Normal FLora

Is found on body surfaces contiguous with the outside environment

• Is semi-permanent, varying with major life changes
• Can cause infection
Chapter 1 • General Microbiology
In a Nutshell
Definitions
Carrier: person colonized by a potential
pathogen without overt disease.
Bacteremia: bacteria in bloodstream
without overt clinical signs.
if misplaced, e.g., fe cal flora to urinary tract or abdominal cavity, or skin
flora to catheter Septicemia: bacteria in bloodstream (mul­
tiplying) with clinical symptoms.
or, if person becomes compromised, normal flora may overgrow (oral
thrush)
• Contributes to health
protective host defense by maintaining conditions such as pH so other
organisms may not grow
serves nutritional function by synthesizing: K and B vitamins
• PATHOGENICITY (INFECTIVITY AND TOXICITY) MAJOR
MECHANISMS
Colonization
(Important unless organism is traumatically implanted.)
Adherence to cell surfaces involves
• Pili/fimbriae: primary mechanism in most gram-negative cells.
• Te ichoic acids: primary mechanism of gram-positive cells.
• Adhesins: colonizing factor adhesins, pertussis toxin, and hemagglutinins.
• lgA proteases: cleaved Fe portion may coat bacteria and bind them to cel­
lular Fe receptors.
Partial adherence to inert materials, biofilms: Staph. epidermidis, Streptococcus mutans
Avoiding Immediate Destruction by Host Defense System:
• Anti-phagocytic surface components (inhibit phagocytic uptake):
- Capsules/slime layers:
Streptococcus pyogenes M protein
Neisseria gonorrhoeae pill
Staphylococcus aureus A protein
• lgA proteases, destruction of mucosal lgA: Neisseria, Haemophilus, S. pneu
• Hunting and Gathering'' Needed Nutrients:
- Siderophores steal (chelate) and import iron.
Antigenic Variation
• Changing surface antigens to avoid immune destruction
• N. gonorrhoeae-pili and outer membrane proteins
• Trypanosoma brucei rhodesiense and T. b. gambiense-phase variation
• Enterobacteriaceae: capsular and flagellar antigens may or may not be
expressed
• HIV-antigenic drift
Ability to Survive lntracellularly
• Evading intracellular killing by professional phagocytic cells allows intra­
cellular growth:
- M. tuberculosis survives by inhibiting phagosome-lysosome fusion.
- Listeria quickly escapes the phagosome into the cytoplasm before phago-
some-lysosome fusion.
• Invasins: surface proteins that allow an organism to bind to and invade nor­
mally non-phagocytic human cells, escaping the immune system. Best stud­
ied invasin is on Yersinia pseudotuberculosis (an organism causing diarrhea).
• Damage from viruses is largely from intracellular replication, which either
kills cells, transforms them or, in the case of latent viruses, may do no
noticeable damage.
• Type Ill Secretion Systems
• Tunnel from the bacteria to the host cell (macrophage) that delivers bacterial
toxins directly to the host cell
• Have been demonstrated in many pathogens: E. coli, Salmonella species,
Yersinia species, P aeruginosa, and Chlamydia
Inflammation or Immune-Mediated Damage
Examples
• Cross-reaction of bacteria-induced antibodies with tissue antigens causes
disease. Rheumatic fever is one example.
• Delayed hy persensitivity and the granulomatous response stimulated by
the presence of intracellular bacteria is responsible for neurological damage
in leprosy, cavitation in tuberculosis, and fallopian tube blockage resulting in
infertility from Chlamydia PID (pelvic inflammatory disease).
