การพิสูจน์ชนิด · Zygomycosis: A history caused byfungi of the order...
Transcript of การพิสูจน์ชนิด · Zygomycosis: A history caused byfungi of the order...
Sporotrichosis Mycetoma Rhinosporidiosis Chromoblastomycosis
Subcutaneous zygomycosis
PhaeohyphomycosisUlocladium sp.
Hyalohyphomycosis
Chromoblastomycosis or dermatitis
verrucosa
A localized chronic
mycosis of skin,
Worldwide esp. tropical zone
Rough, irregular lesions at Foot and leg
lesions
Habit in nature but infect to open wound
Dematiaceous fungi
เชือ้สาเหตุPhialophora verrucosa Fonsecaea pedrosoi
Fonsecaea compacta Cladosporium carionii
Chromoblastomycosis
Fonsecaea compacta Cladosporium carionii
Rhinocladiella aquaspersa พบน้อย
ตุ่มใส และตุ่มหนอง papule / pustule: violaceous papule / pustule: violaceous ุ ุ
นูน ยกตัว VerrucousVerrucous cauliflower-likecauliflower-like
หนอง serous exudate , pusserous exudate , pusหนอง serous exudate , pusserous exudate , pus
สะเกด็แห้ง crust (red to greyish)crust (red to greyish)
localized lesion
Cat: nonpruriticswelling in nasal
planum
Dog: ventral abdominal l ilesion
sclerotic bodies: 5-12 mm
• Brownish round with muriform
5 - 12 um
planate-dividing
specimens: tissue, skin
scraping
l ti
KOH-preparation
hi t th l
Culture
sclerotic bodies at the
lesion
histopathology
Sabouraud
Dextrose Agar
(SDA)
Brown,
Slowly grow
,livaceo
us black
velvet or fluffy
blastoconidia (acropetalous conidiation)
Cladosporium type Rhinocladiella(Acrotheca) type
Phialophora type Fonsecaea type
การพสิูจน์ชนิด
Genus Cladosporium• Cladosporium type
Genus Phialophora• Phialophora type
Genus Rhinocladiella• Rhinocladiella
type
Genus Fonsecaea• All 4 types but
mostly form Fonsecaea type
F. compacta : compact head conidia
( ) ‘Rhi l di ll • two rows of conidia(a) ‘Rhinocladiellatype’
(b) ‘Cladosporium type’
(c) ‘Phialophora type’
• two rows of conidia• role of conidiogenuos cell • one conidial
• Phialides
Zygomycosis: A historycaused byfungi of the order entomophthorales and mucorales
Conidiobolus coronatus: an etiological agent of nasal granulomain horses (1961)
The first case of human infection (1965) from Jamaica
Tropical countries of Africa, Asia and America
Reported in Thailand (1981)
Basidiobolomycosisy• (Entomophthoromycosis basidiobolae)
d b lConidiobolomycosis• (Entomophthoromycosis conidiobolae)
Rhinocerebral zygomycosis in sheep (Morris et al., 2001)
H tH tFungus Fungus
Conidiobolus coronatus
Organ involvedOrgan involved
Orofaciallesion
Sinus and
SourceSource
Soil
HostHost
Horses
Llamas
C. incongruus
C. lamprauges
turbinate bone
Nodular subcutaneous
lesions
Insect
Decaying vegetation
Sheeps
Dogs
Sheep Conidiobolomycosis in Brazil, Silva et al., 2007.
geographical host transmissions source of um
distributions
Africa
Asia
host
human
transmissions
wound contamination
infections
Reptile feces
decayed plantob
olu
s ra
na
ru
America horse insect
bitted
plant
intestinal content of
insects
B
asi
dio
Hor
se • Nasal granuloma with bilateral nasal swellings
• epistaxis• symptoms of
dyspnea Chi
mpa
nzee
• Tumorousswellings on the exterior and interior of the nose
• Both eyebrowsdyspnea C
การตรวจทางห้องปฏบิตักิาร
Tissue sample Splendore hoeppli material observed by H&E
KOH-preparation Histopathology
eosinophilic material coats the grains that are characteristic of the exudatein lesions of mycosis
Large hyphae
8-20 μm
การเพาะเชือ้จากสิ่งส่งตรวจur
e on
SD
A
• grow at room temperature
l b
Hyp
hae • Young
colony: AseptateA i hi
ck-w
alle
d os
pore
sore
• Spherical conidia
• Prominent ill
Cul
tu • glabrous colony
• Aging colony: sparsely septation C
onid
ia: t
hzy
go papillae conidia
Beak-like zygospore
Syncephalestrum sp.
Syncephalastrumracemosum
Cylindrical merosporangia
Fast growing, cottony to fluffy,
becoming dark grey
Soil and dung in tropical and
subtropical regions
Human pathogen, horse normal eye flora (Rosa et al., 2003)
Merosporangia : thin-walled, contain 5-10 globose to ovoid, smooth-walled sporangiospores (merospores).
