LECTURE-MYCOLOGY Cytoplasm and Multicellular None

cytoplasmic with
INTRODUCTION content interdependent
function with
Mycology membrane
bound
- Myco means fungi and logy means study
organelles
- Discovered in 1600
- Grow into two basic forms:
- study of Fungi or mushrooms
a. Yeast
Fungi – Greek: "mykes" b. Molds
• DIMORPHIC FUNGI
- Aka mushroom
▪ Yeast (37°C)
- Grow in irregular masses
▪ Molds (room temp.)
Layman's Terms: ▪ Usual dimorphic fungi are
systemic mycoses
- Mushroom, mildew, puffballs, bracket fungi - may reproduce either asexually (they are
Characteristics of Fungi breaking in or breaking out within their chains
and the cell itself is infectious) or sexually
- Eukaryotic cells which lack chlorophyll (green (fusing of two cells) and it is same as manner of
pigment of plants) infection and reproduction
o 3 principal parts:
▪ Nucleus with chromosomal Morphology
DNA and a RNA rich nucleolus - All fungi, with the exception of yeast, are
within nuclear membrane composed of filamentous or tube-like
▪ Cytoplasm or plasma filaments called "HYPHAE"
membrane - glycoproteins, as
well as ergosterol with lipid Hyphae/Hypha
▪ Rigid cell wall
- Basic structural unit of a mold or fungi
- Most fungi are obligate or facultative aerobes - Tube-like structure
- Absorbs nutrients through the environment - Join to form Mycelium (aggregates of hypha)
- They are chemotrophic (attracted via certain
o Mycelium – the fluffiness or cottony
chemicals) appearance in a bread with mold
- Other fungi like Cryptococcus neoformans
▪ 2 kinds of mycelia:
posses a polysaccharide capsule
• Respiratory mycelia or
- Functions of Cell Wall and capsule
spore generating
o Protection
(infectious)
o Transport of Substances
• Vegetative or
o Contributes to its virulence
metabolically active
o Involved in host response
mycelia
- With complex morphology than the bacteria

Fungi Bacteria
Classification Eukaryotes Prokaryotes
Size Large Small

Structures
Cell wall Rigid – contains With
chitin, mannan, peptidoglycan
B glucan, No
peptidoglycan
Cell membrane Contains sterols None
(ex. Ergosteroll
+ zymosterol)
 - Aseptate/Coenocytic Hypha
o No cross-walls/division or breaks
o Ex. Zygomycetes (Rhizopus and
Mucor)
▪ Zygomycetes – Opportunistic
infections that happens in
patient’s w/ AIDS, HIV,
immunodeficiency, etc. YEAST MOLDS
- Septate Hypha Unicellular, round Multicellular, filamentous
o With cross-walls/division or joints - forms long tubes of
o All fungi except Zygomycetes hypha, occurs as
branching strands of cells
and hypha may have
intracellular division or
crossing (septate or
aseptate)
Reproduce by budding Reproduce by budding
(extrusion of daughter (extrusion of daughter
cells) or by fission cells) or by fission
(splitting of cells) (splitting of cells)
Mycelium
Temp requirement: 35- Temp requirement: 25-
- The mass of growing hyphae 37°C 30°C (room temp.)
- 3 types: Colonial characteristic: Colonial characteristic:
o Aerial Mycelium – responsible for the moist, buttery dry, cottony, velvety
production of spores and infection consistency with alcoholic
o Reproductive Mycelium odor
▪ Portion projecting above the
substrate and capable of
forming the spores
o Vegetative/Thallus Mycelium
▪ Penetrates into the substrate
NOTE:
and absorbs food
- *same lang daw yung aerial mycelium and - Monomorphic - capable of single growth phase
reproductive mycelium - Dimorphic - capable of 2 growth phases
o @RT – MOLD
o @ 37°C-YEAST (Tissue/In vivo/Invasive
phase)
Examples:
▪ Sporothrix schenckii
▪ Blastomyces dermatitides
▪ Histoplasma capsulatum
▪ Paracoccidioides brasiliensis

***Stage infective to man: MOLD

Two Growth Phases of Fungi In man, the Mold becomes Yeast

- Yeast 2 Major Divisions

o Produces creamy colonies resembling
- Meiosis
bacterial colonies
o Nuclear division and reduction
- Mold
resulting in the formation of haploid
o Cottony mycelial mass
nuclei without the cytoplasm
- Not all fungi have yeast or mold phase because
there’s dimorphic, yeast only, mold only or
either or
 o Chromosome number contained with
haploid nuclei is 1/2 the number of
chromosomes in the mother cell
- Mitosis
o Division of nuclear chromosome and
cytoplasm resulting to daughter cells
Asexual Reproduction includes:
- Fragmentation of Hyphae
o Portion of plant body detach and
develop into a new individual
- Budding
o Production of small outgrowth from a
parent cell
o As the bud is formed, nucleus of the
parent cell divides mitotically and
daughter nucleus migrates into the
bud
o The bud is enlarged and forms a new
individual
o If separation does not occur, it results
into a "Pseudohyphae"

Life Cycle of Fungi

- Somatic Phase
o Characterized by feeding or trophic
- Fission
activities and also in the absorption of
o Simple splitting of a cell into a
nutrients
daughter of a cell wall
- Reproductive Phase – 2 general types:
o Asexual or Imperfect State (Anamorph
– only one cell in hypha is the
infectious agent)

- Spore Formation

Spores
o Sexual or Perfect State (Telemorph –
the hypha fuse to become one cell) - For reproduction also conidia (for
reproduction)
- Reproduction in a state of fungi infection,
propagation
- 2 kinds:
o Asexual Spores (conidia)- Imperfect
Fungi (cannot reproduce sexually)
Asexual Reproduction
o Sexual Spores - Perfect Fungi (can
- Production of spores thru differentiation of reproduce sexually)
spore bearing hypha, sometimes called
Asexual Spores
somatic reproduction without nuclear fusion
- No fusion of nuclei (Karyogamy) - Conidia – Arise from the side of the hyphae
- Could be a simple division of unicellular o Microconidia – appear small and
organism or multicellular thallus into new unicellular
individual o Macroconidia – large and multicellular
- Principal asexual structures:
o Conidia - always asexual
o Spores - may be sexual or asexual
- Blastoconidia – Derived from simple budding Sexual Reproduction includes:
- Chlamydoconidia – called chlamydo because of
- Caused by the fusion of 2 cells
semicircular shape, thick-walled spores that
- Plasmogamy – union or fusion of 2 protoplast
are formred during unfavorable conditions;
- Karyogamy – fusion of 2 nuclei
they germinate when the environment
- Meiosis
improves, can arise terminally, laterally,
intercalary (within) Sexual Spores (perfect fungi)
o Terminal Chlamydoconidia - @ tip of
the hyphae - Ascospores
o Intercalary Chlamydoconidia - whithin o Enclosed in a sac-like structure called
the hyphal strand ascus
o Sessile Chlamydoconidia - @ the side o 2-8 spores
or intercalary of the hyphae

- Zygospores
o Derived from the fusion of 2 identical
spores from the same hypha, there’s
- Arthroconidia septum
o "Arthro" - joint (appear jointed)
o Derived from fragmentation of
mycelium
o Barrel-shaped or rectangular spores - Oospores
o Disjunctor cell - empty cell between o Derived from the fusion of 2 cells from
each spore, to easily break the non-identical separate hyphae, it
arthoconidia possesses conjunction
o Fungi with arthroconidia
▪ Coccidioides immitis
▪ Geotrichum candidum – mold
phase of candida albicans but
do not happen to human

- Basidiospores
o Enclosed in a club shaped structure
called basidium

- Sporangiospores
o Asexual spores are enclosed in a
sporangium borne on a specialized
hypha known as a sporangiophore Life Cycle of Fungi – never ending cycle, fast to reproduce
 - Sexual reproduction Classification of Fungi
o From mycelium → fusing of cytoplasm
- 2 Major Phyla:
→ fusing of nuclei → meiosis until the
o Zygomycota – fungi that divides
production of spores → spores will
asexually forming sporangiophores
infect/spread out to other agents →
and rapidly undergoes sexual
germination and (if favorable will go
production
back to sexual reproduction)
o Dikaryomycota – forming a zygote and
- Asexual reproduction
is marked by prolonged sexual cycle
o If not favorable to do sexual
since their haploid do not fuse readily
reproduction it will then produce
asexually Subphyla for Dikaryomycota
Ascocarps - Asacomycotina
o Fungi whose life cycle occur wihtin a
- The fruiting body of an ascomycete fungus
sac (ascus) and sexually produces
- Consists of very tightly woven hyphae and may
"Ascospores"
contain millions of asci, each of which typically
o Includes mostly fungi causing infection
contains 8 ascospores
- Basidiomycotina
- Mostly bowl-shaped, but may take on a
o Fungi whose life cycle commence
number of forms
within a bag termed as basidium with
maturation on the outside of the bag
o They produce sexual spores termed
"Basidiospores"

