LECTURE 3 – MYCOLOGY

VITALES, GIENES
MYV

SYSTEMIC MYCOSES
 Infections that affect internal organs or deep tissues of the body.
 Frequently, the initial site of infection is the Lungs, from which the
organism disseminates hematogenously to other organs of the skin.
 LUNGS – way to get these fungi is through inhalation.
 Generalized symptoms include fever and fatigue. Chronic cough and
chest pain might also accompany these infections.
 Aspergillus, Fusarium, Bipolaris spp as well as some monomorphic
yeasts, Candida albicans and Cryptococcus neoformans.
 It is important to note that any fungus is capable of disseminating from
the primary site of infection in the immunocompromised host.
 The diseases are contracted generally by inhalation of infectious conidia
or spores.
 Aspergillus & Fusarium – causes Keratitis (eyes)
 Bipolaris spp – causes lungs infection
 Candida & Cryptococcus – opportunistic bacteria
 Cyptococcus neoformanis – causes meningitis
 Candida albicans – causes UTI & sepsis
Common Agents of Systemic Mycoses

FUNGUS ECOLOGY CLINICAL DISEASE TISSUE FORM

Blastomyces  Mississippi  Primary lung  Large yeast
dermatitidis and Ohio  Chronic, skin and  Broad
River bone based bud
Valleys  Systemic,
multiorgan
(FAILURE)

Histoplasma  Ohio,  Primary lung  Small oval

capsulatum Missouri,  Asymptomatic yeast in
Mississippi  Immunodeficient histiocytes,
River Host phagocytes
Valleys
 Bird and
Bat guano
(FECES)
 Alkaline
soil

Coccidioides  Semi-arid  Primary lung  Spherules

immitis, regions  Asymptomatic containing
Coccidioides  Mexico,  Progressive endospores
posadasii Central pulmonary,
and South multisystem
Americas
 Soil

Paracoccidioides  Central  Primary lung  hick walled

brasiliensis and South  Granulomatous yeast
America  Ulcerative nasal  Multiple
 Soil and buccal buds
lesions  MARINER’S
 Lymph node WHEEL
involvement
 adrenals
 FUNGUS MACROSCOPIC MICROSCOPIC
Blastomyces  White to dark tan  Oval, pyriform to
dermatitidis  Tenacious young globose
colonies  Smooth conidia
 Older colonies,  Short lateral
glabrous to woolly hypha-like
conidiophores

Histoplasma  White to dark tan  Microconidia, one

capsulatum  Woolly cottony or -celled
granular  Tuberculated
macroconidia,
hypha-like
conidiophores

Coccidioides immitis,  White to dark tan  Alternating one

Coccidioides to dark gray celled barrel
posadasii  Cottony older shaped
colonies  Arthroconidia
 Tend to grow in with disjunctor
concentrix rings cells

Paracoccidioides  White to beige  Sterile hypha

brasiliensis  Glabrous, leathery,  Conidia similar to
flat to wrinkled B. dermatitidis
 Folded or velvety
Blastomyces dermatitidis
 Most prevalent in middle-aged men
 When the primary disease fails to resolve, pulmonary disease may
ensue, with cough, weight loss, chest pain and fever.
 In immunocompromised patients, multiple organ systems may be
involved, and the course may be rapidly fatal.
 GEOPHILIC – mostly found in soil
 Blastomycosis is also known as Gilchrist disease, North American
blastomycosis, and Chicago disease
 Natural disease occurs in dogs and horses, with the disease process
mimicking that seen in human infections.
 The teleomorph is named Ajellomyces dermatitidis