B. ranarum ( haptosporus) asexual spore
Single globose conidia
Elongated conidia
Secondary conidia
germinate
How to differentiate them?
STRUCTURE DIFFERENTIATION
Ejected sporangioles possessing papilla Basidiobolus (-)Conidiobolus (+)( )
Sporangiophores with swollen apices Basidiobolus (+)Conidiobolus (-)
ZygosporesBasidiobolus (+)Conidiobolus coronatus (-)Conidiobolus incongruus (+)
Zygospores with conjugation beaks Basidiobolus (+)Conidiobolus incongruus (-)
Subcutaneous phaeohyphomycosis
Alternaris alternata in mare
Dematiaceous fungi
an opportunistic fungal infection
• Alternaris alternata in mare• Curvularia sp. • Phialemonium curvatum• Ochroconis tshawytschae in salmon, O. humicola in trout• Exophiala jeanselmei• Wangiella dermatitidis
Wound infection
Hosts: Dog cat horse chameleons and trout
• pus • black hyphae
Phaeomycotic abscesses contained
Skin sinus and lung
การวนิิจฉัยทางห้องปฏบิตักิาร
• Dematiaceous septate
hematoxylin and eosin (HE)
periodic acid–Schiffphyphae
• Dark yeast cell• Moniliform hyphae• Sclerotic cell
Staining tools
p(PAS)
Gomori–methenaminesilver (GMS)
Fontana–Massontechniques.
Histopathology
KOH preparation from abscess content (grain)
Culture on SDA for 1 week
Alternaris alternata Curvularia sp. Phialemoniumcurvatum
Ochroconis sp. Exophiala jeanselmei Wangiella dermatitidis
Dematiaceous fungus within the keratin of follicular infundibulum.
Epithelioid-like macrophages, multinucleated giant cells, and lymphocytes
Pyogranulomatous perifolliculitis.
Intrafollicular fungi stained black.
Number of hyaline (non-dematiaceous)
Wound infection
Subcutaneous abscesses
hyaline septate hyphae in grain
Clinical material• Skin and nail scrapings N t h h l
การวนิิจฉัยทางห้องปฏบิตักิาร
p g• urine, sputum • bronchial washings• cerebrospinal fluid• blood; tissue biopsies
Direct Microscopy
Note hyphalelements are often difficult
to detect in H&E stained
sections.
• 10% KOH and Parker ink• Exudates : centrifuged and sediment then examined
with 10% KOH and Parker ink• Tissue sections should be stained
Scedosporiumapiospermum
Fusarium sp. Penicillium sp. Aspergillus sp.
Paecilomyces sp. Acremonium sp. Beauveria sp.Scopulariopsis sp.
Pythiosis: Pythium insidiosum
Pythiuminsidios
um
Ulcerative pyogranulomatous
or fibrogranulomatous
Horse, cattle,
cat and dogs.
Tropical and
subtropical
areas.
Ventral trunk and head Formation of the Pythium
i idi ' i Scanning electron
insidiosum's sporangia and zoospores (a to h) and release of the flagellated zoopores in water (i).
microscopy of Pythium insidiosum'ssporangia.
• Sporangia of Sporangia of Pythium insidiosumon a grass leaf.
Culture plate of Pythium insidiosum (Sabouraud).
Cat pythiosis: tumoral-like mass caused by Pythium insidiosum.
Cutaneous dog pythiosisAtypical case of equine pythiosis. The infection began in the lower extremity and spread, trough lymphatic vessels, to the horse's chest.
Typical cutaneous pythiosis in an equine. Small white masses, known as "kunkers", are observed in the center of the lesion.
Kunkers or leeches:
Granulomatous coagula consist of necrotic Mc, Eoand hyphae
Horse pythiosis with bone involvement (from a chronic case of the disease).
• "Kunkers" removed from the above case. These masses containe viable hyphae of P. insidiosum.
Subcutaneous pythiosis in a Texan boy . Orbital pythiosis is common in children and should be differentiated from orbital zygomycosis and aspergillosis and Thai boy with pythiosis in his carotid artery
• Amputated section of a patient leg that had pythiosis.The picture shows the aneurysm in the femoral artery caused by P. insidiosum from a Thai human patient.
Phylum Chytridiomycota
Diseases of amphibians and
reptiles
Emerging infectious
Abnormal epidermal
sloughing and
Hemorrhage in the skin
diseases
Mass mortality
sloughing and ulceration
A population pyramid to demonstrate the factors thought to heighten the impact of chytridiomycosis in amphibian populations.
Excessive sloughing of skin with thallus of fungi
bInoculum from the suspect tissue Dissecting needle
Observe mycelium on microscopy
Prevent airborne spores
1% formaldehyde for 1 to 5 minutes 70% alcohol
sterile water Culture on agar media
Agar: sweet wort agar, Sabouraud's agar Czapek agar
Low-nutrient: maintaining the Low nutrient: maintaining the strains in collection cultures
spores + mycelium transfer onto agar
Inoculation at a room temperature
4–10°C allow to survive up
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