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Bacteriology > Lab8. Brucella > Flashcards

Lab8. Brucella Flashcards

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
Q

Aeromonas hydrophila

  • pathogenicity
A

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
Q

Aeromonas salmonicida

  • morphology, culture, pathogenicity
A

Morphology:

  • small coccobacillus, no flagella

Culture:

  • optimal temp. 20*C
  • dark brown pigment

Pathogenicity:

  • facultative pathogenic
  • fish (salmonidae): furunculosis
  • infect only fish!
27
Q

Franciella

  • habitat, morphology, staining
A

Habitat:

  • rodents, arthropods (blood sucklin)
  • envirnoment, water

Morphology:

  • 0.5-2 um coccoid rods
  • thin capsule

Staining:

  • gram negative
  • bipolar
28
Q

Franciella

  • culture
A
  • 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
Q

Franciella

  • biochemistry, antigens
A

Biochemistry:

  • catalase +, oxidase -
  • glycerol:

–> +: F. tularensis subsp. tularensis

–> -: F. tularensis subsp. holarctica (palaearctica)

Anigens:

  • uniform
  • cross reactions: Bruceplla spp., or Y. pestis
30
Q

Franciella

  • Resistance, species
A

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
Q

Franciella

  • pathogenicity
A

Facultative intracellular agent

Tulareamia:

  • wild living rodents
  • hare, dog, cat, cattle, swine, sheep, human
32
Q

Bordetella

  • habitat, morphology, staining, culture
A

Habitat:

  • respiratory mucous membrane (mammals, birds)

Morphology:

  • 2-3 um rods

Staining:

  • Gram negative

Culture:

  • obligate aerobic
33
Q

Bordetella

  • biochemistry, species
A

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
Q

Bordetella bronchiseptica

  • habitat, morohology, staining
A

Habitat:

  • mucous membrane, respiratory (swine, dog, cat, horse, rabbit)

Morphology:

  • rods, capsule
  • flagella, fimbria

Staining:

  • gram negative
35
Q

Bordetella bronchiseptica

  • culture
A
  • 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
Q

Bordetella bronchiseptica

  • Biochemistry, antigens
A

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
Q

Bordetella bronchiseptica

  • Resistance, pathogenicity
A

Resistance: medium

Pathogenicity:

  • facultative pathogenic
  • Swine: atrophic rhinitis (with P.mulocida)
  • Rabbit: pneumonia
  • Dog: cennel cough, complication to distemper
  • Cat: bronchitis, bronchopneumonia
38
Q

Bordetella avium

  • habitat, morphology, culture,
A

Habitat:

  • poultry respiratory tract
  • ucous membranes of birds

Morphology:

  • capsule, flagella, fimbria

Culture:

  • simple
39
Q

Bordetella avium

  • Biochemistry, pathogenicity
A

Biochemistry: cytoxins

  • demonecrotoxin
  • tracheatoxin
  • osteotoxin

Pathogenicity:

  • avian bordetellosis (turkey coryza –> similar as the one in hens)

–> turkey, chicken

40
Q

Moraxella

  • habitat, morphology, staining, culture
A

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
Q

Moraxella

  • resistance, species
A

Resistance: weak

Species:

  • M. lacunata: human kerato-conjuctivitus
  • M. catarrhalis: sinusitis, midde ear infection (children)
  • M. bovis
  • M. ovis
42
Q

Moraxella bovis

  • morphology, culture
A

Morphology:

  • rods in pairs
  • fimbria: important virulence factor

Culture:

  • fastidious, blood agar
  • beta hemolysis - virulence factor
43
Q

Moraxella bovis

  • biochemistry, antigens, resistance, pathogenicity
A

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
Q

Moraxella ovis

  • habitat, morphology, pathogenicity
A

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

Bacteriology (10 decks)

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