SEMINAR NOTES: PRELIMS 3 Species of Staphylococcus according to

BACTERIOLOGY pigment formation:

1. S. aureus - golden yellow pigment
GRAM POSITIVE AEROBIC COCCI 2. S. citreus - lemon yellow pigment
– these are natural inhabitants of skin & 3. S. albus - white pigment
mucous membrane in human
– can be found throughout the
environment via dust & inanimate 3.1 STAPHYLOCOCCUS AUREUS
objects – pathogenic organism that can:
– the infection are spread through direct (a). Produce toxins
contact w/ infected individual or (b). Multiply in tissues that can cause
contaminated objects inflammation
– PYOGENIC- due to accumulation
of neutrophil, bacterial cells & fluid at CULTURAL CHARACTERISTICS
the infection site – Cultivate easily in broth (uniform
turbidity)
FAMILY MICROCOCCACEAE – On plates - smooth, circular & opaque
- this group are gram (+) aerobic, colonies w/ oil like or butyrous
facultative anaerobic catalase (+) appearance
except Stomatococcus – Beta hemolytic on BAP
– Enzymes produced - urease, gelatinase,
PLANOCOCCUS lipase,hyaluronidase, staphylokinase,
- bacteria from the marine environment, beta lactamase/penicillinase,
rarely affect man leukocidin, hemolysin, coagulase

STOMATOCOCCUS Surface Characteristics that Establish &

- rare opportunistic pathogen that Course Of Infection:
causes endocarditis & bacteremia 1. Protein A
- no growth w/ the presence of 0.5% 2. Capsular polysaccharide
NaCl, 3. Peptidoglycan & teichoic acid
- Encapsulated
- Weakly catalase (+) and coagulase (-) TOXINS PRODUCED BY S. aureus:
1. Enterotoxin A & B ( heat stable CHON)
STAPHYLOCOCCUS can cause food
- frequently associated w/ human poisoning, vomiting & diarrhea
infections 2. Enterotoxin F - known as Toxic Shock
Syndrome Toxin (TSST-1)
GENERAL CHARACTERISTICS: 3. Exfoliatin - an epidermolytic toxin
- arrange in grape like cluster, in causing scalded skin syndrome
special conditions it may appear
singly or by pairs PATHOGENESIS:
facultative anaerobes ferment glucose - can cause infection in multiple organs
anaerobically (skin & its appendages)
- non-motile, non-spore former & non- - affects the skin such as folliculitis,
encapsulated boils, furuncles, carbuncles, acne, post-
- strongly catalase (+), modified oxidase operative infections, bacteremia,
(-), easily stained w/ aniline dye & endocarditis and impetigo.
produce pigment at RT
LABORATORY DIAGNOSIS
CATALASE TEST - used to differentiate 1. Gram’s stain smear - gram (+),
Staphylococcus & Monococcus from rounded, usually in clusters
Streptococcus able to reduce nitrate to nitrites 1. Has jet black colonies on Tellurite Agar
& grows in 7.5 - 10 % NaCL – due to tellurite

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 2. Mannitol (+) using MSA (indicator- isolated as contaminant or as a normal
Phenol Red, contains 7.5-10% NaCl), flora
indicated by a yellow color
3. Nitrite & Dnase (+) GENERAL CHARACTERISTICS:
4. Alpha Hemolysis Test (+) 1. gram (+) in tetrads or in cuboidal
5. Sensitive to lysotaphin & Novobiotin packets
6. Resistant to Polymyxin 2. Strict aerobes
7. Thermostable Nuclease (+) 3. 3. Utilize glucose oxidatively/non-
sacharrolytic
COAGULASE TEST - test to differentiate 4. BAP - gamma hemolytic
pathogenic from non-pathogenic Staphylococci 5. 5. Nitrate (-)
- Slide Coagulase – to detect the bound 6. 6. Grows at 5% but not at 7.5% NaCl
coagulase/clumping factor 7. Grows on 5% Sheep BA & Chocolate
- Tube Coagulase – demonstration of free Agar but not on McConkey
coagulase 8. Grows on broth