Laboratory Diagnosis
 Large, spherical, refractile yeast cells
 8 to 15 um in diameter, with a double contoured wall and bude
connected by a broad base.
 In the mold phase, conidia are borne on short lateral branches that are
ovoid to dumbbell shaped and vary in diameter from 2 to 10 um
 In culture at 22 degree C, colonies can be white, tan or brown and may
be fluffy to glabrous.
 Frequently, raised areas termed as SPICULES are seen in centers of the
colonies.
 When grown at 37 degree C, on suitable media, this produces
characteristic broad based, budding yeast cells.
Histoplasma capsulatum
 Histoplasmosis is acquired by the inhalation of the microconidia of
Histoplasma capsulatum var capsulatum
 The microconidia are phagocytized by macrophages in the pulmonary
parenchyma. – FATAL, DISSEMINATED DISEASE.
 Other manifestations of the disease are mediastinitis, pericarditis, and
mucocutaneous lesions.
 Histoplasmosis is also known as reticuloendothelial cytomycosis, CARE
DSE., DARLING’S DSE. And SPELUNKER’S DSE.
 This organism resides in soil with high NITROGEN content,particularly in
areas heavily contaminated with bat and bird guano ( FECES).

Laboratory Diagnosis
 When smears are stained with Giemsa or Wright stain, the yeast cells
are commonly seen within monocytes and macrophages, occurring in
significant numbers.
 Grows as a white to brownish mold. Conversion of the mold to yeast
form using BHI agar incubated at 37 degree C, is confirmatory for H.
capsulatum.
 The small cells when found in tissue, resembles the blastoconidia of
Candida glabrata, but they can be differentiated by fluorescent
antibody techniques or culture.
 As the colony matures, large echinulate or tuberculate macroconidia
characteristic for the species are formed.
Coccidioides immitis
 Is probably the most virulent of all human mycotic agents
 The inhalation of only a few arthroconidia produces
primary coccidioidomycosis.
 Clinical infections include asymptomatic pulmonary disease and allergic
manifestations. Allergic manifestations can manifest as toxic erythema,
erythema nodosum (desert bumps), erythema multiforme (valley
fever), and arthritis (DESSERT RHEUMATISM).
DESSERT RHEUMATISM – found in mediterranian area
 In symptomatic patients, fever, respiratory distress, cough, anorexia,
headache, malaise and myalgias can manifest for 6 weeks or longer.
 The disease might then progress to secondary coccidioidomycosis,
which can include nodules, cavitary lung disease, and/or progressive
pulmonary disease.
 Misdiagnosed as allergic reaction.

Laboratory Diagnosis
 After inhalation, the barrel shaped arthroconidia, which measure 2.5 to
4 um x 3 to 6 um, round up as they convert to spherules.
 Colony appears to enlarge in a circular bloom, and as the culture ages,
the vegetative hyphae also fragment into arthroconidia.
 Direct smear examination of secretions may reveal the spherules
containing the endospores.
 Microscopic examination of the culture shows fertile hyphae arising at
right angles to the vegetative hyphae, producing alternating hyaline
arthroconidia.
Paracoccidioides brasiliensis
 Affects lymph nodes involvement & ulcerative granulomatous lesions
 Primary route of infection is pulmonary
 Ulcerative granulomatous lesions of the buccal, nasal, and occasionally
gastrointestinal mucosa. A striking lymph node involvement is also
evident.
 Is the causative agent of Paracoccidioidomycosis, also known as South
American blastomycosis, Brazilian blastomycosis, Lutz-Splendore
Almeida disease, Paracoccidioidal granuloma. A chronic, progressive
fungal disease endemic to South and Central America
 Tendency to go to adrenal glands.

Laboratory Diagnosis
 The typical budding yeast measures 15 to 30 um in diameter with
multipolar budding at the periphery, resembling a “mariner's wheel”.
 Many buds of various sizes can occur or there can be only a few buds
giving the appearance of a so -called “MICKEY MOUSE CAP” to the yeast
cell.
 Flat colonies are glabrous to leathery, wrinkled to folded, floccose to
velvety, pink to beige brown with a yellowish brown reverse, resembling
those of B. dermatitidis.
 On BHI blood agar at 37 degree C, the mycelial phase
rapidly converts to yeast phase.
Penicillium marneffei
 Is unique among the Penicillium spp., being dimorphic.
 The only true pathogen in the genus.
 Infections are usually disseminated, with multiple organ involvement.
 The fungus can be isolated from cutaneous lesions, which are frequently
present in infected patients.
 Respiratory pathogen
 Common host: rodents; particularly in bamboo rats
 Yeast: red pigment
 Mold: green to red colony
 Cells of Penicillium marneffei resembles Histoplasma capsulatum
 They can be detected through Wright’s stain on lesions and biopsy
specimen.

