Staphylococcus 1 & 2

Staphylococci (Gram positive Cocci)

- Cell wall has: Peptidoglycan, bactoprenol, PBP (transpeptidase), lysozymes,
teichoic acid, lipoteichoic acid (only in G+)
o Negatively charged, poorly immunogenic, causes inflammation
- Non motile (but can bind to motile), does not form spores
- Catalase positive, facultative aerobe and anaerobe
- 18-40 degrees, 7.5-10% NaCl
- Catalase test: catalase dismutates H2O2 into H2O and O2

Staphylococcus aureus (Gram positive Cocci)

- Appear as colonies on agar plate; golden pigment
- Reservoirs: humans, multidrug resistant strains, opportunistic pathogen (normal or pathogenic), nares, skin
- Carriers and sources of infection: direct contact, fomites, shed from skin, food poisoning
- Ferments mannitol  turns phenol red indicator yellow on plate as it ferments mannitol
- Coagulase positive species: enzyme that clots plasma; converts fibrinogen to fibrin; protects from phagocytosis
- Virulence Factors: protein A, lipoteichoic acid (LTA), FnBP, cytolytic toxins, enzymes
o Protein A  cell wall component, binds Fc part of IgG, opsonization of staphylococcus reduced
o Lipoteichoic acid  adherence, inflammation, septic shock
o FnBP  fibronectin binding protein, cell wall component, fibronectin binding and cell aggregation
o Cytolytic toxins  hemolysins (damage membrane), leukocidins (pore formation; act on PMN)
o Enzymes  hyaluronidase (destroys ground substance), fibrinolysin/staphylokinase (dissolves fibrin),
lipases, nuclease
- Pyogenic Infection (abscesses, cutaneous and deep): local infection, caused by host compromise; forms pus
o Cutaneous  localized, can present as folliculitis, furuncle (boils), carbuncles (interconnected abscess),
cellulitis, impetigo (spreading superficial crusty lesion), wound infection
 Impetigo: contagious, superficial, purulent infection, can be pustules or crusted lesion,
pyoderma
o Deep  bacteria can pass into systemic circulation, cause bacteremia; can lead to osteomyelitis,
arthritis, acute endocarditis, and affect brain
- Toxemias (staphylococcal toxin diseases)
o Scalded skin syndrome: caused by toxic protease; primary hosts; toxin is exfoliatin; outer layers of skin
peel off, cause blisters; mainly in young children
o Toxic shock syndrome: TSST-1 toxin (superantigen); fever, rash (diffuse, but esp. on hands and feet),
chills, vomiting, diarrhea, muscle pains, hypotension, desquamation
o Food poisoning: SEB enterotoxin (superantigen), toxin ingested, heat resistance (survives cooking),
resistant to gastric juices/enzymes, short incubation period (0.5-8h), nausea, vomiting, cramps, diarrhea
- Combination of Infection and Toxemia/Antibiotic resistance
o MRSA (methicillin resistant)  PBP2a transpeptidase is resistant to methicillin; treat w vancomycin
 Community acquired: skin infections, can lead to pneumonia, if untreated - sepsis
 Hospital acquired: resists more than community acquired
 Secondary infection: children w flu can catch staph. Aureus, and leads to pneumonia
o VRSA (vancomycin resistant) 
treat with linezolid
Staphylococcus epidermidis (Gram positive cocci)
- Responsible for prosthetic infections
- Catalase positive, coagulase negative
- Virulence factor  capsule or slime layer (binds to tissue and foreign bodies) forms biofilm
- Novobiocin sensitive
- MSSE (methicillin sensitive)  bacteria can secrete beta-lactamase, sensitive to methicillin, nafcillin also works
- MRSE (methicillin resistant)  vancomycin w rifampin (treat like MRSA)

Staphylococcus saprophyticus (Gram positive cocci)

- Catalase positive, coagulase negative
- UTI; cystitis (inflammation of bladder); #2 cause of UTI behind E.coli, young women at risk
- Pathogen attacks normal flora of GI and vaginal tracts, and perineum
- Virulence factors  biofilm, urease (degrades urea into C02, increases pH and helps bacteria grow)
- Novobiocin resistant, treat with trimethoprim-sulfamethoxazole (TMP-SMX)

