MLAB 1331 – MYCOLOGY

CHARACTERISTICS OF FUNGI

DIVERSE GROUPS OF ORGANISMS , GENERALLY CLASSIFIED AS MOLDS OR YEASTS; SOME ARE PATHOGENS AND OTHERS ARE SAPROBES.

• CONTAIN TRUE NUCLEI- FUNGI ARE EUKARYOTIC
• NUCLEAR MEMBRANE
• HAVE NO CHLOROPHYLL
• ALL FUNGI HAVE A CHITIN CELL WALL
• CELL MEMBRANE
• SEXUAL OR ASEXUAL REPRODUCTION
• AEROBES
• MOISTURE NECESSARY FOR GROWTH

4 PHYLA

• ZYGOMYCOTA – ORGANISMS THAT PRODUCE SPARSELY SEPTATE HYPHAE
  ASEXUAL REPRODUCTION BY SPORANGIOSPORES AND SEXUAL REPRODUCTION BY ZYGOSPORES
• ASCOMYCOTA – CHARCTERISIZED BY PRODUCTION OF SEXUAL SPORES ASCOSPORES, CONIDIA IS FORMED BY ASEXUAL REPRODUCTION; THEY FORM SEPTATE HYPHAE
• BASIDIOMYCOTA – REPRODUCE SEXUALLY BY FORMING BASIDIOSPORES ON A STRUCTURE CALLED BASIDIA; PLANT PATHOGENS AND ENVIRONMENTAL ORGANISMS
• DUETEROMYCOTA – LACK A SEXUAL REPRODUCTIVE CYCLE; CONIDIA IS THEIR ASEXUAL REPRODUCTIVE STRUCTURES

PRACTICAL CLASSIFICATION OF FUNGI

·         SUPERFICIAL OR CUTANEOUS MYCOSES –FUNGAL INFECTIONS INVOLVING HAIR,
SKIN OR NAILS WITH NO DEEP INVASION OF TISSUES; ALL INFECT KERATINIZED TISSUES ; INCLUDE THE DERMATOPHYTES

·         SUBCUTANEOUS MYCOSES – FUNGI THAT INFECT SUBCUTANEOUS TISSUE WITHOUT DISSEMINATING TO OTHER SITES; INCLUDE CHROMOBLASTOMYCOSIS, MYCETOMA AND PHAEOHYPHOMYCOTA

·         SYSTEMIC MYCOSES – INFECTIONS INVOLVE THE LUNGS BUT MAY DISSEMINATE AND INVOLVE ORGAN SYSTEMS; INCLUDE BLASTOMYCES, COCCIDIOIDES, HISTOPLASMA AND PARACOCCIDIOIDES

·         OPPORTUNISTIC MYCOSES – INFECTIONS OCCUR IN THE IMMUNOSUPPRESSED AND IMMUNOCOMPROMISED PATIENT; INCLUDE ASPERGILLUS, ZYGOMYCETES, CANDIDA AND CRYPTOCOCCUS

YEAST versus MOLD

• YEAST – SINGLE VEGETATIVE CELL
• REPRODUCE BY BUDDING
• OVAL TO ROUND CELLS
• REPRODUCTIVE STRUCTURES - CONIDIA
• GROW AT 35-37°C
• COLONIES - SMOOTH, CREAMY, BACTERIAL-LIKE
• MOLD –FUZZY OR WOOLY APPEARANCE
• HAVE MYCELIUM – AERIAL (SUPPORTS REPRODUCTION) AND VEGETATIVE (ABSORB NUTRIENTS)

HYPHAE – AERIAL OR VEGETATIVE TYPES OF HYPHAE

• VEGETATIVE HYPHAE
• GROW BELOW THE SURFACE OF THE AGAR OF SUBSTRATE
• AERIAL HYPHAE
• GROW ABOVE THE SURFACE OF THE AGAR OF SUBSTRATE
• CONTAIN REPRODUCTIVE STRUCTURES
• OTHER TYPES OF HYPHAE
  1.             ANTLER
  2.             RACQUET
  3.             SPIRAL
  4.             RHIZOIDS

OTHER CHARACTERISTICS

• SEPTATE – HYPHAE  SHOW FREQUENT CROSS-WALLS BETWEEN CELLS
• SPARSELY SEPTATE HYPHAE HAVE FEW CROSSWALLS AT IRREGULAR INTERVALS
• PIGMENTATION TO AID IN IDENTIFICATION
• HYALINE HYPHAE LIGHTLY OR COLORLESS
• DEMATIACEOUS HYPHAE ARE DARK PIGMENTED DUE TO MELANIN

DIMORPHIC FUNGI

• A FEW FUNGI HAVE BOTH FORMS
• AT 35 - 37⁰C - THEY ARE YEAST
• AT 25 - 30⁰C - THEY ARE MOLD
• THESE ARE CAUSES OF SERIOUS FUNGAL DISEASE

REPRODUCTIVE STRUCTURES OF FUNGI

• MOST FUNGI REPRODUCE ASEXUALLY BY:
• CONIDIOGENESIS - CONIDIA
• ARTHROCONIDIA
• SEXUAL REPRODUCTION  - TWO COMPATIBLE NUCLEI MUST UNITE FOLLOWED BY MEIOSIS; PRODUCES ASCOSPORES OR BASIODIOSPORES
• BLASTOCONIDIA - BUDDING
• POROCONIDIA
• STALK WITH A "PORE"
• PHIALOCONIDIA
• GROWING FROM A VASE SHAPED "PHIALIDE"
• MACROCONIDIA  
• MICROCONIDIA
• ARTHROCONIDIA
• SPORANGIOSPORES

OTHER FUNGAL ROLES

• INDUSTRY
  1. DECOMPOSITION OF ORGANIC MATTER
  2. USED TO SYNTHESIZE ORGANIC ACIDS

·         FOOD PROCESSING

1.        BEER AND WINE FERMENTATION

2.        CHEESE

3.        BREADMAKING

• MEDICINE
  1. VITAMINS
  2. ANTIBIOTICS
     
      
• HARMFUL EFFECTS
  1. DESTRUCTION OF MAN-MADE GOODS
  2. FOOD SPOILAGE
  3.  ANIMAL AND HUMAN DISEASES
  4. SOURCES OF ALLERGENS AND TOXIN

