Lab8. Brucella Flashcards
Brucella
- Taxonomy
- Habitat
- Morphology
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
Brucella
- staining
- gram negative
- will not stain very well because of the lipid content of the cell wall, therefore better to use Köster-staining - Köster-staining
- 3% sfranin 3-5minutes, steam once (because of the lipids)
- 0.5% H2SO4
- washing
- methylene blue 2 min - Modified Ziehl-neelsen (stamp) staining
Brucella
- Culture
- 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)
Brucella
- Biochemistry
- Antigens
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
Brucella
- Groups
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)
Brucella
- Resistance
Medium,
Highly sysceptible to acidic pH:
- inactivated in fermented dairy products and meat
- 30 day long survival in bedding –> shedding by the infected host
Brucella
- Pathogenicity
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
Burkholderia
- habitat, morphology, staining
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
Burkholderia
- culture
- species
Culture:
- not fastidious, obligate aerobic
- pigment production (except B. mallei)
Species:
- B. mallei (comes from B. pseudomallei in newly research)
- B. pseudomallei
- close relationship
Burkholderia mallei
- habitat, morphology, staining, culture:
Habitat:
- infected host
Morphology:
- 1.5-5 um rod
- no flagella
Staining:
- gram negative
- irregular staining –> Bipolar
Culture:
- glycerol supports growth
- no pigment production
Burkholderia Mallei
- Biochemistry, Antigens
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.
Burkholderia Mallei
- resistance, pathogenicity
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
Burkholderia Pseudomallei
- Habitat, morphology, culture:
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
Burkholderia Pseudomallei:
- pathogenicity
- malioidosis: generalized disease –> abcesses on the body
- swine, cattle, sheep, goad, dog, cat, human
Pseudomonas:
- Habitat, morphology, staining
Habitat:
- mucous membranes
- soil, water, plants, environment
Morphology:
- 1.5-5 um rod, flagella
Staining:
- gram negative
Pseudomonas
- Culture, pathogenicity, species
Culture:
- not fastidious: nutrient agar. enterobacterium selective media
- obligate aerobic
- pigment production
Pathogenicity:
- saprophytes - plant pathogens - animal pathogens
Sepcies:
- Pseudomonas aeruginosa
- Pseudomonas angulliseptica
Pseudomonas aeruginosa
- habitat, morphology, culture
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
Pseudomonas aeruginosa
- Biochemistry, antigens
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
Pseudomonas aeruginosa
- resistance
- 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
Pseudomonas aeruginosa
- pathogenicity
- 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
Pseudomonas anguilliseptica
- Biochemistry, pathogenicity
Fish pathogen
Biochemistry:
- inactive
Pathogenicity:
- salmon, eel: septicaemia, keratitis, haemorrhages
Aeromonas
- habitat, morphology, staining, culture
Habitat:
- water, sewage, gut
Morphology:
- 1-4 um rods
- generally flagellated
Staining:
- gram negative
Culture:
- simple, aerobic, facultative anaerobic
Aeromonas
- biochemistry, groups, species
Biochemistry: catalase +, oxidase +
Virulence factors:
- toxins (enterotoxins, haemolysin)
- extracellular enzymes
Groups, species:
- “mesophilic aeromonas” species: eg. A. hydrophila
- “Psychrofilic aeromonas” species: eg. A. salmonicida
Aeromonas hydrophila
- morphology, culture, biochemistry
Morphology:
- 1-4 um rods, flagella
Culture:
- simple
- hemolysis
- optimal temperature 28*C
biochemistry:
- soemtimes gas is produces
- resembles E. coli
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
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!
Franciella
- habitat, morphology, staining
Habitat:
- rodents, arthropods (blood sucklin)
- envirnoment, water
Morphology:
- 0.5-2 um coccoid rods
- thin capsule
Staining:
- gram negative
- bipolar
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
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
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
Franciella
- pathogenicity
Facultative intracellular agent
Tulareamia:
- wild living rodents
- hare, dog, cat, cattle, swine, sheep, human
Bordetella
- habitat, morphology, staining, culture
Habitat:
- respiratory mucous membrane (mammals, birds)
Morphology:
- 2-3 um rods
Staining:
- Gram negative
Culture:
- obligate aerobic
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
Bordetella bronchiseptica
- habitat, morohology, staining
Habitat:
- mucous membrane, respiratory (swine, dog, cat, horse, rabbit)
Morphology:
- rods, capsule
- flagella, fimbria
Staining:
- gram negative
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)
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
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
Bordetella avium
- habitat, morphology, culture,
Habitat:
- poultry respiratory tract
- ucous membranes of birds
Morphology:
- capsule, flagella, fimbria
Culture:
- simple
Bordetella avium
- Biochemistry, pathogenicity
Biochemistry: cytoxins
- demonecrotoxin
- tracheatoxin
- osteotoxin
Pathogenicity:
- avian bordetellosis (turkey coryza –> similar as the one in hens)
–> turkey, chicken
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
Moraxella
- resistance, species
Resistance: weak
Species:
- M. lacunata: human kerato-conjuctivitus
- M. catarrhalis: sinusitis, midde ear infection (children)
- M. bovis
- M. ovis
Moraxella bovis
- morphology, culture
Morphology:
- rods in pairs
- fimbria: important virulence factor
Culture:
- fastidious, blood agar
- beta hemolysis - virulence factor
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)
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