Laboratory Diagnosis
 Yeast -like cells can be detected in Wright-stained smears from skin
lesions or biopsy specimens.
 Cells resemble those of H. capsulatum, oval to cylindric, measuring 3 to
6 um long, and may have a cross wall.
 The mold form is described as having sparse green aerial and reddish
brown vegetative hyphae and production of a red diffusible pigment.
AGENTS OF OPPORTUNISTIC MYCOSES
 Surgical wounds are ideal points of inoculation, allowing saprobes to
become opportunistic agents of disease.
 Skin and nail bed infections as well as severe respiratory infections can
be caused by a variety of fungi in patients with AIDS.

MUCORALES

Cunninghamella
 Can be recovered from the sinuses or other organs during disseminated
disease.
 Sporangiophores are erect, branching into several vesicles that bear
sporangioles and can be covered with long spines.
 They form a cottony colony that is initially white but becomes gray with
age.

Lichtheimia
 Have a predilection for vascular invasion, causing thrombosis and
necrosis of the tissues.
 Usually found in Diabetic patients suffering from ketoacidosis.
 From the sinuses, infections rapidly spreads to the orbits, face, palate,
and brain.
 This presentation is known as Rhinocerebral zygomycosis.
 Lichtheimia hyphae are broad and ribbon like, with few septations.
Sporangiospores are smooth and ovoid.
 Internodal rhizoids are short, thin, projections that anchor the growing
cells to the substratum is also present.
Mucor
 Have been implicated in rhinocerebral zygomycosis in addition to
disseminated disease.
 Sporangiospores are formed in sporangia on erect sporangiosphores.
The sporangia remain frequently intact as opposed to Rhizopus spp., in
which the sporangia typically collapse.
 They grow rapidly and form cottony, dirty white colonies that become
mousy brown with age.

Rhizopus
 It is typically involved in diabetic patients with ketoacidosis, presenting
as Rhinocerebral mucormycosis
 At the base of sporangiophores are Brown rhizoids.
 The sporangia are typically fragile and are not easily retained when
making slide culture preparations, resulting in an umbrella -shaped
structure at the end of the conidiophores.

Syncephalastrum
 Is rarely implicated in human disease but has been documented in
cutaneous infections.
 Microscopically, erect sporangiophores are noted. Each has a large
columella on which merosporangia (containing stacks of
sporangiospores) are formed.
 SEPTATE AND HYALINE SAPROPHYTES

Beauveria
 Beauveria bassiana is a rare, human isolate, uncommonly associated
with Keratitis
 This fungus is a known insect pathogen, and is found worldwide on
vegetation and in the soil.
 Abundant, single celled, tear shaped sympoduloconidia are formed on
sympodulae, which taper extremely from a rather swollen base.
 Conidiophores may cluster in some isolates to form radial tufts.

Aspergillus
 Aspergillus fumigatus os the specie most commonly isolated.
Other pathogenic species include A. flavus, A. terreus, A. niger.
 Nuetropenia is the single most predictive factor for developing
aspergillosis.
 Aspergillus spp., are the most frequent cause of disease in bone marrow
transplant recipients in addition to other cancer and transplantation
patients.
 Another frequent presentation is the so -called “fungus balls” in the
lungs of agricultural workers who routinely are in contact with fungal
conidia from environmental sources.
 Can be uniseriate or biseriate.
 Most known pathogens in this group form green to tan colored species
Chrysosporium
 Have been recovered from skin and nails lesions
 Microscopically simple, wide based, single celled conidia are produced
on non-specialized cells
 Colonies are hyaline with a moderate growth rate that with age, can
develop light shades of pink, gray or tan pigment.