FUNGI IMPERFECTI

Types of Ascocarps - Include fungi that are not completely classified

due to the absence (not recognized) of sexual
- Cleistothecium – ascocarp is entirely enclosed state
without opening - Asexual spores – These include Candida,
- Perithecium – flask-shaped fruiting body with Torulopsis and Epidermophyton
an opening through which the ascospores can
escape
- Apothecium – Cup-shaped asci are formed on
the inside of the cup

- Gymnothecium – resembles cleistothecium

except the outer walls of the ascocarp are
loosely organized so the asci are released
through the openings in the walls

- Ascostroma – asci are produced in locules

(compression of cavities) in hard masses of
supporting hyphae called a stroma (mattress)
LABORATORY-MYCOLOGY Hyaline, septate molds

- Dermatophytes:
Lab Identification of Clinically Significant Fungi
o Trichophyton and Microsporum
- Some patients who are immunosuppressed o Stains: KOH and calcofluor white
and have chronic conditions sometimes get o Hyaline, septate hyphae and
infected with fungi. arthroconidia
o e.g. Patient have prolonged fever and o Smooth, club-shaped, thin-walled
tried antibacterial and yet fever isn’t macroconidia, sometimes have small
still resolved. Viral infections are self- Ars conidia
limiting infections and fever shouldn’t o Subculture onto cornmeal or potato
progress long but then after 3-5 days dextrose agar is necessary to induce
of antibacterial treatment; patient’s sporulation
state doesn’t improve. So that is when - Trichophyton
to consider fungi to be involve and is o T. RUBRUM:
needed to be identify by the lab what ▪ pyriform conidia
fungi is infecting the patient. ▪ cherry red colonies (vroom
o Specimens for fungi – sputum, CSF, vroom Ferrari)
blood, urine, stool ▪ urease and hair perforation
test negative
Basis of identification
o T. MENTAGROPHYTES:
• Microscopic morphology – for micro ▪ numerous microconidia in
• Culture characteristics – for micro grape-like clusters and cigar-
• Biochemical tests – for micro shaped macroconidia
• Immunodiagnostics – for serology ▪ rose-brown colonies
▪ urease and hair perforation
Morphologic classification test positive
- Microsporum
• Hyaline, pauciseptate molds
o Large, spindle-shaped, thick-walled
o Pauciseptate – loosely septated
macroconidia EXCEPT M. nanum w/c
• Hyaline, septate molds – intensely septated
produces 2-celled macroconidia
• Dematiaceous, septate molds – there’s
o M. audouinii: antler or racquet hyphae
melanin deposition in hypha
o M. canis: thick-walled, echinulate
o Dematiaceous – there’s melanin,
macroconidia with round ends,
brown in color
colonies with lemon-yellow or yellow
• Yeasts – unicellular, eukaryotic cell
orange fringe at periphery
Hyaline, pauciseptate molds ▪ Fringe – like ridges at the
periphery
- Zygomycetes: o M. gypseum: large, ellipsoidal,
o Rhizopus, Mucor and Absidia multisegmented, echinulate
▪ They are aseptate but some macroconidia with rounded ends
clinical specimen
demonstrates some
septations
o Stains: KOH (lyses keratin) and
calcofluor white (stains cell wall –
chitin of fungi)
o Branching, broad aseptate hyphae and
sporangium
o Rapid growth of grayish hyphae dotted
with brown to black sporangia
Opportunistic molds: o S. schenkii: small, round to oval to
cigar-shaped yeast cells
- Attack patients who are immunocompromised
surrounded by amorphous pink
- Aspergillus (#1 opportunistic molds), Fusarium,
material, most likely any
Geotrichum (are not common), Acremonium,
carbohydrate
Penicillium (used for the production of
II. Examination from culture
penicillin), Paecilomyces and Scopulariopsis
• B. dermatitidis: "lollipop" appearance
- Hyaline, septate hyphae with dichotomous
of conidia on short conidiophores
branching
• C. immitis: septate hyphae with right-
angle branches and racquet forms
• S. schenckii: "flowerette arrangement"
of conidia on conidiophores
dichotomous branching
- Aspergillus Culture
o A. fumigatus: foot cell at the base of
• C. immitis: "cobweb" appearance of colonies
conidiophores
• B. dermatitidis: "prickly state" – tufts of
o A. niger: darkly pigmented spores, one
hyphae project upward from colonies, hard
of the deadliest Aspergillus species
mycelia
▪ Common manifestations:
• YEPA w/NH4OH: optimal recovery of H.
pneumonia, unresolving with
capsulatum, B. dermatitidis and C. immitis
antibacterial, prolonged fever,
from contaminated specimens
sometimes respiratory arrest
o YEPD Agar (Yeast Extract Peptone
o Immunodx: galactomannan assay
Dextrose Agar) – they provide optimal
o A. fumigatus: blue-green powdery
recovery of H. capsulatum, B.
colonies
dermatitidis and C. immitis
o A. flavus: yellow-green colonies
o A. niger: yellow colonies with black Dematiaceius, septate molds
dots, turns black with age
- Superficial mycosis agents (cutaneous type of
Dimorphic fungi (Systemic) – molds and yeast phase mycosis)
o Exophiala werneckii and Piedraia
- Blastomyces dermatitidis
hortae
- Coccidioides immitis
o KOH: spindle-shaped asci with 2 to 8
- Histoplasma capsulatum
aseptate ascospores
- Paracoccidioides brasiliensis
- Mycetoma agents (mycotic mycetoma)
- Penicillium marneffei
o Pseudallescheria
- Sporothrix shenckii
o Acremonium
I. Direct examination (systemic)
o Exophiala jeanselmei
• Saline preparation:
o Curvularia
o B. dermatitidis: large, thick-walled
o Madurella
yeast cells w/ a broad based single
- Chromoblastomycosis agents (subcutaneous
bud
type of mycosis)
o C. immitis: nonbudding, thick
o Cladosporium, Phialophora and
walled spherule (“ghost
Fonsecaea
spherules” or “mickey mouse
o KOH: sclerotic bodies
head”)
o C. carrionii: long chains of elliptical
• Wrights or Giemsa stain:
conidia arising from tall branching
o H. capsulatum: small, round to
conidiophores
oval intracellular (found in
o P. verrucosa: phialides with distinct
macrophage) yeast cells
cup or flask shaped collarette
o P. brasiliensis: multiply budding
yeast cell ("mariner's wheel")
• PAS stain:
 Griseofulvin

- For dermatophytes that are unresponsive to

azole treatment
- From Penicillium
- Antimitotic
- For dermatophytoses unresponsive to azole
- Phaeohyphomycosis therapy
o Infections caused by other - Side effects: headache, GI disturbances and
dematiaceous fungi photosensitivity
Yeasts Antimetabolite agents

• Candida – germ tube, most know Candida - Flucytosine (5-Fluorocytosine)

albicans - Azole antifungals
• Cryptococcus
Flucytosine (5-Fluorocytosine)
• Trichosporon
• Malassezia - Fluorinated uracil
- DNA and protein synthesis inhibitor
Antifungal agents
- Combined with amphotericin B to treat
Mechanism of action Candidiasis and Cryptococcosis

