Ma. Isabella M.

Arenas
3BMT

Staphylococci
GENERAL CHARACTERISTICS

Catalase (+)
o Catalase test: Bacterial colony + H2O2 Positive result: the formation
of bubbles
Gram (+)
Spherical (cocci) cells singly, in pair or in clusters
Non-motile and non- spore forming
Aerobic or facultatively anaerobic ok with or without O2
o Except S. saccharolyticus (obligately aerobic)
Differentiated by coagulase test
o Positive result: formation of clit in plasma
o Staphylocoagulase the active enzyme in this test
o S. aureus coagulase (+)
o CoNS Coagulase Negative Staphylococci
Members of the Family Bacillaceae
o Resemble Micrococceae Family
Planococcus, Stomatococcus, Micrococcus
Catalase (+), coagulase(-), gram (+)

Staphylococcus aureus

Most clinically significant species of Staphylococci

Responsible for a number of infections both relatively mild and life-threatening

VIRULENCE FACTORS

Enterotoxins
- Heat-stable exotoxins that cause a variety of symptoms including diarrhea and vomiting
- Produced by 30-50% of Staphylococcus aureus isolates
Enterotoxins A, B, D food poisoning
Enterotoxins B, C, sometimes G, I Toxic Shock Syndrome (TSS)
Enterotoxin B Staphylococcal pseudomembranous enterocolitis
Toxic Shock Syndrome Toxin-1
- Causes nearly all cases of menstruating associated Toxic Shock Syndrome
Exfoliative Toxin (epidermolytic toxin)
- Causes skin to slough off
- Known to cause Staphylococcal Scalded Skin Syndrome (SSS)/Ritter disease
- Implicated in bullous impetigo
Cytolytic toxins hemolysins and leukocidins
Alpha hemolysin lyses erythrocytes, damage platelets and macrophages, causes
severe tissue damage
Beta hemolysin acts on sphingomyelin in RBC membrane
Gamma hemolysin Panton-Valentine Leukocidin (PVL) lethal to
polymorphonuclear neutrophils (PMNs)
 Enzymes coagulase, protease, hyaluronidase, lipase
Coagulase produced by S.aureus
Hyaluronidase hydrolyzes hyaluronic acid present in intracellular ground
substance that makes up connective tissues, permitting the spread of bacteria
during infection
Lipase act on lipids present on the surface of the skin particularly fats and oil
secreted by sebaceous glands
Protein A bind to Fc portion of IgG

CLINICAL INFECTIONS

Skin and Wound Infections

Purulent abscess
Folliculitis relatively mild and inflammation of a hair follicle or oil gland
Furuncles (boils) extension of folliculitis; large, raised, superficial abscesses
Carbuncles larger, more invasive lesions from multiple furuncles, which may
progress into deeper tissues
Bullous Impetigo superficial bacterial infection and highly contagious; usually
seen in children
Cellulitis caused by a fast-spreading infection in the dermis and the
subcutaneous tissues
Toxic Shock Syndrome multisystem disease characterized by a sudden onset of fever, chills,
vomiting, diarrhea, muscle aches, and rash
Toxic Epidermal Necrolysis it is commonly drug induced; with multiple causes
Food Poisoning

Staphylococcus epidermidis

CoNS sp.
Infections caused by S.epidermidis are predominantly hospital acquired.

VIRULENCE FACTORS

Biofilms key component in bacterial pathogenesis and is a complex interaction between

host, indwelling device and bacteria
Poly-gamma-DL-glutamic acid for adherence; protective advantage against host defences

CLINICAL INFECTIONS

Hospital-acquired Urinary Tract Infections

Prosthetic Valve endocarditis
Septicemia in immunocompromised patients

Staphylococcus saprophyticus

CoNS sp.; Novobiocin resitstant

Associated with Urinary Tract Infections in young women
The species adheres more effectively to the epithelial cells lining the urogenital tract than
other CoNS; it is rarely found on other mucous membranes or skin surfaces

Staphylococcus lugdunensis

CoNS sp.
 Can cause both community associated and hospital acquired infections
Can be more virulent and can clinically mimic S.aureus infections
Has been known to contain the gene meCA that encodes Oxacillin resistance

CLINICAL INFECTIONS

Infective endocarditis it is particularly aggressive; frequently requiring valve replacement,

and infections have a high mortality rate

CULTURE, INCUBATION AND ISOLATION

Staphylococci grow easily on routine laboratory culture media, particularly Sheep Blood Agar (SBA)

Selective media for the isolation of staphylococci:

Chapman-stone Agar
Columbia-Colistin-Nalidixic Agar (CAN)
Phenylethyl alcohol (PEA)
Mannitol Salt Agar
High Slat concentration (7.5%)
Differential: Mannitol fermentation
o Yellow: S. aureus
o Red/Pink: S. epidermidis

Colonies produced after 18-24 hours of incubation at 35-37C are medium sized and appear cream-
colored, white, or rarely light gold and butterfly-looking

Rare strains of Staphylococci are fastidious requiring COz, hemin or menadione for growth

Small colony variants grow on media containing blood, forming colonies about 1/10th the size of wild
type strains.