LABORATORY SAFETY

• UNIVERSAL PRECAUTIONS
• NEVER SMELL A FUNGUS CULTURE  - AIRBORNE CONIDIA MAY BE INHALED
• BIOLOGICAL SAFETY CABINET CLASS II IS NEEDED
• USE SCREW TOPPED TUBES INSTEAD OF PETRI DISHES FOR CULTURES
• AUTOCLAVE ALL CULTURES

CLINICAL SITES AND SPECIMENS

·         RESPIRATORY TRACT SPECIMENS

·         CEREBROSPINAL FLUID

·         BLOOD

·         HAIR, SKIN AND NAIL SCRAPINGS

·         URINE

·         TISSUE

·         BONE MARROW

·         STERILE BODY FLUIDS

CULTURE MEDIA AND INCUBATION REQUIREMENTS

• PRIMARY RECOVERY MEDIA – SABOURAUD DEXTROSE AGAR  (2% DEXTROSE, PH OF 6.8) , MYCOSEL, BRAIN HEART INFUSION WITH OR WITHOUT ANTIBIOTICS (TABLE 53-6)

• DIFFERENTIAL TEST MEDIA – CORNMEAL AGAR WITH TWEEN 80, COTTONSEED CONVERSION AGAR, ASCOSPORE AGAR, UREA AGAR AND FERMENTATION AGAR AND BROTH (TABLE 53-6)

• INCUBATED AT ROOM TEMPERATURE OR  PREFERABLY AT 30°C FOR 30 DAYS OR 4 WEEKS BEFORE REPORTING NEGATIVE WITH HUMIDITY, SOME YEAST WILL GROW AT 2-3 DAYS, SOME WILL GROW AT 3-5 DAYS

• BECAUSE OF BACTERIAL CONTAMINATION AND RAPIDLY GROWING FUNGI THE NEED OF ANTIFUNGAL OR ANTIBACTERIAL AGENTS IS REQUIRED TO BE ADDED TO THE MEDIA

• LOW PH INHIBITS GROWTH OF BACTERIA

IDENTIFICATION OF FUNGI

• DIRECT TESTS PROVIDE RAPID REPORT TO PHYSICIAN, SPECIFIC MORPHOLOGICAL CHARACTERISTICS OF FUNGI AND IT CAN PROVIDE EVIDENCE OF INFECTION IN SPITE OF NEGATIVE CULTURE

DIRECT EXAMS

1.                    KOH – POTASSIUM HYDROXIDE 10-20% SOLUTION OF KOH, USED TO DETECT FUNGUS ELEMENTS IN SKIN, HAIR, NAILS AND TISSUES.

2.                    KOH WITH CALCOFLUOR WHITE       

3.                    INDIA INK

4.                    TISSUE STAINS

5.                    ACID FAST

6.                    GRAM STAIN

(TABLE 53-8 LISTS FUNGI FEATURES IN DIRECT EXAMS)

EXAMINATION AND IDENTIFICATION OF THE GROWN CULTURE

• THE GROWTH RATE – ALL GROWTH RATES VARY
• COLONY MORPHOLOGY - VISUALLY OBSERVE FOR GROWTH, COLOR, COLONY SIZE
• MICROSCOPIC MORPHOLOGY - MICROSCOPIC EXAMS FOR FUNGAL ELEMENTS TO OBSERVE SEPTATE VS NONSEPTATE HYPHAE, HYALINE OR DEMATIACEOUS HYPHAE AND THE TYPES, SIZE, SHAPE AND ARRANGEMENTS OF CONIDIA

1. TEASE MOUNT – USES A TEASING NEEDLE TO REMOVE PORTION OF MYCELIUM, ADD A DROP OF LACTOPHENOL COTTON BLUE STAIN (LPCB) ON A SLIDE AND OBSERVED FOR CONIDIA STRUCTURES.
2. CELLOPHANE TAPE PREP –
3. MICROSLIDE CULTURE –

LPCB DYE - THE COMBINATION OF LACTIC ACID, PHENOL AND THE BLUE DYE KILLS, PRESERVES AND STAINS THE ORGANISMS

VIRULENCE FACTORS

• THE SIZE OF THE ORGANISM
• THE ABILITY TO GROW AT 37°C AT A NEUTRAL PH
• CONVERSION OF THE DIMORPHIC FUNGI
• POSSIBLY TOXIN PRODUCTION

YEAST

• DO NOT PRODUCE TRUE HYPHAE
• ARE NOT DIMORPHIC
• REPRODUCE BY BUDDING
• YEAST ARE PART OF THE NORMAL HUMAN MICRO FLORA
• IMMUNOCOMPROMISED PATIENTS ARE MORE SUSCEPTIBLE
• YEAST IDENTIFICATIONS MUST PERFORMED FOR:
  1.             ALL YEAST FROM STERILE BODY FLUIDS
  2.             YEASTS FROM ALL IMMUNOCOMPROMISED PATIENTS
  3.             ISOLATES FROM RESPIRATORY SECRETIONS SHOULD BE SCREENED FOR
                 C. NEOFORMANS
  4.             YEASTS ISOLATED IN LARGE AMOUNTS FROM ANY CLINICAL SOURCE
  5.             YEASTS RECOVERED FROM SEVERAL SUCCESSIVE SPECIMENS
   

CANDIDA ALBICANS

• OFTEN FOUND AS NORMAL FLORA
• OPPORTUNISTIC PATHOGEN
• CAUSES THRUSH
• INTRAVASCULAR DEVICES (CATHETERS)
• THE MOST COMMON CAUSATIVE AGENT OF HUMAN MYCOSIS  

• CLINICAL DISEASES RANGE FROM SUPERFICIAL SKIN INFECTIONS TO DISSEMINATED INFECTIONS
• PRIMARILY SEEN IN IMMUNOCOMPROMISED, IMMUNOSUPPRESSED, AIDS PATIENTS
• PATIENTS WHO ARE RECEIVING LONG TERM IV THERAPY
• CHEMO- THERAPY

SPECIMENS

• SPUTUM - USUALLY SENT TO LAB FOR BACTERIAL OF AFB CULTURE
• YEAST ARE VISIBLE ON USUAL MEDIA AND BY USUAL STAINING
• THE AMOUNT OF YEAST IS IMPORTANT
• URINE - A FEW COLONIES MAY NOT BE SIGNIFICANT
• PREDOMINANT YEAST MAY BE INFECTION
• CSF, BLOOD AND OTHER BODY FLUIDS
• ANY YEAST FOUND IN THESE STERILE BODY FLUIDS WOULD BE SIGNIFICANT
• CORNMEAL AGAR WITH TWEEN 80 AND TRYPAN BLUE USED TO ID C. ALBICANS BY CHLAMYDOSPORE PRODUCTION
   

