STAPHYLOCOCCUS Catalase (+)- differentiates Staph from Strep, gram +, cream colored, white rarely light gold and

buttery-looking Coagulase test- (+) stapyhlocoagulase-active enzyme Cutaneous infection- folliculitis,boils (superficial abscess) ,carbuncles (resultimg from furuncles), impetigo & purulent abscesses Toxin induced- food poisoning, scalded skin syndrome (SSS) & toxic shock syndrome (TSS)- tampon Coagulase-Negative staphylococci (CoNS)- x produce coagulase (S.epidermiditis & S.saprophyticus) S.epdermiditis- nosocomial S.saprophyticus- UTI(female adolescents&young women) Staphylokinase- lysis fibrin clots Leukocidin- lethal to Polymorphonuclear leukocytes Lipases- acts on fats and oils secreted by sebaceous glands allowing colonization of the organisms Beta hemolysis- most staphy Tube coagulase test- presence of free coagulase Slide Coagulase test- screening type, clumping factor Staphylococcus aureus Yellow pigment- nutrient agar Virulence Factors: Enterotoxins- heat stable, grp A-E & G-J Toxins- creamy sauces, mayonnaise Toxic Shock Syndrome Toxin 1- enterotoxin F Exfoliative Toxin- epidermolytic toxin -causes SSS/ ritter disease Cytolytic Toxins- a-hemolysin-damage platelets and macrophages, severe tissue damage b-hemolysin- sphingomyelin, hot-cold lysine Enzymes- coagulase, protease, hyaluronidase (spreading factor), lipase Protein A- antiphagocytic structure Infections-folliculitis,furuncles,carbuncles,bullous impetigohighly contagious skin infection, SSS, TSS, food poisoning (enterotoxins A,D,B), osteomyelitis, stye-eye infection Catalase + (bubble formation), coagulase + (gel), beta hemolysis Staphylococcus epidermidis Skin flora, most common cause of UTI Nosocomial infections:implant,catheter,pacemakers etc Non hemolytic, coagulase (-) Virulence factor: slime layer Staphylococcus saprophyticus UTIs in young women Presence in urine- significant Coagulase (-) Novobiocin susceptibility test- 5 ug/novobiocin disk S. sapro- Resistant S. epi Susceptible MRSA- methicillin resistant S. aureus MRSE- methicillin resistant S. epidermidis VRSA- vancomycin resistant S. aureus

VRSE- vancomycin resistant S. epidermidis CA MRSA- Community-associated HA MRSA- hospital acquires BORSA- borderline oxacillin-resistant S. aureus PBP- penicillin binding protein Staphylococci catalase +, gram +, spherical cells (.5-1.5um) Non motile, non spore forming, aerobic& facultatively anaerobic except for S. saccharolyticus (obligate anaerobe) Staphe bunches of grapes Colonies after 18-24 hrs medium sized (4-8um) appear cream-colored, whote, rarely light gold and buttery looking. Bullous impetigo caused by S. aureus, highly contagious. Hemolysins cytolytic toxin affect RBS Enterotoxin heat stable exotoxin cause diarrhea and vomiting Coagulase virulence marker of S. aureus Epidermolytic toxins can cause Ritters disease Staphylokinase lysis fibrin clots Stye eye infection by s. aureus Slime virulence/adhernace of s. epidermis Boils superficial abscess Novobiocin antibiotic disc used for s testing s. sapro and s. epi Hyaluronidase spreading factor Leukocidins lethal PMN Catalase test differentiate staphy from strepto Lipases acts on fats and oils secreted by sebaceous glands S. epidermidis most common cause of hospital acquired UTI Also prostethic valve endocarditis Adherence polygamma Dl glutamic acid S. saprophyticus associated w/ UTIs in young women S. lugdunensus + clumping, - tube coagulase Microscopic exam Grow on SBA. Produce round, smooth, white creamy colonies 18-24 hrs At 35-37 C. S aureus hemolytic zones around colonies and may exhibit pigment production (yellow) extend incubation S. epidermidis small-med, non hemo, white-gray S. sapro larger, 50% produce yellow pigment, S. haemo medium, with moderate/weak hemo Streptococcus S. pyogenes Grp. A S. agalactiae Grp B S. bovi, equi, C S, bovis - D Gram + resembling strep: Aerococcus, lactobacillus, leukonostic, pediococcus Hemolyis type S. pneumonia alpha; all are beta