Fusarium
 Are frequently seen in Mycotic keratitis
 Patients presents with high fever, possibly disseminated skin lesions,
and in some, fungemia.
 Macroconidia are banana- or -canoe shaped and are formed singly, in
small clusters, or clustered together in mats termed sporodochia. They
are typically multi -celled.
 Is a rapidly growing hyaline fungus that can develop various colors with
age, ranging from rose to mauve to purple to yellow.

Geotrichum
 Has been implicated in pulmonary disease in immunocompromised
patients.
 Microscopic evaluation reveals abundant arthroconidia formed from the
vegetative hypha that occurs singly or may be branched.
 Colonies are white to cream, yeastlike and can be confused with
Trichosporon spp.
Paecilomyces

 Phialides of Paecilomyces are generally longer and more obviously

tapered, and they may be singly formed or arranged in a verticillate
pattern, on which long chains of spindle -shaped or somewhat
cylindrical conidia are formed.
 They grow rapidly and usually form flat, granular to velvety colonies in
shades of tan, brownish gold, or mauve.

Penicillium

 Rarely cause infections, most reports of disease involve chronic fungal

sinusitis.
 Conidiophores are erect, sometimes branched, with metulae bearing
one or several phialides on which oval to ovoid conidia are produced in
long, loose chains.
 This commonly seen fungus is a rapid grower with colonies usually in
shades of green or blue green.
Scopulariopsis
 Are commonly isolated from nail specimens and have been implicated
in pulmonary disease in immunocompromised patients
 Conidia are formed from annellides which increase in length as conidia
are formed.
 The truncate based conidia tend to remain in chains on the annellides.
 Are moderate rapid growers and forms colonies covered by tan to buff
conidia. Some species are phaeoid.

Trichoderma
 Causes pulmonary and skin infections
 Rapidly growing and form hyaline hyphae that gives rise to yellow green
to -green patches oc conidia formed on clusters of tapering phialides.
 Colonies are intensely green and granular, with abundance of conidia.
SEPTATE AND PHAEOID SAPROPHYTES

Alternaria
 Primarily implicated in chronic fungal sinusitis.
 Can be found systemically in those suffering from immune suppression.
 Multi -celled conidia have angular cross walls and taper toward the
distal ends.
 They are phaeoid, rapidly growing fungi with colonies ranging from
shades of gray to brown to black.

Aureobasidium (water loving)

 This organism may be recovered from blood, tissues and abscesses.
Usually seen in wet conditions, such as shower tiles and water lines.
 Young cultures are off - white to pink, but with age they become black,
with the production of darkly pigmented arthroconidia.
Chaetomium (cellulose loving)
 Infections with this organism have been reported in the brains of
patients with central nervous system disease.
 These fungi are found in the environment amd have a predilection for
cellulose products.
 These perithecia are pineapple shaped and are ornamented with
straight or curled hairs.
 At maturity, the pigmented, lemon shaped ascospores are released
within the perithecium.
 Some species produce a diffusible pigment that turns the agar
completely red.

Cladosporium
 Infections are typically confined to the sinuses or following traumatic
inoculation.
 They form brown to olive to black hyphae and conidia
 Branched conidium bearing cells may dislodge, and the three scars on
each of these cells give them the appearance of a shield.
 Slow to moderate growers, with granular to velvety to fluffy colonies,
ranging in color from olive to brown or black.
Curvularia
 These isolates are usually implicated in chronic sinusitis in
immunocompetent patients.
 This genus is among the easiest to identify because of the frequently
seen crescent -shaped conidia with 3 to 5 cells of unequal size and an
enlarged central cell.
 These fungi form a rapidly growing phaeoid colony that is cottony and
dirty gray to black.

Phoma
 Produce pycnidia, which as appear as black fruiting bodies that are
globose and lined inside with short conidiophores.
 Large numbers of hyaline conidia are generated in the pycnidium and
flow out of a small apical pore.
 They specie produce a moderately rapid growing gray to brown colony.