3.2. STAPHYLOCOCCUS EPIDERMIDIS LABORATORY DIAGNOSIS

- normal skin flora but it can also cause - Modified Oxidase Test - (+) dark blue
bacteremia, UTI, stitch abscess, color in 2 minutes
meningitis & endocarditis
STREPTOCOCCI
GENERAL CHARACTERISTICS: - gram (+) spherical to ovoid usually
1. Seen in clusters arranged in chains, non-motile, non-
2. Colistin Nalidixic Acid (+) spore former & non-encapsulated,
3. Mannitol (-) - white colonies facultative anaerobes.
4. Cell wall- present of glycerol
5. Sensitive to Novobiocin and growth is GENERAL CHARACTERISTICS:
aided w/ biotin 1. Capnophilic - need CO2 for initial
6. Dnase (-) isolation
7. Maltose (-) 2. Catalase (-)
8. Coagulase (-) 3. BAP - produce tiny, discrete, mucoid,
9. BAP - non-hemolytic, produce white, slightly opaque to grayish transluscent
creamy colonies, and raised growth on & pinpoint colonies
BAP 4. Insoluble in Bile, do not ferment inulin
(to differentiate them from
3.3. STAPHYLOCOCCUS SAPROPHYTICUS Pneumococci)
- Resembles to S. epidermidis 5. Susceptible to Vancomycin
- Common cause of mild & acute UTI,
bacteriura (sexually active women). CLASSIFICATION:
- Associated w/ Pyelonephritis & Cystitis a. Smith & Brown
(indwelling w/ catheters) 1. Alpha hemolytic - shows
incomplete hemolysis
GENERAL CHARACTERISTICS: (green/brown color surrounding
1. Coagulase (-) colonies - Methemoglobin)
2. Mannitol (-) and Nalidixic acid - S. pneumoniae
3. Dnase (-) - S. mitis & mutans
4. Maltose (+) (viridans streptococci)
5. BAP - non-hemolytic 2. Beta hemolytic - shows complete
6. Resistant to Novobiocin (5 mg) hemolysis on BAP (clear zone
around the colonies)
MICROCOCCI - S. pyogenes
- an opportunistic pathogen found in 3. Gamma hemolytic - do not show
immunocompromised host & maybe hemolysis on BAP
- S. faecales

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 b. Academic or Bergey’s Anthony’s Fire
c. Lancefield
Diagnostic Tests for SCARLET FEVER:
NOTE: 1. Dick’s Test
 Cell Surface Antigen produced by 2. Schultz Charlton Test
Beta – hemolytic
 Streptococcus are M protein & 2. GROUP B
Hyaluronic acid - It is the normal flora of GIT, pharynx,
and vaginal tract (S. agalactiae).
TYPES OF STREPTOCOCCI - Can cause neonatal sepsis, pneumonia
1. GROUP A and meningitis.
- most pathogenic to man (S. pyogenes - - In adult, it can cause post partum
infectious agent of bacterial endometritis, UTI, bacteremia, skin &
pharyngitis) soft tissue infections, pneumonia,
- “flesh eating bacteria” endocarditis, meningitis, arthritis and
- Susceptible to Bacitracin & Penicillin osteomyelitis.
- Oxygen labile
- Antibodies to Streptolysin O may GENERAL CHARACTERISTICS:
indicate a recent infection 1. 1. Resistant to Bacitracin & Bile
Only group w/c is PVR (+) Esculin (-)
- growth on BAP may enhance w/ the 2. The only group that produces CAMP
addition of SXT (Sulfamethoxasole) & factor - responsible for hydrolysis of
Trimethoprim Sodium Hippurate ---- CAMP TEST
- it can elaborate about 20 extracellular 3. 3. Latex Agglutination or Coagulation
products including enzymes like:
1. 1. Streptokinase 3. GROUP C
2. 2. Hyaluronidase - mostly are pathogens of animals
3. 3. Dnases - causes severe pharyngitis which may
4. 4. NADase be followed by BACTEREMIA
5. 5. Erythrogenic/pyrogenic toxin 1. S. equisimus
6. 6. Streptolysin O and S 2. S. equi
3. S. equisimilis
BILE ESCULIN TEST – is used to differentiate: 4. S. dysgalactiae
a. Enterococci from other group D 5. S. zooepidermicus
Streptococci
b. Streptococcus faecalis from Listeria 4. GROUP D
c. Group D Streptococci from other - the only group that is soluble in BILE
Streptococci ESCULIN MEDIA and grows in the
d. Group A from group B Streptococci presence of 40oC bile, grows at 45oC
and can stand up to 60oC.
LABORATORY TESTS FOR GROUP A BETA - TWO GROUPS:
HEMOLYTIC STREPTOCOCCI o ENTEROCOCCI - occur as part
1. Bacitracin Disk Test - to differentiate of the normal flora of the skin,
beta hemolytic Streptococci from other GIT,GUT and URT. But frequent
Streptococci cause of UTI’s wound infections,
2. PYR (Pyrrolidonyl Beta Naphthylamide) endocarditis
Test - to identify gram (+) cocci  Grows at 6.5% NaCl
3. Latex agglutination or Coagulation -  2. Resistant to Penicillin
using fluorescent onoclonal antibodies  3. Bile Esculin Test (+)
4. ASO Test - to identify group A  PYR Test (+)
Streptococcal infections
5. Physical manifestation of SCARLET
FEVER - can cause Streptococcal
pharyngitis, Cellulitis, Erysipelas or St.