GERM TUBE TEST FOR CANDIDA ALBICANS

• MOST BASIC AND EASIEST TEST TO PERFORM ON YEAST
• USED TO IDENTIFY THE PATHOGENIC CANDIDA FROM THE NON PATHOGENIC
• PROCEDURE PAGE 792
• TRUE GERM TUBES LACK CONSTRICTION AT THEIR BASE WHERE THEY ARE ATTACHED TO THE MOTHER CELL.
• IF A CONSTRICTION IS OBSERVED, IT IS NOT A TRUE GERM TUBE
• HUMAN INFECTIONS OF CANDIDA AND CRYPTOCOCCUS
   

CRYPTOCOCCUS NEOFORMANS

• CAUSES CRYPTOCOCCOSIS, A MULTISYSTEM DISEASE
• USUALLY ENTERS BODY THROUGH RESPIRATORY SYSTEM
• DISSEMINATED CRYPTOCOCCOSIS VERY COMMON IN AIDS PATIENTS
• IN ALL CASES THE INFECTION IS FIRST PULMONARY THEN DISSEMINATES TO OTHER BODY SITES CAUSING ENDOCARDITIS, HEPATITIS, RENAL INFECTIONS AND PLEURAL EFFUSIONS

• CRYPTOCOCCUS NEOFORMANS IS FOUND WORLDWIDE
• PIGEON EXCRETA APPEARS TO BE THE CHIEF SOURCE FOR DISTRIBUTION
• ORGANISM MAY REMAIN VIABLE FOR LONG PERIODS
• CRYPTOCOCCUS NEOFORMANS HAS A POLYSACCHARIDE CAPSULE
• CAPSULE THAT CAN BE SEEN IN INDIA INK PREP
• PRODUCES MUCOID COLONIES
• NIGER SEED AGAR AND UREA AGAR TO IDENTIFY IT
   

INDIA INK FOR CRYPTOCOCCUS NEOFORMANS

• A DROP OF THE SPECIMEN IS PLACED ON A SLIDE WITH A DROP OF INDIA INK AND EXAMINED UNDER MICROSCOPE
• THE CAPSULE WILL SHOW CLEAR AROUND THE BUDDING YEAST  
• INDIA INK EXAMINATION IS QUICK AND SIMPLE BUT MAY BE MISINTERPRETED
• WBC’S LOOK SIMILAR
• NOT ALL C.NEOFORMANS FORM CAPSULES
• A LATEX AGGLUTINATION TEST IS AVAILABLE FOR C. NEOFORMANS

OTHER YEAST IDENTIFICATION SYSTEMS

·         COMMERCIAL YEAST ID SYSTEMS ARE UTILIZED
THEY ARE RAPID AND RESULTS ARE AVAILABLE IN 72 HOURS
ID BASED ON A DATABASE OF NUMEROUS YEAST BIOTYPES
EXAMPLES: API-20, UNI-YEAST TEK, MICROSCAN YEAST PANEL, VITEK BIOCHEMICAL CARD, CHROMAGAR AND RAPID YEAST PLUS

·         CONVENTIONAL METHODS BESIDES GERM TUBE INCLUDE CORNMEAL AGAR MORPHOLOGY, CARBOHYDRATE UTILIZATION, NIGER SEED AGAR, RAPID UREASE TEST, AND SERODIAGNOSIS

TRICHOSPORON BEIGELLII

• SUPERFICIAL MYCOSES
• CAUSES WHITE PIEDRA
• LIGHT BROWN, SOFT NODULES AROUND BEARD AND MUSTACHE HAIRS
• CULTURE ON SAB AT ROOM TEMPERATURE
• CREAM COLORED WRINKLED COLONY WITHIN 5 DAYS
• FINDINGS- HYPHAL ELEMENTS WITHIN SHAFT NODULE AND BUDDING BLASTOCONIDIA AND ARTHROCONIDIA PRESENT IN CULTURE
   

MALASSEZIA FURFUR

• SUPERFICIAL MYCOSES
• CAUSES TINEA VERSICOLOR
• CAUSES SCALY PATCHES OF DIFFERENT COLOR
• REDDISH BROWN, BROWN AND WHITE
• IDENTIFIED BY KOH – BUDDING YEASTS, WITH SEPTATE AND BRANCHING SHORT CURVED HYPHAL ELEMENTS
• YELLOW FLUORESCENCE UNDER A WOODS LAMP
• DISTRIBUTED WORLDWIDE

ZYGOMYCETES: THE HYALINE, ASEPTATE OR SPARSELY SEPTATE MOLDS

• LARGE RIBBON LIKE HYPHAE THAT ARE IRREGULAR
• SPORANGIA-SACLIKE FRUITING STRUCTURES
• SPORANGIOSPORES – YELLOW OR BROWN SPORES PRODUCED BY SPORANGIA
• WORLDWIDE DISTRIBUTION, FOUND ON DECAYED VEGETABLE MATTER OR SOIL
• INFECTION ACQUIRED BY INHALATION OF SPORES
• DISEASE - CAUSES ZYGOMYCOSIS
• COLONIES FLUFFY, WHITE TO GRAY OR BROWN HYPHAL GROWTH
• INCLUDES THREE SPECIES – ABSIDIA, MUCOR AND RHIZOPUS

ABSIDIA

• UNCOMMON ISOLATE
• CHARACTERIZED BY PRESENCE OF RHIZOIDS THAT ORIGINATE BETWEEN SPORANGIOSPORES
• SPORANGIA ARE PYRIFORM AND HAVE A FUNNEL SHAPED AREA AT JUNCTION OF THE SPORANGIUM
• MYCELIA IS USUALLY ASEPTATE

MUCOR

• WHITE FLUFFY AT ROOM TEMPERATURE
• ASEPTATE
• SINGLE OR BRANCHING SPORANGIOPHORES THAT AT TIP HAVE A  ROUND SPORANGIUM FILLED WITH SPORANGIOSPORES
• NO RHIZOIDS ARE PRESENT
• MAY CAUSE OTOMYCOSIS AND ALLERGIES

RHIZOPUS

• FUNGUS WITH ERECT SPORANGIOPHORES TERMINATED BY DARK SPORANGIA AND SPORANGIOSPORES
• AT BASE OF SPORANGIOPHORE ARE BROWN RHIZOIDS
• UNBRANCHED SPORANGIOPHORES FORM AT THE RHIZOID NODES
   

