Microbiology Lab Diagnostic Flowcharts

by Shishin Yamada
last updated: September 23, 1998

Gram Positive Lab Flowchart

 Gram Negative Lab Flowchart

© "First Aid for the USMLE Step 1" by Appleton and Lange.
 Bacterial Cocci
*Many thanks to Jason Collins for citing reference material for these charts
(from "The Microbiology Companion" book by ALERT and ORIENTATED publishers)

Species Gram Diagnostic Diseases

Reactions
Staphylococcus + Catalase + Abcesses
–aureus (coagulase +, beta) pale yellow Food poisoning
–epidermidis (coagulase –) colonies, cocci Toxic shock syndrome
–saprophyticus (coagulase –) in grape-like Pneumonia
clusters

Streptococcus + Catalase – Pharyngitis, rheumatic fever

–pyogenes (group A, beta) off-white small Pyoderma, acute glomeronephritis
–agalactiae (group B, beta) colonies, cocci Pneumonia
–bovis (group D, alpha) in grape-like Meningitis
–pneumoniae (alpha) clusters Subacute bacterial endocarditis
–viridans (alpha)

Enterococcus + Catalase – Endocarditis

–enterococcus faecalis white medium UTI
colonies, cocci
in long chains

Neisseria – Oxidase + Menigitis

–meningitidis small, kidney- Urethritis
(ferments maltose) bean shaped PID
–gonorrhoeae diplococci
(cannot ferment maltose)

*KEY:
Catalase = Mix with 3% hydrogen peroxide. Catalase + if forms bubbles.
Coaguase = Check for coagulase enzyme. Coagulase + if plasma clots, clumps or forms precipitate.
Oxidase = Determines presence of cytochrome-c in bacteria. Oxidase + if paper turns dark purple quickly.

Alpha = alpha hemolysis (very small clear area of hemolysis around colonies on blood agar)
Beta = beta hemolysis (large clear area of hemolysis around colonies on blood agar)
Gamma = NO hemolysis (no signs of hemolysis around colonies on blood agar)

Fermentation of specific sugars means that they change the pH of the tube over 48 hours. Fermentation is + for
when the upper third of the tube turns yellow, while the bottom two-thirds remains red.
 Gram Positive Rods
*Many thanks to Jason Collins for citing reference material for these charts
(from "The Microbiology Companion" book by ALERT and ORIENTATED publishers)

Species Spores Diagnostic Reactions Diseases

Bacillus YES Only AEROBIC Gram + Anthrax exotoxin
–anthracis rod that forms spores. Food poisoning
(non-motile, catalase +)
–cereus Colonies grow in long
(motile!) chains.

Clostridium YES Only ANAEROBIC G+ Botulism

–botulinun rod that forms spores. Food poisoning
(blocks Ach at NM junction) Wound infections
–tetani When inhibits Gas gangrene
(Inhibits GABA, glycine) neurotransmitters, often see Pseudomembranous colitis
–perfringens flaccid paralysis (Ach) or
(food poisoning, necrosis) spastic paralysis (GABA,
–difficile glycine)
(inflammatory lesion of colon)

Corynebacterium NO Selective medium: Diptheria (URI).

–diptheriae Loeffler's agar; since CDEF = Corynebacterium
difficult to separate Diptheriae blocks Elongation
Factor; so inhibits protein synth in
all eukaryotic cells

Listeria NO Catalase + DOES NOT produce exotoxins

–monocytogenes Beta hemolytic Opportunistic meningitis
Tumble-weed motility
LPS-like component
Intracellular reproduction
in mononuclear phagocytes
 ENTERIC Gram Negative Rods
*Many thanks to Jason Collins for citing reference material for these charts
(from "The Microbiology Companion" book by ALERT and ORIENTATED publishers)

Species Oxidase Growth on Glucose Diseases

Enterics
–Escherichia coli
(metallic-green on EMB)
–Shigella
(non-motile, no flagella)
–Salmonella
(S. typhi makes a little H2S,
swims like a salmon in body
for systemic infection)
–Klebsiella pneumonia
–Enterobacter aerogenes
–Proteus vulgaris
(H2S gas, "swarms")
–Yersinia pestis
MacConkey Metabolism
– + Ferments Traveller's diarrhea (ETEC)
glucose Dysentery (EIEC)
Diarrhea and vomiting (EPEC)
Hemolytic colitis (EHEC)
Very bloody diarrhea (Shigella)
Enterocolitis/gastritis (Salmonella)
Nosocomial/pneumonia (Kleb)
UTI (Enterobacter, Proteus)
Bubonic plague (Yersinia)
Many other shitty effects!

Pseudomonoadaceae + + Oxidizes Pneumonia, meningitis

–Pseudomonas aeruginosa glucose UTI, Burn infections
(smells like grapes, procyanin)

Vibrionaceae + + Ferments Cholera ("rice-water" stools)

–Vibro cholera glucose Diarrhea
(comma-shaped)

Campylobacteraceae + + Inert to Diarrhea (C. jejuni)

–Campylobacter jejuni glucose Gastritis (H. pylori)
(curved or S-shaped)
–Helicobacter pylori
(curved or S-shaped)
 RESPIRATORY Gram Negative Rods
*Many thanks to Jason Collins for citing reference material for these charts
(from "The Microbiology Companion" book by ALERT and ORIENTATED publishers)

Species Lab findings Diseases

Haemophilus influenza Coccobacillus URI
Pneumonia
Growth requires factor V (NAD) and Meningitis
factor X (heme). Found in old fashioned
(not modern) chocolate agar

Legionella pneumophila Stains very lightly Legionaire's disease

Pneumonia with fever, headache,
diarrhea, vomiting

Bordetella pertusis Selective medium: Bordet-Gengou agar Whooping cough (only humans)

ZOONOTIC (associated with an animal)

Gram Negative Rods
Species Lab findings Diseases
Francissella tularensis From rabbits, e.g. Lawnmover-vs-rabbit Caseating granulomas can form
nest case. Spread by ticks. throughout body. Intracellular
growth in monocytes and PMNs
allow it to evade antibodies and
complement-mediated lysis.