Susceptibility to: (sensitive) Vancomycin all Bacitracin S. pyogenes, s. pneumoniae SXT S. pneumoniae, Viridans Optochin- S. pneumoniae Hydrolysis type: (+) Hippurate - S. agalactiae PYR S. pyogenes, Enterococcus CAMP S. agalactiae Types of hemolysis Alpha partial lysis of blood cells around colony -greenish discoloration of area around colony Beta complete lysis; clear area around colony *Bacitracin differentiate group A from other; conc. - 0.04u *SXT sulfamethoxazole trimethoprim - improves accuracy of GRP A + D (resistant to SXT) *Vancomycin gram + are sensitive *Optochin ethyl hydrocouprein hydrochrolide For differentiation of S. prenumoniae from other *Hippurate differentiate group B from other B hemolyitic S. agalactiae has enzyme: HIPPURASE (4 hydrolysis Na) Detected by Ninhydrin (reacts w/ it = PURPLE color) *PYR separate grp A and enterococcus strep from other Principle: S. pyogenes able to hydrolyze the substrate PYR Enterococcus substrate L-pyrro L- puyroglutenic acid B naph Group A Strep/ S. pyogenes Antigenic structure: M protein attached peptidoglycan of CW Virulence F: M protein (best defined VF) Hemolysin resp for hemolysis on BAP Streptolysin O O labile, active only in reduced form - lyses wbc, platelet, ++ -Highly antigenic - measured in ASO test - resp for SUBSURFACE hemolysis Streptolysin S = oxygen stable, non antigenic -lyses WBC, resp for SURFACE hemolysis DNases Dnase B most common Streptokinase acts on plasminogen Hyaluronidase spreading factor Erythrogenic toxin a protein exotoxin. Cause red spreading rash Clinical Infections: Bacterial Pharyngitis strep throat Pyodermal inf: Impetigo localized skin disease, very young children Erysipelas = subcutaneous tissues, elderly Cellulitis serious, life threatening, may lead to GANGRENE Scarlet fever caused by erythrogenic toxin, diffuse red rash Post-strep inf:

Rheumatic fever complication of Pharyngitis. Fever + inflammation of heart, bv, joints and subcutaneous tissues AGN acute glomerulonephritis, occur fter cutaneous or pharyngeal inf. More common in children - result of immunologic mechanisms Antimicrobial therapy: Grp A strepto susceptible to Penicillin (Erythro for allergic) Lab diagnosis: S. pyogenes =- BAP; small, transparent and smooth colonies with a well defined area of B-hemolysis Gram + cocci with short chains Transport media not rqd cause S. pyogenes is resistant to drying BAP primarily isolation medium Selective media SBA + SXT (for better recover of b-hemo strep) Biochem methods: BACITRACIN + PYR HYDROLYSIS Group B strep/ S. agalactiae Antigenic structure Group B spec. antigen acid stable polysaccharide loc CW Virulence: Capsule important, prevents phagocytosis, Sialic acid (MOST SIGNIFICANT) Neonatal meningitis found babies GRP B STREP cause madstitis cattle 1 early and 2 late infection Early 3 days after birth,mortality rate is high GRP B STREP IN VAJ most important det. Factor Late 1 weeka dn 3 months after birth. Lab diagnosis: Grp B strep grow BAP- grayish, white mucoid colonies Biochem testing. CAMP/HIPPURATE Group C S. equi, equi subsp. Group D S. canis Group D a. enterococcal found in intestinal tract. B. non entero E NON-E Bile esculin + + NaCl + Penicillin R S Enterococcus E. faecalis most common to isolate S. pneumonia aka pneumococcus Antigenic structure: concains an antigen C substance Key antigen: Capsular antigen Capsule : antigenic, QUELLENG RXN. Causes otitis,baceremia, Lobar pneumona most common Complications: Pleural effusion (EMPYEMA_ If resistant to penicillin give ertythromycin or chicamphenicol Microscopy: gram + diplococcic; lancet shapes. CAPSULES S. pneumonia requires increased CO2. (facultative anaerobe)