• Immune complexes damage the kidney in post streptococcal acute glomeru­
lonephritis.
• Peptidoglycan-teichoic acid (large fragments) of gram-positive cells:
- Serves as a structural toxin released when cells die.
- Chemotactic for neutrophils.
• Physical Damage
• Swelling from infection in a fixed space damages tissues; examples:
meningitis
and cysticercosis.
• Large physical size of organism may cause problems; example: Ascaris
lumbricoides blocking bile duct.
• Aggressive tissue invasion from Entamoeba histolytica causes
intestinal
ulceration and releases intestinal bacteria, compounding problems.
TOXINS
Toxins may aid in invasiveness, damage cells, inhibit cellular processes,
or trigger im­
mune response and damage.
 Structural Toxins
• Endotoxin (Lipopolysaccharide = LPS)
- LPS is part of the gram-negative outer membrane.
- Toxic portion is lipid A: generally not released (and toxic) until death of
cell. Exception: N. meningitidis, which over-produces outer membrane
fragments.
- LPS is heat stable and not strongly immunogenic so it cannot be con­
verted to a toxoid.
- Mechanism
0 LPS activates macrophages, leading to release of TNF-alpha, IL-1,
and IL-6.
0 IL-1 is a major mediator of fever.
0 Macrophage activation and products lead to tissue damageDamage to the endothelium
from bradykinin-induced vasodilation
leads to shock.
° Coagulation (DIC) is mediated through the activation of Hageman
factor.
• Peptidoglycan, Te ichoic Acids
 Exotoxins
• Are protein toxins, generally quite toxic and secreted by bacterial cells
(some gram +, some gram -)
• Can be modified by chemicals or heat to produce a toxoid that still is
immunogenic, but no longer toxic so can be used as a vaccine
• A-B (or "two") component protein toxins
B component binds to specific cell receptors to facilitate the internaliza­
tion of A.
A component i.s the active (toxic) component (often an enzyme such as
an ADP ribosyl transferase).
Exotoxins may be subclassed as enterotoxins, neurotoxins, or cytotoxins.
• Cytolysins: lyse cells from outside by damaging membrane.
C. perfringens alpha toxin is a lecithinase.
- Staphylococcus aureus alpha toxin inserts itself to form pores in the
membrane.
 ENDOSPORES
• Organisms: Bacillus and Clostridium
Function
• Survival not reproductive ( 1 bacterium � 1 spore)
• Resistance to chemicals, dessiccation, radiation,
freezing, and heat
Mechanism of resistance
• New enzymes (i.e., dipicolinic acid synthetase, heat-
resistant catalase)
• Increases or decreases in other enzymes
• Dehydration: calcium dipicolinate in core
• Keratin spore coat
Exponential Growth by Binary Fission
• Lag Phase
• Initial Phase (only 1 lag phase)
• Detoxifying medium
• Turning on enzymes to utilize medium
• For exam, n u m ber of cells at beginning
equals number of cells at end of lag
phase.
Log Phase
• Rapid exponential growth
• Generation time = time it takes one
cell to divide into two. This is deter­
mined during log phase.
Stationary Phase
• Nutrients used up
• Toxic products like acids and alkali
begin to accumulate.
• Number of new cells equals the n u m­
ber of dying cells.
• CULTURE OF MICROORGANISMS
• Obligate intracellular pathogens (viruses, rickettsias, chlamydias,
etc.) :
Tissue cultures (cell cultures), eggs, animals, o r not a t all
• Facultative intracellular or extracellular organisms: Inert lab media
(broths
and agars)
- Selective medium (S): A medium that selects for certain bacteria by