DERMATOPHYTES: HYALINE, SEPTATE, MONOMORPHIC MOLDS
 

• INFECTIONS INVOLVING SUPERFICIAL AREAS – HAIR, SKIN, NAILS (DERMATOMYCOSES)
• BREAKDOWN KERATIN AS A SOURCE OF NITROGEN
• THEY ADAPT TO GROW ON HAIR, NAILS  AND SKIN THAT ARE KERATINIZED (CONTAIN SCLEROPROTEIN KERATIN)
• INCAPABLE OF PENETRATING SUBCUTANEOUS TISSUE
• CUTANEOUS MYCOSES REFERRED TO AS  "TINEA" OR "RINGWORM"
• CATEGORIZED BY AREA OF THE BODY AFFECTED
• LATIN NOUN TO DESIGNATE AREA
  • TINEA CORPORIS (BODY)
  • TINEA CRURIS (GROIN)
  • TINEA CAPITIS (HEAD)
  • TINEA BARBAE (BEARD)
  • TINEA MANUUM (HAND)
  • TINEA PEDIS (FEET)
  • TINEA UNGUIUM (NAIL)

·         THEY ARE DISTRIBUTED WORLDWIDE, SOIL, ANIMALS AND HUMANS

LABORATORY SPECIMENS

• SKIN SCRAPINGS
• KOH PREPS OBSERVED FOR BRANCHING SEPTATE HYPHAE
• NAIL SCRAPINGS
• SAME AS SKIN
• HAIR – ENDOTHRIX (WITHIN HAIR SHAFT) AND ECTOTHRIX  (OUTSIDE HAIR SHAFT)
• DETECTION TESTS – CALCOFLUOR WHITE OR KOH
• GROWTH RATE MAY HELP DISTINGUISH SPECIES
• GENERA: TRICHOPHYTON, MICROSPORUM AND EPIDERMOPHYTON

TRICHOPHYTON

·         WIDELY DISTRIBUTED AND MOST IMPORTANT AND COMMON CAUSE OF INFECTIONS OF HAIR, SKIN AND NAILS

·         MOST ARE ANTHROPOPHILIC – HUMAN –LOVING

·         FEW ARE ZOOPHILIC – INFECTING ANIMALS

·         HAIRS INFECTED  WILL NOT FLUORESCE UNDER ULTRAVIOLET OF WOOD’S LAMP

TRICHOPHYTON MENTAGROPHYTES

·         CAUSES INFLAMMATORY TINEA PEDIS, TINEA CORPORIS, TINEA UNGUIUM, TINEA BARBAE AND TINEA CAPITIS

·         GROWTH RATE 7-10 DAYS

·         WHITE GRANULAR AND FLUFFY COLONIES; YELLOW PERIPHERY WITH REVERSE BUFF TO REDDISH BROWN

·         ROUND GRAPE-LIKE CLUSTERS OF MICROCONIDIA ON HAIR  

·         MACROCONIDIA ARE THIN-WALLED, SMOOTH, MULTISEPTATE; NUMEROUS OR RARE

·         FEW SPIRAL HYPHAE

·         POSITIVE UREA AGAR

TRICHOPHYTON RUBRUM

• FLUFFY WHITE WITH PINK PERIPHERY AND DEEP RED REVERSE ON SAB
• MICROCONIDIA TEARDROP, MOST COMMONLY BORNE ALONG SIDES OF HYPHAE
• MACROCONIDIA USUALLY ABSENT, WHEN PRESENT ARE SMOOTH, THIN-WALLED AND PENCIL-SHAPED
• NEGATIVE UREA AGAR; GROWTH ON CORNMEAL DEXTROSE AGAR
• CAUSES TINEA: CORPORIS, PEDIA,CRURIS AND UNGUIUM  

TRICHOPHYTON TONSURANS

• WHITE, TAN TO YELLOW OR RUST POWDERY COLONY, WRINKLED WITH HEAPED OR SUNKEN CENTER; REVERSE YELLOW TO TAN TO RUST RED
• CAUSES BLACK DOT TYPE OF TINEA CAPITIS
• MICROCONIDIA ARE TEARDROP OR CLUBSHAPED WITH FLAT BOTTOMS; VARY IN SIZE; USUALLY LARGER
• MACROCONIDIA ARE RARE AND BALLOON FORMS FOUND WHEN PRESENT
• GROWTH ENHANCED BY PRESENCE OF THIAMINE

MICROSPORUM  AUDOUINII

• SLOW GROWING ( 10-14 DAYS)
• DOWNY WHITE TO SALMON-PINK COLONY:  REVERSE TAN TO SALMON-PINK
• RECOVERY SITE-HAIR
• FLUORESCE YELLOW GREEN WITH WOOD’S LAMP (ULTRAVIOLET)
• STERILE HYPHAE, TERMINAL CHLAMYDOSPORES, FAVIC CHANDELIERS
• MACROCONIDIA RARELY SEEN – BIZARRE-SHAPED IF SEEN
• MICROCONIDIA RARE OR ABSENT
•  TINEA CAPITIS IN CHILDREN  

MICROSPORUM CANIS

• WOOLLY WHITE WITH WHITE TO BUFF OVER  YELLOW –ORANGE: BROWN CENTER, YELLOW OR ORANGE REVERSE
• NUMEROUS ROUGH, THICK-WALLED, SPINDLE-SHAPED MACROCONIDIA
• INFECTED HAIRS FLUORESCE
• CAUSES TINEA CORPORIS AND TINEA CAPITIS
• USUALLY CONTRACTED FROM CATS AND DOGS
•  

MICROSPORUM GYPSUM

• GROWS WITHIN A WEEK
• POWDERY BUFF COLORED COLONY
• THIN WALLED ELLIPTICAL MACROCONIDIA WITH 4-6 CELLS
• CAUSES TINEA CORPORIS AND TINEA CAPITIS

EPIDERMOPHYTON FLOCCOSUM

• GROWS WITHIN 10 DAYS
• VELVETY YELLOW COLONY, TAN REVERSE
• NUMEROUS CLUB SHAPED, SMOOTH, THIN-WALLED MACROCONIDIA WITH 2-4 CELLS
• CAUSES EPIDEMIC TINEA PEDIS
• ALSO TINEA CORPORIS AND TINEA CAPITIS