• Polyene macrolide agents – attacks the chitin Azole

• Antimetabolite agents
o for bacteria: tmp smx
trimethoprim/sulfamethoxazole
• Echinocandins
• Chemicals
Polyene macrolide agents
- Attacks chitin/cell wall
- Penicillin
- Conjugated double bond and one to three ring
structures
- Amphotericin B, nystatin and griseofulvin
Amphotericin B
- From Streptomyces nodosus
- Binds to cell membrane ergosterols and alters
its selective permeability
- Side effects: renal insufficiency
- Resistant species: Pseudallescheria boydii,
Aspergillus terreus and Fusarium (bacteria)
Nystatin
- From Streptomyces noursei
- For oral or vulvovaginal candidiasis
o Candidiasis in the case of HIV/AIDS
(marker for AIDS):
▪ CD4/CD8 cell count ratio
▪ 2°/secondary infection – oral
candidiasis
- Not for injection
- Imidazoles, triazoles, ketoconazole,
– fluconazole, etc.
- 6-carbon ring structures with conjugated
double bonds and chloride residues
- 5-carbon ring structures with at least 2
nitrogen molecules
- Interfere with ergosterol synthesis and disrupt
cell membrane
- They are like macrolides in bacteria
Echinocandins
- Glucan synthesis inhibitors
- Caspofungin, micafungin and anidulafungin
o Used for systemic infections
o Caspofungin – used for systemic
candidiasis, systemic cryptococcosis
Chemicals
- Selenium sulfide: sporicidal (anti dandruff)
o Malassezia furfur (superficial mycosis)
– causes dandruff
- Potassium iodide: for sporotrichosis
MYCO (2nd week)
Superficial Mycoses
• Skin parasites
• Contained mainly to the horny non-living
layer of the skin and extrafollicular parts of
the hair
• Infections are innocuous
• Tineas – for skin infections
• Piedras – for hair infections
• AN.
- Superficial cutaneous is divided by 4:
superfilical, cutaneous, subcutaneous,
systemic, and opportunistic
- Only the epidermis of the skin (upper level
lang lahat)
- Innocuous – not harmful, commensal but
di sya maganda sa skin
- Tineas – ringworm
Tinea versicolor
• Synonyms:
▪ Ptyriasis versicolor
▪ Dermatomycosis furfuracea
▪ Tinea flava
▪ Chromophytosis
▪ “liver spot”
• Mild chronic fungal infection
• Affects the dermis of the skin (only upper part
of dermis)
• Asymptomatic
• Manifestation: appearance of hypopigmented
or hyperpigmented, finely scaly, coalescing
macules
▪ Hypopigmented – white macular
lesions on dark skinned patients
▪ Hyperpigmented – brown macular
lesions on fair skinned patients
• Involves: trunk of the body and upper arm
(seen in clavicle, deltoids)
• Lesion: non-inflammatory, pruritic, with
branny or fufuraceous scales (Makati na nag
sscales)
• Causative agent: Malassezia furfur (dandruff)
• Microscopically: spaghetti
(hyphae) and meatballs
(sphores) appearance
NOTE: it is septated
• Laboratory Diagnosis
Specimen sources:
▪ Skin scrapings – from discolored areas
of skin
▪ Blood or tissue samples – for
suspected disseminated infection
• Under Lab Diagnosis
1. Macroscopic Examination
▪ Wood’s Lamp Examination –
doubtful lesions maybe made
readily visible which shows a
golden yellow to light green
fluorescence in the skin
NOTE: UV light siya na shina-shine
don sa lesions.
2. Direct Microscopic Examination of Skin
Scrapings
▪ Staining of the Specimen with
alkali stain (Ex. Crystal violet,
Iodine or Methylene Blue)
▪ Treatment of the specimen with
10% KOH on Methylene blue or
Lactophenol Cotton Blue (LCB)
Malassezia furfur in 10% KOH
NOTE:
Ang ginagawa sa skin scrapings: 10% KOH
and calcufluor white
KOH – keratin remover
3. Culture Media
▪ Sabouraud Dextrose Agar supplemented
with a layer of olive oil or other vegetable
oil plus antibacterial antibiotics (general
culture media for all fungi species)
▪ Malt Extract Agar with addition of lipids
and antimicrobial agents (di masyado
nagamit)
▪ Incubation Temperature: 35oC – 37oC
 ▪ Macroscopic : • Laboratory Diagnosis
- Bacteria-like colonies appear after 1-2
1. Direct Microscpic Examination of
weeks
- Shiny or pasty white to cream colored
Specimen
which later turns dull and beige ▪ Wet Mount Preparation
▪ Microscopic: - Presence of dark brown
- Resemble bowling pins or medicine mycelial fragments
capsules and bottles
(diagnostic)
NOTE: problem with myco is matagal tumubo compared to bacte
• Mode of Transmission: ▪ KOH Preparation
▪ Direct Skin contact - fungi are interspersed
throughout epithelial cells as
• Treatment:
▪ Whitfield’s ointment filaments and with small
spherical spores (blastopores
▪ 3% Salicylic Acid in 70% Alcohol
– product of one cell)
▪ Selenium Sulfide Lotion
- Long brownish hypha that
▪ Imidazole, Ketoconazole,
branch freely and with
Fluconazole – ginagamit lang pag
closely spaced septa
di umeffect yung salicylic acid and
selenium
▪ Miconazole cream for folliculitis
▪ Locally manufactured Vinegar
Tinea nigra palmaris
◾ Synonyms: Tinea Nigra appearance in KOH cleared tissue
▪ Keratomycosis nigricans palmaris 2. Culture Media
▪ Cladosporosis epidermica ▪ Modified Sabouraud Dextrose
▪ Ptyriasis nigra Agar
▪ Microsporosis nigra ▪ Littman Oxgall Agar
• Palmaris means black discoloration ▪ Incubation Temperature : 25 ° C –
• Superficial, asymptomatic 30 °C ( mold)
fungal disease ▪ Macroscopic
• Characterized by - After 7 days, young colonies
development of a single, appear shiny, moist, dark
sharply demarcated gray, yeast-like colonies which
(borders) brown to black become very dark olive green
non-scaly macules to olive black
(papules na malalaki) - Front side: Velvety or woolly
• Pigmentation - confined to the stratum texture as short mycelia
corneum of the palmar surface of the hand develop
or on the plantar surface of the foot - Reverse side: jet black
• Causative agent:
▪ Exophiala werneckii –eto lang daw
tandaan
▪ Cladosporium werneckii
▪ Phaeoannellomyces werneckii
Tinea nigra on Sabouraud Dextrose Agar
• Specimen sources
▪ Skin scrapings from darkly pigmented
cutaneous lesions
 o Causative agent: Trichosporon
beigelii
▪ Microscopic
o Treatment:
- Young olive-black elliptical
✓ Shaving or clipping of
yeast-like cells,
hair
typically two-
✓ Oral Itroconazole
celled
- Annelloconidia
are elliptical
to subglobose,
usually unicellular with
smooth hyaline to olive brown
walls
-
• Treatment:
▪ Whitfield’s Ointment
▪ 5-10% Salicylic Acid, Tincture of Iodine
(if di kaya gamutin, try yung
ketoconazole…)
▪ Ketoconazole, Econazole, Miconazole

Piedra

• Superficial hair infection with nodular masses

of fungal elements surrounding the hair shaft
• Hair appears normal except for the
appearance of nodules
• Base of hair shaft and the hair follicle are
unaffected
• Black Piedra
▪ Black brown crust
▪ Outside the hair
shaft
▪ Causative agent:
Piedra hortae
▪ Diagnosis
o Direct Observation
o Culture
▪ Treatment:
o Shaving or clipping hair
o Topical or Oral Azoles
o Terbinafine
▪ White Piedra
o Characterized by
presence of light
brown nodules on
beard
CUTANEOUS MYCOSES • for maintenance & dissemination of species
• Anthropophilic fungi:
Outline
▪ Examples:
• Introduction to Dermatophytes Microsporum audouinii
• Ecology of Dermatophytes Trichophyton rubrum - red
• Classification Trichophyton schoenleinii
• Clinical manifestations Trichophyton tonsurans
• Laboratory diagnosis Trichophyton violaceum - violet
• Mycology Classification of Dermatophytes
Cutaneous mycoses
• No living tissue Microsporum
• Host Rxn to fungus
• Keratinase: hair, skin, nails
• Seen in deeper parts, usually in dermis
because may keratinase na sila.
Dermatophytosis
• Disease : Dermatophytosis (ringworm)
• Causative organisms: Dermatophytes
• Microsporum, Trichophyton, Epidermophyton
Cutaneous mycoses
• Disease
▪ Candidiasis of skin, mucous Trichophyton
membranes & nails
o Dermatomycosis
• Causative organisms
▪ Candida albicans & related species
o Soil fungi (Scytalidium,
Fusarium, etc.)
o Systemic fungi (Histoplasma,
etc)

Ecological Groups of Dermatophytes

Geophilic Epidermophyton (club-shape)

• “earth-loving”
• inhabit soil where they decompose
keratinaceous debris
• Dead animals
• Free-living soil saprophytes

Zoophilic

• Parasitic on animals

Anthropophilic fungi

• primarily parasitic to man

• man as exclusive host
Clinical Manifestations of Dermatophytes • Onychomycosis- non dermatophyte

Tinea capitis Laboratory Diagnosis

• Scalp, eyebrow, eyelashes 1. Skin scraping specimen

• Microsporum & Trichophyton
• Capitis meaning head

Tinea favosa

• Scutulum
• Mass of mycelia & epithelial debris
• Cup shaped crusts
• Favosa = hair 2. Direct Examination
Tinea corporis • Wet mount
• KOH
• Non-hairy skin – 10% to 30%
• Rings with scaly centers – with Parker Superquink blue-
• Rxn vs fungus black ink (required kasi mabilis
• Epidermophyton floccosum, Trichophyton, nya pianpasok ang fungal
Microsporum elements)
Tinea imbricata – gentle warming

