BACTERIAL EYE PATHOGENS Dr. WILLIAM J. BENJAMIN Eye Physiology & Ocular Prosthetics Laboratory University of Alabama at Birmingham University of Alabama at Birmingham School of Optometry Presented at the UAB School of Optometry as Part of MIC 200: MICROBIOLOGY and IMMUNOLOGY Otb 30 & N b 3 2008 October 30 & November 3, 2008
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BACTERIAL EYE PATHOGENS
Dr. WILLIAM J. BENJAMINEye Physiology & Ocular Prosthetics Laboratory
University of Alabama at BirminghamUniversity of Alabama at BirminghamSchool of Optometry
Presented at the UAB School of Optometry as Part ofMIC 200: MICROBIOLOGY and IMMUNOLOGY
O t b 30 & N b 3 2008October 30 & November 3, 2008
SPREAD OF VIRULENT PROPERTIESPlasmid Exchange Between BacteriaPlasmid Exchange Between Bacteria
INCREASED PURULENCE: PABA in Puru-lence a Contraindication to Sulfonamides
HOW ANTIBACTERIALS WORK
INHIBIT CELL WALL SYNTHESISCell Lysis or Alters Cell ShapeIron Acquisition Proteins in Cell WallElectron Transport System in Cell WallEXAMPLES: Penicillins CephalosporinsEXAMPLES: Penicillins, Cephalosporins
Bacitracin, VancomycinINHIBIT DNA / RNA SYNTHESIS
Bind DNA Polymerase or DNA GyraseEXAMPLE: Rifamycin
HOW ANTIBACTERIALS WORK
INHIBIT CELL MEMBRANE FUNCTIONLeakage of Cell ContentsIntracellular Build-up of Toxic Byproducts“Starve” Cell of Nutrients, MetabolitesEXAMPLE: PolymixinsEXAMPLE: Polymixins
INHIBIT PROTEIN SYNTHESISAt the Ribosome (30S)At the Ribosome (30S)Blocks tRNA from Forming ProperlyEXAMPLE: Aminoglycosides
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
Staphylococcus aureus
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
STERILE SALINE; NO ANESTHETICDACRON SOAKED IN CALCIUM ALGINATEDACRON SOAKED IN CALCIUM ALGINATEDACRON OR COTTON TIP APPLICATORSSWAB INNER LID MARGIN
Lower Preferred, but …..STREAK MARKED AGAR PLATE & DISPOSESWAB PALP CONJ & C-d-SSWAB PALP CONJ & C-d-S
Lower Preferred, but …..STREAK MARKED AGAR PLATE & DISPOSE
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
INCUBATE AT 35 DEGREES FOR 24-48 Hrs.OR SEND TO MICRO LABORATORYOR SEND TO MICRO LABORATORY
DO NOT WAIT TO START ANTIBIOTICSKEY AGAR PLATES FOR THE O.D. OFFICEBlood Agar: Use for EverythingChocolate Agar: If Suspect Neisseria, etc.Sabouraud’s Agar: Fungi; 25 Deg 2-10 DaysSabouraud s Agar: Fungi; 25 Deg, 2-10 DaysCULTURE TRANSPORT PACKS: Not Good,
but sometimes necessary
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
OCULAR PATHOGENS REQUIRINGCHOCOLATE AGAR + COCHOCOLATE AGAR + CO2
CULTURE TRANSPORT PACKS: Not Good, but sometimes necessaryy
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
CULTURING OF BACTERIA FROM THE CONJUNCTIVA,
AND EYELIDS
CULTURING OF BACTERIA FROM THE CONJUNCTIVA AND
EYELIDS
CULTURING OF BACTERIACULTURING OF BACTERIA FROM CORNEAL ULCERS
STERILE SALINE; ANESTHETIC REQUIREDKIMURA SPATULA but not CYTOBRUSHDACRON SOAKED IN CALCIUM ALGINATEDACRON OR COTTON TIP APPLICATORSSCRAPE CENTER AND LEADING MARGINSCRAPE CENTER AND LEADING MARGIN
CORNEAL PATHOGENSCORNEAL PATHOGENSWITH SPECIAL AFFINITY FOR
CORNEAL EPITHELIUM
Streptococcus pneumoniaeStreptococcus pneumoniaePrefers Corneal Epithelial Cells in generalBe sure to scrape the “leading edge”
Pseudomonas aeruginosaPrefers Basal Corneal Epithelial CellsBe sure to scrape the center at bottomBe sure to scrape the center at bottom
Staphylococcus aureus
CULTURING OF BACTERIACULTURING OF BACTERIA FROM CORNEAL ULCERS
STREAK AGAR PLATEDISPOSE OR FLAME: Be sure to let coolINCUBATION AT 35 DEGREES FOR 24-48 Hr.