OPPORTUNISTIC MYCOSES: HYALINE, SEPTATE, MONOMORPHIC MOLDS

ASPERGILLUS

• FILAMENTOUS UBIQUITOUS FOUND IN NATURE
• CAUSE ASPERGILLOSIS
• ALMOST ANY ORGAN OR SYSTEM CAN BE INVOLVED; BEING THE MOST COMMON ONE IN INVASIVE INFECTIONS
• PRODUCE MYCOTOXINS WHICH CAN CAUSE CANCER
• RAPID GROWER MOLD ON SABS
• SOMETIMES IDENTIFIED ON BASIS OF COLOR
• MICROSCOPIC: HYPHAE ARE HYALINE AND SEPTATE
• UNBRANCHED CONIDIOPHORES WITH FOOT CELL, LARGE VESICLE AT TIP
• VESICLE SUPPORTS SHORT, FLASK-SHAPED PHIALIDES (STERIGMATA)
• EACH PHIALIDE PRODUCES CHAINS OF ROUND PHIALOCONIDIA

ASPERGILLUS FUMIGATUS

• THE MOST PATHOGENIC SPECIES; "FARMERS LUNG" – PULMONARY DISEASE
• EYE AND EAR INFECTIONS
• COLONIES SURFACE BLUE-GREEN TO GRAY; REVERSE IS WHITE TO TAN
• SEPTATE HYPHAE, SHORT OR LONG CONIDIOPHORES  WITH CHARACTERISTIC OF “FOOT CELL” AT BASE
• THE TIP OF CONIDIOPHORE HAS DOME SHAPED VESICLE WITH PHIALIDES ON UPPER HALF OF VESICLE

ASPERGILLUS NIGER

• COMMONLY SEEN IN CLINICAL LABORATORY
• CAUSE OF FUNGUS BALL AND OTITIS EXTERNA
• COLONIES ARE BLACK GROWTH, REVERSE WHITE TO YELLOW
• MICROSCOPIC: SEPTATE HYPHAE, LONG CONIDIOPHORES WITH LARGE SPHERICAL VESICLE

ASPERGILLUS FLAVUS

• YELLOW TO GREEN COLONIES  WITH REVERSE BEING GOLD TO RED-BROWN
• FELT-LIKE GROWTH
• LOOSE UNCOMBED LOOK
• MICROSCOPIC: VESICLES ARE GLOBOSE AND PHILIADES ARE PRODUCED DIRECTLY FROM VESICLE

FUSARIUM

• FILAMENTOUS AND FOUND ON PLANTS AND SOIL
• RAPID GROWTH ON SAB MEDIA
• COLONIES ARE FLUFFY TO COTTONY AND APPEAR IN COLORS FROM PINK , PURPLE, YELLOW , GREEN AND OTHER SHADES; REVERSE IS COLORLESS, TAN TO BROWN
• MICROSCOPIC: HYALINE SEPTATE HYPHAE, CONIDIOPHORES, PHIALIDES AND MICROCONIDIA (SINGLE CELLED)
• MACROCONIDIA CAN BE SINGLE OR MULTICELLED AND ARE SICKLED OR BOAT SHAPED WITH SEPTATION
• CAUSES MYCOTIC KERATITIS AND HAS BEEN ISOLATED FROM BURN PATIENTS
• GROWTH ON POTATOE DEXTROSE AGAR IS KEY TO ID

GEOTRICHUM

• FOUND WORLDWIDE IN SOIL, WATER, AIR, PLANTS, DAIRY PRODUCTS AND CEREAL
• CAUSES GEOTRICHOSIS, ACQUIRED BY INGESTION OR INHALATION AND ORAL THRUSH
• GROW RAPIDLY AND COLONIES ARE WHITE, DRY POWDERY TO COTTONY; RESEMBLE GROUND GLASS
• MICROSCOPIC: SEPTATE HYPHAE THAT PRODUCE NUMEROUS ARTHROCONIDIA THAT ARE RECTANGLE TO CYLINDRICAL TO BARREL-SHAPED

ACREMONIUM

• INFECTIONS IN IMMUNOCOMPROMISED PATIENTS AND BONE MARROW RECIPIENTS
• COLONY IS WHITE, PALE GREY ON SURFACE AND REVERSE IS EITHER UNCOLORED OR A PINK TO ROSE PIGMENT
• CONIDIA IN CLUSTERS AT TIP OF THE CONIDIOPHORES
• HYALINE, SEPTATE HYPHAE WHICH PRODUCE SINGLE, UNBRANCHED, TUBE-LIKE PHIALIDES

PENICILLIUM SPECIES

• COLONY  SURFACE VELVETY AND WOOLLY DUE TO CONIDIA
• RAPID GROWING AND FILAMENTOUS
• WHITE AND TURN BLUE/GREEN WITH AGE
• MICROSCOPIC: HYPHAE SEPTATE AND HYALINE THAT PRODUCE BRUSHLIKE CONIDIOPHORES - PENICILLUS
• THE METULAE, SECONDARY BRANCHES FORMED ON CONIDIOPHORES CARRY FLASK SHAPED PHIALIDES
• CONIDIA ARE  ROUND, UNICELLULAR AND SEEN AS UNBRANCHING CHAINS AT THE TIPS OF PHILIADES

PAECILOMYCES

• CAUSES PAECILOMYCOSIS AND  CORNEAL ULCERS, KERATITIS AND ENDOPHTHALMITIS DUE TO LONG CONTACT LENS WEAR
• VERY RAPID GROWING AND THERMOPHILLIC
• COLONIES ARE FLAT AND POWDERY, COLOR IS WHITE TO YELLOW TO PINK OR VIOLET
• MICROSCOPIC: SEPTATE HYALINE HYPHAE, CONIDIOPHORES, PHIALIDES, CONIDIA AND CHLAMYDOSPORES ARE OBSERVED
• PENICILLUS TYPE PHIALIDES THAT ARE LONG, DELICATE AND TAPERED

SCOPULARIOPSIS SPECIES

• FILAMENTOUS THAT IS FOUND IN SOIL, PLANT MATERIAL, FEATHERS AND INSECTS
• ASSOCIATED WITH ONYCHOMYCOSIS, PULMONARY INFECTIONS AND FUNGUS BALL
• RAPID GROWER; COLONIES APPEAR WHITE AND LATER TURN BROWN AND POWDERY
• MICROSCOPIC: SEPTATE HYPHAE, CONIDIOPHORES, ANNELIDES AND CONIDIA ARE SEEN
• BRANCHING OR PENICILLUS-TYPE "ANNELLOPHORES"
• LEMON SHAPED
• CONIDIA ARE LARGE, HAVE A FLAT BASE AND ARE ROUGH WALLED
   

