Lab8. Brucella Flashcards

(44 cards)

1
Q

Brucella

  • Taxonomy
  • Habitat
  • Morphology
A

Taxonomy:

is not fully settled, you can see different naming, eg. B. abortus, B. melitensis serovar Abortus

  • Monospecies system: B. melitensis, whti the species biovars

–> B. melitansis biovar abortus 1, 2, 3

  • Multispecies system: B. abortus 1, 2, 3
  • Both system are in use

Habitat:

  • infected host: mucous membranes of the genitals, lymph nodes, udder, joint, tendon sheat, bursae

Morphology:

  • 0.6-1.5 um coccoid rods
  • lipids and proteins in the cell wall
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2
Q

Brucella

  • staining
A
  1. gram negative
    - will not stain very well because of the lipid content of the cell wall, therefore better to use Köster-staining
  2. Köster-staining
    - 3% sfranin 3-5minutes, steam once (because of the lipids)
    - 0.5% H2SO4
    - washing
    - methylene blue 2 min
  3. Modified Ziehl-neelsen (stamp) staining
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3
Q

Brucella

  • Culture
A
  • Obligate aerobic
  • Fastidious

–> bloog agar, serum agar

–> Vitamin (B vitamins), yeast extract

  • CO2
  • S-R colonies (at the isolation):

–> S colonies: B. melitensis, B. abortus, B. suis

–> R colonies: B. canis, B. ovis

  • selective culture:

–> polymyxin, bacitracin (inhibition of Gram +), cycloheximide (inhibition of molds and fungi), etc

  • (lab. animals)
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4
Q

Brucella

  • Biochemistry
  • Antigens
A

Biochemistry:

  • Catalase +, Oxidase +, Urease +

Anitgens:

lipopolysaccharide, peptides, lipids

close antigen relationship:

  • “S colony type”:

–> B. melitensis-, B. abortus-, B.suis- cross reaction

–> A and M polysaccharides: B. melitensis-, B. abortus-, B. suis

  • “R colony type”:

–> B. ovis-, B. canis- cross reaction

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5
Q

Brucella

  • Groups
A

Species differentiation:

  • phage susceptibility (Tbilisi phage)
  • oxidative metabolism: the O2 consumption of the bacteria
  • natural host
  • genome structure

Biotype differentiation:

  • CO2 demand
  • H2S
  • fuchsin/thionin tolerance
  • agglutination (A-M-R)
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6
Q

Brucella

  • Resistance
A

Medium,

Highly sysceptible to acidic pH:

  • inactivated in fermented dairy products and meat
  • 30 day long survival in bedding –> shedding by the infected host
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7
Q

Brucella

  • Pathogenicity
A

Facultative intra-cellular bacterium: survives within macrophages

Diseases:

  • generalised diseases,
  • acute: abortion, epidymitis, orchitis
  • Subacute: arthritis, bursitis, tenosynovitis
  • B. melitensis: goat, sheep, cattle human (primarily small ru)
  • B. abortus: cattle, sheep, human (abortion primarily in 3rd semester)
  • B. suis: swine, hare, reindeer, rodents (biotype 1,2,3: swine, biotype 4: reindeer)
  • B. pinnipediales: seal, walrus
  • B. ceti: whale, dolphin
  • B. ovis: sheep
  • B. canis: dog
  • B. neotomae: desert rat
  • B. microti: vole
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8
Q

Burkholderia

  • habitat, morphology, staining
A

formerly Pseudomonas. Zoonotic!

Habitat:

  • mucous membrane (B.mallei)
  • Soil, water, plants (B. pseudomallei)

Morphology:

  • 1.5-5 um rod
  • flagella (except B. mallei)

Staining:

  • gram negative
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9
Q

Burkholderia

  • culture
  • species
A

Culture:

  • not fastidious, obligate aerobic
  • pigment production (except B. mallei)

Species:

  • B. mallei (comes from B. pseudomallei in newly research)
  • B. pseudomallei
  • close relationship
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10
Q

Burkholderia mallei

  • habitat, morphology, staining, culture:
A

Habitat:

  • infected host

Morphology:

  • 1.5-5 um rod
  • no flagella

Staining:

  • gram negative
  • irregular staining –> Bipolar

Culture:

  • glycerol supports growth
  • no pigment production
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11
Q

Burkholderia Mallei

  • Biochemistry, Antigens
A

Biochemistry:

  • catalase +, oxidase +, oxidative

Antigens: Mallein (PPD)

  • cell wall extract
  • glycoprotein

–> Mallein = antigen extract of the growth culture

–> PPD = Purified antigen extract. used to test in allergic tests –> injected into eyelids in the animals, in the conjuctiva or in the skin.

