SEMINAR NOTES: PRELIMS 3 Species of Staphylococcus according to
BACTERIOLOGY pigment formation:
GRAM POSITIVE AEROBIC COCCI
– these are natural inhabitants of skin &
mucous membrane in human
– can be found throughout the
environment via dust & inanimate
objects
– the infection are spread through direct
contact w/ infected individual or
contaminated objects
– PYOGENIC- due to accumulation
of neutrophil, bacterial cells & fluid at
the infection site
FAMILY MICROCOCCACEAE
- this group are gram (+) aerobic,
facultative anaerobic catalase (+)
except Stomatococcus
PLANOCOCCUS
- bacteria from the marine environment,
rarely affect man
STOMATOCOCCUS
- rare opportunistic pathogen that
causes endocarditis & bacteremia
- no growth w/ the presence of 0.5%
NaCl,
- Encapsulated
- Weakly catalase (+) and coagulase (-)
STAPHYLOCOCCUS
- frequently associated w/ human
infections
GENERAL CHARACTERISTICS:
- arrange in grape like cluster, in
special conditions it may appear
singly or by pairs
facultative anaerobes ferment glucose
anaerobically
- non-motile, non-spore former & non-
encapsulated
- strongly catalase (+), modified oxidase
(-), easily stained w/ aniline dye &
produce pigment at RT
CATALASE TEST - used to differentiate
Staphylococcus & Monococcus from
Streptococcus able to reduce nitrate to nitrites
& grows in 7.5 - 10 % NaCL
CAYETANO COMPILED NOTES: SEMINAR
1. S. aureus - golden yellow pigment
2. S. citreus - lemon yellow pigment
3. S. albus - white pigment
3.1 STAPHYLOCOCCUS AUREUS
– pathogenic organism that can:
(a). Produce toxins
(b). Multiply in tissues that can cause
inflammation
CULTURAL CHARACTERISTICS
– Cultivate easily in broth (uniform
turbidity)
– On plates - smooth, circular & opaque
colonies w/ oil like or butyrous
appearance
– Beta hemolytic on BAP
– Enzymes produced - urease, gelatinase,
lipase,hyaluronidase, staphylokinase,
beta lactamase/penicillinase,
leukocidin, hemolysin, coagulase
Surface Characteristics that Establish &
Course Of Infection:
1. Protein A
2. Capsular polysaccharide
3. Peptidoglycan & teichoic acid
TOXINS PRODUCED BY S. aureus:
1. Enterotoxin A & B ( heat stable CHON)
can cause food
poisoning, vomiting & diarrhea
2. Enterotoxin F - known as Toxic Shock
Syndrome Toxin (TSST-1)
3. Exfoliatin - an epidermolytic toxin
causing scalded skin syndrome
PATHOGENESIS:
- can cause infection in multiple organs
(skin & its appendages)
- affects the skin such as folliculitis,
boils, furuncles, carbuncles, acne, post-
operative infections, bacteremia,
endocarditis and impetigo.
LABORATORY DIAGNOSIS
1. Gram’s stain smear - gram (+),
rounded, usually in clusters
1. Has jet black colonies on Tellurite Agar
– due to tellurite
 2. Mannitol (+) using MSA (indicator-
Phenol Red, contains 7.5-10% NaCl),
indicated by a yellow color
3. Nitrite & Dnase (+)
4. Alpha Hemolysis Test (+)
5. Sensitive to lysotaphin & Novobiotin
6. Resistant to Polymyxin
7. Thermostable Nuclease (+)
COAGULASE TEST - test to differentiate
pathogenic from non-pathogenic Staphylococci
- Slide Coagulase – to detect the bound
coagulase/clumping factor
- Tube Coagulase – demonstration of free
coagulase
3.2. STAPHYLOCOCCUS EPIDERMIDIS
- normal skin flora but it can also cause
bacteremia, UTI, stitch abscess,
meningitis & endocarditis
GENERAL CHARACTERISTICS:
1. Seen in clusters
2. Colistin Nalidixic Acid (+)
3. Mannitol (-) - white colonies
4. Cell wall- present of glycerol
5. Sensitive to Novobiocin and growth is
aided w/ biotin
6. Dnase (-)
7. Maltose (-)
