections

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11
Q

Brucella Target site

A

Reticuloendothelial system & genital tract Abortion & epididymitis/orchitis

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12
Q

Brucella Pathogenic species

A

*B. melitensis, B. abortus, B. suis* B. canis, B neotomae, B. ovis B. cetaceae, B. pinnipediae

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13
Q

Brucella Reportable?

A

YES REPORTABLE

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14
Q

Brucella Virulence factors

A

Cell Wall Erythritol Outer membrane proteins Superoxide dismutase & catalase T4SS

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15
Q

Brucella Erythritol role

A

4 carbon sugar alcohol *Allantoic fluid factors* (stimulates growth) Preferential location for reproduction & mammary glands

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16
Q

Brucella Omp (Outer membrane protein)

A

Stimulate delayed type hypersensitivity

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17
Q

Brucella Superoxide dismutase and catalase function

A

Defense against oxidative killing

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18
Q

Brucella T4SS function

A

Intramacrophage survival

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19
Q

Brucella Resistance factors

A

Survives freezing & thawing Survives in milk & other fluids Pasteurization kills it

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20
Q

Brucella Diversity

A

Genome very stable Different species vary in host preference & degree in virulence

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21
Q

Brucella What type of colonies are more virulent? Smooth or rough?

A

Smooth more virulent than rough Depends on polysaccharide side chain in LPS

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22
Q

What factors change Brucella host preference and virulence?

A

CO2 requirements H2S production Urease production Susceptible to different concentrations of certain dyes Susceptibility to bacteriophage (Tbilisi phage)

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23
Q

How does Brucella abortus differ from other strains?

A

B. abortus is susceptible to Tbilisi phage, others are resistant

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24
Q

Brucella Zoologic & Geographic reservoirs

A

Obligate parasite Host preference (some broad range) Survival time in environment depends on temperature & moisture

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25
Q

B. abortus

A

Cattle Smooth colonies, virulent Bison, camels, yaks

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26
Q

B. suis

A

Swine Smooth colonies, virulent Hares, reindeer

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27
Q

B. melitensis

A

Goat & Sheep Smooth colonies, virulent Camelids

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28
Q

B. neotomae

A

Wood Rat Smooth colonies, virulent

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29
Q

B. ovis

A

Sheep

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30
Q

B. canis

A

Dogs

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31
Q

Brucella Modes of transmission

A

Ingest cow/goat milk In utero (direct) Venereal Urine (dogs) Insects (maybe, minor)

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32
Q

Describe Brucella’s venereal transmission

A

Male sex gland infections Dissemination occurs without lesions Common in swine, sheep, dogs

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33
Q

Brucella Pathogenesis

A

Penetrates intact mucosa–> Macrophage–> survives –> Regional lymph node –> hematogenous dissemination & localization in reticulo-endothelial system & repro tract –> Abortions

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34
Q

How do Brucella species survive inside macrophages?

A

T4SS, Superoxide dismutase, catalase

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35
Q

How do Brucella species cause abortions?

A

Interference with fetal circulation / placentitis Endotoxin Fetal stress / Inflammation

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36
Q

What type of inflammation is shown in the picture?

A

Granulomatous inflammation!

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37
Q

What can cause Brucella infections to move extra-genitally? (Disseminate)

A

Prolonged bacteremia

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38
Q

Brucella Pathology

A

Grossly visible lesions in placenta Intercotyledonary thickening with yellow gelatinous fluid Necrotic cotyledon with thick brown exudate Aborted edematous fetus Male genitalia

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39
Q

Describe Brucellosis in males

A

Palpable enlargement of epididymus Increase neutrophils in semen (acute cases) Orchitis Scrotal dermatitis in dogs Results: decrease fertility, sometimes sterility

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40
Q

Describe orchitis from Brucellosis in males

A

Necrosis in testicular parenchyma with pus, prostatitis, fibrinopurulent, seminal vasculitis

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41
Q

What is shown in the picture? What species caused it?

A

Necrotic cotyledons in aborted material. B. melitensis

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42
Q

Brucella Disease Pattern Female reproductive tract

A

Abortion (most commonly) Retained placenta Mastitis (goats)

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43
Q

Brucella When will it cause cattle to abort?

A

~5month gestation

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44
Q

Brucella When will it cause swine to abort?

A

At any time

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45
Q

Brucella When will it cause dogs to abort?

A

~50 days gestation

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46
Q

Brucella Repeat abortions or lasting immunity?

A

Females usually abort once Acquired immunity

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47
Q

What does this picture show?

A

Brucella melitensis Stillbirth fetus with necrotic placenta

48
Q

What do these pictures show?

A

Orquitis - Swelling & irregular conformation of tail of epididymis of ram infected with Brucella ovis.