Pithomyces
 Conidia are somewhat barrel shaped, formed singly on simple short
conidiophores
 Conidia have both transverse and longitudinal cross walls and are often
echinulate.
 Produce rapidly growing phaeoid colonies.
Ulocladium
 Conidiophores bear dark, multi -celled conidia on sympodial
conidiophores
 Conidia have angular cross walls, and in some species, echinulate
surfaces.
 They are rapidly growing phaeoid fungi, forming colonies ranging in
color from brown to olivaceous black.

AGENTS OF YEAST INFECTIONS

 Yeast fungi can be classified into one of the 2 groups, yeasts and yeast -
like fungi.
 Isolates that reproduce sexually, either by forming ascospores or
basidiospores, are truly yeasts.
 Most isolates that are not capable of sexual reproduction, or whose
sexual state has not yet been discovered are correctly termed as yeast -
like fungi.
 Normal color of a yeast colony ranges from white to cream or tan, with
a few species forming pink to salmon colored colonies.
 Texture of the yeast colonies also varies.

CANDIDA
 Normal biota of the skin, mucosa and digestive tract.
 Clinical disease ranges from superficial skin infections to disseminated
diseases.

Candida albicans
 Is recovered as normal biota from a variety of sites, including mouth,
skin and vagina.
 One of the most widely recognized manifestations of C. albicans
infection is Thrush, an infection of the oral mucosa.
Candida glabrata
 Infections associated with these tend to be aggressive and difficult to
treat with traditional antifungal therapy.
 The second most common Candida species to cause disease and may
account for 21% of all urinary yeast isolates.
 This organism has different sugar assimilation patterns, notably rapid
assimilation of Trehalose, from those of C. albicans and therefore easily
differentiated.
 Other notable species of Candida are Candida krusei and Candida
tropicalis.
 Candida parapsilosis has become a major cause of outbreaks among
nosocomial infections.

Cryptococcus
 Cryptococcus neoformans is the most notable pathogen in this
genus, a major cause of opportunistic infections among AIDS patients.
 Commonly found in soil contaminated with pigeon droppings and
most likely acquired by inhalation.
 Cryptococcus gattii, is an emerging pathogen, particularly in the Pacific
Northwest of the US
 Cryptococcus spp express a capsule that produces the characteristic
Mucoid colony. This can be detected surrounding the budding yeast
using India Ink or Nigrosin.
 All species of this genus are urease positive and the nitrate reaction
varies.
 Production of Phenoloxidase is a feature differentiating C. neoformans
from many other Cryptococcus spp.
 A key laboratory characteristic of C. gattii is that it will use Glycine as a
sole carbon and nitrogen source in the presence of canavanine,
whereas C. neoformans will not.
 Canavanine glycine bromothymol blue agar is commercially available
for this purpose.
Rhodotorula
 Are noted for their bright Salmon pink color
 They are closely related to the cryptococci because they bear a capsule
and are urease positive.
 Some species are also nitrate positive.

Pneumocystis Infection
 P. carinii was originally classified with the protozoa.
 P. carinii is the specie most commonly found in rats, and P. jirovecii is
the specie most often found in Humans.
 Pneumocystis spp initially was identified as the causative agent in
interstitial plasma cell pneumonia seen in malnourished or premature
infants.
 They are a non-filamentous fungus with 3 life stages. Trophozoite, 1 to
5 um in size and is irregularly shaped, the pre cyst 5 to 8 um in size; and
the cyst, which is a thick walled sphere of about 8 um in size containing
up to 8 intracystic bodies.

Laboratory Diagnosis
 transbronchial biopsy, tracheal aspirate, pleural fluid and induced
sputum.
 Sputum, is the least productive specimen.
 Giemsa and Gomori methenamine silver are used. Using the latter, the
cyst wall stains black.
 Cysts often have a punched out ping pong ball appearance.
 With the Giemsa stain, the specimen appears round and the cyst wall is
barely visible.