• Concentric rings
• Trichophyton concentricum
• Happens in periorbital area

Tinea barbae

• Bearded areas of face and neck

Tinea cruris
3. Nail Specimen
• Jock itch • Clean with 70% alcohol
• Moist groin area • Scrape off outer surface, discard
• E. floccosum, T. rubrum • Scrape deeper portion
▪ Collect whole nail or clippings
Tinea pedis
▪ Collect debris (use paper)
• Athlete’s foot ▪ Materials: paper/envelope, scalpel
• Toe webs & soles, even nails 4. Specimen analysis
• Id reaction, circulating fungal antigens • Direct microscopy
o False negative = 5 - 15%
Tinea manuum
o 50% onychomycosis nail fail to yield
• Interdigital areas & palmar surfaces pathogen in culture
• Also seen in retropalmar 5. Hair Specimen
• Tweezers
Tinea unguium
• Scissors
• Invasion of nail plate by dermatophytes • Paper/envelope
• Thickened, discolored & brittle
Pattern of Hair Invasion Microsporum Canis

Ectothrix • Zoophilic
▪ cats and dogs
• formation of arthroconidia
• Invades
on the outside of hair
▪ Hair
shaft
▪ Skin
• cuticle of hair is destroyed
▪ rarely nails
• Hair invasion by a dermatophyte
• distribution
▪ Microsporum canis
▪ worldwide
▪ M. gypseum
• Lab diagnosis
▪ Trichophyton equinum
▪ Culture
▪ T. verrucosum
o White cottony
• Wood’s UV light
growth
▪ infected hairs
o Golden yellow
fluoresce
reverse colony – lagi
▪ Bright greenish
natinginan ditto kesa sa white
yellow
• microscopic:
• formation of
▪ spindle shaped, one
arthroconidia within
end pointed, other end
hair shaft
blunt
• cuticle of hair remains
▪ thick walled verrucose
intact
macroconidia
• do not fluoresce under
▪ 6 to 12 cells
Wood’s UV light
• ALL AGENTS ARE Microsporum gypseum
ANTHROPOPHILIC
• Geophilic
• Trichophyton
• usually produces a single inflammatory skin or
tonsurans, T. violaceum
scalp lesion
• Culture Media
• Distribution: worldwide
▪ Non-selective - Sabouraud’s dextrose
• lab diagnosis – culture
agar (SDA)
▪ flat, spreading suede-
▪ Selective
like to granular
- SDA with chloramphenicol &
▪ cinnamon growth
cycloheximide (Mycosel or
▪ yellow brown pigment
Mycobiotic agar)
on reverse of colony
- Dermatophyte test medium
• microscopic:
• Incubation:
▪ symmetrical ellipsoidal
▪ Room temperature
▪ thin walled verrucose
▪ At least 2 weeks
macroconidia
• Identification
▪ distal end slightly
▪ Gross color & texture
rounded, proximal
▪ Microscopic characteristics
(point of attachment) is
▪ Confirm / compare with
blunt
o Written descriptions
▪ 4 to 6 cells
o Drawings
o Photographs
Trichophyton mentagrophytes • lab diagnosis
▪ Varying sizes and
• zoophilic: mice, cats,
shape of
horses, sheep, rabbits
microconidia
• inflammatory skin or scalp
▪ Long clavate to
lesions in humans
broad pyriform
• ectothrix
▪ Occasional
• distribution: worldwide
clavate
• lab diagnosis - culture macroconidia
▪ flat, white to cream color; powdery to ▪ Partial requirement for thiamine
granular surface
• Microscopic
▪ spherical microconidia
- forming dense clusters, “en-
grappe”
▪ spiral hyphae
- smooth thin- walled clavate
With thiamine (left), w/o thiamine (right)
multiseptate macroconidia
• lab diagnosis Trichophyton concentricum
▪ positive for in-vitro hair perforation
• Anthropophilic
(mostly used)
• chronic non-inflammatory Tinea corporis
▪ positive urease production
• Tinea imbricata – concentric scaling of skin
Trichophyton rubrum • Not invade hair
• Lab diagnosis
• anthropophilic
▪ Slow growing deeply
• chronic infections of the skin, nails, rarely
folded thallus
scalp
▪ Cream to orange
• ectothrix or endothrix hair infection
brown in color
• distribution: worldwide ▪ Reverse buff to brown
• lab diagnosis – culture
• Microscopic – “antler tips” hyphae,
▪ white, suede-like to
chlamydoconidia
downy
▪ wine red pigment on Trichophyton schoenleinii
reverse side
• Anthropophilic
▪ scanty to moderate numbers of
• Cause favus
slender clav to pyriform microconidia
• Chronic scarring form of tinea capitis
▪ arranged “en-thyrse”
• Saucer shaped crusted lesions or scutula
▪ negative for in-vitro hair perforation &
• Permanent hair loss
urease production
• Lab diagnosis -Culture
Trichophyton tonsurans ▪ Waxy or glabrous
▪ Deeply folded honeycomb-like thallus
• Anthropophilic
with sub- surface growth
• Causing inflammatory or chronic non-
• Lab diagnosis
inflammatory finely scaling lesions of skin,
▪ Microscopic
nails and scalp
• Distribution: Worldwide
 o Favic chandeliers – mostly
used. yung parang mga branch
sa picture
o No macroconidia
o No microconidia

Epidermophyton floccosum

• anthrophophilic
• does not invade hair in vivo
• distribution: worldwide
• Culture:
▪ greenish-brown or
“khaki”
▪ colored
▪ suede-like surface
▪ raised & folded center,
with flat periphery
▪ yellowish brown reverse pigment
• Microscopic
▪ smooth thin-walled macroconidia
often in clusters growing directly
from hyphae
▪ no microconidia
▪ numerous chlamydoconidia
MYCOLOGY Etiologic agents:

Subcutaneous Fungal Infections - Fonsecaea

- Cladosporium
Subcutaneous mycoses - Phialophora
- “Inoculation mycoses” Types of sporulation:
- chronic, localized infections of the skin and
subcutaneous tissue - Acrotheca – conidia at the side
- traumatic implantation of the etiologic agent - Cladosporium – conidia in chains
- soil saprophytes of regional epidemiology - Phialophora – conidia in cluster
- ability to adapt to tissue environment and
elicit disease is extremely variable
Subcutaneous mycoses
Disease Causative organisms
Sporotrichosis Sporothrix schenkii
Chromoblastomycosi Fonsecaea,
Phialophora,
Cladosporium, etc
Eumycotic mycetoma Pseudallescheria, Acrotheca Type
Madurella,
Acremonium, - Conidia borne at the ends and sides of
Exophiala, etc conidiophore
Subcutaneous Basidiobolus ranarum, - Fonseceae pedrosoi, Fonseceae compacta
zygomycosis Conidiobolus o May exhibit all 3 types of conidiation
(Entomophthoromycosis) coronatus
Subcutaneous Rhizopus, Mucor, Cladosporium Type
zygomycosis Rhizomucor, Absidia,
(Mucormycosis) Saksenaea etc
Rhinosporidiosis Rhinosporidium
seeberi
Lobomycosis Loboa loboi

Chromoblastomycosis
- Chronic, slowly progressive & localized
infection Phialophora Type
- Tissue proliferation around area of
inoculation Phialophora verrucose
- flask-shaped or elliptical phialides with
flaring collarettes
- phialospores on top of the phialide

- Caused by dematiaceous fungi associated

with decaying vegetation or soil
Lab diagnosis:
- Crusted, verrucose, wart-like lesion
KOH of skin scrapings / crusts:
Distribution:
- brown pigmented, planate-dividing, rounded
- Worldwide but more common in bare-footed sclerotic bodies
populations in the tropics
 - Eumycotic mycetoma
o Madurella grisea
o Madurella mycetomatis
o Pseudallescheria boydii
o Curvularia

Culture: o Leptosphaeria
slow growing, suede-like, olive black in color

Mycetoma
- Human & animal infection characterized by:
o draining sinuses
o granules vary in:
o Pseudallescheria
▪ Size
▪ Color
▪ Hardness

o Exophiala
o Acremonium

o infection characterized by:

▪ tumefaction
▪ destruction of bone
▪ distortion of foot or hand
▪ hyperplasia at openings of
sinus tracts
- caused by traumatic implantation of spores
- involves cutaneous & subcutaneous tissues,
fascia & bone of foot or hand
Distribution:
- Worldwide
- more common in bare-footed population
o living in tropical or subtropical Lab diagnosis:
regions
- collect granules from sinuses, place sterile
Etiologic agents: gauze overnight
- culture granules to grow etiologic agents
- Actinomycotic mycetoma - Presence of sulfur - tissue biopsy; H & E staining:
granules, do not stain with fungal stain o Madurella mycetomatis - black
o Nocardia grains
▪ Nocardia asteroids
▪ Nocardia brasiliensis
o Actinomadura (Actinomyces)
▪ Actinomadura madurae o Streptomyces pelletierii - red grains
o Streptomyces o Streptomyces somaliensis - white
grains
 Etiologic agent:
- Sporothrix schenckii
o Dimorphic fungus
o Found in soil & decaying vegetation
o @ RT – MOLD
▪ Flowerette Conidia
▪ Old culture – Sleeve
formation
o @37oC – YEAST
▪ Cigar shaped yeast cells
Lab diagnosis:
- Stained tissue biopsy
Sporothricosis
o Yeast-like cells, may bud
- Rose gardener’s disease o “cigar bodies”
- Infection of the cutaneous or subcutaneous
Sporothricosis – lab diagnosis at 25°C:
tissues & adjacent lymphatics
- Culture on SDA:
o moist & glabrous
o wrinkled & folded surface
o color – white to cream to black
- Microscopic:
o Daisy-like microconidia
o short conidiophores at right angle
from thin hyphae
Sporothricosis – lab diagnosis at 37°C:
- Culture on BHI agar:
o Yeast colonies
- Microscopic:
o Budding yeast cells
- Characterized by nodular lesions which may
suppurate & ulcerate
- Traumatic implantation of the fungus into
the skin, rarely by inhalation into lungs

Subcutaneous zygomycosis
Entomophthoromycosis
- Caused by Conidiobolus
- Caused by Basidiobolus
Distribution: Members of subdivision Zygomycota
- Worldwide - Non-septate hyphae
- particularly tropical & temperate regions - Reproduce by zygospores (sexual)
Splendore-Hoeppli phenomenon in tissue
- Eosinophilic infiltration around hyphae
Entomophthoromycosis Caused by Conidiobolus
- Chronic inflammatory or granulomatous
disease restricted to the nasal submucosa
- Characterized by polyps or palpable
subcutaneous masses
Distribution:
- Worldwide, especially tropical rain forests of
Africa
Etiologic agent:
- Conidiobolus coronatus
- Present in soil & decaying leaves
Lab diagnosis:
- H & E stain of tissue biopsy
o Broad sparsely septate hyphae
surrounded by eosinophilic sheath
- Culture on SDA:
o Flat, cream colored, glabrous,
radially folded colony covered by
fine, powdery, white surface
mycelium
- Microscopic:
o Spherical conidia with hair-like
appendages (villae) & prominent
papillae, marking site of former
attachment to the sporangiophore

Subcutaneous mycoses
Rhinosporidiosis
- Rhinosporidium seeberi
Lobomycosis
- Loboa loboi
Treatment:
- Amphotericin B
- Surgical removal of infected tissue,
amputation
MYCOLOGY
Systemic Mycoses

- Systemic → blood infection, system based,

CNS infection, septicemia due to fungi
- Systemic mycoses that causes systemic shock
→ syndrome of severe vasodilation therefore
severe hypotension leading to decrease tissue
perfusion leading to multi-organ failure then
eventually death
o TYPES OF SHOCK:
▪ Hypovolemic (common) – low
volume of plasma
▪ Septic (common)
▪ Euvolemic etc.
- Septicemia – symptomatic blood infection due
to infectious agents (bacteria, myco, viro,
para)
- Bacteremia – presence of bacteria in blood
The Dimorphic Fungi
- They exist as yeast at 37°C and exist as mold
at room temp.
o Histoplasma capsulatum
o Histoplasma duboisii
o Coccidiodes immitis
o Blastomyces dermatitidis
o Paracoccidiodes brazielensis
Factors that Influence Dimorphism
- Temperature (yeast → 37°C ; mold → RT)
- Oxidation-Reduction Potential (due to
neutrophil present)
- Availability of Sulfahydryl Groups (because
fungi are sulfur-loving organisms, they’re more
likely on liver, blood)
- CO2 Tensions (fungi are aerobic)
Transformation from mycelial to tissue phase and
vice versa will result in changes:
Features of Systemic Mycosis
Host:
- Human host is a dead end in their life cycle
- Fungi have the ability to elicit disease process
in a NORMAL HOST
- In immunocompromised host - high risk of
disseminated disease
Mode of Transmission:
- Inhalation of large inoculum (fungi can easily
get an access to blood when it is inhale rather
than when it is eat because we have simple
squamous epithelium on both sides of
capillaries and alveoli)
Primary infection:
- asymptomatic or short lived
Host Response:
- persons with cell-mediated defects are at risk
for dissemination of disease or reactivation of
dormant infections
Etiologic agents:
- dimorphic, with mold and tissue phases
- geographically restricted
COCCIDIOIDOMYCOSIS

- Metabolism (will slow) - caused by Coccidioides immitis

- Physiology (will change) - Synonyms:
- Metabolic Rate o San Joaquin Valley Fever
- Cell Wall Composition o Valley Fever
- RNA polymerase regulators o Dessert Fever
o such as histine o Dessert Rheumatism
o lipid content o Cocci
o The Bumps
NOTES:
Life Cycle of Coccidioides immitis
Difficult to treat when morphing and it needs to be
isolated in one way. For example, isolate it in yeast - Yeast → mold → yeast → mold (basta mold
phase (what’s present in the blood or body) yun yung yung nasa outside world)
titingnan ng treatment.

LIFE CYCLE OF DIMORPHIC FUNGI

- Yeast → mold → yeast → mold

Where do you find C. immitis? Clinical Types of Coccidioidomycosis

- This fungus is found in the soil in the Western Primary

hemisphere: 40 degrees N (California) to 40
degrees S (Argentina) - Pulmonary: asymptomatic/symptomatic
- Cutaneous (rare)
Natural habitat
Secondary
- The fungus is found in dry, alkaline dessert
soil, like those in San Joaquin Valley in - Pulmonary: Benign chronic/progressive
California - Cutaneous
- Meningeal
Ecology of C. immitis - Disseminated

Clinical manifestations

- Fever, malaise, cough, chest pain (flu-like

symptoms)
- Erythema nodosum in women (pre-cutaneous
manifestation)
- Erythema multiforme in children (pre-
cutaneous manifestation)

Primary Pulmonary coccidioidomycosis

Factors that predispose to serious disease include:
- Exist as spherule in the lungs
- Amount of inoculum: infective dose is small
- Infective stage → arthroconidia (there’s joints
or juncture cells)
o “Very few arthroconidia are required
for infection. Outbreaks have followed
windstorms and earthquakes.”
o (MMWR Vol. 43# 10, p194, 1994)

- Right lobe consolidation. The appearance is

nonspecific and could be produced by other
- With cell-mediated immune defects, as in AIDS pathogens.
o “Primary exposure or reactivation of o Consolidation – suggestive imaging
earlier disease may lead to finding indicates that there is pus or
dissemination.” fluid build up in that particular section
- Race of the lungs.
o “For many years, the high incidence of
Laboratory Diagnosis
serious disease in FILIPINOS and
AFRICAN Americans was attributed to Specimens:
environment exposure.”
o “However, a violent windstorm that - Sputum
blew arthroconidia into urban areas - Biopsy
north of California resulted in - Etc.
disseminated rates, much higher in
Direct Microscopy: KOH/FAT
these races than seen with other
infected persons.” Culture:
- Pregnancy / Gender
o “Hormone receptors are present on - Needs BSL-2 (because the spores/conidia are
fungal surfaces and growth is aerosol)
stimulated by testosterone, - media without antibiotics
progesterone and 17B estradiol.” - slants used/bottles
o “Estrogen inhibits the transition from (HAZARDOUS)
mycelial to yeast phase.” NOTES:
- The absence of toxic erythemas during primary
infections Aerosol – > time, lighter, hospital
- Other factors such as presence of:
o HLA-A9 Droplet - < time, heavier, community
o Type “B” Blood

- Multinucleated giant cell – characteristic
astrocyte and microglia
- Macrophage that responds to Coccidioides but
the problem is that the spherule is very
resistant reason why it spherulyse
- Spherule contains lots of spores and when it
bursts it will then infect other cells
BLASTOMYCOSIS (North American
Blastomycosis)
- Caused by Blastomyces dermatitidis
- “Acute or chronic infection of lung, skin, bone
or genitourinary tract, characterized by
formation of pyogranuloma.”
o Pyogranuloma – granulomatous lesion
with pus
o Granuloma – build up of dead
neutrophils that does not look like a
pus, more like wound that do not heal
like leishmania infection
Etiologic Agent
- Blastomyces dermatitidis
o Dimorphic (mold at RT, yeast at body
temp.)
Where is it found?

- Largely confined to Canada and USA-states

around the Mississippi and Ohio rivers.
- Occasional cases seen in Africa.