8. Coagulase (-)
9. BAP - non-hemolytic, produce white,
creamy colonies, and raised growth on
BAP
3.3. STAPHYLOCOCCUS SAPROPHYTICUS
- Resembles to S. epidermidis
- Common cause of mild & acute UTI,
bacteriura (sexually active women).
- Associated w/ Pyelonephritis & Cystitis
(indwelling w/ catheters)
GENERAL CHARACTERISTICS:
1. Coagulase (-)
2. Mannitol (-) and Nalidixic acid
3. Dnase (-)
4. Maltose (+)
5. BAP - non-hemolytic
6. Resistant to Novobiocin (5 mg)
MICROCOCCI
- an opportunistic pathogen found in
immunocompromised host & maybe
CAYETANO COMPILED NOTES: SEMINAR
isolated as contaminant or as a normal
flora
GENERAL CHARACTERISTICS:
1. gram (+) in tetrads or in cuboidal
packets
2. Strict aerobes
3. 3. Utilize glucose oxidatively/non-
sacharrolytic
4. BAP - gamma hemolytic
5. 5. Nitrate (-)
6. 6. Grows at 5% but not at 7.5% NaCl
7. Grows on 5% Sheep BA & Chocolate
Agar but not on McConkey
8. Grows on broth
LABORATORY DIAGNOSIS
- Modified Oxidase Test - (+) dark blue
color in 2 minutes
STREPTOCOCCI
- gram (+) spherical to ovoid usually
arranged in chains, non-motile, non-
spore former & non-encapsulated,
facultative anaerobes.
GENERAL CHARACTERISTICS:
1. Capnophilic - need CO2 for initial
isolation
2. Catalase (-)
3. BAP - produce tiny, discrete, mucoid,
slightly opaque to grayish transluscent
& pinpoint colonies
4. Insoluble in Bile, do not ferment inulin
(to differentiate them from
Pneumococci)
5. Susceptible to Vancomycin
CLASSIFICATION:
a. Smith & Brown
1. Alpha hemolytic - shows
incomplete hemolysis
(green/brown color surrounding
colonies - Methemoglobin)
- S. pneumoniae
- S. mitis & mutans
(viridans streptococci)
2. Beta hemolytic - shows complete
hemolysis on BAP (clear zone
around the colonies)
- S. pyogenes
3. Gamma hemolytic - do not show
hemolysis on BAP
- S. faecales
 b. Academic or Bergey’s
c. Lancefield
NOTE:
 Cell Surface Antigen produced by
Beta – hemolytic
 Streptococcus are M protein &
Hyaluronic acid
TYPES OF STREPTOCOCCI
1. GROUP A
- most pathogenic to man (S. pyogenes -
infectious agent of bacterial
pharyngitis)
- “flesh eating bacteria”
- Susceptible to Bacitracin & Penicillin
- Oxygen labile
- Antibodies to Streptolysin O may
indicate a recent infection
Only group w/c is PVR (+)
- growth on BAP may enhance w/ the
addition of SXT (Sulfamethoxasole) &
Trimethoprim
- it can elaborate about 20 extracellular
products including enzymes like:
1. 1. Streptokinase
2. 2. Hyaluronidase
3. 3. Dnases
4. 4. NADase
5. 5. Erythrogenic/pyrogenic toxin
6. 6. Streptolysin O and S
BILE ESCULIN TEST – is used to differentiate:
a. Enterococci from other group D
Streptococci
b. Streptococcus faecalis from Listeria
c. Group D Streptococci from other
Streptococci
d. Group A from group B Streptococci
LABORATORY TESTS FOR GROUP A BETA
HEMOLYTIC STREPTOCOCCI
1. Bacitracin Disk Test - to differentiate
beta hemolytic Streptococci from other
Streptococci
2. PYR (Pyrrolidonyl Beta Naphthylamide)
Test - to identify gram (+) cocci
3. Latex agglutination or Coagulation -
using fluorescent onoclonal antibodies
4. ASO Test - to identify group A
Streptococcal infections
5. Physical manifestation of SCARLET
FEVER - can cause Streptococcal
pharyngitis, Cellulitis, Erysipelas or St.
CAYETANO COMPILED NOTES: SEMINAR
Anthony’s Fire
Diagnostic Tests for SCARLET FEVER:
1. Dick’s Test
2. Schultz Charlton Test
2. GROUP B
- It is the normal flora of GIT, pharynx,
and vaginal tract (S. agalactiae).
- Can cause neonatal sepsis, pneumonia
