Day 5: Introduction Bacteriology and Diagnosis Flashcards
HC12, 13
HC12: Microbiome
Human-associated microorganisms
> 4* 10^13
> 500-1000 different species
There are different microbiota on different parts of the body, why?
- Temperature
- Moisture
- more location specific factors
Microbiota as source health and disease
- Nutrition and metabolism of foods
> breakdown of indigestivle polysaccharides
> Production vitamin B and K - Maturation and instruction of immune system: and tolerance
- Colonization resistance: protection against invaders
- Potenital disease causing: risk infection
Balance microbiota should be towards…
Commensals and not pathogens
Pathogen
Microorganism causing disease
Commensal
Microorganism in symbiotic relationship with host
Commensialism
Presence microorganisms multiplying in host without damage and reaction from the host (no advantage or disadvantage for host)
Infection
Presence (invasion) of microorganisms multiplying in the host, with damage to and reaction of the host
Name 4 types of opportunistic commensals
- Neisseria meningitidis: meningococcus
- Streptococcus pneumoniae: causing pneumonia
- Staphylococcus aureus: normally harmless in skin, can get into bloodstream when invasion
- Group A Streptococcus: infection of throat which can get into blood: sepsis
Carrier
Host that harbors potential pathogen without disease
Pathogenicity
Ability to produce disease in host organism
Virulence
The degree of pathogenicity of the microbe
Virulence factor
Microbial component that contributes to disease-making ability
Different clinical pictures for infection S. aureus
- Different factors of bacterium
- Scalded skin syndrome
- Food poisoning by the excreted toxins
> bacteria can be killed by cooking food but toxins survive - same causative agent
Course of infection is determined by
1: Route of contamination and invasion of host
> name entries for S. aureus
- Skin or mucous membrane > SSSS (staphylococcus scalded skin syndrome)
- Transcutaneous: vectors, insects
- Directly in bloodstream: needles
- Airways > SSSS
- Gastrointestinal > food poisoning
Pathogenicity: invasiveness and toxigenesis
- Invasiveness
> ability to invade tissues
> colonization, bypass host defense - Toxigenesis
> ability to produce toxins
> exotoxins, endotoxins
> > Virulence factors
Course of infection determined by
1 Route of contamination and invasion of host
2 Properties of the microorganism
> name virulence factors
- Adhesins
- Invasins
- Capsule (polysaccharide): prevent phagocytosis or decrease complement activation
- Toxins
- Enzymes
- Pili: protein strands which point out for attachment
Different toxins in SSSS and S. aureus food poisoning
SSSS
> S. aureus secretes exfoliative toxins A and B
> disrupt desmosomes by binding them and unstick epithelial cells of skin
> bacteria from local infection
Food poisoning
> S. aureus enterotoxin A
> potent gastrointestinal exotoxins
Tropism types
Cellular, tissue and host tropism
Virulence factors S. aureus: toxins
- Cytolytic toxins: tissue destruction/abscess formation
- Enterotoxins (A-E): toxicity
- Exfoliative toxin: blistering: loss of desmosomes
Virulence factors S. aureus: enzymes
- Coagulase: blood clotting
> reacts with prothrombin in blood > complex: staphylothrombin > enables enzyme to act as protease and convert fibrinogen to fibrin > blood clotting - Hyaluronidase, lipase, fibrinolysis: distribution pathogen in tissues
- Catalase: prevent oxidative burst, catalyse hydrogen peroxide
> > diversity within one bacterial species
Enterotoxin A from S. aureus (food poisoning) is resistant to:
Heat and low pH.
> gastrointestinal
Sources SSSS and S. aureus food poisoning
Infected wound
Contaminated processed food
Course of infection determined by?
1 Route of contamination and invasion of the host
2 Properties of the microorganism
3 Properties of the host
> age
> healthy or reduced resistance
> vaccination
Normal vs hampered immune response
Immunosuppressed individuals (AIDS, cytostatica) have lower burden for colonization for the opportunistic bacteria
Neisseria meningitidis and streptococcus clinical pictures are …
Similar
> completely different causative agents
> both the throat as natural habitat
N. meningitidis character
- Gram negative diplococcus (coccus form)
- Humans are only natural host
> can cause meningitis and sepsis
Vaccination campaign N. meningitidis
Target: polysaccharide capsule
> first against serotype C (based on capsule polysaccharide group C)
> now MenACWY
> serologic reactivity polysaccharide: groups A, B , C, Y, W
Gram positive bacterium
Purple on gram staining
> one cell membrane and thick peptidoglycan cell wall layer around it
How are the gram positive cocci genuses staphylococcus and streptococcus distinguished
- Staphylococcus: Catalase positive
- Streptococcus: Catalase negative
Staphylococcus species classification
- Coagulase positive: S. aureus
- Coagulase negative: CONS like S. epidermidis
Streptococcus species classification
- Group alpha-hemolytics: make blood plate green because make hydrogen peroxide. Common in throat and mouth.
- Group beta-hemolytics: make toxins to destroy blood cells in vitro
- Group gamma-hemolytics: no hemolytic activity on blood plate
Gram negative
Two cell membranes with thin peptidoglycan layer between them (in periplasmic space)
> pink on gram stain
Name of bacteria
Genus name Species name
Typhi is the result of ..