Natural habitat

- Natural habitat remains an enigma

- Evidence indicate it favors environment:
o with high nitrogen content
o acid pH
o abundant moisture,
o and perhaps enriched with animal
excreta
- “It has been isolated in more than one
occasion from an abandoned kitchen near
Augusta, Georgia. It was isolated from
decaying debris collected near the boiler lid.”
- Isolated once from nature, from a beaver dam

Mode of Transmission

- Inhalation of spores (microconidia)

Clinical manifestations

- Usually pulmonary manifestation

- Symptomatic in the sense of atypical
Blastomycosis in AIDS
pneumonia (dry coughing at night)
- Some patients develop fever, some don’t - Low prevalence in AIDS
- Acute Respiratory Distress Syndrome (ARDS) – - Seen in patients with CD4 below 200/ul
drowning on its own breath like what happens - If it occurs in AIDS, it is a late manifestation
to COVID - May occur as infection localized in the lungs or
- Excessive pneumonia leading to low tissue as a disseminated disease
perfusion (respiratory distress)
NOTES:

- AIDS complicates everything (Uropathogenic E.

coli)
- In aids patients can cause sepsis (meningitis,
endocarditis, etc.)

Laboratory Diagnosis

- Direct Microscopic examination: KOH/FAT

Other forms of blastomycosis:

o Multinucleated giant cell - response to
Cutaneous blastomycosis
an abnormal foreign body, will recruit
- Blastomyces dermatitidis neutrophils and then it will recruit
o As the name implies, the infection, macrophages which will then ingest
although acquired by inhalation, is that foreign body, they’re like suicide
often seen as ulcers in the skin cells that keeps the foreign body
(dermis) inside their cytoplasm forever.
o It may also infect the bone
 Serology (super low sensitivity, not frequently used)

- Serum complement fixation

- Immunodiffusion test
- Radio immunoassay
- ELISA

“None have been useful because of low sensitivity and

specificity rates.”

NOTE:

- Serology - Used more on opportunistic

Cryptococcus

Epidemiology
Culture
- Blastomyces dermatitidis is an endemic fungus
- Requires 5 days to 4 weeks for growth; can be
that causes acute and chronic infections in
as short as 2-3 days
humans and other animals.
- Particularly found in the southeastern, south
central and midwestern part of the United
States
- Outside the US it has been reported in
Canadian provinces bordering the great lakes,
Africa, India, the Middle East and Central and
South America
- Risk factors for acquiring infections
o Associated with activities in proximity
to water ways
o Outbreaks have been associated with
campers and canoeists where
presumably, the soil has been
disturbed while gathering firewood.
- Exposure to dust and excavations

PARACOCCIDIOMYCOSIS (South American

Blastomycosis)
- Caused by Paracoccidioides brasiliensis
- “Chronic granulomatous disease that
characteristically produce a primary infection,
often inapparent, and then disseminates to
form ULCERATIVE granulomata of the buccal,
nasal, skin, adrenal glands and occasionally
the gastrointestinal tract.”
o Granulomata – plural of granuloma

Etiologic Agent

- Paracoccidiodes brasiliensis
o Dimorphic
Where is it found?

- It is endemic in Brazil and restricted in Latin

America from Mexico (23 degrees N) to
Argentina (23 degrees S)

Natural Habitat

- “Remains to be elucidated, but has been

isolated in acid soil.”

Mode of Transmission

- Inhalation of spores HISTOPLASMOSIS (Great "mimic" in mycosis"

caused by Histoplasma capsulatum)
Clinical Manifestations
Histoplasma capsulatum – common infection site is
Primary Benign disease: bone marrow
- 1° Pulmonary - Pulmonary infection resembles tuberculosis.
- Pulmonary: re-infection with allergic - It was first recognized as a disease among
manifestation patients who were X-ray positive, but
tuberculin negative.
Acute and chronic progressive-disease:
- Affects the RES cells (present in lungs, spleen,
- Disseminated disease liver, bone marrow and contains macrophage);
- Acute and chronic progressive pulmonary lesions are not confined to the lungs.
disease
NOTE:

TB-like infection

- Histoplasma capsulatum
- Paragonimus westermani

Etiologic Agent

- Dimorphic
- Diagnostic: thin-walled, single budding cells
inside macrophages

Diagnosis

- This disease has a long latency period. 10-20

years may pass between infection and
manifestation of the infection in the non-
endemic areas of the world.
- Probing for history of travel to endemic areas
is important in the diagnosis

Laboratory Diagnosis
Where is it found?
- Demonstration of multiple-budding cell from
clinical specimens - Histoplasma capsulatum is found along the
major river valleys in North and South America
- some areas of Africa and
- Far east-at least 50 countries in the temperate
and tropics including the Philippines

Ecology of the agent

- Grows in soil with HIGH nitrogen content;

prefers rotting guano of bats and birds mixed
with soil.

NOTE:

Cryptococcus neoformans - bird droppings / pigeon

droppings born
Environment favored by Histoplasma capsulatum NOTE:

- Open environment Cell-mediated immune response (CD4, CD8) - clears

o Nitrogen rich
o 22-29°C
o rainfall of 35-50 inches
o 67-87% relative humidity
o Guano from birds, particularly Sturnus
vulgaris or the starlings
o Bird not infected but organism can be
isolated from its feathers
- Closed environment
histoplasma
Clinical Syndromes in Histoplasmosis
- More Common
o Acute primary pulmonary (pneumonia)
- Less Common
o Disseminated histoplasmosis (in adults
with defective CMIR)
o Chronic pulmonary
o Mediastinal granulomatosis
o Fibrosing mediastinitis
- Bone involvement (bone marrow) is RARE

Pathogenesis: Mode of Transmission

- Inhalation of spores
- Risk factor: exposure to bird or bat droppings

“Massive exposure and infection has followed activities

such as earth Day exercises where school yards were
cleared and soil under tree roosts were disturbed.”

Histoplasma capsulatum

- If not removed from the reticuloendothelial

system, the organisms may become
disseminated and settle in many sites such as
the spleen, kidneys, brain, bone marrow and
lymph nodes.

o Looks like miliary TB

o Miliary TB – small lesions or peklat, TB-
like but tuberculin negative, AFB
negative

Features of Infections

- Most infections are self-limiting unless the

exposure is massive
- Persons at risk for disseminated infections and
Histoplasmosis in AIDS
high mortality include:
o Defects in CMIR (large increase since
the onset of HIV infections)
o Malnutrition
- In some cases, onset of a cell-mediated defect
can lead to reactivation of organisms latent
from a previous asymptomatic infection.
- Causes disseminated disease in 95% of cases
with AIDS
- 90% when CD4 count is below 200/ul
- Localized pulmonary disease when CD4 is
higher than 300/ul
- CNS involved in 10-20% of cases
Laboratory Diagnosis

- Specimens (these can demonstrate yeast cells)

o Sputum
o Buffy coat of blood (WBC layer)
o Sediments of biopsy materials
o Gastric lavage
o Urine
- Culture (needed for confirmation)
o Media - Direct Microscopic Examination
▪ SAB o (Wright-Giemsa methods or Grocott
▪ BAP without antibiotic silver methenamine stain)
▪ Smith and Goodman's medium
- for heavily contaminated
specimens
▪ Yeast extract with ammonium Diagnostic of
hydroxide H. capsulatum
▪ Culture: at 25°C, mycelial
form

- Complement Fixation Test

- Animal inoculation
- Skin testing
o Histoplasmic skin test

NOTE:

Commonly seen in lab is the bone marrow of chronic

disease

- Serology
o Double diffusion test (there’s
homologous antibodies)
▪ Rationale: nag didiffuse
papunta sa antigen yung
antibody. So if they are
homologous for example Test
Control (anti-histoplasma) and
Patient (unknown anti-
histoplasma), you’re going to
measure it against your
histoplasma antigens (has
precipitin) → your patient Ab
will then diffuse towards the
histoplasma Ag. When the Ab
binds to Ag, matatanggal yung
precipitin ng histoplasma Ag
resulting in a line of similarity.

Epidemiology

- Causes mild respiratory infections; results in

long lasting immunity
- common in areas with plenty of bats and birds
like the starling
o Population of the Sturnus vulgaris in
USA started with 60 birds from Europe
 brought to New York in 1890 by a
Shakespearean officionado
- In USA important infections among AIDS
patients

AFRICAN HISTOPLASMOSIS (Histoplasma

duboisii)
- Di daw nya itatackle kasi wala sa Jawetz!
MYCOLOGY - Responsible for the “moldy” description of the
earth
Opportunistic Systemic Mycosis “Aspergillus species produce large quantities of spores
Opportunistic mycosis (not common in the lab) which are easily dispersed by wind.”