SYSTEMIC MYCOSIS: HYALINE SEPTATE DIMORPHIC MOLDS

SYSTEMIC MYCOSES

• SERIOUS FUNGAL INFECTIONS THAT CAN INVADE THE ENTIRE SYSTEM
• CAN ALSO CAUSE CUTANEOUS MYCOSES
• BLASTOMYCES DERMATITIDIS
• COCCIDIOIDES  IMMITIS
• HISTOPLASMA  CAPSULATUM
• PARACOCCIDIODES BRASILIENSIS
• PENICILLIUM MARNEFFEI
• SPOROTHRIX SCHENCKII

BLASTOMYCES DERMATITIDIS

• CAUSES BLASTOMYCOSIS
• CHRONIC GRANULOMATOUS INFECTION OF THE LUNGS
• INFECTION IS BY INHALING THE MOLD PHASE; SYMPTOMS APPEAR FIRST AS RESPIRATORY IN LUNGS AND DESSIMINATES TO OTHER ORGANS
• COMMONLY FOUND IN NORTH AMERICA, MAINLY IN  MISSISSIPPI, OHIO AND MISSOURI RIVER VALLEY
• CUTANEOUS FORM OCCURS FROM ABSCESSES OF SKIN AND SUB SKIN
• BLASTOMYCOSIS MORE COMMON IN MEN

• IN DIRECT EXAMINATION APPEARS AS LARGE, SPHERICAL, THICK-WALLED YEAST CELLS WITH SINGLE BUD THAT IS CONNECTED TO PARENT CELL BY BROAD BASE
• DIRECT TESTS: KOH PREP, GRAM STAIN AND SPECIAL STAINS (GMS & PAS)

• AT 25°C (MOLD) – SLOW TO MODERATELY RAPID GROWER, COLONY IS DOWNY TO WOOLLY, SURFACE WHITE  OR TAN AND REVERSE IS PALE TO BROWNISH
• MICROSCOPIC: SEPTATE,  HYALINE HYPHAE; PYRIFORM (PEAR SHAPED) CONIDIA  ON LONG OR SHORT CONIDIOSPHORES, RESEMBLING LOLLIPOPS
• YEAST PHASE (37⁰ TISSUE PHASE) – TEXTURE CREAMY AND YEASTLIKE, APPEARS WRINKLED, WAXY, FLAT TO HEAPED; COLOR IS WHITE TO BEIGE
• MICROSCOPIC: BUDDING YEAST CELLS, DOUBLE CONTOURED REFRACTILE WALLS AND BROAD BASE
   

COCCIDIOIDES IMMITIS

• TRUE CAUSE OF SYSTEMIC MYCOSES- COCCIDIOIDOMYCOSIS
• FOUND IN SOUTHWEST (DESERT) UNITES STATES, SEMIARID REGIONS OF MEXICO AND CENTRAL AND SOUTH AMERICA
• FOUND ALL OVER BECAUSE OF EASE OF TRAVEL
• INFECTION FROM INHALATION OF INFECTIVE ARTHROCONIDIA
• 60% ARE ASYMPTOMATIC OR MILD RESPIRATORY INFECTION
• ABOUT 2% OF PULMONARY CASES DISSEMINATE
• INFECTIOUS ARTHROCONIDIA ARE PRODUCED FROM SEPTATE HYPHAE THAT GROW IN THE SOIL IN SEMI-ARID HOT CLIMATE
  
   
• MOST INFECTIOUS OF ALL FUNGI -CONSIDERED A BIOHAZARD FOR LABORATORY WORKERS; MUST HANDLE UNDER A BSC
• COLONIES GROW RAPIDLY; GROW AT 25 OR 37° C ON SABOURAUD DEXTROSE AGAR
• COLONIES ARE MOIST, SMOOTH, MEMBRANOUS AND GRAYISH WITH A COBWEB-LIKE GROWTH; LATER IT PRODUCES WHITE COTTONY AERIAL MYCELIUM
• MICROSCOPIC:
• THICK WALLED, BARREL-SHAPED ARTHROCONIDIA, SOME ARE ELONGATED CELLS ; HYPHAE ARE HYALINE, SEPTATE AND THIN AND IN ROPELIKE STRANDS; RAQUET FORMS ARE SEEN IN YOUNG CULTURES
• YEAST PHASE 37⁰ – LARGE, ROUND, THICK-WALLED SPHERULES FILLED WITH ENDOSPORES
   

HISTOPLASMA CAPSULATUM

• FOUND IN NATURE; SOIL CONTAMINATED WITH BAT EXCREMENT OR BIRD DROPPINGS
• ENDEMIC IN TENNESSEE-OHIO-MISSISSIPPI RIVER BASINS
• HISTOPLASMOSIS - MAJOR DISEASE OF HUMANS AND ANIMALS
• RANGES FROM ACUTE TO BENIGN PULMONARY INFECTION OR FATAL DISSEMINATED DISEASE
• COMMON FUNGAL PATHOGEN IN AIDS PATIENTS
• COMPROMISED PATIENTS MAY HAVE SYSTEMIC COMPLICATIONS
• INFECTIOUS MICROCONIDIA OR HYPHAE  ARE INHALED

• DIRECT EXAMINATION: INTRACELLULAR IN MONONUCLEAR CELLS AS SMALL, ROUND TO OVAL YEAST CELLS (GIEMSA OR WRIGHT’S STAINS)
• MOLD PHASE: COLONIES GRANULAR TO COTTONY, WHITE AND BECOMES BUFF BROWN WITH AGE (SAB)
• MICROSCOPIC: HYPHAE SEPTATE AND HYALINE: LARGE ROUND MACROCONIDIA WITH SMOOTH SPINY OR FINLIKE TUBERCULATE EDGES
• MICROCONIDIA IS ROUND, PYRIFORM AND SMOOTH
• TISSUE PHASE: COLONIES ARE CREAMY, MOIST, WHITE TO TAN ON BHIA
• MICROSCOPIC: SMALL, OVAL TO SPHERICAL  SINGLE BUDDING YEAST
   

PARACOCCIDIODES BRASILIENSIS

• HIGHEST PREVALENCE IN BRAZIL, VENEZUELA AND COLOMBIA SOURCE AND MEANS OF INFECTION IS UNCERTAIN
• CAUSES PARACOCCIDIOIDOMYCOSIS
• ORIGIN IS PULMONARY BY INHALATION OF ORGANISM FROM ENVIRONMENT
•  INFECTION MAY DISSEMINATED TO OTHER TISSUES 
   
