Staphylococcus aureus

HPR: Normal flora, opportunistic

MOT: Auto-inoculation, direct contact (person-to-person)

POE: Skin

INC: 2-4 days (depends on host)

VFs: see below

STRUCTURAL FEATURES

Capsule: Karakawa type 5 or 8

Peptidoglycan TLR-2, TLR-4  cytokines, IL1β, IL2 = pyrogenic

Protein A: molecular mimic for FCγ receptor

Lipoteichoic Acid, Lipo-phosphate ribitol (an LPS-like material):

mimics endotoxic shock.

Plasminogen

↓ Bradykinin

Plasmin  Kininogen ------ Kinin

↓ ↑ ↓clot dissolution

Fibrinogen ------ Fibrin  multimerizes clot

X

VII Citrate inhibits these factors to keep blood from clotting

COAGULASE PROMOTES THE BOTTOM HALF OF THIS PATHWAY.

Coagulase does not need factors X and VII to clot, so S. aureus is not inhibited by citrate.

EXOENZYMES

1) Coagulase, which is a clumping factor (fibrinogen is the substrate)

A) Keeps out PMNs by coagulating blood

B) Fibrin also coats bacteria to hide it from PMNs, and forms an

obstruction to keep PMNs from being able to get through to them.

2) Catalase: Deactivates the toxicity of H2O2 H2O + O2

3) Lipase/ Phospholipase (degrades fatty acids and sebum)

Promotes sebaceous gland colonization  causes abscess in sebaceous

gland, which hardens because of coagulase  carbuncle

4) β Lactamase: degrades β lactams

5) Hyaluronidase: degrades hyaluronic acid/ neuraminic acid

-Causes Necrotizing Fascitis: necrosing tissue and moving through fascia layers.

6) Staphylokinase: mimics bradykinin (dissolves clots)

NOTE: Coagulase and Staphylokinase cannot function at the same time.

EXOTOXINS

1) Hemolysins

α hemolytic: 34 KDa transposition-mediated (“Jumping Genes”).

Lyses RBC’s, Platelets, WBC’s

β hemolytic: Degrades RBC’s, Platelets, Fibroblasts.

γ hemolytic: “Detergent”-like lysis of RBC’s, Platelets, Endothelial cells.

Leukocidin: WBC lysis. “S” subunits bind GM ganglioside,

“F” subunits bind sphingomyelin to cause WBC lysis.

Pantone Valentine Leukocidin (PVL): Creates mini-cytokine

storm to contribute to shock.

2) Exfoliative Toxin (ET)

ETA (30KDa) and ETB (25 KDa) has a trophism for squamous epithelium

and their capillaries; they injure and lyse them. “Scalded Skin Syndrome”.

3) Toxic Shock Syndrome Toxin (TSST- 1,2): A Super Antigen

TSST + Mg  MHC II / Macrophage  T cell = hypersecretes cytokinins.

4) Enterotoxins: A, B, C1, C2, D,E

NAD+ Dependent G-Protein ribosylase (like cholerae)

5) Quorum Sensing Regulon and VF gene regulation

This regulon is found on the chromosome of S. aureus. It contains the genes shown below as block arrows.

(requires RNA II) P1 P3 (requires RNA III)

Agra C D B SK Hα Hβ PVL ETA Cat

Transcript Sensor Encodes 8-AA peptide

Activator for Octap. = Octapeptide

STIMULATES GENES FOR EXOTOXINS AND EXOENZYMES

AGR = Accessory Gene Regulator

P1 = Promoter P1 (Controls AGR genes, requires RNA II)

RNA II = Regulator for AGR genes

RNA III = Regulator for exotoxins and exoenzymes

Inoculation  Coagulase to coat itself, hide, and allow it to replicate 

High cell density ↑waste, ↓nutrients, immune challenge 

triggers P1 activation  expresses RNA II  Expression of AGR genes.