Hemolysis on Blood Agar Plate: contains 5% sheep blood that produce hemolysins that lyse RBC
- 𝛂-Hemolysis partial; surrounded by a green, opaque zone
- 𝝱-Hemolysis  complete; a clear zone surrounding the colonies
- S. aureus
- γ-Hemolysis  no hemolysis
Streptococcus 1 & 2

Streptococci (gram positive cocci)

- Catalase negative, non-motile, facultative anaerobe, arrange in chain/pairs, fastidious
- S. pyogenes (strep throat, skin), s. agalactiae (neonatal sepsis), s. pneumoniae (pneumonia, otitis media,
meningitis), viridans group (caries, endocarditis), enterococcus faecalis, nonenterococcus s. bovis

Streptococci classification
- Brown classification: based on hemolysis
o Alpha (partial), beta (complete), gamma (none) hemolysis
- Lancefield classification: based on serology (surface antigen), looks at C carbohydrate in cell wall
o Group A (GAS): s. pyogenes
o Group B (GBS): s. agalactiae
o Group D: enterococcus faecalis, s. bovis
o Non groupable: s. pneumoniae, s. mutans viridian (major pathogen in caries)

Streptococci pyogenes (Group A strep, GAS)

- C carbohydrate in cell wall, beta hemolytic, capsule has hyaluronic acid (non-antigenic), habitat virulence,
transmitted by aerosols, droplets, or fomite coming on contact with broken skin
- Virulence factors:
o M protein  ANTIPHAGOCYTIC, AUTOIMMUNE RHEUMATIC FEVER, INHIBITS OPONIZATION,
ADHERENCE. Responsible for pharyngitis, skin infection, rheumatic fever, nephritogenic. 3 repeat
regions (A – interacts with fibrinogen, B – reacts with cardiac proteins, C – inhibits complement)
- Model for GAS inflammasome activation
o Lipoteichoic acid (LTA), part of gram-positive bacteria, is recognized by PRR, and induces inflammasome
activation, releasing IL-1beta
o Soluble M protein undergoes clathrin-mediated endocytosis, causing potassium efflux and
inflammasome assembly. Drives inflammatory responses.
- S. pyogenes exotoxin
o Streptococcal pyrogenic exotoxins (SPE): erythrogenic toxin, induces fever, rash, toxic shock like disease
 SpeA – bacteriophage transduced; lysogenic conversion, superantigen
 SpeB – protease; degrade antibodies, cytokines, C3b; destroys tissues, causes necrotizing
fasciitis
o Cytolytic toxins: pore forming, affect RBCs,
 Streptolysin O (SLO): oxygen sensitive, antigenic, ASO test for rheumatic fever
 Streptolysin S (SLS): oxygen stable, beta hemolysis
- Invasive Factors (exoenzymes)
o Streptokinase (fibrinolysin)  breaks down clots, used with Streptodornase (DNAase, also helps break)
o C5a Peptidase  inactivates C5a, reduces inflammation, chemotaxis affected
o Hyaluronidase  hydrolyses hyaluronic acid
o Protein F  binds fibronectin, clumps bacteria
- Suppurative Diseases – soft tissue infections
o Pharyngitis – strep throat; fever, swollen and sore throat; all at risk, esp. children 5-15 y/o; penicillin
used to prevent rheumatic fever
o Scarlet fever – rash on upper chest spreading to extremities, strawberry tongue, erythrogenic toxin, tx
with penicillin
o Impetigo – streptococcal pyoderma; crusty amber scab; face and limbs; children at risk; tx with
mupirocin (topical) or penicillin (if systemic)
o Erysipelas- fiery red infection on skin of face or lower limb
o Cellulitis
 o Necrotizing fasciitis – invasive GAS disease, tissues destroyed, tx with early and aggressive debridement
o Puerperal fever – childbirth fever, new mothers, uterine infection
- Non-suppurative complications
o Rheumatic fever – autoimmune, starts with pharyngitis; fever, joint pain, rheumatic heart disease;
preventable; abx prophylaxis
o Poststreptococcal glomerulonephritis – kidney disease, starts w pharyngitis or impetigo; hematuria,
proteinuria, hypertension, ASO titer elevation
- S pyogenes lab ID
o Catalase negative, group A antigen, hemolysis, bacitracin sensitive, positive PYR test
- Treatment  drainage and debridement, penicillin or clindamycin abx