inclusion of special nutrients and/or antibiotics.

- Differential medium (D): A medium on which different bacteria can
be
distinguished by differences in colonial morphology or color.
STAINS
• GRAM-POSITIVE COCCI
• Staphylococcus
• Streptococcus
 GENUS: STAPHYLOCOCCUS
• Genus Features
• Gram-positive cocci in clusters
• Catalase positive (streptococci are catalase negative)
Species of Medical Importance
• S. aureus
• S. epidermidis
• S. saprophyticus
Staphylococcus aureus
Distinguishing Features
• Small, yellow colonies on blood agar
• �-hemolytic
• Coagulase positive (all other Staphylococcus species are negative)
• Ferments mannitol on mannitol salt agar
Reservoir
• Normal flora
- Nasal mucosa (25% of population are carriers)
- Skin
Tr ansmission
• Hands
• Sneezing Surgical wounds
• Contaminated food
- Custard pastries
- Potato salad
- Canned meats
• Predisposing Factors for Infection
• Surgery/wounds
• Foreign body (tampons, surgical packing,
sutures)
• Severe neutropenia ( <500/μL)
• Intravenous drug abuse
• Chronic granulomatous disease
• Cystic fibrosis
 Pathogenesis
• Protein A binds Fe component of IgG, inhibits
phagocytosis
• Enterotoxins: fast acting, heat stable
• Toxic shock syndrome toxin-I (TSST-1): superantigen (see
Chapter 6 of
Immunology for further explanation of a superantigen)
• Coagulase: converts fibrinogen to fibrin clot
• Cytolytic toxin (a. toxin); pore-forming toxin
• Exfolatins: skin-exfoliating toxins (involved in scalded
skin syndrome [SSS ] )
and bullous impetigo
• TReatment
• Gastroenteritis is self-limiting.
• Nafcillin/oxacillin are drugs of choice because of
widespread antibiotic
resistance.
• For methicillin-resistant Staphylococcus aureus
(MRSA): vancomycin
• For vancomycin-resistant Staphylococcus aureus
(VRSA) or vancomycin­
intermediate S. aureus (VISA): quinupristin/dalfopristin
GENUS: STREPTOCOCCUS
Genus Features
• Gram-positive cocci in chains
• Catalase negative
• Serogrouped using known antibodies to the cell wall carbohydrates
(Lancefield groups A-0 )
S. pneumoniae serotyped via capsule
- S. pyogenes serotyped via M protein
Species of Medical Importance
• S. pyogenes
• S. agalactiae (group B streptococci; GBS)
• S. pneumoniae
• Viridans streptococci
• Enterococcus fa ecalis!Enterococcus faecium
Streptococcus pyogenes (Group A Streptococcus; GAS)
Distinguishing Features
• � hemolytic
• Bacitracin sensitive
• Pyrrolidonyl arylamidase (PYR) positive
• Reservoir
• Human throat
• Skin
Transmission
• Direct contact
• Respiratory droplets
Pathogenesis
• Hyaluronic acid: is non-immunogenic
• M-protein: antiphagocytic, Ml2 strains associated with acute
glomerulonephritis
• Streptolysin 0: irnmunogenic, hemolysin/cytolysin
• Streptolysin S: not irnmunogenic, hemolysin/cytolysin
• Spreading Factors
• Streptokinase: breaks down fibrin clot
• Streptococcal DNAse: liquefies pus, extension of lesion

• Hyaluronidase: hydrolyzes the ground substances of

the connective tissues
• Exotoxins A-C (pyrogenic or erythrogenic exotoxins)
Phage-coded (i.e., the cells are lysogenized by a phage.)
Cause fever and rash of scarlet fever
0 Superantigens
Treatment
• Beta lactam drugs
• Macrolides are used in the case of penicillin allergy.
Prevention
• Prophylactic antibiotics should be considered in patients for at least 5 year
post acute rheumatic fever.
• Beta lactams and macrolides
Streptococcus agalactiae (Group B Streptococci; GBS)
Distinguishing Features
• p hemolytic
• Bacitracin resistant
• Hydrolyze hippurate
• CAMP test positive (CAMP factor is a polypeptide that "complements" the
sphingomyelinase of S. aureus to create an enhanced hemolytic pattern in
the shape of an arrowhead)
Reservoir
• Human vagina ( 1 5-20% of women)
• Gastrointestinal tract
Transmission--newborn infected during birth (increased risk with prolonged labor
after rupture of membranes)
Pathogenesis
• Capsule
• p hemolysin and CAMP factor
Diseases-neonatal septicemia and meningitis; most common causal agent
Treatment-ampicillin with an aminoglycoside or a cephalosporin
Prevention
• Prophylaxis during delivery in women with a positive vaginal or rectal cul­
ture of GBS, a history of recent infection with GBS, or prolonged labors
after membrane rupture
• Ampicillin or penicillin drugs of choice
• Clindamycin or erythromycin for penicillin allergies
.S agalactiae
• G r am (+) , catalase (-) , 13 hemolytic,
bacitracin resistant, CAMP test (+)
• Neonatal meningitis and septicemia:
#1 cause, especially in prolonged
labors