Young culture: round, glistening, wet, mucoid, dome shaped Old: coin with raised rim Viridans: (DO NOT USE LANCEFIELD CLASS) Most common cause of SBE Virulence factor: Dextran: allows organism toadhere to damaged v. endothelium Biochem rxn: bile insoluble, optochin resistant, 6.5 nacl no gro

Cerebrospnal meningitis - organisms brought to to cns by bloodstream N. meningitidis - glucose + maltose N. gonorrhoeae glucose only Ceftriaxone for penicillin allergic Prophylaxis Rifampicin, ciproflaxin CORYNEBACTERIUM closely related to mycobacteria. Nocardia C. diptheriae premier pathogen - facultative anaerobe,multiplication occurs 15-40 C 8 amino acids needed; ferments glucose and maltose - produce acid, not gas, not urease, reduces nitrate to nitrite Stained smear = bacilli appear in PALISADES, V and L forms Loeffler coagulated serum cells are PLEOMORPHIC - club shaped swellings Iron content for full toxin production Virulence factor: Diptheria toxin major virulence factor * only toxin producing c. diptheriae causes infection - nontoxic until exposure to trypsin Fragment a & b for cytotoxicity Fragment a for toxicity Fragment b binds to receptors on eukaryotic cells Less iron for toxin production More iron for optimal growth Clinical infections: Respiratory , Cutaneous - occurs unimmunized populations Natural host: HIMAS Site affected: UPPER RESPIRATORY TRACT The infecting toxigenic strain of c. diptheriae produces toxin locally causing tissue necrosis and exudates formation - forms a very tough gray to white PSEUDOMEMBRANE attaches tissues Treatment: Antitoxin produced in horses ANTIBIOTICS penicillin Lab diagnosis: Gram +, non sporulating, non motile Highly pleomorphuc Appears in V and L form (Chinese characters Stain irregulary in methylyne blue Metachromatic granules: babes-ernst granules -> accumulated polymerized polyphosphates Represents accumulation of food reserves Cultural char: Nutrient agar Loeffler serum agar/ pai slant w/ blood or serum mas better BAP shows very small zone of hemolysis Tinsdale agar sheeps blood, bovine, cystine, potassium tellurite Colonies: black and brown useful differentiate c. diptheriae and c. ulcerans and c. pseudotuberculosis Colonies on tinsdale medium (produces brown halo)

NEISSERIA: N. meningitidis , N. gonorrhoeae (pathogenic) Catalase and oxidase (+) Gram (-), kidney/coffee-bean shape with adjacent sides flattened non-encapsulated, non-motile, non-spore-former Aerobic and facultative anaerobe Highly susceptible to drying and exposure to unfavorable pH or sunlight grow on BAP except N. gonorrhoeae - requires added nutrients of CAP or special enrichments Thayer Martin medium Isovitalex, Vancomycin (gram + inhibit) , Colistin (gram inhibit), Nystatin (fungal inhibit) Modified Thayer Martin +hemoglobin solution, trimethoprim lactate (inhibits swarming of Proteus). Transgrow Medium + glucose, 2% agar, trimethoprim lactate and CO2 Martin Lewis Medium - anisomycin for nystatin New York City Medium Amphotericin B for nystatin Gonorrhea Neisser 1879; gonos sed; rhoia flow Pelvic inflammatory disease inflammation of uterus, fallopian tube, ovary Opthalmia neonatorum infection eye newborn, Prevented by credes prophylaxis (1% Silver nitrate) Smear: gram (-), diplococci, intracellular w/ PMN 5 % CO2 Penicillin drug of choice N. meningitidis Small, g (-), diplococcus found in and out cells VF: LPS capsule inhibits phagocytosis LPS Endotoxin complex major virulence NASOPHARYNX portal of entry Pili help attachment epithelial cells If ma reach sng organisms ang blood stream -> bacteremia Fulminant meningococcemia - more severe Waterhouse-Friderichsin syndrome - intravascular coagulation