Octapeptide can activate itself = AUTOCRINE = Self-Stimulating

S. aureus uses to ESTABLISH (SAR)METASTASIZE (AGR)

CoagulaseStaphylokinase

Capsule ONCapsule OFF

Protein A Toxins ON

Adhesins ONAdhesins OFF

Enzymes OFFEnzymes ON

Establish new colony

Non-Infectious Food Poisoning

Shigella / Vibrio / ETEC / S. aur. / Bacillus / Clostridium
Toxins / Shigatoxin / Choleragen / Entero-
toxin / Ent B / Ent Tox / Botulinum
INC / 12-72 h / 12-24 h / 24-72h / 1-6 h / 3-18 h / 6-12 h
Mech. / Protein synthesis / NAD G-rib / G-ribo / G-ribo / G-ribo / Ntrans inhibt
Duration / 2-4 d / 2-5 d / 2-4 d / 1-2 d / 1-2 d / 2-5 d
Diarrhea / ++++ / ++++ / ++++ / - / - / +/-
Vomiting / - / - / - / ++++ / ++++ / +/-

Pathogenesis:

Furuncle (sebaceous glands) or Carbuncles (hair follicles)  abscess /boils

Cellulitis/ soft tissue infections.

Scalded Skin Syndrome ETA/ETB dermatitis (similar to scarlet fever by Strep)

Necrotizing Faciitis

Toxic Shock

Enterointoxication (enterotoxin-mediated diarrhea). This is Dz, not infection.

Pneumonia (even though there’s no IgA protease) = 50% mortality rate =

MORE LETHAL than URI pathogens.

Pyemea (see below)

Osteomyelitis (diaphyseal = middle part of long bones). Requires 6-8 weeks of iv antibiotics

Renal Abscess infarcts (seeds from renal artery, forms abscess, clots blood beyond that site)

Endocarditis (heart valve vegetation)  destruction of valve  valvular regurgitation septic embolism

Staphylococcus epidermidis

Has fewer virulence factors, needs immunocompromised conditions

It has Protein A, but no coagulase

Pathogenesis:

Surgical wounds

Artificial Heart Valves

Prosthetic devices (hips, etc)

Catheters

NOTE: S. epidermidis is similar to Strep’s viridans group

S. aureus is similar to Strep A

ENDOTOXIC SHOCK

GRAM NEGGRAM POS

GI InfectionIngestion of organismIngestion of toxin

LPS/LOSPeptidoglycanEnterotoxin ABDC12E

CD14 and LPS-BP TLR 2,4Needs NAD+

Macrophages IL 1β, IL 2G-protein ribosylate

Cytokine Storm Fever↑ CAMP

TNFαLipoteichoic Acid Na+ and H2O leave

TSST 1, 2 Diarrheas

Hypovolemia

Streptococcus: Mostly opportunistic pathogens with various virulence factors

GROUPSPECIESPOE

Apyogenesnasopharyngeal

Bagalactiaeoral mucosa

C-G (viridans)mutans/salivariusoral mucosa

mitisoral mucosa

Enterococcus faecalisGI

Enterococcus faeciumGI

Nonpneumoniaeairway

Virulence Factors

St. pyogenes(frequent co-infection with St. pneumoniae)

1) Capsule

2) Protein M (mimics Fcγ R)

3) hemolysins

4) Streptolysin S (SLS) = O2 labile can function with or without oxygen

5) Streptolysin O (SLO) = O2 stable

6) Pyrogenic exotoxin: Chromosome-encoded by viral lysogeny (some strains)

7) Neuraminidase (like Hyaluronidase in Staph aureus)

8 DNAse

9) +/- IgG protease

10) Streptokinase (fibrinolysis): used for therapeutic medicine post heart-attack

St. pneumonia (frequent co-infection with St. pyogenes)

1) Special Capsule to deal with alveolar macrophages: Quelling Reaction (swells).