OR SEND TO MICRO LABORATORY50% of Microbial Ulcers WON’T GROW OUT50% of Microbial Ulcers WON T GROW OUT
CULTURING OF BACTERIACULTURING OF BACTERIA FROM CORNEAL ULCERS
50% of Microbial Ulcers WON’T GROW OUTTherefore, many attempts are “Culture-Negative”
CULTURING OF BACTERIACULTURING OF BACTERIA FROM CORNEAL ULCERS
50% of Microbial Ulcers WON’T GROW OUTUlcers should not be called “Sterile”
CULTURING OF BACTERIACULTURING OF BACTERIA FROM CORNEAL ULCERS
50% of Microbial Ulcers WON’T GROW OUTSuch an Ulcer is a “Culture-Negative Ulcer”
but is not necessarily a “Sterile Ulcer”but is not necessarily a Sterile Ulcer
CULTURING OF BACTERIACULTURING OF BACTERIA FROM CORNEAL ULCERS
Culture-Negative Macular Keratitis turned Ulcer
Started on TobramycinStarted on Tobramycin Ophthalmic Drops every 2
hours as dual Macular Keratitis with Anterior
Chamber Reaction
Intensified to dual Ulcers overnight with greater ACRovernight with greater ACR and Corneal Edema, VA ↓
Fortified Tobramycin and F tifi d V iFortified Vancomycin
alternating every half hour
CULTURING OF BACTERIA:CULTURING OF BACTERIA: The Hospital Micro Laboratory
CULTURING OF BACTERIA:CULTURING OF BACTERIA: The Hospital Micro Laboratory
IDENTIFICATION OF BACTERIAIDENTIFICATION OF BACTERIA FROM CULTURES
GRAM STAIN FOR (+) or (–), CELL MORPHOL.DIFF QUIK or GIEMSA STAIN SMEARS FOR
CLINICAL PEARLS ABOUTCLINICAL PEARLS ABOUT BACTERIAL SUSCEPTIBLITY
Sodium sulfacetamide, a bacteriostatic agent, with 84% in vitro coverage of all isolates, and an average cost of $13.11, appeared to be most cost-effectivecost of $13.11, appeared to be most cost effective for empirical coverage of bacterial conjunctivitis.The fluoroquinolones, although more expensive and of recent introduction gave only similar coverageof recent introduction, gave only similar coverage.
PERHAPS RESISTANCE FADED AWAY AFTER 20PERHAPS RESISTANCE FADED AWAY AFTER 20 YEARS OF DISUSE FOR SODIUM SULFACETAMIDE!
Will there be a 20-year recycling effort?
MICROORGANISMS THAT CAN INFECT (PENETRATE ?) THE
INTACT CORNEA
Neisseria gonorrhoeaeStreptococcus pneumoniaeCorynebacterium diphtheriae (+ other sp.)Haemophilus sp (H aegyptius H influenzae)Haemophilus sp. (H. aegyptius, H. influenzae)Listeria sp. ( L. monocytogenes)
BACTERIA THAT CAUSE MAJORITY OF
CONJUNCTIVITIS IN CHILDREN
Streptococcus pneumoniae (< 5 years old)Haemophilus aegyptius (< 5 years old)Staphylococcus aureus (> 5 years old)
INSTANCES WHEN MICRO LABINSTANCES WHEN MICRO LAB STUDIES ARE MANDATORY
NEONATAL CONJUNCTIVITISHYPERACUTE CONJUNCTIVITISMEMBRANEOUS CONJUNCTIVITISCORNEAL ULCERS not obviously HerpeticCORNEAL ULCERS not obviously HerpeticPOSTOPERATIVE EYE INFECTIONSSEVERE LONG-STANDING CONJUNCTIVITISSEVERE LONG STANDING CONJUNCTIVITISSUSPECTED FUNGAL EYE INFECTION
MICROORGANISMS CAUSING CONJUNCTIVAL MEMBRANES orCONJUNCTIVAL MEMBRANES or
UBIQUITOUS; SPORE FORMING G(+) RodEXOTOXIN: Highly Virulent PathogenPENETRATING EYE TRAUMAMETASTATIC INFECTION OF DRUG ADDICTMETASTATIC INFECTION OF DRUG ADDICTPOST-TRAUMATIC ENDOPHTHALMITISCORNEAL RING ABCESS WITHIN 24 Hrs.CORNEAL RING ABCESS WITHIN 24 Hrs.EYE IS BLIND WITHIN 72 Hrs. OF INFECTION