Streptococcus agalactiae (Group B strep, GBS)

- Short chain, beta hemolytic, catalase negative, capsule, found in upper
resp. and GI tracts, bacitracin resistant
- Neonatal meningitis, pneumonia, bacteremia  transmitted from
neonate to mother during pregnancy or birth; screening prevention;
intrapartum penicillin abx treatment
- CAMP test  GBS produces CAMP factor; enhances hemolytic activity;
appears as larger clear area on test plate
- Hippurate test  purple color indicates positive test for GBS

Streptococcus pneumoniae (non-groupable)

- Alpha hemolysis; highest in children; endogenous (nose  distal); children,
elderly, abnormal resp tract, splenectomy are at risk
- Causes meningitis (most dangerous), Otitis media, pneumonia, sinusitis
- Virulence factors
o Capsule  antigenic
o Cytolytic toxins  pneumolysin, released after autolyin acts,
destroys membranes and hemoglobin, local ifxn
o IgA protease  colonize oropharynx and spread
- Lab ID  alpha hemolytic, green around colony, sensitive to antibiotics,
optochin sensitive, bile soluble, capsular swelling test, can perform multiple tests at same time
- Treatment and prevention: penicillin, cephalosporins, erythromycin, vancomycin (only for critically ill)
o Vaccine: 1. BCR bind to polysaccharide, 2. B cell internalize and degrades conjugate, 3. B cells display
toxoid derived peptides on surface MHC II, 4. Helper T cells and recognize and activate the B cell, 5. B
cell produce anti polysaccharide antibody

Streptococci Viridans (non-groupable)

- S. mutans, viridans streptococci in in oral flora; s. sanguinis in subacute bacterial endocarditis (SBE); prevention
of SBE in dental patients at risk; antibiotics before and after surgery; prophylaxis for high-risk patients

Enterococcus - E. faecalis and E. faecium (Group D antigen)

- D antigen: a glycerol teichoic acid, gamma hemolysis
- Normal microbiota of human intestines, found in feces, avirulent usually, but nosocomial infections (fomites)
- At Risk: hospitalized for prolonged periods and dysbiosis after antibiotic treatments
- Most infections are endogenous; urinary tract, blood
- Causes UTI, Peritonitis (usually polymicrobial), wound infections, bacteremia (w or w/o endocarditis)
- Lab ID  Gram stain, 6.5% salt
- Treatment  MDR: almost all antibiotics, VRE strains: inherently vancomycin resistant
- Prevention: Restriction of antibiotic use, appropriate infection-control practices

Nonenterococcus - S. bovis (Group D antigen)

- γ-hemolysis, causes septicemia and infective endocarditis in elderly and immunocompromised
 - Virulence factor: capsule & pili
- Opportunistic bacteria, strongly associated to colorectal cancer (CRC), 80% of patients may have CRC
Microbiology of Dental Caries

Caries  Localized destruction of the tissues of the tooth by bacterial fermentation of dietary carbohydrates. A
multifactorial plaque-related chronic infection of the enamel, cementum or dentin.

WD Miller 1890  showed carb containing food result in acid formation in vitro (chemo-parasitic theory); caries caused
by multiple species, but not specific one (non-specific plaque hypothesis); prevent by removing bacteria (brush/floss)

Polysaccharide production by S. mutans – extracellular enzymes that split sucrose

- Glucosyltransferases  form soluble and insoluble glucans
- Fructosyltransferases  form soluble fructans

Acidogenicity and Aciduricity

- Sugar can drop pH from 6.5 to 5.5 in 5 mins
- Critical pH – 5.5, when saliva and plaque aren’t supersaturated with calcium and phosphate, losing enamel
- S. mutans survives by maintaining glycolysis at low pH, repairs DNA and protein damage due to acid, and keeps
intracellular pH more alkaline than environment

Acquired Pellicle
- S. mutans has cell surface protein adhesin (Ag I/II) that binds to pellicle. Allows for 1 and 2 colonizers

Lactobacillus (Gram positive Rods)