49
Q

Brucella Extragenital manifestations in Swine

A

Arthritis Lumbar spondylitis Tissue necrosis Posterior paralysis

50
Q

Brucella Extragenital manifestations in Dogs

A

Meningoencephalitis Osteomyelitis Discospondylitis Anterior uveitis Ocular manifestations

51
Q

Brucella Extragenital manifestations in Cattle

A

Hygromas - Chronic (false bursa, on joint sacs)

May be secondary to abortion

52
Q

Brucella Extragenital manifestation in Horses

A

Poll evil or fistulous withers

53
Q

Brucella Extragenital manifestation in Humans

A

Undulant Fever Malta Fever

54
Q

What body system does undulant fever affect?

A

Reticulo-endothelial system

55
Q

Common signs of Undulant Fever

A

Mild lymphadenopathy, splenomegaly, hepatomegaly 2-3 weeks post-exposure Fever, chills, night sweats, fatigue, muscle & joint pain, backaches, depression, insomnia, arthritis, osteomyelitis, endocarditis

56
Q

How do humans get Undulant fever?

A

Handling infected tissues Often: aborted fetuses, placentas, post-abortion uterine fluieds, milk, milk products Human-human rare

57
Q

What human professions are at risk for Brucellosis?

A

Veterinarians, ranchers, slaughterhouse workers

58
Q

What Brucella species can affect humans?

A

B. melitensis** B. suis B. abortus B. canis

59
Q

Which Brucella species do not affect humans, generally?

A

B. ovis B. neotomae (always questionable)

60
Q

What predisposes an animal to a Brucellosis infection?

A

Younger more resistant Sexually mature animals MORE susceptible, (infected for life possible) Higher herd size & density

61
Q

What helps SPREAD Brucellosis in swine herds?

A

New boars Confine breeding swine in common pens Feral swine

62
Q

What helps CONTROL Brucellosis in swine herds?

A

Separate the breeding pairs Quarantine boars Limit exposure to feral population Eliminate infected boars Minimize exposure to aborted tissue

63
Q

How is Brucellosis often spread in sheep herds?

A

Dissemination during breeding season Older rams more likely infected Homosexual rams spread it

64
Q

What are common differentials to Brucellosis in older rams?

A

Actinobacillus seminis Histophilus somni

65
Q

How is Brucellosis spread in dogs?

A

Close confinement/ Kennels Urine!

66
Q

Brucella Immune mechanisms in pathogenesis

A

Antibodies = protective & detrimental Phagocytes disseminate the organism

67
Q

Describe the role of antibodies in a Brucella infection

A

Low levels= complement High levels = opsonization IgA autoantibodies in dogs

68
Q

What class of immunoglobulin are used in low level antibody production?

A

Mainly initially IgM and low IgG (Cause complement-mediated lysis)

69
Q

What class of immunoglobulin are used in high-level antibody production?

A

High levels of IgG Survive inside phagocytes Interfere with complement and opsonize bacteria

70
Q

Brucella Immunologic aspects Mechanisms of Resistance & recovery

A

Effective immunity = **cell mediated** Macrophages & NK cells

71
Q

What cytokines are released in cell mediated immunity against Brucella infections?

A

IL-2, -12, -18 IFN-g TNF

72
Q

When will an animal develop the most effective immune response to Brucella?

A

**Better response if infected prior to sexual maturity**

73
Q

Brucella Vaccinations in cattle

A

B. abortus 45/20 (killed, 2 doses) B. abortus 19 & RB51 (live-attenuated, 1 dose, 4-8mo old)

74
Q

What can happen if you vaccinate a full grown bull for B. abortus?

A

Orchitis

75
Q

What are some problems with the B. abortus vaccines in cows?

A

Abortions Serological false positives

76
Q

Brucella vaccines for Goats and Sheep

A

B. melitensis Rev 1 (Live attenuated) B. melitensis H38 (killed) -Sometimes protects against B. ovis

77
Q

Are there vaccines for B. suis or B. canis?

A

No

78
Q

Can vaccines for B. melitensis be used in eradicated countries?

A

Cannot be used in countries free of B. melitensis because of serological test interference

79
Q

What is important about submitting suspected Brucella specimens?

A

Biosafety level 3 practices (prevent aerosolization) Submit to reference labs!

80
Q

What samples should be collected for Brucella lab diagnosis?

A

Abscess material, semen, vaginal fluids, milk samples, blood (dogs, bacteremia of B. canis) Necropsy & Abortion material

81
Q

What necropsy material is collected for Brucella lab diagnosis?

A

Spleen, liver, udder, lymph nodes, epididymis, testicle, accessory male sex organs

82
Q

What abortion material is collected for Brucella lab diagnosis

A

Abomasal fluids Lungs of aborted fetus Placenta

83
Q

What direct exam methods are used for Brucella Lab diagnosis?