2) IgA, IgG, IgM protease

3) pneumolysin (like SLO, O2 stable)

4) Peurpera Producing Principle(PPP): targets subdermal capillary endothelial cells, causes

embolism infarcts (like N. meningitis)

St. mutans

1) Capsule

2) Dextram Sucrase: cleaves sucrose into (glucosedextrams) and (fructoselevans)

Enterococci: Capsule +/-

Pathogenesis:

ACCESS-TISSUE INTERACTION-IA-VFs-CLINICAL MANIFESTATIONS

St. pyogenes

1) Impetigo/erysipelas: Skin infections, especially around face

2) Acute purulent pharyngitis (Strep Throat)

3) Acute Necrotizing Fasciitis (using enzymes)

4) Scarlet Fever

5) Endocarditis

Non-infectious, post-infectious sequelae: From our Ab response to the Ags; confusion between MHC I and II, causes autoimmunity from pyogenes challenge.

1) Rheumatic Fever

2) Rhematoid Arthritis

3) Rheumatoid Glomerulonephritis

St. pneumoniae

1) Meningitis (#1 cause of adult meningitis)

2) Lobar pneumonia (with rusty seropurulent exudates, high fever): #1 cause of death in US due to infection

St. mutans

1) Dental caries

2) Endocarditis

Enterococcus: normal GI flora

MDR, especially VRE (Vancomycin resistant enterococcus)

SPORE FORMERS: Bacillus (aerobic) and Clostridium (anaerobic)

Bacillus cereus: Produces enterotoxins in food, which is then ingested. No sepsis.

Causes rapid onset of vomiting, but not as fast as S. aureus.

B. anthracis:

Strict aerobes

Found in soil

Long bacilli with ROUNDED ends.

Infections are from spore exposure (from soil), esp around livestock.

SPO regulon activates MEDIAL spores. Occurs when there is lack of nutrients, uv light, noxious agents (like Antobiotics), or dessication (↓water).

Growth Cycle:

SPO H2O

------------

Poor growth conditions Medial endospore Germination

HPR: Obligate pathogen, “zoonotic agent” (transmitted around other animals, esp. livestock)

MOT: Livestock/ soil aerosol or direct contact

POE: Inhalation or skin contact

INC: 4-10 days (virulent = ↓↓MOI)

VFs:

1) Polyglutamyl capsule (glutamic acid): prevents opsonization, C’, phagocytosis

2) Anthratoxin: Three component exotoxin

a) Protective Ag (PA)

b) Edema Factor (EF) Joined by a protein

c) Lethal Factor (LF)

3) FIP

4) Superoxide Dismutase (SOD) Can tolerate macrophages

5) Catalase

Px of Anthratoxin: Endospores bind to the host cell membrane (alveolar or dermal epithelial cell, or macrophage). then it germinates from body moisture, temp, and host stress. It produces anthratoxin, which causes ↑Furin response in the host. Furinis an enzyme which cleaves the linking protein of anthratoxin, releasing the toxin componants). This is exploitation of the host cell protease.

PA binds to the host’s VWF I receptor, and serves as a docking site for the toxin.

EF is a bacterial Adenyl Cyclase

LF is a metaloprotease.

Px of Bacillus anthracis

1) Edema and local lymphadenopathy (esp inhalation  mediastinal lymph nodes)

2) Papule (vesicular) goes from blue  purple  black (necrosis of cell). These

papules form in both the skin and lung forms of anthrax.

3) Eschar (temporary cap on vesicle, but organisms are not being killed).

 Leads back to #1 above, and cycle repeats.

Inhalation Anthrax = “Wool-handler’s Disease”

Kindlization: 1) kill vegetative cells and germinate spores 2) kill new veg. cells.

NOTE: Anthrax is a category “A” bioterrorism agent

Clostridium

Gram + bacilli

Strict anaerobes or microaerophilic

Soil bug

SPO

“Boxcar” (square ends) =(vegetative) --- “Drumstick” (terminal endospore)

Three main groups (Except C. difficile is normal flora)

A) Histolytic

1) C. perfringes

2) C. sporogenes

3) C. histolyticum

4) C. septicum

B) C. botulinum

C) C. tetani

Histolytic

HPR: Obligate pathogen

MOT: contact with soil

POE: crushingtrauma or compound Fx (dirty injuries with devascularization)

INC: 2-4 days

VFs: 1) Lipase

2) Protease

3) DNAse

4) SOD

5) Catalase

6) AA Decarboxylases (to use our AA to make their own AA’s)

gas production(CO2, CH4, H2S, H2)

AAdeCO2ase removes the carboxyl groups from amino acids  1° amines

1° amines: Putresceine and cadaverine

Liquifaction occurs from phospholipases, proteases, and DNAses.