• MOLD PHASE: COLONIES FILAMENTOUS, LEATHERY, FLAT TO WRINKLED, WOOLLY; COLOR IS WHITE CREAM TO TAN OR BROWN AND REVERSE IS YELLOW/BROWN
• MICROSCOPIC: HYALINE, SEPTATE HYPHAE THAT DOES NOT SPORULATE; PYRIFORM CONIDIA GROWING FROM SIDES OF HYPHAE OR ON SHORT CONIDIOPHORES
• YEAST PHASE: COLONIES ARE HEAPED, WRINKLED, MOIST, AND YEASTLIKE; WITH AGE THEY ARE COVERED WITH SHORT AERIAL  MYCELIUM
• MICROSCOPIC: TYPICAL BUDDING YEAST WHICH SURROUND THE WHOLE SURFACE OF THE MOTHER CELL RESEMBLING A STEERING-WHEEL OR MARINER’S WHEEL

SPOROTHRIX SCHENCKII

• CAUSES SPOROTRICHOSIS (ROSE GARDENER’S DISEASE)
• LYMPHOCUTANEOUS ULCERATED LESIONS
• NODES APPEAR AT SITE OF PUNCTURE
• PRIMARY LESION MAY HEAL BUT CHRONIC INFECTION MAY DEVELOP ALONG THE TRACK OF THE LYMPHATIC SYSTEM
• MOLD PHASE:  AT ROOM TEMPERATURE A MOIST, LEATHERY TO VELVETY WITH A WRINKLED SURFACE
• MICROSCOPIC: HYPHAE DELICATE, SEPTATE WITH BRANCHING AND ONE CELL CONIDIA; THESE APPEAR IN CLUSTERS AT END OD CONIDIOPHORE
• YEAST PHASE: AT 37⁰  COLONIES ARE  YEASTLIKE AND CREAMY; ARE SOFT WHITE, CREAM TO BEIGE COLORED  
• MICROSCOPIC: SINGLY OR MULTIPLY BUDDING, SPHERICAL , OVAL OR CIGAR SHAPED  YEAST CELLS
   

SEPTATE DEMATIACEOUS MOLDS

SUPERFICIAL INFECTIONS – TINEA NIGRA AND BLACK PIEDRA; THESE DISEASES OCCUR PRIMARILY IN TROPICAL AREAS OF THE WORLD

EXOPHIALA WERNICKII

• CAUSES TINEA NIGRA - SKIN – BLACKISH BROWN PATCHES ON THE PALMS OF THE HANDS OR SOLES OF FEET
• SLOW GROWER
• COLONIES OLIVE TO BLACK SHINY AND YEASTLIKE: LATER COLONIES BECOME FILAMENTOUS WITH VELVET-GRAY AERIAL HYPHAE
• DEMATIACEOUS HYPHAL FRAGMENTS AND BUDDING YEAS

PIEDRA HORTAE

• CAUSES BLACK PIEDRA - BLACK NODULES ATTACHED TO OUTSIDE OF SCALP AND HAIR  SHAFT
• SLOW GROWER
• COLONIES APPEAR DARK BROWN TO BLACK, PRODUCE AERIAL MYCELIUM
• SOME ISOLATES PRODUCE A RED TO BROWN PIGMENT
•  PRESENT HIGH SEPTATE DEMATIACEOUS HYPHAE

MYCETOMA – CHRONIC GRANULOMATOUS INFECTION THAT INVOLVES LOWER EXTREMITIES BUT CAN OCCUR IN ANY PART OF BODY

MYCETOMA TYPE I – ACTINOMYCOTIC (BACTERIAL) MYCETOMA CAUSED BY NOCARDIA, ACTINOMADURA AND STREPTOMYCES

MYCETOMA TYPE II – EUMYCOTIC MYCETOMA CAUSED BY FUNGI

·         PUS CONTAINS GRANULES OF COMPACT MYCELIAL MASSES

·         GRANULES EXAMINED FOR COLOR (LIGHT OR DARK), SIZE, TEXTURE (SOFT OR HARD)

·         SHOULD BE CULTURED ON FUNGAL MEDIA

·         DIRECT EXAM STAINS LOOK FOR HYPHIAL ELEMENTS

PSEUDALLESCHERIA BOYDII

• WHITE GRAIN MYCETOMA
• SUBCUTANEOUS AND GROW RAPIDLY
• INITIALLY WHITE, FLUFFY COLONY; LATER TURNS DARK GRAY OR SMOKEY BROWN
• REVERSE OF COLONY BECOMES BROWN TO BLACK
• SEPTATE HYALINE HYPHAE
• BOTH ASEXUAL AND SEXUAL REPRODUCTION

ACREMONIUM

• WHITE GRAIN MYCETOMA
• COLONY IS WHITE, PALE GREY ON SURFACE AND REVERSE IS EITHER UNCOLORED OR A PINK TO ROSE PIGMENT
• CONIDIA IN CLUSTERS AT TIP OF THE CONIDIOPHORES
• HYALINE, SEPTATE HYPHAE WHICH IS VERY FINE AND NARROW
• SIMPLE, UNBRANCHED ERECT CONIDIOPHORES

EXOPHIALA JEANSELMEI

·         BLACK GRAIN MYCETOMA

• PRODUCES OLIVACEOUS TO BLACK COLONIES
• YEASTLIKE WHEN YOUNG
• VELVETY APPEARANCE AT MATURITY
• CONIDIA AGGREGATE IN MASSES AT TIPS OF CONIDIOPHORE
• AERIAL HYPHAE
• SLOW GROWERS

CRUVULARIA

·         BLACK GRAIN MYCETOMA

·         ARE TWISTED AT THE ENDS WHERE CONIDIA ARE ATTACHED

·         CAUSES SINUSITIS

·         DEMATIACEOUS AND SEPTATE HYPHAE

·         SUBCUTANEOUS, HEART VALVES, EYE AND LUNGS

·         FLUFFY OR DOWNY, OLIVE TO GRAY TO BLACK COLONY

MADURELLA MYCETOMATIS

• BLACK GRAIN MYCETOMA
• SLOW GROWER
• BROWN DIFFUSIBLE PIGMENT IS CHARACTERISTIC OF THIS FUNGUS
• COLONIES VARY FROM WHITE TO OLIVE BROWN
• DEMATIACEOUS HYPHAL FRAGMENTS