Aspergillus fumigatus
- Fungal or fungus-like disease occurring in an
animal/human with a compromised immune - Accounts for over 90% of all infections
system - Grows in a wide range of temperature
- These are resident normal flora that become - can thrive up to 50°C
pathogenic only when the host’s immune
- Inhibited by Cycloheximide
system is altered and upsets the balance of
bacterial flora in the body Summary of clinical syndromes associated with
genus Aspergillus
Characteristics
- Allergy (to conidia/or transient growth of
- Host: compromised
organism in body orifices)
- Portal of entry: various (pulmonary,
- Colonization of pre-formed cavities (like in
cutaneous, etc.)
sinuses)
- Prognosis: Recovery depends on the severity of
- Invasive, inflammatory, granulomatous,
impairment of host defenses
necrotizing diseases of lungs and other tissues
- Immunity: No specific resistance to infection (ex. Aspergilloma)
(because patient is immunocompromised)
- Toxicity due to ingestion of contaminated food
- Host response: Depends on degree of (diarrhea due to Aspergillus toxin)
impairment – necrosis to pyogenic to
- RARELY systemic and fatal disseminated
granulomatous
diseases
- Morphology in tissue: No change in morphology
(except Pneumocystis carinii) Predisposing factors in Aspergillosis
- Distribution: ubiquitous
- Immunocompromised host (old age, AIDS
Common Opportunistic Fungi causing systemic patient, chemotherapy patient, cancer
mycosis patient, pregenetic diseases)
- Presence of other disease, etc or
- Pneumocystis carinii comorbidities
- Aspergillus spp
- Cryptococcus neoformans Mode of Transmission
- Candida albicans
- Respiratory
- Mucor/Rhizopus spp
- Direct Inoculation
ASPERGILLOSIS - Ingestion

“Designates a variety of pathologic conditions caused RESPIRATORY

by fungi belonging to the genus Aspergillus.”
- Initial and most usual sites of infections:
Etiologic Agents: (cavities)
o Respiratory tract (paranasal sinuses –
- Aspergillus fumigatus manifestation is paranasal sinusitis
- Aspergillus flavus that does not resolve with antibiotics;
- Aspergillus niger lungs – manifestation is Aspergilloma)
- Aspergillus terreus ▪ DISSEMINATE to other sites
- Aspergillus nidulans - Pathogenesis (Aspergilloma)
- Aspergillus clavatus o Spores disseminated from humidifiers
- Aspergillus restrictus and Air conditioner filters and ducts
that have accumulated moisture, and
- Aspergillus amsteloidami
from environment (after inhalation it
Red colored fonts – most common isolates will be deposited in the lungs. In the
CT-scan of lungs there’s a ball like that
Aspergillus species spores inside cannot be seen)
▪ Aspergilloma : Tomogram of
- About 300 species are found in the soil, dust
lung cavity containing fungus
and decaying matter
ball outlined by air space
- 8 are commonly associated with disease
NOTES: INGESTION

Air condition organism in bacte
haemophila or pneumoniae
– Legionella - Food contaminated with fungi producing
toxins:
- Aspergillus flavus – grows on peanuts and
grains and produce aflatoxin (most severe food
borne toxicity)

Other clinical manifestations: DISSEMINATED ASPERGILLOSIS

Allergy:

- Asthma
- Farmer’s lung

DIRECT INOCULATION

- Into subcutaneous tissues (mycetoma) Laboratory diagnosis

- Into the blood (drug abuse, valve replacement,
arterial catheterization etc) Microscopic Examination

NOTES: - Demonstration of organism in the tissues

(biopsy or autopsy materials)
Mycotic – caused by fungi o septate hyphae
o dichotomous hyphae
Actino mycotic – caused by Actinomyces species
o spores
There’s also mycetoma in Aspergillus
Culture

- Isolation of organism from tissues

- SAB agar used
- Incubated at room temperature (because it is
mold)
- Identification: Base on the characteristic
spores produced by the species

Traumatized tissue (black) on the pic – caused by - fan like appearance

Aspergillus niger
Treatment Mucor

For invasive aspergillosis: Microscopy:

- Amphotericin B - Non septate hyphae

- Itraconazole - Having branched sporangiophores with
sporangium at terminal ends
For aspergilloma: “individualized treatment”
Rhizopus
- Lobectomy may benefit some (can also be
tumor removal depend on patient’s Microscopy:
assessment)
- Shows non septate hyphae
NOTES: - Sporangiophores in groups (they are joined in
one stem)
Aspergilloma – surrounded by fibrous tissues
- They are above the Rhizoids
Omas – fibrous tissues or tumor like tissues surrounded
Mucormycosis: Clinical forms, probable routes of
by fibrous web (in aspergilloma – needed to remove
infection and underlying conditions
because treatment cannot perforate fibrous tissue)
Rhinocerebral/Rhino-orbital
Epidemiology
- Airborne
- Profuse sporulation and airborne
dissemination of spores promote human - Diabetes (common comorbidity/underlying
contact condition associated with this condition)
- Normal individuals get rid of spores by regular Pulmonary
“clearing mechanism” - phagocytosis
- Susceptibility is dependent on host’s particular - Airborne
altered condition - Leukemia, lymphoma
- Certain cases showed no apparent cause of - Immune suppression
implantation
Gastrointestinal
- Prevalence increases with the advances in
medicine, i.e. immunosuppression, - Ingestion
transplantation, cardiac surgery, etc - malnutrition, Amebic colitis
MUCORMYCOSIS (SYSTEMIC SYGOMYCOSIS Skin
OR PHYCOMYCOSIS)
- Traumatic
Mucormycosis (aseptate fungi) - Diabetes, wounds

- Opportunistic mycosis occurring worldwide Rhinocerebral mucormycosis: Bloodstained nasal

- Affects immunologically or physiologically discharge with left sided ptosis and proptosis (not
compromised hosts blood and there’s erythema in paranasal sinuses)
- Shows minimal pathogenicity for normal
NOTE:
persons
- Caused by saphrophytic molds found in the Left sided ptosis – aka eye/eyelid drooping
environment
Left and right ptosis – seen in Myasthenia gravis
- Cellulitis causes extensive tissue destruction
(cutaneous manifestation)
- Spread from nasal mucosa to turbinate bone,
paranasal sinuses, orbit, and brain (respiratory
manifestation – sinusitis)
- Rapidly fatal if untreated

Includes infections caused by: (what’s inside

Rhinocerebral mucormycosis: View through nasal
mucormycosis)
speculum showing fungal material arising from nasal
- Rhizopus turbinates
- Rhizomucor
- Absidia

NOTE:

Aseptate/non-septate hyphae → mucormycosis

Rhinocerebral mucormycosis: Gross brain specimen
viewed from base with acute superficial necrosis of
temporal lobe and thrombosis of left internal carotid
artery (this is already cerebral mucormycosis)
Cryptococcus
- found worldwide in the soil.
- It flourishes in bird guano and is often found in
large numbers in pigeon roots, old barns and in
soil beneath trees used as bird roosts.

NOTE:

C. neoformans – bird feces

Rhinocerebral mucormycosis with infarction of the Two varieties of Cryptococcus

hard palate C. neoformans C. gattii
Distribution Worldwide tropical &
subtropical
Reservoir Pigeon feces Eucalyptus
trees
Host • AIDS Normal
• Sarcoid host
• Lymphoma
Rhinocerebral mucormycosis: Advance case with • Corticosteroids
necrosis of nasal and maxillary tissue with black • CLL, ALL
eschar. Note periorbital edema and serosanguinous • Organ
discharge from eye. transplantation
Infection in Yes Rare
AIDS
Characteristics

