Microbiology Flashcards
What is a pathogen?
An organism that causes disease
Define commensal. What is it also called?
- An organism that colonises the host but causes no disease under normal circumstances
- Also called colonisation
What is an opportunist pathogen?
A microbe that only causes disease if the host defence is compromised
Define virulence/pathogenicity.
The degree to which an organism is pathogenic
What are the categories for describing bacteria?
- Gram +ve or gram -ve
- Shape: cocci (circular) or bacilli (rods)
- Cocci can be individual, clusters or chains
- Bacilli can be chains, curved or spiral rods
What do we mean by gram positive and gram negative bacteria? What colours do these stain? Which dyes are responsible for these colours?
- Gram positive: has a thick layer of peptidoglycan, stains PURPLE. This is because it is stained by crystal violet
- Gram negative: has a thin layer of peptidoglycan (high lipid content), stains PINK. This is because it is stained by safranin

What are the stages for the gram staining method?
Mnemonic: Come In And Stain = Crystal violet, Iodine, Alcohol, Safranin
- Fixation of clinical materials to microscope slide (heat/methanol)
- Application of crystal violet. Wash off excess
- Application of iodine. Wash off excess
- Decolorisation step: use acetone or ethanol, this will distinguish between gram +ve and -ve bacteria. This is because gram +ve will be purple + rest will decolourise
- Application of safranin to stain gram -ve pink

What type of organism would you stain with Ziehl-Neelsen stain?
Used to identify acid-fast (resist decolorisation by acids during staining procedures) organisms, mainly mycobacteria, e.g. TB
What types of bacteria release endotoxin?
Gram negative bacteria. An example of endotoxins are lipopolysaccharides
What types of bacteria release exotoxins?
Gram positive and gram negative bacteria
What can gram positive cocci be split into? Which test can be used to distinguish between these?
- Gram +ve cocci can be split into staphylococcus (clusters) and streptococcus (chains)
- A CATALASE test can be used - this detects the presence of catalase enzyme using hydrogen peroxide (H2O2 + staphylococci = gas bubbles)
- +ve catalase test = staphylococcus, -ve catalase test = streptococcus

Which test can be performed to distinguish between staphylococci? What do positive and negative tests indicate?
- COAGULASE test. This looks at whether a fibrin clot is produced
- Coagulase +ve = staphylococcus aureus. This is because it produces coagulase that converts soluble fibrinogen to insoluble fibrin
- Coagulase -ve = all others (staphylococci epidermis, staphylococcus saprophyticus)

Which test can we perform to distinguish between streptococci?
Blood agar haemolytic. Blood agar can undergo 3 types of haemolysis depending on the organism: beta haemolysis = complete lysis (clear area), alpha haemolysis = partial lysis (green), gamma haemolysis = none

If alpha haemolysis is shown after haemolysis in blood agar, which test do we perform to differentiate? What do positive and negative results indicate?
- OPTOCHIN test
- Optochin +ve = STREPTOCOCCUS PNEUMONIAE (clear demarcated ring around optochin disc)
- Optochin -ve = streptococcus viridans

What is the appearance of strep pneumoniae under a microscope?
Gram positive diplococci
If beta haemolysis is shown after haemolysis on blood agar, which test is performed? What do the different groups indicate?
- LANCEFIELD TEST. This groups bacteria based on the carbohydrate composition of the bacterial antigen on their cell wall. Groups into A, B, C, D and G
- A: streptococcus pyogenes
- B: streptococcus agalactiae
- G: streptococcus dysgalactiae

If gamma haemolysis is shown after haemolysis on blood agar, which test is performed? What do the results indicate?
- Lancefield test
- D: streptococcus bovis, enterococcus

How does the haemolysis test work?
The haemolysis test uses hydrogen peroxide to test the reaction with haemoglobin
Useful table that summarises gram positive cocci. But learn microbiology by the condition, not the organism.

List some important gram positive bacilli.
- Listeria monocytogenes
- Propionibacterium acne - acne
- Clostridium difficile - diarrhoea from antibiotic overuse

What kind of bacteria is MacConkey agar used with? What does it contain? How does it work?
- MacConkey agar is used to differentiate gram negative bacilli
- MacConkey agar contains bile salts, lactose and neutral red (pH indicator)
- Identifies bacteria that can ferment lactose
- Ferment lactose -> produce acid -> pH drop < 6.8 = pink, pH>6.8 = normal colour

What can gram negative bacilli be split into? What is the appearance on MacConkey agar?
- Lactose fermenting and non-lactose fermenting
- Lactose fermenting = pink: E. coli, klebsiella
- Non-lactose fermenting = colourless/yellow: shigella, salmonella, pseudomonas, proteus
What is XLD agar used for?
It is a very selective growth medium used to isolate salmonella and shigella. Salmonella shows black dots.
The MacConkey agar appears yellow. Which test can be performed to decide which gram negative bacilli it is?
- OXIDASE test. This is used to determine if a bacterium produces certain Cytochrome C oxidases
- Positive result = black/purple = pseudomonas or neisseria (this is a COCCI and not a BACCILI)
- Negative result = colourless = shigella, salmonella, proteus