Some salmonella species
Clostridium species character
Can form spores and survive most extreme envirnoments
Infection and transmission > results
Transfer microorganisms from one place to another
> possible multiplication
> possible damage
> possible reaction of the host
Ways of transmission
- Direct: person-to person or droplet (saliva)
- Indirect: fecal-oral (contaminated food), airborne (aerosol), fomites (infected objects)
- Vector: insect bite
Zoonosis
Infection through contact with animals
> both directions possible
> like ebola virus disease and salmonellosis
HC13: Biofilms
Bacteriën gaan zo organiseren dat ze een eigen taak hebben (op plastic of geïnfecteerde protehese)
Diversiteit en volhardendheid van bacteriën
- Biofilms
- Resistentie
- Lastig te behandelen
- Problemen zoals opnieuw opereren
Gramkleuring bacteriën
Positief: violet
Negatief: roze
Uitspraak bacterienamen
Klemtoon op twee na laatste lettergreep
Rangschikking bacteriën
Ketens: streptococcus
Tros: staphylococcus
Vorm bacteriën
Kok of staaf
Indeling bacteriën op metabolism
- Facultatief
- Strikt aeroob / non fermenters
- Anaeroben
Gramkleuring Clostridioides difficile en Escherichia coli
C. difficile > positief
E. coli > negatief
Hoe kweken anaerobe bacteriën
Apparatuur en reagentia om luchtdichte potten mt kweekplaten zuurstofvrij te maken
Betrokkenheid van bacteriën is afhankelijk van locatie: in de keel
- Vergroenende streptococcen (non hemolytisch)
- Hemolytische streptokok
- anaeroben
- gram negatieve flora
- gisten
Darmen bacteriën
- Gram negatieve staven
- Anaeroben
- Enterokokken
- Gisten
Gewicht feces is …. aan bacteriën
1/3
> hoog dragerschap en kolonisatie
Diagnose bacterie: de bron
Patientenmateriaal
> bloed, urine, feces, wond uitstrijk, sputum, neuskeelspelsel, BAL (longspoeling), catheter tip, IUD, heupkop
Determinatie bacterie
species genus
Resistentiebepalingen staan in een …
antibiogram
> voor welke antibiotica resistentie en gevoeligheid
Diagnose op welke vlakken?
- Determinatie
- Resistentiebepaling
- Virulentiefactoren: hoe pathogeen
- Clusteranalyse: verwantschap
Diagnostiek: microscopie
- Directe kleuring patientenmateriaal
> of kleuring bacteriële isolaten na kweek - Kleuringen
> kleuring
> fluorescentie
> immuunfluorescentie met specifieke antilichamen
» techniek afhankelijk van celwand
Gramkleuring inhoudelijk
Fixeren en dan kleuring
> crystal violet > joodoplossing > tegenkleuring
» werkt goed doordat het de biologie van de bacterie matcht
> kleur spoelt weg bij gramnegatief: geen binding peptidoglycaan
Gramnegatieve staven vaak bij ..
Urineinfecties
Locatie van de kweek beïnvloedt de ..
gebruikte kweekmedia
Determinatie bacterie
Massaspectrometrie
> MALDI-TOF
> Laser: ionisatie > door buis heen
> tijd tot detector geeft massa
> call voor een bepaalde bacterie door software
Oude determinatie: bonte rij
Pak kolonie en doe in buizen
> in elke buis, bepaald substraat en indicator
> in stoof
> kleurtje: beslisboom gebruiken in boek en classificatie
Geautomatiseerde bonte rij
Alles in well plates
> kleurencode in lab en afgelezen
> biochemische eigenschappen gebruikt voor determinatie
Antibiogram maken
Kaart met read gemaakt
> zuigt vacuum
> wells met antibioticum en verschillende concentraties op well
> in draaimolen
> gaan bacteriën groeien of niet
> scannen en detectie
> kaart is vooraf gemaakt met de antibiotica erin
MIC bepaling
Bepalen minimal inhibitory concentration
> gebruik plaatjes met antibioticum en gradient
> minimale concentratie voor resistentie
Resistentie mechanismen
- Intrinsieke resistentie
- Verandering antibioticum target
- Enzymproductie
- Lagere permeabiliteit celwand
- Efflux mechanismen
Waarom antibioticum met ijzermineraal erin?
Verleiden bacterie om het op te nemen
Toepassing verkregen resultaat antibiogram
> naar arts en RIVM
hoge incidentie resistentie: minder antibioticum geven > adviezen op RIVM
gaat per centrumL infecties en resistenties verschillen per medisch centrum
Empirische antibioticakuur
Starten met antibiotica zonder kennis van veroorzaken micro-organismen, in afwachting van kweekuitslagen, op basis van RIVM cijfers
Serologie
Detectie antigen / antilichamen in patientenmateriaal
Agglutinatietesten
- Antistoffen tegen deel antigenen bacterie samenbrengen
- Soms ter determinatie
- Meerdere antigeen en antistof complexen moeten bundelen
Lateral flow assay
Corona zelftest of zwangerschapstest
> membraan met antilichaam voor toxines van bepaalde bacterie/virus
PCR test
Voor virale infecties handig, lastig te kweken, of lastige bacterien
Sequentie analyse
Determinatie
> resistentiemechanismen voor specifieke genen
> clusteranalyse: verwantschapsonderzoek