A

Gram Stains Modified Gram Stains Modified Ziehl-Neelsen Macchiavello stains

84
Q

Brucella Lab Dx Describe Gram Stains (fetal contents vs. semen)

A

Fetal contents reveal high # gram negative cocci Few numbers of bacteria in semen

85
Q

Brucella Lab Dx Modified Gram Stain

A

Use carbol fuschin instead of safranin

86
Q

Brucella Lab Dx Modified Ziehl-Neelsen (with good differentials)

A

0.5% Acetic acid to destain Stain Red DDx:: Coxiella burnetii and Chlamydophila abortus

87
Q

Brucella Lab Dx Macchiavello stains

A

Mainly from fetal stomach content & placenta Stain red

88
Q

Brucella Lab Dx Microscopy

A

Gram stain & immunofluorescence

89
Q

Brucella Lab Dx Isolation - samples used?

A

Tissues Fluids Milk (cream & sediment)

90
Q

Brucella Lab Dx Isolation - Agars used?

A

Serum dextrose Tryptose Columbia agar w/ 5% serum Brucella agars

91
Q

Brucella Lab Dx Isolation - Mediums used?

A

Farrell’s medium Thayer-martin modified medium

92
Q

Brucella Lab Dx Isolation - Mediums - Conditions for culture

A

Aerobic environment, 37C, 10% CO2, 10-21 days Bluish color under obliquely transmitted light

93
Q

What is the most sensitive method for Brucella laboratory diagnosis?

A

Animal inoculation

94
Q

Describe animal inoculation in Brucella lab diagnosis

A

Necessary when low # are present Guinea pigs are most sensitive Test 3-6 weeks post inoculation

95
Q

Describe the colonies on blood agar

A

Brucella Gram negative, coccobacilli, non-hemolytic Catalase & oxidase positive Urease positive (except B. ovis) No fermentation of glucose or lactose

96
Q

Brucella species and biovar determinations?

A

Urease production CO2 requirements H2S production Oxidation of metabolic substrates Agglutination in mono-specific antisera Growth in presence of varying concentration of thionin Basic fuschin Phage typing

97
Q

Molecular techniques in Brucella Lab diagnosis

A

PCR, real-time PCR, RLFP PCR Strain 19 - deletion in ery locus Strain RB51 - Insertion in wbo A gene

98
Q

What makes strain 19 different from other Brucella strains.

A

Deletion in the ery locus

99
Q

What makes strain RB51 different?

A

Insertion sequence in wbo A gene

100
Q

What does the Brucellin Test test?

A

Type 4 Hypersensitivity

101
Q

What tests for B. ovis best?

A

Agar gel immunodiffusion

102
Q

Brucella Immuno Dx Rose-Bengal plate test

A

Antigen suspension, pH=3.6, allows IgG1 agglutination Positive results need confirmation

103
Q

Brucella Immuno Dx Brucella milk ring test

A

Purple ring develops in top with cream (picture)

104
Q

Brucella Immuno Dx Confirmatory tests

A

CFT - complement fixation test Indirect ELISA

105
Q

Brucella Immuno Dx For canine brucellosis

A

Rapid slide agglutination test

106
Q

Brucella Immuno Dx Which species share antigens?

A

Antigens shared with Y. enterocolitica O:9 and E. coli O:157

107
Q

Brucella Control & Prevention

A

Depends on animal species Brucella Species Husbandry Availability/Efficacy of vaccines

108
Q

Brucella Treatment Livestock

A

No treatment in livestock High failure Cost Problem maintaining infected during eradication program

109
Q

Brucella Treatment Rams with B. ovis

A

Prior to palpable epididymal lesions, abscesses or fibrosis: Tetracycline or dihydrostreptomycin After these lesions - difficult to treat

110
Q

Brucella Treatment Dogs

A

Neuter affected animals Antibiotics (tetracycline & dihydrostreptomycin) Cull males in breeding program Don’t treat breeding colonies

111
Q

Brucella Prevention Immunization alone

A

Reduces # abortions No eradication

112
Q

Brucella Prevention Bovine Brucellosis Immunization THEN test & slaughter (Tests in dairy vs. beef)

A

Vaccinate young cattle, test @ sexual maturity Seropositives culled & slaughtered -Ring test in dairy -Blood test in beef cattle

113
Q

Brucella Prevention Testing & removal without immunization

A

Depopulation of entire swine, sheep, goat population Re-stock

114
Q

How often Rams palpated?

A

Rams tested twice per year by palpation

115
Q

Brucella Prevention Dogs (# tests needed for “free” status)

A

Dogs tested before breeding Need 3+ seronegative test results for “free” status

RUSVM Bacteriology Gram Negatives (26 decks)

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