IgA -ase, OUTER MEMBRANE ADHESINSPILI (FIMBRIAE)IRON ACQUISITION PROTEIN IN CELL WALLQUICKLY KILLED BY HEAT DRYING SUNQUICKLY KILLED BY HEAT, DRYING, SUNMUTATED VERSIONS HAVE RESISTANCE:
G(–) ROD, SUBSPECIES OF H. influenzaeRARELY CAUSES SYSTEMIC ILLNESSACUTE CONJ’ITIS WITH BRIEF INCUBATIONMOST COMMON IN WARM MONTHS SOUTHMOST COMMON IN WARM MONTHS, SOUTHMUCOPURULENT DISCHARGE; Kids < 5 yrsINFERIOR LIMBUS ULCERATIONSINFERIOR LIMBUS ULCERATIONSSOME SCARRING OF INFERIOR CORNEA
PILI (FIMBRIAE)ENDOTOXINCAPSULEGROWTH ENHANCED AROUND Staph spGROWTH ENHANCED AROUND Staph. sp.CAN INFECT INTACT CORNEAL EPITHELIUMREQUIRES CHOCOLATE AGAR + CO2REQUIRES CHOCOLATE AGAR CO2
MORAXELLA LACUNATA
MORAXELLA LACUNATA
G(–) DIPLOBACILLUS with PILI (FIMBRIAE)ANGULAR BLEPHAROCONJ’ITIS (Temporal)CORNEAL ULCERATION with HYPOPYONRARE IN YOUNG CHILDRENRARE IN YOUNG CHILDRENMUST NOW DIFFERENTIATE FROM Staph.REQUIRES CHOCOLATE AGAR + CO2REQUIRES CHOCOLATE AGAR CO2
STREPTOCOCCUSSTREPTOCOCCUSPNEUMONIAE
STREPTOCOCCUSSTREPTOCOCCUSPNEUMONIAE
G(+) DIPLOCOCCUS with PILI (FIMBRIAE)OPHTHALMIA NEONATORUMADULT DACRYOCYSTITISACUTE CATARRHAL CONJ’ITIS; Kids < 5 yrsACUTE CATARRHAL CONJ ITIS; Kids < 5 yrsCORNEAL ULCERATIONS with HYPOPYONSHOWS PREFERENTIAL ADHERENCE TOSHOWS PREFERENTIAL ADHERENCE TO
CORNEAL EPITHELIAL CELLSCONJ. MEMBRANE or PSEUDOMEMBRANE
STREPTOCOCCUSSTREPTOCOCCUSPNEUMONIAE
WELL ORGANIZED CAPSULE (SLIME COAT)BIOFILM (GLYCOCALYX)IgA -ase FibrinolysinHyaluronidase StreptokinaseHyaluronidase StreptokinaseHemolysin LeukocidinPNEUMOLYSIN: Inhibit Chemotaxis of PMNsPNEUMOLYSIN: Inhibit Chemotaxis of PMNsCAN INFECT INTACT CORNEAL EPITHELIUM
PSEUDOMONAS AERUGINOSA
PSEUDOMONAS AERUGINOSA
UBIQUITOUS G(–) RODMOST VIRULENT CORNEAL PATHOGENCAN NOT PENETRATE INTACT EPITHELIUMCORNEAL ULCERATIONS AND KERATITISCORNEAL ULCERATIONS AND KERATITISCAN PERFORATE CORNEA IN 24 Hrs.CAN FEED ON FLUORESCEIN IN SOLUTIONCAN FEED ON FLUORESCEIN IN SOLUTIONCAN FEED ON CAMPHOR, NAPHTHALENECAN GROW IN DISTILLED WATER
PSEUDOMONAS AERUGINOSA
PILI (FIMBRIAE); BIOFILMPOLAR FLAGELLUMEndotoxin Elastase ProteasesExotoxins Phosphatase CollagenaseExotoxins Phosphatase CollagenaseHEMOLYSIN: Unusual for a Gram (–)BLUE-GREEN PURULENCE from BG PYACINBLUE GREEN PURULENCE from BG PYACINSHOWS PREFERENTIAL ADHERENCE TO
BASAL CORNEAL EPITHELIAL CELLS
STAPHYLOCOCCUS AUREUS
STAPHYLOCOCCUS AUREUS
G(+) COCCI, NORMAL OCULAR FLORACOMMON & CHRONIC EYELID INFECTIONSHORDEOLUM: Nearly always the causeACUTE & CHRONIC CONJ’ITIS; Kids > 5 yrsACUTE & CHRONIC CONJ ITIS; Kids > 5 yrsKERATITIS AND OCCASIONAL ULCERATIONPOST-OPERATIVE ENDOPHTHALMITISPOST OPERATIVE ENDOPHTHALMITISMEIBOMIAN GLAND DYSFUNCTION: Most
common, with Propionibacterium acnes
STAPHYLOCOCCUS AUREUS
CELL WALL BINDING PROTEIN BINDS TO CORN EPITHELIAL SURFACE FIBRONECTINCORN. EPITHELIAL SURFACE FIBRONECTINCatalase Wax Esterase HemolysinCoagulase Cholesterol EsteraseCoagulase Cholesterol EsteraseExotoxins Cytotoxins LeukocidinsHyaluronidaseGROWS SLOWLY EVEN AT 5 DEGREES AND CAN CONTAMINATE REFRIDGERATED SOLN
STREPT. VIRIDANS
STAPH EPIDERMIDISSTAPH. EPIDERMIDIS
LINE-UP: CAN YOU IDENTIFY THE PERPETRATOR ?THE PERPETRATOR ?