- Positive correlation with caries; acidogenic (makes lactate and acetate), aciduric (initiate and grow at low pH)
- Negative correlation with caries; low tooth surface affinity, low # in early plaque, # decreases in saliva after
caries
- Thus, not involved in initiation of caries, but involved in progression of lesion

Actinomyces (Gram positive Rod)

- Aciduric bacteria, anaerobic, gingival crevice, part of root caries, gives scaffold for other bacteria to progress

Veillonella parvula (Gram negative Cocci)

- Found in plaque, anaerobic, asaccharolytic, consumes lactic acid (increases pH), helpful bacteria

Prevention  less sugar, sealants, reducing cariogenic flora, fluoride

Bacillus, Clostridium, Corynebacterium, Listeria

Bacillus (Gram positive Rod)

- Aerobic, spore forming, exotoxin important in pathogenesis; two types – anthracis and cereus

Bacillus anthracis (aerobic, spore forming, non-fastidious, non-hemolytic, capsule)

- Causes anthrax, spores present in soil, picked up by animals, produces 3 exotoxins (inhale, cutaneous, intestine)
- Capsule  nontoxic, protects organism against bactericidal components, helps establish infection
- Exotoxins (need protective antigen in order to have effect, all 3 produces edema + necrosis + lethal)
o Protective antigens (PA)– responsible for binding to host
o Edema antigens (EF) – protective antigen + adenylyl cyclase (activated by calmodulin)
o Lethal toxin (LF) – protective antigen + zinc metalloprotease (zinc dependent)
- Mechanism of anthrax toxins
o Protective antigen binds to anthrax toxin receptor (ATR)
- Genetics of B. anthracis
o Ames strain has pX01 which makes toxin and pX02 which makes capsule
o Sterne strain has only pX01, used for vaccine
- Discomfort, low fever, flu and GI symptoms, sudden onset hyper acute illness, dyspnea, cyanosis, high fever,
disorientation, then shock coma and death
- Diagnosed by characteristic papule, eschar with surrounding edema, history of animal exposure
o Cutaneous always easy to diagnose
o GI and pulmonary harder; don’t notice until fulminant phase, high mortality
- Lab ID  gram positive rods, not fastidious, rapid growth, non-hemolytic, non motile, hydrolysis neg, catalase
positive,
- Tx: penicillin, cipro. Host defenses – vaccines eliciting humoral and cell mediated response

Bacillus cereus (Gram positive rod)

- Spore forming, aerobic, nonfastidious, virulence not well defined, motile, positive lab tests
- Produces heat stable enterotoxin, necrotic toxin, heat liable toxin, cereolysin, phospholipase C
- Opportunistic pathogen, food poisoning, causes gastroenteritis (increases cAMP, superantigen)
- Emetic form (heat stable) causes nausea and vomiting, <6h incubation, 8-10 hour duration, rice
- Diarrheal form (heat liable) is slower onset shits, >6h incubation, 20-36 hour duration, meat veggies
- Tx: vancomycin, clindamycin, ciprofloxacin, gentamicin, NO penicillin or cephalosporin

Clostridium (Gram positive Rod)

- Strict anaerobic, spore forming, exotoxin important in pathogenesis, several strains, similar, different pathogenic
islands; deadly  tetani, botulinum, perfringens/gangrene, less deadly  difficile. Underdeveloped countries
- All motile except perfringens, all spore forming, all have multiple exotoxin except tetani
- Found in soil, poorly canned food. Prevent by keeping shit clean, there is vaccine now
- Tx: penicillin, metronidazole, vancomycin, chloramphenicol

C. tetani
- Tetanospasmin, heat liable neurotoxin, causes spasms and convulsions and tetanolysin, a hemolysin
- Transmitted from soil via puncture wounds, germinate, and produce toxins that attack inhibitory cells
- Results in cramping and twitching around wound, auditory hyperacuity, pain in neck, lockjaw, fixed smile,
seizures. Lab ID done by culture
- Presents as generalized tetanus (most common), cephalic (high mortality), localized/wound (good prognosis),
neonatal tetanus (high mortality)