Laboratory diagnosis
- Microscopic: KOH of tissues (and with other
stains)
- Findings: broad, irregularly shaped, non-
septated hyphae with right angle branching
- Culture
o Care in handling the specimen, since
organism is found in the environment
o 3-5 days incubation at room
temperature
o To identify: LPCB mount of growth
Treatment
- Early diagnosis highly essential for effective
curve
- High doses of IV Amphotericin B (for patients
with disseminated infections)
- Surgical interventions (for sinusitis, nasal sinus
drainage)
- Control of Diabetes as a basic requirement for
better clinical outcome
CRYPTOCOCCUS
“A subacute or chronic infection most frequently
involving the tissues of the CNS, but occasionally
producing lesions in the skin, bones, lungs or other
internal organs.”
- Encapsulated, yeast-like
- Reproduce by budding
- Most common infection in AIDS patients
- Size of capsule varies with surrounding growth
conditions
- Capsule basis for 4 serotypes
o A: neoformans, most common
o D: neoformans, rare linked with
primary cutaneous
o B and C: gattii
- C. neoformans, grows at 37°C on SAB or malt
agar (distinguishes it from non-pathogenic
species)
- In culture, colonies are visible after 72 hours
(white or tan colored, mucoid, smooth
colonies)
- Canavanine-glycol-bromothymol blue agar:
o C. gattii grows to produce color
change to cobalt blue
o C. neoformans does not grow,
indicator remains yellow
Pathogenesis
- Inhalation of infected particles from pigeon
feces and from flowering Eucalyptus
camaldulensis trees
- Primary Pulmonary infection:
o Asymptomatic
o Dormant
o Disseminated (with large inoculum)
Clinical syndromes
- Isolated pulmonary cryptococcosis (IPC)
- Disseminated cryptococcosis
o Chronic meningitis (usually seen)
o Systemic disease
o Cutaneous cryptococcosis
o Cryptococcosis and AIDS
Laboratory diagnosis
Microscopic examination
- India Ink mount (sputum, CSF sediments) (50%
sensitivity -less satisfying)
o In chronic meningitis – manifest as
seizures, WBC count of CSF do not
change and yet do not see infectious
agents. In CSF → there should be india
ink or negative staining which colors
the background and what you see is
conidia. Report immediately esp.
when the manifestation is meningitis
in immunocompromised patient.)
Culture
- (75% sensitivity): in SAB without cyclohexidine
- Demonstration of cryptococcal antigen by
Latex agglutination (for disseminated
infections)
- Serology: Detection of capsular antigen by
antibodies (use of rabbit’s anti - C. neoformans
capsule antigen) – 95% sensitivity (usual na
ginagawa)
- PCR
Treatment
Fluconazole
- First choice for mild to moderate pulmonary
disease and CNS or chronic pulmonary,
negative or positive for HIV infections
Amphotericin B
- Second choice mild to moderate pulmonary
disease, and CNS or chronic pulmonary,
negative or positive for HIV infections
Epidemiology
C. neoformans exist worldwide
- Found frequently in accumulated, old pigeon
droppings; has been isolated in Fort Santiago
- Fresh or wet droppings rarely contain the
organisms
- Pigeons are not clinically affected
- Has been isolated from droppings of other
birds such as parakeets and canaries
Cryptococcosis is considered an AIDS- defining illness
Cryptococcus gattii
- Found in Australia, South East Asia, Zaire,
Brazil Venezuela and Southern California
- Appears to related to the distribution of the
red gum tree (Eucalyptus camaldulensis)
which harbors the organism.
- Infectivity is correlated with the flowering of
the eucalyptus tree
- Does not cause outbreaks or clusters of
infections
CANDIDIASIS
Candidemia and Disseminated Candidiasis
“The genus Candida is associated with a number of
clinical syndromes usually seen among patients with
abrogated immune response, debilitating diseases, as
well as on those taking corticosteroids, anti-cancers
and on long antibiotic therapy.” (because residual
bacterial flora is eradicated)
The Genus Candida
- Over a hundred species
- Only 10 are commonly associated with disease
- Distribution:
- Saprophytes in human and animal digestive
tract
o Candida albicans (important)
o Candida albicans var stellatoidea
- Saprophytes in soil, air, water, dairy products,
fermenting/rotting vegetable products
o Candida tropicalis and pseudotropical
(also found in the respiratory tract)
o Candida parapsilosis (also in the skin)
o Candida krusei
o Candida guilliermondi
o Candida lusitaniae
o Candida viswanatii
“Candida albicans is the most common species
associated with human illness”
Candida albicans
- Exist primarily in the yeast form
o In tissue:
▪ It may be found in both yeast
and mold form
(pseudohyphae) – germ tube
o In agar medium: (looks like
Staphylococcus)
▪ Surface – oval budding cell or
yeast (white colonies)
▪ Lower layer – pseudohypha
Characteristics of Candida albicans
- Gram positive, coccus in shape (there’s
budding)
- Reproduce by budding, formation of
chlamydospores and by sexual reproduction
- Colonies are white and smooth
- Presumptive identification is based on
formation of germ tube within 90 minutes of
placing the organism in serum
HOW GERM TUBE IS MADE
Culture suspected of Candida albicans
1. Get 1 colony then put it on citrated plasma
2. Incubate it at 37°C for 90 minutes to 24 hours
3. Get the citrated plasma after incubation
4. Put it on slide then put coverslip
5. Place it on microscope to see whether there’s
a formed germ tube or lollipop like (because
this is part of the metabolism of Candida
albicans)
- Yeast cell will form germ tube (the pre-
requisite of hypha and pseudohypha)
- Germ tube is diagnostic of Candida albicans
Pathogenesis
- Candida as part of the normal flora, does not
cause disease - “Host must remain normal”
- Factors important to prevent infections:
o undisturbed normal flora
Factors which predispose to infection
- damage skin and mucous membranes within
GIT tract
- abnormal cell-mediated immunity
- presence of other diseases such as AIDS,
cancer, diabetes
- presence of:
o Central venous line
o Indwelling catheter
- Infancy – Old Age – Pregnancy
- Immunosuppression
- Diabetes mellitus
- Zinc and iron deficiencies
Candidemia and Disseminated Candidiasis
- May present as asymptomatic or fulminant
sepsis
- Must be considered when patient has these
risks factors:
o Malignancy
o Chemotherapy-induced neutropenia
o Organ transplantation
o GI surgery
o Indwelling catheter
o Burns
o Exposure to broad spectrum
antibiotics for a duration of >3 weeks
Disseminated candidiasis: typical skin lesions are
pinkish-red nodules. Patient with leukemia.
Oral candidiasis, severe infection, plaques of white
exudates on the tongue and palate. Severe infection in
a patient with AIDS earliest clinical sign of HIV
infection and may persists for months.
NOTE:
Oral thrush – AIDS earliest clinical sign, AIDS defining
illness
Laboratory diagnosis of Candidemia

- Culture: recovery of C. albicans from culture

- Microscopic: Demonstration of organism in the
biopsy materials by histological examination
(only done by pathologist)
- Serology available is not reliable because of
high false-negative and false-positive

Treatment

“Fluconazole seem to be a better alternative to

amphotericin B.”

Prevention and Control

“More on elimination of risks factors rather than the

elimination of the organism since Candida species are
ubiquitous.”
Pathogenesis
PNEUMOCYSTIS CARINII
“Pneumocystis carinii has long been considered a
member of the protozoan kingdom.”

- Still on debate whether it is parasite or fungi

Historical Background

- 1988 its ribosomal RNA sequence were found

to be closely parallel fungal rRNA
- Recently: gene sequence analysis suggests it is
a fungus
- Similarity of mitochondrial RNA with
ustomycetous red yeast fungi was striking - Patient with Pneumocystis carinii infection
manifests as Pneumocystis carinii pneumonia
Characteristics - Doesn’t know the treatment
- What you see on biopsy is fungal elements but
- Obligate extracellular organism
treatment is sometimes anti-protozoal or anti-
- Has not been cultured in vivo fungal
- Have different mammalian hosts - Disease progresses very fast
- Genetically diverse (different strains)
o i.e. isolate from one host will not Infection in AIDS patients
grow in another host
- They get sick more slowly than other patients
- Identified as most important opportunistic
and may survive longer without treatment
fungal infection in the Era of AIDS (aims for
- They die of the infection (harder to treat)
pneumonia)
- Many animals harbor on lungs in Rats, Ferrets, - Signs and symptoms: (more likely atypical
and Rabbits pneumonia)
o Triad of:
- Causes the disease in human if
▪ Shortness of breath with
immunocompromised
exertion
- Pneumocystis carinii – found in rats
▪ Non-productive cough
- Pneumocystis jiroveci in human species ▪ Fever

Laboratory diagnosis

- Imaging
 - Microscopic Examination: using various
staining methods
o Specimens:
▪ bronchoalveolar-lavage (BAL)
▪ transbronchial-biopsy
(spherule)
▪ Induced sputum
o Basis for diagnosis in > 90% of patients.
o Considered the “Gold Standard” in the
diagnosis
- Stains preferred:
o Giemsa (usually use)
o Toluidine blue
o Methenamine silver
o Calcofluor white (usually use)
o H&E
- X-ray of Chest supports the diagnosis
- Culture – not yet possible (it cannot be culture
in vitro)

Predisposing factors

- Corticosteroid therapy
- Transplant recipients
- Antineoplastic therapy
- A major cause of death in AIDS patients - when
retroviral treatment is not started
- Infections of the other organs is on raise,
spleen, lymph nodes, and bone marrow

Immunity

- In the absence of immuno suppressiom, P.

jiroveci does not cause disease
- Cell-mediated immunity plays a dominant role
in resistance to infection
- Infection not seen until CD4 counts drop to
<400 microliters

Drugs used in the management of PCP

- Trimethoprim – Sulphamethoxazole: for acute

infections
- Pentamidine: very effective
- Steroids (prednisone)
- Dapsone
- Clindamycin
- Atavoquone

Epidemiology

- New information suggest that PCP occurs as a

result of transmission from human host and not
a reactivation of dormant / latent infection
- Evidence for the person-to-person
transmission – underscored by the peak in
infections occurring about 4 months after
winter viruses
- Spread maybe from aerosolized secretions of
persons with viral infections
- Clustered outbreaks also support person to
person transmission