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12
Q

Burkholderia Mallei

  • resistance, pathogenicity
A

Resistance:

  • weak: dies if dehydrated in the environment
  • survives in water

–> sheds in the mucous discharge from resp.tract and can survive there

Pathogenicity:

  • malleus/glanders (horse, donkey)
  • cat, dog
  • human
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13
Q

Burkholderia Pseudomallei

  • Habitat, morphology, culture:
A

Habitat:

  • soil, water, mud
  • tropics, subtropics

Morphology:

  • 1.5-5 um rod, flagella

Culture:

  • simple: nutrient agar
  • Glycerol agar: large colonies
  • Broth: layer
  • Pigment production: yellow, brown, orange
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14
Q

Burkholderia Pseudomallei:

  • pathogenicity
A
  • malioidosis: generalized disease –> abcesses on the body
  • swine, cattle, sheep, goad, dog, cat, human
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15
Q

Pseudomonas:

  • Habitat, morphology, staining
A

Habitat:

  • mucous membranes
  • soil, water, plants, environment

Morphology:

  • 1.5-5 um rod, flagella

Staining:

  • gram negative
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16
Q

Pseudomonas

  • Culture, pathogenicity, species
A

Culture:

  • not fastidious: nutrient agar. enterobacterium selective media
  • obligate aerobic
  • pigment production

Pathogenicity:

  • saprophytes - plant pathogens - animal pathogens

Sepcies:

  • Pseudomonas aeruginosa
  • Pseudomonas angulliseptica
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17
Q

Pseudomonas aeruginosa

  • habitat, morphology, culture
A

Habitat:

  • environment, water, sewage, gut
  • surface of soil
  • mucous membrane of the host

Morphology:

  • 1 polar flagellum
  • polar fimbria

Culture:

  • simple 5-42 *C
  • Beta-hemolysis
  • odour
  • pigments: piocianin, fluurescein (yellow, blue) –> both turn green
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18
Q

Pseudomonas aeruginosa

  • Biochemistry, antigens
A

Biochemistry:

  • catalase +, oxidase +

virulence factors:

  • toxin (exotoxin A)
  • extracellular enzyme (protease, phospholipase C)
  • Production of exo-polysaccharide (biofilm formation)

Antigens:

  • O antigens:

–> 27 O antigens

–> several components

  • H antigens
  • Phage typing
  • Pyocin typing –> antibiotic protein of the bacteria
19
Q

Pseudomonas aeruginosa

  • resistance
A
  • resistane: low against heat
  • quaterner NH4 salts: desinfectants
  • amphoterid disinfectants

–> both of them cause big problem in food industry (hygiene)

  • susceptibility:

–> dehydration, heat, most disinfectants

20
Q

Pseudomonas aeruginosa

  • pathogenicity
A
  • facultative pathogenic
  • local suppuration, lesions
  • cattle, mare: mastitis, metritis
  • Sheep: dermatitis
  • Dog: Otitis externa
  • Fur animals: septicaemia
  • Human: middle ear infection, secondary agent. Nosocomial (hospital) infections
21
Q

Pseudomonas anguilliseptica

  • Biochemistry, pathogenicity
A

Fish pathogen

Biochemistry:

  • inactive

Pathogenicity:

  • salmon, eel: septicaemia, keratitis, haemorrhages
22
Q

Aeromonas

  • habitat, morphology, staining, culture
A

Habitat:

  • water, sewage, gut

Morphology:

  • 1-4 um rods
  • generally flagellated

Staining:

  • gram negative

Culture:

  • simple, aerobic, facultative anaerobic
23
Q

Aeromonas

  • biochemistry, groups, species
A

Biochemistry: catalase +, oxidase +

Virulence factors:

  • toxins (enterotoxins, haemolysin)
  • extracellular enzymes

Groups, species:

  • “mesophilic aeromonas” species: eg. A. hydrophila
  • “Psychrofilic aeromonas” species: eg. A. salmonicida
24
Q

Aeromonas hydrophila

  • morphology, culture, biochemistry
A

Morphology:

  • 1-4 um rods, flagella

Culture:

  • simple
  • hemolysis
  • optimal temperature 28*C

biochemistry:

  • soemtimes gas is produces
  • resembles E. coli
25
Aeromonas hydrophila - pathogenicity
facultative pathogenic fishes, amphibians, reptiles: - septicaemia farm animals, humans: - colonisation, sometimes wound infection, local suppuration - asymptomatic colonisation - food born infection - septicaemia in immunosuppressed humans
26
Aeromonas salmonicida - morphology, culture, pathogenicity
Morphology: - small coccobacillus, no flagella Culture: - optimal temp. 20\*C - dark brown pigment Pathogenicity: - facultative pathogenic - fish (salmonidae): furunculosis - infect only fish!
27
Franciella - habitat, morphology, staining
Habitat: - rodents, arthropods (blood sucklin) - envirnoment, water Morphology: - 0.5-2 um coccoid rods - thin capsule Staining: - gram negative - bipolar
28
Franciella - culture
- aerobic, fastidious - additives: --\> protein --\> cystein, cystin: (the blood agar needs to contain sulphur-containing molecules) --\> yeast extract Media: - Francis-blood agar: cystein - glucose - rabbit serum - Cystein - glucose - egg yolk
29
Franciella - biochemistry, antigens
Biochemistry: - catalase +, oxidase - - glycerol: --\> +: F. tularensis subsp. tularensis --\> -: F. tularensis subsp. holarctica (palaearctica) Anigens: - uniform - cross reactions: Bruceplla spp., or Y. pestis
30
Franciella - Resistance, species
Resistance: weak. Good survival in cold Species: - F. tularensis subsp. tularensis: North America - F. tularensis subsp. holarctica (palaearctica): Northern hemisphere - F. tularensis subsp. mediasiatica: Central Asia - F. tularensis subsp. novicida: Sproadic USA, Asia
31
Franciella - pathogenicity
Facultative intracellular agent Tulareamia: - wild living rodents - hare, dog, cat, cattle, swine, sheep, human
32
Bordetella - habitat, morphology, staining, culture
Habitat: - respiratory mucous membrane (mammals, birds) Morphology: - 2-3 um rods Staining: - Gram negative Culture: - obligate aerobic
33
Bordetella - biochemistry, species
Biochemistry: - Catalase +, oxidase + - cannot utilise carbohydrates - toxins are produced Species: - B. pertussis: human pertussis (cough in children) - B. parapertussis: human parapertiussi, ovine pneumonia - B.bronchiseptica - B. avium
34
Bordetella bronchiseptica - habitat, morohology, staining
Habitat: - mucous membrane, respiratory (swine, dog, cat, horse, rabbit) Morphology: - rods, capsule - flagella, fimbria Staining: - gram negative
35
Bordetella bronchiseptica - culture
- simple: no special demands - growth on media containing only citrate as carbon source - selective isolation: from mucous membrane into mixed culture --\> vrystal violet, bile salts (MacConkey agar), penicillin, nitrofurantion ( --\> to increase selective capacity)
36
Bordetella bronchiseptica - Biochemistry, antigens
Biochemistry: cytotoxins are produced - dermonecrotoxin: osteoblast are damaged - trachetoxin: ciliated epithelial cells are damaged - Adenylate cyclone haemolysin: phagocytosis is inhibited Antigens: - several O, K, H and fibria antigens
37
Bordetella bronchiseptica - Resistance, pathogenicity
Resistance: medium Pathogenicity: - facultative pathogenic - Swine: atrophic rhinitis (with P.mulocida) - Rabbit: pneumonia - Dog: cennel cough, complication to distemper - Cat: bronchitis, bronchopneumonia
38
Bordetella avium - habitat, morphology, culture,
Habitat: - poultry respiratory tract - ucous membranes of birds Morphology: - capsule, flagella, fimbria Culture: - simple
39
Bordetella avium - Biochemistry, pathogenicity
Biochemistry: cytoxins - demonecrotoxin - tracheatoxin - osteotoxin Pathogenicity: - avian bordetellosis (turkey coryza --\> similar as the one in hens) --\> turkey, chicken
40
Moraxella - habitat, morphology, staining, culture
Habitat: - mucous membranes (upper respiratory way, conjuctiva) Morphology: - 1-3 um rods, capsule - paired rods, paired cocci: some can form cocci, some can form rods, always occur in pairs Staining: - gram negative Culture: - fastidious - blood agar - serum agar
41
Moraxella - resistance, species
Resistance: weak Species: - M. lacunata: human kerato-conjuctivitus - M. catarrhalis: sinusitis, midde ear infection (children) - M. bovis - M. ovis
42
Moraxella bovis - morphology, culture
Morphology: - rods in pairs - fimbria: important virulence factor Culture: - fastidious, blood agar - beta hemolysis - virulence factor
43
Moraxella bovis - biochemistry, antigens, resistance, pathogenicity
Biochemistry: - catalase +, oxidase + - haemolysine: citotoxin, damage the cornea - extracellular enzymes: --\> proteases --\> hyaluronidase --\> fibrinolysine Antigens: - fimbria 7 groups: --\> virulence factors --\> important in vaccine production Resisntace: low Pathogenicity: - cattle infectious kerato-conjuctivitis (pink eye)
44
Moraxella ovis - habitat, morphology, pathogenicity
Habitat - sheep, calf: conjuctiva, nasal cavity Morphology: - cocci in pairs - fimbria on some of them Pathogenicity: - facultative pathogenic - keratoconjuctivitus of sheep, goats and young calves