C. botulinum
- Nerve toxin, very potent, within hours, blurred vision, difficulty swallowing, breathing, flaccid paralysis
 - 8 exotoxins; heat liable neurotoxin, A B E most toxic, blocks Acetylcholine release at NMJ
- Food poisoning; weak, dizzy, blurred vision, dry mouth, abd. pain, no fever, neurotoxin, no lab ID
C. perfringens
- Gas gangrene, spores, exotoxins (lectinases, hemolysins, collagenases, proteases, lipases), causes massive tissue
damage, (skin and tissue dies as blood flow stops), intense pain, foul smell, turns black, cut off tissue
- Treated with abx, hyperbaric chamber, amputation

C. difficile (pseudomembranous colitis, PC)

- Abx therapy destroys normal flora, toxin A (enterotoxin) causes GI symptoms, toxin B (cytotoxin) kills cells
- Abdominal pain, watery diarrhea, diagnosed by colonoscopy, cytotoxic and immunoassays do lab ID

Corynebacterium (Gram positive small Rod)

- Non spore forming, exotoxin important in pathogenesis, non acid fast, non motile, catalase positive

Cornebacterium diphtheria
- Causes diphtheria, colonizes mucous membranes of resp. tract; toxicity due to tox exotoxin
- Tox exotoxin found on bacteriophage; normally under control of DtxR repressor gene; low iron turns DtxR off
- Also produces diphthin, a protease that inactivates IgA
- Two subunits  A subunit (catalytic region; creates biochemical rxn) and B subunit (receptor binding region;
binds toxin to cell + translocation region; brings toxin into cell)
- DT monomer binds to HB-EGF-like receptor  undergoes endocytosis  A domain cleaved and inhibits
elongation factor 2 (EF-2)  protein synthesis inhibited  cell death
- Part of resp. system/tract; can be commensal or toxic depending on tox genes.
- Presents as muscle weakness, sore throat, low fever, pseudomembrane on tonsils; later is damage, bleeding,
difficulty breathing; complications include heart failure, systemic spread
- Elek test, abx treatment wth penicillin, erythromycin, gentamicin; vaccinations

Listeria (Gram positive short Rod)

- Non spore forming, exotoxin not important in pathogenesis, aerobic, catalase positive, food poisoning
- Motile at room temp, capable of growth at low temp, cause listeriosis
- Adherence and invasion; exotoxins (hemolysin kills blood and zinc metalloprotease causes GI issues)
- Move in macrophages and move within cells to avoid clearance, have actin tail

L. monocytogenes
- Infection from food, high # monocytes, mortality >25%, neonatal infection (early onset in utero, late onset after
birth as meningitis), adult infection (meningitis, stiff neck, headache, coma). Abx with penicillin, ampicillin,
gentamicin, but no vaccine

Actinomyces (A. israelii)

- Anaerobic gram positive bacilli; form filaments with branch, lack mitochondria, not acid fast, sulfur granules
- Low virulence potential, occurs after disruption of mucosal barriers by trauma, surgery or infection, upper resp.
- No major toxins, presence of sulfur granules confirms actinomyces; intact mucosa first line of defense
- May present as acute pyogenic infection; more commonly as chronic, suppurative, granulomatous infection
- Characterized by multiple abscesses and interconnecting sinus tract
- Cervicofacial actinomycosis (most common) associated with poor oral hygiene, invasive procedure, trauma
- Swelling, red, hard lump on face or upper neck, fever, weight loss, minimal pain, draining sores
- A viscosus – involved with gingivitis, produces succinate, a growth stimulating factor for P. gingivalis. Surgical
removal of tissue as antibiotics not effective against diseased tissue

Nocardia (gram positive rod)

- Aerobic, filaments, contain mycolic acid, weakly acid fast, nocardia asteroids causes nocardia pulm. infxn
- Nocardosis: occurs in ppl w weak immune system; starts in lungs (pulmonary), can cause cutaneous and brain
- Diagnosed by bacterial culture; identified with acid fast stain
 - Long term abx therapy required; sulfonamides for 6 months
- Eg N. asteroides
Gram Negatives, Enterics and E. coli

Lactose present  E. coli, Klebsiella, Enterobacter

No lactose present  Shigella (no H2S), Yersinia (no H2S), salmonella (H2S), proteus (H2S)

Large intestine has most bacteria, mostly anaerobes and facultative anaerobes