and meningitis.
- In adult, it can cause post partum
endometritis, UTI, bacteremia, skin &
soft tissue infections, pneumonia,
endocarditis, meningitis, arthritis and
osteomyelitis.
GENERAL CHARACTERISTICS:
1. 1. Resistant to Bacitracin & Bile
Esculin (-)
2. The only group that produces CAMP
factor - responsible for hydrolysis of
Sodium Hippurate ---- CAMP TEST
3. 3. Latex Agglutination or Coagulation
3. GROUP C
- mostly are pathogens of animals
- causes severe pharyngitis which may
be followed by BACTEREMIA
1. S. equisimus
2. S. equi
3. S. equisimilis
4. S. dysgalactiae
5. S. zooepidermicus
4. GROUP D
- the only group that is soluble in BILE
ESCULIN MEDIA and grows in the
presence of 40oC bile, grows at 45oC
and can stand up to 60oC.
- TWO GROUPS:
o ENTEROCOCCI - occur as part
of the normal flora of the skin,
GIT,GUT and URT. But frequent
cause of UTI’s wound infections,
endocarditis
 Grows at 6.5% NaCl
 2. Resistant to Penicillin
 3. Bile Esculin Test (+)
 PYR Test (+)
 o NON-ENTEROCOCCI -associated
w/ UTI, endocarditis, septecemia
- S. bovis
- S. equines
 Growth is inhibited by
6.5% NaCl
 Sensitive to Penicillin
 PYR (-)
OTHER LANCEFIELD GROUPS:
5. GROUP F
- under the group is S. anginosus -
causes Cellulitis
6. GROUP G
- found as normal flora of the human
GIT, vaginal tract & oropharynx
7. GROUP N
- produce normal souring of milk,
found in dairy products
Lactic Streptococci - S. lactis &
S. cremoris
ACADEMIC OR BERGEY’S CLASSIFICATION
- this is a physiologic classification
1. PYOGENIC - do not grow at 10oC
and 450C - S. pyogenes
2. VIRIDANS - grows at 450C but
not at 100C, they are alpha
hemolytic on BAP
- S.mutans,
- S. mitis,
- S. sanguis,
- S. salivarius,
- S. constellatus
- S. intermedius
NOTE: they are normal flora of
human oropharynx & important
agents of subacute bacterial
endocarditis
3. ENTEROCOCCI - grows at 100C
and 450C
4. LACTIC GROUP - grows at 100C
but not at 450C
STREPTOCOCCUS PNEUMONIAE/
DIPLOCOCCUS PNEUMONIAE
- normal flora of the URT but can cause
otitis media, pericarditis, endocarditis
& meningitis
CAYETANO COMPILED NOTES: SEMINAR
GENERAL CHARACTERISTICS:
1. Lancet/bullet shaped arranged in pairs
(diplococcus)
2. Non-motile, non-spore former
3. With definite capsule of polysaccharide
CULTURAL CHARACTERISTICS:
1. Isolated on BAP under anaerobic
incubation (candle jar or CO2 chamber)
2. Shows alpha hemolysis on BAP
3. Young colonies - “dome shaped”
4. Old colonies (after 48 hrs -colonies
collapse) – “nail head or checker
appearance”.
BIOCHEMICAL CHARACTERISTICS:
1. BILE SOLUBILITY TEST - to
differentiate S. pneumoniae from other
alpha hemolytic Streptococci
S. peumoniae (+) while S. faecalis (-)
2. OPTOCHIN DISK TEST- same objective
but different species
(+) S. pneumoniae and S. mitis
3. POSITIVE TO NEUFELD QUELLUNG
REACTION - (+) Capsular swelling
4. Ferments INULIN
5. Susceptible to Quinidine
6. FRANCIS TEST - skin test
Antigenic Compounds Associated with
S. Pneumoniae that Contribute to its
Virulence:
a. capsular swelling
b. Pneumolysin
c. M protein
d. capsular polysaccharide
DIFFERENCES BETWEEN PNEUMOCOCCI
and STREPTOCOCCI
PNEUMOCOCCI STREPTOCOCCI
BILE
SOLUBLE INSOLUBLE
SOLUBILITY
DOES NOT
INULIN FERMENTS
FERMENT
FERMENTATION INSULIN
INULIN
CAPSULAR
+ -
SWELLING
QUINIDINE AND
OPTOCHIN
SUSCEPTIBLE RESISTANT
 GRAM POSITIVE ANAEROBIC COCCI
- These are normal flora of the bowel,
female genital tract, oral cavity, skin &
respiratory tract but can be associated
w/ abscess of the liver, brain & wound
infections