CHROMOBLASTOMYCOSIS – CHRONIC FUNGAL INFECTIONS ACQUIRED VIA TRAUMATIC INOCULATION OF AN ORGANISM, PRIMARILY SKIN AND SUBCUTANEOUS TISSUE. CHARACTERIZED BY THE DEVELOPMENT OF PAPULE THAT SLOWLY FORMS ITCHY WARTY NODULES ON SKIN (FEET AND LEGS)

                                                                          CLADOSPORIUM

• MICROSCOPIC : DARK HYPHAE
• SHORT CHAINS OF DARK 1-4 CELLED BLASTOCONIDIA WITH REPEATED FORKING
• DARK HYPHAE
• CONIDIOPHORES WITH SPARSELY BRANCHING LONG CHAINS OF CONIDIA
• MAY FIRST APPEAR AS BLACK MOLD

PHIALOPHORA

• FOUND IN SOIL AND DECAYING FOOD
• COLONIES ARE WOOLY TO VELVETY AND MAY BE HEAPED AND GRANULAR
•  COLOR IS INITIALLY WHITE AND LATER BECOMES DARK; REVERSE IS IRON GREY
   BLACK
• SEPTATE HYPHAE, PHIALIDES AND CONIDIA ARE OBSERVED; THE HYPHAE ARE BRANCHED AND HYALINE TO BROWN

FONSECAEA

• FILAMENTOUS FOUND IN ROTTEN WOOD AND SOIL; IT PRODUCES A SPECIFIC
  STRUCTURE (SCLEROTIC BODY) ONLY IN TISSUES
• PRODUCES FLAT TO RAISED AND FOLDED VELVETY TO COTTONY COLONIES ON POTATOE DEXTROSE AGAR
• FRONT AND REVERSE, THE COLONIES ARE OLIVACEOUS TO BROWN-BLACK IN COLOR
• MICROSCOPIC: SEPTATE, DARK BROWN HYPHAE AND SUB ERECT CONIDIOPHORES THAT HIGHLY BRANCH
• MICROSCOPICALLY DIFFERENT IN DIFFERENT SPECIES

PHAEHOHYPHOMYCOSIS - INFECTIONS CAUSED BY DEMATIACEOUS FUNGI THAT FORM PIGMENTED HYPHAE AND YEASTLIKE CELLS IN TISSUE; CAN BE SUBCUTANEOUS, LOCALIZED OR SYSTEMIC

ALTERNARIA

• SUBCUTANEOUS
• LIGHT GRAY RAPID GROWER ON SABS, DARKENS WITH AGE
• MICROSCOPIC:  HYPHAE,  SEPTATE AND DEMATIACEOUS (GOLDEN BROWN)
• CHAINED MACROCONIDIA WITH HORIZONTAL AND VERTICAL SEPTA
• CONIDIOPHORES WITH A CHAIN OF LARGE CONIDIA RESEMBLING A DRUMSTICK
   

DRECHSLERA

·         RAPID GROWER, VELVETY - GRAYISH-BROWN TO BLACK

·         MICROSCOPICALLY, THE HYPHAE IS SEPTATE AND DEMATIACEOUS

·         CONIDIA ARE GENICULATE – SERIES OF BENT KNEE STRUCTURES

·         DEMATIACEOUS, MULTICELLED CONIDIA

CURVULARIA

• SUBCUTANEOUS, HEART VALVES, EYE AND LUNGS
• MICROSCOPICALLY: SEPTATE WITH DEMATIACEOUS HYPHAE
• MULTICELLUAR  MACROCONIDIA WITH TRANSVERSE SEPTATIONS
• CONIDIOPHORES ARE TWISTED WHERE CONIDIA ARE ATTACHED
• CONIDIA ARE CURVED (BOOMERANG) WITH CENTRAL SWOLLEN CELL

EXOPHIALA

• IMMATURE COLONIES APPEAR AS BLACK YEASTS
• MATURE COLONIES APPEAR HAIRLIKE OR VELVETY, GRAY-BROWN TO BLACK MYCELIUM COVERING THE SURFACE
• MICROSCOPICALLY DEMATIACEOUS HYPHAL FRAGMENTS AND BUDDING YEASTS
• THEY PRODUCE CONIDIOGENOUS CELLS WITH ANNELLOPHORES

EXSEROHILUM

• HYPHAE ARE SEPTATE AND DEMATIACEOUS
• CONIDIOPHORES ARE GENICULATE AND ARE PRODUCED SYMPODIALLY
• ELONGATED, MULTICELLED CONIDIA WITH PROMINENT PROTRUDING HILUM
• CONIDIA ARE LONGER AMD PENCIL-SHAPED

PNEUMOCYSTIS CARINII – ATYPICAL FUNGI

• OPPORTUNISTIC FUNGI THAT AFFECTS IMMUNOCOMPROMISED HOSTS, PARTICULARLY AIDS PATIENTS; PRESENTS ITSELF AS  PNEUMONIA
• PREVIOUSLY BELIEVED TO BE A PROTOZOAN
• DIFFERS FROM FUNGI IN THAT CELL MEMBRANE CONTAINS CHOLESTEROL
• CLASSIFIED BETWEEN ASCOMYCETES AND BASIDIOMYCETES

• DISTRIBUTED WORLDWIDE
• TRANSMITTED THROUGH REPIRATORY ROUTE; WITH THE CYST BEING THE INFECTIVE STAGE; INTRACYSTIC BODIES ARE RELEASED IN THE LUNG AND MULTIPLY ON CELLS LINING LUNGS
• CLINICAL SPECIMENS TO ISOLATE, RESPIRATORY SECRETIONS SUCH AS BRONCHOALVEOLAR LAVAGE OR TRANSBRONCHIAL BIOPSY AND INDUCED SPUTUM

• DOES NOT GROW IN CULTURE
• DIRECT EXAMINATIONS ARE GIEMSA STAIN, CALCOFLUOR WHITE OR METHENAMINE SILVER STAIN
• MICROSCOPIC OBSERVATIONS: TROPHOZOITE FORM ARE THE PREDOMINANT FORM SEEN (1-4 um ); IRREGULARLY SHAPED AND VISIBLE IN GIEMSA STAIN
• CYSTS ARE ALSO OBSERVED AND ARE SPHERICAL, CONCAVE, UNIFORM IN SIZE (4-7 MICRONS ) CONTAIN INTRACYSTIC BODIES
• COMMERCIAL KITS FOR MONOCLONAL ANTIBODIES DIRECTED AGAINST P. CARINII STAIN BOTH TROPHOZOITES AND CYSTS