Streptococci
GENERAL CHARACTERISTICS:

Gram (+) cocci

Cells appear more elongated than spherical
Arranged in pairs or in chains
Catalase-negative
Weak false positive when taken from media containing blood
May be considered as aerotolerant anaerobes
o Grow in the presence of O2 but unable to use O2 for respiration
Most but not all behave like facultative anaerobes
Fermentative metabolism of carbohydrates
o Major end product: lactic acid
o No gas produced
Capnophilic

Streptococcus Pyogenes

Flesh-eating bacteria
Common Term: Group A Streptococcus
 Lancefield Group AG: A

VIRULENCE FACTORS

M-protein attached to the peptidoglycan and extends to cell surface; functions in

adherence and resistance to phagocytosis
Fibronectin-binding protein Aka Protein F; mediates adherence to epithelial cells (oral
mucosal cells)
Lipoteichoic acid secures attachment of strep to oral mucosal cells; protein F and
lipoteichoic acid mediate adhesion to host epithelial cells
Hyaluronic acid capsule weakly immunogenic; prevents opsonized phagocytosis by
neutrophils and macrophages

CLINICAL INFECTIONS

Acute Pharyngitis
o Most common along with tonsillitis
o Due to S.pyogenes and some by Group C and G
o Strep throat 5 to 15 years old
o Spread by droplets and close contact
Pyodermal Infections
o Impetigo: localized skin disease, small vesicle to weeping lesions
o Erysipelas: elderly, acute spreading lesion
o Cellulitis: develop deeper invasion by strep
o Scarlet Fever: diffuse red rash
Necrotizing Fasciitis
o Type 1 (Polymicrobial infection)
o Type 2 (Group A streptococci)
o Type 3 (Gas gangrene/Clostridial myonecrosis)
o Flesh eating bacteria syndrome
Streptococcal Toxic Shock Syndrome
o Entire organ system shuts down->death
Post Streptococcal Sequelae
o Rheumatic Fever: follows S.pyogenes pharyngitis; chronic sequel is Rheumatic heart
disease; Pathogenicity: antigenic cross reactivity, direct toxicity and actual invasion
o Acute glomerulonephritis: after cutaneous or pharyngeal infection

Streptococcus agalactiae

Common Term: Group B Streptococcus

Lancefield Group AG: B

VIRULENCE FACTORS

Capsule prevents phagocytosis; ineffective after opsonisation

o Sialic acid most significant component of the capsule, critical virulence determinant

CLINICAL INFECTIONS

Pneumonia or meningitis with bacteremia invasive disease in the Newborn

o Early onset: < 7 days old
o 80% of cases
 o Vertical transmission from mother to child
o Obstetric complications, prolonged rupture of membranes, premature birth
o Late onset: > 7 days old 3 months
o Meningitis

Group C and G Streptococci

Uncommon human pathogens

Large colony forming isolates are classified with pyogenic streptococci
Small colony performing beta haemolytic isolates belong to the S.anginosus group

CLINICAL INFECTIONS:

S. dysgalactiae subsp. equisimilis isolates were received from:

o Upper Respiratory Tract
o Vagina
o Skin of humans

Group D Streptococci

Nonenterococcus

CLINICAL INFECTIONS

Opportunistic infections

Streptococcus pneumoniae

Common Term: Pneumococcus

Lancefield AG: none

VIRULENCE FACTORS

Capsular Polysaccharide
o Antigenic, identified with appropriate antisera in Neufield Test
o Capsule swells when reacting with capsular serum
o After opsonisation, organism is avirulent
Hemolysin
Ig A protease
Neuramidase
Hyaluronidase

CLINICAL INFECTIONS

#1 cause of bacterial pneumonia

o Not usually a primary infection, rather a result of disturbance of normal defense
barriers
o Lobar Pneumonia: infecting organism in alveoli stimulate outpouring of fluid which
stops only when the fluid reaches the fibrous septa that separate lung lobes
Sinusitis, otitis media, bacteremia, meningitis, endocarditis, peritonitis, sterile pleural effusion
(empyema)

CULTURE, INCUBATION AND ISOLATION

Brain-heart infusion agar, TSA with 5% sheep RBCs, chocolate agar

 Poor growth on Nutrient Agar
Small and transparent colonies

Enterococci
GENERAL CHARACTERISTICS:

Gram (+), often occur in pairs or short chains

Facultative anaerobes
Non-spore forming
Antigenic Structure:
o Group D antigen
o Consists of gram (+) cocci that are natural inhabitant of the intestinal
tracts of humans and animals
o Most enterococci are nonhemolytic or alpha haemolytic
o Sometimes it exhibit a pseudocatalase reactionweak bubbling in
catalase test

VIRULENCE FACTORS

Virulence factors are not completely understood

E.faecalis

Extracellular surface protein (extracellular serine protease) and gelatinase play a role in the
colonization of species and adherence to heart valves and renal epithelial cells
Also produces cytolysin (two subunit toxin)

CLINICAL INFECTIONS

Nosocomial (most common UTI post catheterization)

Bacteremia often observed in hemodialysis patients, immunocompromised patients with a
serious underlying disease
Prolonged hospitalization risk factor for acquiring enterococcal bacteremia
Endocarditis in elderly with prosthetic valves or valvular heart disease
Enterococci account 5-10% of those with bacterial endocarditis
Burn patients
Sepsis

CULTURE, INCUBATION AND ISOLATION

Produced acid in CHO broth

Bile esculin agar