All of the above in red leads to GAS GANGRENE. Frequently seen in diabetic and decubitus ulcers. Soil grown garlic used on arrowheads = Bioterrorism.

TREATMENT: Amputation

C. botulinum“sausage”

HPR: Obligate pathogen, or just intoxification (need not to be present)

MOT: Contact with soil or food thereof

POE: 1° = Ingestion

a) Infection: usually neonate (honey, etc, can contain spores)

b) Intoxication: adults

2° = Intravenous Drug Abuse (Dirty drugs and paraphenelia)

INC: 6-8 hours (intox)

4-20 days (infect)

VFs: Botulinum toxin

Px: Descending Symetric Paralysis

1) Ptosis in eyes

2) Gag/swallow reflex

3) Diaphragm paralysis (CAUSE OF DEATH) and weak limbs

TREATMENT: No organisms present, so no antibiotics

C. tetani

HPR: Obligate pathogen

MOT: Contact with soil

POE: wound, deep puncture

INC: 2-10 days

VFs: Tetani toxin

Px: Non-descending. Starts with major muscles (esp masseters = “Lock-Jaw”) and spinous muscles  arched back, breaks spinal cord.

Cause of death is from suffocation (paralysis of diaphragm)

“Rhizous Sardonis” = Sardonic smile, eyebrows up, can’t blink.

TREATMENT

Tetanus Toxoid (Ag, but not toxogenic). Not anamnestic (need vaccine every five years). No actual damage to cleft; so one can recover. Supportive treatment with intubation, wound care, antibiotics).

Phage-Mediated Transduced Toxins: A, B, E. Pt serum is injected into several mice

Anti-A anti-toxinAnti BAnti ENone

The mouse that lives is the one that has the same anti-toxin as the toxin type that isaffecting the human.

BOTLINUM TOXINTETANUM TOXIN

Targets pre-synaptic cleft Targets post-synaptic cleft

Prevents release of acetylcholine Inhibits acetylcholinesterase

 no neurotransmission Inhibits acetylcholine re-uptake

 FLACCID Paralysis  neurotransmitters hang out in cleft,

constantly triggering nerve stimulation

CHRONIC CHOLINERGIC ACTIVATION

= TRISMIC PARALYSIS

Corynebacterium

Lipid in cell envelope (like mycobacterium)

Unusual cell wall and membrane

Unusual gram +, pleomorphic

Acid fast (like M. turberculosis)

Divide by snapping fission = Chinese letter bacteria

C. diphtheriae

HPR: Obligate pathogens, humans are reservoir

MOT: droplet nuclei

POE: Inhalation  nasopharyngeal colonization

INC: 2-6 days

VFs: 1) Heat labile K Ag

2) Capsule with proteins, not just carbohydrate

3) Heat stable O Ag (like Gram neg’s): Lipo-arabinogalactan (lipid-sugars)

serves as an LPS orthologue

4) Diphtheria toxin: Two subunits with disulfide bond. ADP dependent

EF-2 ribosolase (like Shigatoxin, but trophism for nasal epithelium

instead of GI)

DIPHTHERIA TOXIN

A –-S—S-—B

Nasal Epithelial Cell

EF-2 (Elongating Factor-2; for protein syn)

(A)a

ADPTransaminase

EF-2 ribosolase

Px: Nasopharyngeal colonization 

Mucoid pseudomembrane and Necrotic NPG Epi cells death by suffocation

 possible systemic intoxification. The organisms don’t metastasize, just the toxin.

 altered CNS status, cardiomyopathy, nephropathy

The pseudomembrane is grey material (from nectrotic tissue) behind the uvula,

not made of pus. The cell dies from not being able to make protein.

This happens to unvaccinated adults and children < 1yr in other countries.