Test for toxigenicty Diagnosis dpends on showing that the isolates produces Diphtheria toxin Listeria monocytogenes Recovered from soil,water, vegetation, animal products Listeriosis- pregnant, neonatal, elderly, immunocompromised Virulence factor- listeriolysin O (damage phagososme membrane prevents killing of organism), catalase, superoxide dismutase,phospholipase C. surface protein (p60)- induces phagocytosis MOT- ingestion of contaminated food Infections: Pregnant- seen third trimester, spontaneous abortion & stillborn neonates. Flulike (fever,headache,myalgia) Newborn- extremely serious, high fatality rate Early onset listeriosis- shortly after birth,UTI=sepsis, aspiration of amniotic acid Late- days-weeks after birth, meningitis Immunosuppressed host- chemo patients, young children, older adults (immunocompromised), high fatality rate, CNS infection & endocarditis Healthy individuals- eats contaminated food (cheese, coleslaw, chicken,) Treatment- penicillans, aminoglycosides, macrolides Ampicillin- drug of choice Neomycin susceptible Lab Dx- gram +, culture (small, round,smooth, translucent), Beta hemolysis, catalase +, motile in room temp, wet mount (tumbling), motility medium (umbrella pattern), Positive- hippurate & bile esculin hydrolysis, CAMP test (Rhodococcus equi instead of S.aureus, block type) Erysipelothrix rhusiopathiae Pleomorphic, long filaments,test tube brush appearance X spore forming, x branching, catalase (-), H2S (+) Infections: Erysipeloid- localized skin disease, 2-7 days (incubation) Septicemia- associated with endocarditis Diffuse cutaneous disease Lab DX- thin, rod shaped, gram +, Nutrient broth- 1% glucose, 5% CO2, 35 celsius Catalase -, x motile, urease (-) Bacillus Aerobic, Spore forming (endospore), X branching, catalase + B.anthracis- (fried rice bacillus) cause anthrax-herbivores (cows&sheep) biological terrorism and warfare MOT (humans) direct contact with infected animals Endospore, square ends, box car bacillus, bamboo-rod Virulence factor: capsule (glutamic acid)- X phagocytosis 3 component protein exotoxin/anthrax toxin Protective Antigen (PA)- promotes entry of EF, Edema factor (EA)- active A subunit, impairs neutrophil, massive edema Lethal factor (LF)- inactivates protein Plasmids- pX01(encodes toxin) pX02 (encodes genes necessary for synthesis of capsule)

Clinical Infetcions: Cutaneous Anthrax- black eschar, painless round black lesion w/ rim of edema, lesion= malignant pustule, mortality=20% Inhalation/Pulmonary Anthrax=woolsorters disease -spores are inhaled, mortality=90 % Gastrointestinal Anthrax- ingestion of the spore, mortality=100% Medusa Head- colony morphology Prevention- penicillin, doxyxyclin,ciprofloxacin, levofloxacin, vaccine (Protective antigen (PA)) Bacillus cereus Motile, x capsulated, penicillin resistant Infective- spores, has toxins, Diarrheal- ingestion of meat and poultry products Emetic- ingestion of fried rice Enterotoxins- heat labile , heat stable Eye infection- most common non gastrointestinal infection (endophthalmitis, panophthalmitis keratis w/ abscess formation Treatment- vancomycin, clindamycin Bacillus subtilis- common laboratory contaminant