Diagnosis, Therapy, and Prevention of Bacterial Diseases 63

A
B
Gram-Positive
Cocci
Staphylococcus
Clusters
Catalase
Rods
+
Streptococcus
Pairs/chains
Catalase
Bacillus (spores)
Nocardia (branching)
Listeria (motile)
Corynebacterium
Aerobic
S. pneumoniae (optochin S; capsule)
Viridans streps (optochin R; no capsule)
hemolysis
(partial, green)
S. pyogenes (bacitracin S)
S. agalactiae (bacitracin R)
Enterococcus spp.
hemolysis
(complete, clear)
Enterococcus spp.
Peptostreptococcus
(no) hemolysis
Clostridium
(spores)
Actinomyces
(branching)
S. aureus
Coagulase +
S. epidermidis (novobiocin S)
S. saprophyticus (novobiocin R)
Coagulase
Anaerobic
Gram-Negative
Rods
Aerobic*
Cocci
(in pairs)
Neisseria
Bacteroides
Anaerobic
N. meningitidis
Maltose
Brucella
Legionella
Francisella
Bordetella
Haemophilus
(requires factors
X and V)
Campylobacter
(seagull shape)
Intracellular
growth
Pseudomonas
Vibrio (comma shape)
Oxidase Oxidase
N. gonorrhoeae
Fastidious growth
requirements
Simple growth
requirements
Maltose +
Escherichia
Klebsiella
Enterobacter
Serratia
Citrobacter
Lactose
Salmonella (H2S)
Shigella
Proteus (swarming)
Lactose +
+

Intracellular
growth +
8-1: Laboratory algorithms for identifying medically important gram-positive (A) and gram-negative (B) bacteria. Catalase, coag-
ulase, and oxidase: +, organismproduces the enzyme; , it does not. Maltose and lactose: +, organismferments the sugar; , it
does not. S, sensitive to indicated drug; R, resistant to drug. *Facultative aerobic
Mycobacterium and Nocardia: waxlike outer layer; visualized with acid-fast stains
Mycoplasma: no cell wall
Rickettsia, Coxiella, Chlamydia: intracellular bacteria
Treponema: thin, spiral organisms visualized by dark-field microscopy and fluorescent antibody staining
Borrelia: thin, spiral organisms that take Giemsa or Wright stain
BOX 8-1 GRAM-RESISTANT ORGANISMS
64 Microbiology and Immunology
2. Jests for enzymes
Catalase test. + reaction = lillles (gas formation)
a. Listingiisles stalylococci (ositive) from stretococci (negative)
Coagilase test. + reaction = reciitate/gel formation
a. Listingiisles Staphylococcus aureus (ositive) from Staphylococcus epidermidis
(negative)
Oxiuase test. + reaction = llie color
a. Positive for Pseudomonas, Neisseria, anu Vibrio secies (ositive reaction)
D. Immunologic tests (see Chapter 3)
1. Letection of sirface antigens is isefil in iuentifying some lacteria anu uistingiisling
letween strains (serotyes) of a secies tlat lave one or more iniqie antigens.
2. !or examle, serotyes of Escherichia coli ossess uifferent O antigens. Jle O157.H7
serotye are enterolemorrlagic E. coli strains.
E. Antibiotic sensitivity assays
1. Serial uilition assay
Qiantitative uetermination of urig concentration tlat inlilits growtl or kills an
organism in ciltire
a. Minimal inlilitory concentration (MC). lowest concentration of a urig tlat
inlilits growtl of an organism
TABLE 8-2 Significant Colony Characteristics
CHARACTERISTIC BACTERIA IDENTIFIED
Hemolysis in blood agar (diagnostic feature of
streptococci)
Green zone around colony ( hemolysis) Viridans streptococci (catalase negative; Optochin resistant)
Streptococcus pneumoniae (catalase negative; Optochin sensitive)
Clear zone around colony ( hemolysis) Staphylococcus aureus (catalase positive; coagulase positive)
Streptococcus pyogenes (catalase negative; bacitracin sensitive)
Streptococcus agalactiae (catalase negative; bacitracin resistant)
Swarming Proteus mirabilis, Proteus vulgaris
Pigment Formation*
Yellow colonies S. aureus
Blue-green colonies Pseudomonas aeruginosa
Red colonies Serratia marcescens
*Only certain species formpigments.
TABLE 8-1 Media for Isolating or Identifying Selected Bacteria
MEDIUM BACTERIA COMMENTS
Selective/Differential Media
Mannitol salt agar Staphylococcus aureus High salt inhibits growth of other bacteria
MacConkey agar Gram-negative enteric rods Pink/purple colonies (lactose positive): Escherichia coli,
Klebsiella species
Colorless colonies (lactose negative): Salmonella, Shigella,
Proteus species
Eosin methylene blue agar Gram-negative enteric rods Red-black colonies: E. coli
Purple colonies: Klebsiella species
Colorless colonies: Salmonella, Shigella species
Hektoen enteric agar Gram-negative rods Yellow-red colonies: E. coli, Klebsiella species
Green colonies: Shigella species
Green colonies, black center (H
2
S): Salmonella species
Special Media
Chocolate agar Neisseria and Haemophilus
species
Mediumsupplies factors V (NAD) and X (hemin)
Lwenstein-Jensen agar Mycobacteria Slow growing; colorless or buff-colored colonies:
Mycobacterium tuberculosis, Mycobacterium leprae
Tellurite agar Corynebacterium diphtheriae Gray-to-black colonies
Thayer-Martin medium Neisseria gonorrhoeae Selective chocolate agar medium
Buffered charcoal yeast
extract agar
Legionella species
4
3
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1
cells in pairs
oxidase +ve
Neisseria
Moraxella
Gram-positive
cocci rods
cells in clusters
catalase +ve
Staphylococcus
cells in pairs/chains
catalase -ve
Streptococcus
coagulase +ve
Staph. aureus
coagulase -ve
Staph. epidermidis
Staph. saprophyticus
-haemolytic
group A
(Strep. pyogenes)
group B
group C
group D
enterococci
group G
-haemolytic
Strep. pneumoniae
(optochin S)
viridans streps
(optochin R)
non-hemolytic
Strep. milleri
anaerobic streps
some enterococci
aerobic
Corynebacterium
Bacillus (spores)
Listeria
anaerobic
Clostridium
(spores)
Actinomyces
(branching)
lactose fermenters
Escherichia
Klebsiella
(mucoid)
lactose
non-fermenters
Salmonella
Shigella
Proteus
(urease +ve)
others
Pseudomonas
(oxidase +ve)
Vibrio
(comma shape)
Gram-negative
cocci rods
aerobic anaerobic
simple growth
requirements
fastidious growth
requirements
Bacteroides Fusobacterium
Legionella Haemophilus
(X & V factors)
Bordetella
Brucella
Campylobacter
(seagull shape, 43C)
organisms that stain
poorly or not at all
with Grams stain
Mycobacterium (acid fast)
Treponema
Leptospira
Borrelia
Mycoplasma (no cell wall)
Rickettsia, Coxiella, Chlamydia
(intracellular bacteria)
spiral
Figure 31.16 Identifying bacteria. The preliminary investigation of the bacteria of medical importance has traditionally been made on the basis of a few key characteristics (see text). Further identification may
then be made on the basis of biochemical and serologic tests.