Tx:

1) Tracheostomy and intubaation

2) surgical debridement of membrane

3) Anti-diphtheria antitoxin

4) Antibiotics

Vaccine: Diphtheria toxoid: this is inactive because it’s oxidized by treatment with aldehyde. Our body responds to the protein in the inactive toxin.

Mycobacterium

Unusual G+ pleomorphic bacillus (gram variable)

“Beaded” bacillus from fission scar which doesn’t always break off

Acid-Fast

Slow growth (germination time = days instead of minutes)

M. tuberculosis TB

M. Leprae Hansen’s Diseas

M. avium intracellulare (MAI)  common cause of death in HIV

M. tuberculosis

HPR: Obligate pathogen

MOT: Droplet nuclei

POE: inhalation  lung  alveolar macrophages  survives there

INC: days-months

VFs: NO EXOTOXINS OR ENDOTOXINS

VFs:

1) Mycolate: Trehalose -6, 6’-deiycolate (polysaccharide + 60% lipid)

2) Sulfolipids (makes up 60% of the organism)

3) Intracellular Parasite

Px: Intracellular Parasite  prevents phagolysosomal fusion by coating the interior of phagosome to prevent fusion of lysosomes  eventual macrophage lysis  re-phagositized by other macrophages  cycle repeats  granulomatous lesion = accumulation of macrophages and monocytes (unlike pus, which is PMNs)

Host tries to isolate the lesion by forming calcified nodule = Ghon complex around macrophages with living organisms inside them, causing caseative (cheese-like) necrosis

REACTIVATION

GRANULOMATOUS

Aveolar MФ survival in MФ eventual MФ lysis Calcification

1° INFECTION (ACUTE) CHRONIC INFECTION

TB TEST

Mycolate and Sulfolipids can be extracted from culture to make a

Purified Protein Derivitive (PPT) injected into arm for TB test.

Positive reaction shows evidence of previous exposure (if test was recently negative, person gets one pill a day x9 months).

Then, do chest x-ray to see if there are Ghon complexes (tubercles). If so, person gets three pills a day for 12 months.

1.8 billion people have TB, out of a possible 6 billion (more common than malaria).

M. avium intracellulare (MAI)

Can be killed by type I immunity, but if the person has no CD 4+ cells, can be fatal.

M. leprae

HPR: Obligate pathogen

MOT: droplet nuclei

POE: inhalation  alveolar MФ IP  trophism for cool parts of the body

(toes, fingers, nose, eyebrows, earlobes)

INC: days - months

VFs: see M. tuberculosis

TWO FORMS OF LEPROSY

1) tuberculoid: trophism for distal Schwann cells (which make the myelin that coats nerves). Infects the cell, but doesn’t kill it  demyelenates  anesthesia in those tissues. It’s disfiguring (face skin thickens = stone face; body hair falls out)

 trauma occurs without pain  death from 2° bacterial infections.

2) Lepromatous: (seen in those with lower immunity)

 metastasis (brain, lung, spleen, liver, bone)

OTHER GENUS/SPECIES TO KNOW: Nocardia asteroides, Actinomyces israelii, Listeria monocytogenes, Erysipelothrix

ANSWERS TO SOME QUESTIONS

What does Karakawa type 5 or 8 capsule MEAN?

Karakawa was an expert on S. aureus capsules; types 5 and 8 are the most common serotypes associated with human disease; 5 vs. 8 differ in carbohydrate composition.

Can you explain the terms "Jumping genes" and "transposition-mediated"?

That would take a while – would suggest you read the following essay:

Essentially, transposons, or jumping genes, are genes that can move within a genome, and thereby become more or less active.

Regarding leukocidin: what is a "GM ganglioside" and "sphingomyelin"?

Two constituents of mammalian cell membranes not found in prokaryotic membranes; not lipids, but not sterols – sort of in the middle / hybrids.

If leukocidins mainly lyse WBC's, how does PVL cause a mini-cytokine storm?

Leukocidins can delay / deflect PMN attack, but can’t prevent it; PVL evokes hypercytokine response from PMNs and macrophages that do show up.

1