PEPTOCOCCUS
- known as anaerobic Staphylococci,
appears in clusters, Catalase (+), a rare
cause of human infections.
- P. niger

PEPTOSTREPTOCOCCUS
- Known as Anaerobic Streptococci.
- P. anaerobius - large coccobacilli in
chains, resistant to polyanethol
sulfonates (SPS), kanamycin & colistin.
- P. tetradius - appears in tetrads

Peptostreptococcus specie which is usually

appeared in tetrads:
- P. anaerobius
- P. tetradius
- Peptococcus

CAYETANO COMPILED NOTES: SEMINAR

 GRAM NEGATIVE AEROBIC COCCI
- The most important gram negative cocci
are Neisseroa and Branhamella, which
is now referred to as Moraxella
NEISSERIA
1. Non-motile, Aerobic, Capnophilic
2. All species are catalase (+) except
N.elongata
3. Fastidious organism requiring blood,
serum, cholesterol & oleic acid to
counter act growth of inhibitors
4. 4.Oxidase (+) and Cytochrome Oxidase
(+)
1. NEISSERIA MENINGITIDIS
- Bean shaped cocci
- Non-spore former, non-motile w/
virulent polysaccharide capsule
- Grows well on Chocolate agar & Thayer
Martin
- Ferments glucose and maltose
- Lactose, sucrose & fructose (-)
- Dnase (-), Beta lactamase (-)
- Do not reduce Nitrates to Nitrites
- Neufeld Quellung Reaction(+)
PATHOGENESIS: human host
MODE OF TRANSMISSION: via respiratory
tract
1. Bacterial Meningitis
2. Meningococcemia
 Waterhouse Friderichsen
Syndrome – severe form which is
characterized by high fever,
shock & adrenal insufficiency
3. Meningococcal meningitis - signs are
fever, headache, stiff neck & increased
levels of PMN’s in spinal fluid
VIRULENCE FACTORS OF MENINGOCOCCI
1. Polysaccharide capsule
2. Endotoxin
3. IgA Protease
4. Pili
SPECIMEN FOR DIAGNOSIS: Blood and CSF
CAYETANO COMPILED NOTES: SEMINAR
2. NEISSERIA GONORRHEA/GONOCOCCI
- Kidney or bean shaped diplobacilli
- Non-motile, non-spore former & no
polysaccharide capsule
- Chocolate Agar Cysteine media for
growth
- Requires immediate incubation in CO2
for growth
- Not inhibited by Vancomycin or
lincomycin, colistin and nystatin or
anisomycin
- Ferments only glucose
- Maltose, lactose, sucrose & fructose (-)
MEDIA USED FOR CULTIVATION
1. Thayer Martin Agar
2. Modified Thayer Martin
3. Transgrow
4. Martin Lewis Media
5. Cystine Tryptose Agar
6. New York City Media
7. Chocolate Agar
PATHOGENESIS
1. Gonorrhea – gonococci that causes
disease only in humans (may cause
sterility in males and females)
2. Ophalmia Neonatorium – causes
disease in newborns (gornorrheal
conjunctivitis acquired transplacental)
3. Ectopic Pregnancy and Sterility – long
term effect of infection
MODE OF TRANSMISSION: sexual contact
DIRECT METHODS FOR IDENTIFICATION OF
N. GONORRHEA
- Coagulation
- Fluorescent Antibody Test (FAT)
- ELISA
- Chemiluminescent probe
MISCELLANEOUS NEISSERIA SPECIES
1. Neiserria cineria - occasional cause of
bacteremia, conjunctivitis & nosocomial
pneumonia but normal flora of
oropharynx & ganital tract. May utilize
glucose & therefore maybe confused w/
N. gonorrhea
2. N. cineria - grows on MHA but not on
Modified Thayer Martin & susceptible
to colistin
 3. N. gonorrhea - grows on Modified
Thayer Martin but not on MHA,
resistant to colistin
4. Neisseria mucosa - normal flora of the
respiratory tract, rare cause of of
meningitis, endocarditis & cellulitis
5. Neisseria polysaccharea - able to
produce large amount of polysaccharide
on 1-5% sucrose agar,can utilize
glucose & maltose
GRAM NEGATIVE ANAEROBIC COCCI
- classified under this group are
Veilonella, Megasphaera &
Acidoaminococcus
VEILONELLA
- these are gram (-) cocci arranged in
pairs, short chains or irregular clumps,
rare cause of human infections

GROWS ON MHA BUT NOT IN THAYER MEGASPHAERA

MARTIN AND SUSCEPTIBLE TO COLISTIN
1. N. mucosa ACIDOAMINOCOCCUS
2. N. gonorrhea
3. N. polysaccharea
4. N. cinerea

BRANHAMELLA
- Also referred to as MORAXELLA

1. MORAXELLA CATARRHALIS
1. Morphologically & biochemically
resembles to Neisseria
2. Non-motile, encapsulayed organism w/
pili that serves as adhesion
3. Grows in ordinary (Nutrient agar at 35-
370C) media at RT unlike N.
meningitidis & N. gonorrhea
4. Does not ferment glucose, maltose &
lactose
5. Reduces nitrates to nitrites
6. Dnase & beta lactamase (+), Oxidase (+)
& catalase (-)
7. Non-hemolytic on BAP
8. Colonies are non-pigmented, opaque
gray & smooth

PATHOGENESIS
- normal flora of oro & nasopharynx but
can also cause bronchitis, otitis
media,sinusitis & pneumonia
-

CAYETANO COMPILED NOTES: SEMINAR