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 o NON-ENTEROCOCCI -associated
w/ UTI, endocarditis, septecemia GENERAL CHARACTERISTICS:
- S. bovis 1. Lancet/bullet shaped arranged in pairs
- S. equines (diplococcus)
 Growth is inhibited by 2. Non-motile, non-spore former
6.5% NaCl 3. With definite capsule of polysaccharide
 Sensitive to Penicillin
 PYR (-) CULTURAL CHARACTERISTICS:
1. Isolated on BAP under anaerobic
OTHER LANCEFIELD GROUPS: incubation (candle jar or CO2 chamber)
5. GROUP F 2. Shows alpha hemolysis on BAP
- under the group is S. anginosus - 3. Young colonies - “dome shaped”
causes Cellulitis 4. Old colonies (after 48 hrs -colonies
collapse) – “nail head or checker
6. GROUP G appearance”.
- found as normal flora of the human
GIT, vaginal tract & oropharynx BIOCHEMICAL CHARACTERISTICS:
1. BILE SOLUBILITY TEST - to
7. GROUP N differentiate S. pneumoniae from other
- produce normal souring of milk, alpha hemolytic Streptococci
found in dairy products S. peumoniae (+) while S. faecalis (-)
Lactic Streptococci - S. lactis & 2. OPTOCHIN DISK TEST- same objective
S. cremoris but different species
(+) S. pneumoniae and S. mitis
ACADEMIC OR BERGEY’S CLASSIFICATION 3. POSITIVE TO NEUFELD QUELLUNG
- this is a physiologic classification REACTION - (+) Capsular swelling
4. Ferments INULIN
1. PYOGENIC - do not grow at 10oC 5. Susceptible to Quinidine
and 450C - S. pyogenes 6. FRANCIS TEST - skin test
2. VIRIDANS - grows at 450C but
not at 100C, they are alpha Antigenic Compounds Associated with
hemolytic on BAP S. Pneumoniae that Contribute to its
- S.mutans, Virulence:
- S. mitis, a. capsular swelling
- S. sanguis, b. Pneumolysin
- S. salivarius, c. M protein
- S. constellatus d. capsular polysaccharide
- S. intermedius
NOTE: they are normal flora of DIFFERENCES BETWEEN PNEUMOCOCCI
human oropharynx & important and STREPTOCOCCI
agents of subacute bacterial
endocarditis PNEUMOCOCCI STREPTOCOCCI
3. ENTEROCOCCI - grows at 100C BILE
SOLUBLE INSOLUBLE
SOLUBILITY
and 450C DOES NOT
4. LACTIC GROUP - grows at 100C INULIN FERMENTS
FERMENT
but not at 450C FERMENTATION INSULIN
INULIN
CAPSULAR
+ -
STREPTOCOCCUS PNEUMONIAE/ SWELLING
QUINIDINE AND
DIPLOCOCCUS PNEUMONIAE OPTOCHIN
SUSCEPTIBLE RESISTANT
- normal flora of the URT but can cause
otitis media, pericarditis, endocarditis
& meningitis

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 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

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

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 3. N. gonorrhea - grows on Modified GRAM NEGATIVE ANAEROBIC COCCI
Thayer Martin but not on MHA, - classified under this group are
resistant to colistin Veilonella, Megasphaera &
4. Neisseria mucosa - normal flora of the Acidoaminococcus
respiratory tract, rare cause of of
meningitis, endocarditis & cellulitis VEILONELLA
5. Neisseria polysaccharea - able to - these are gram (-) cocci arranged in
produce large amount of polysaccharide pairs, short chains or irregular clumps,
on 1-5% sucrose agar,can utilize rare cause of human infections
glucose & maltose

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
-

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