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LABORATORY DIAGNOSIS OF COMMON FUNGAL DISEASES Prof. Louella A. Dancel, RMT, MSc. University of Perpetual Help System DALTA   42 nd  PAMET ANNUAL CONVENTION Manila Hotel – December 2,2006
Clinical specimens submitted for Fungal Isolation 2004-2006* Clinical specimen  2004 2005 2006 Total Respiratory 29 35 34 98 Body fluids 37 17 34 88 Tissues 9 0 23 32 Skin 1 0 0 1 Nails 29 31 20 80 Hair 0 1 0 1 CSF 8 5 8 21 Others 5 20 24 49 TOTAL 118 109 153 380
Fungal Isolates 2004-2006 * Isolate 2004 2005 2006 C.albicans 5.1% 13.8% 11.8% C.tropicalis 1.8% 3.7% 5.2% C.parapsilosis 5.1% 6.4% 3.9% C.glabrata 0.91% 0.91% 3.9% C.famata 1.8% 4.6% 2.0% Aspergillus spp. 0.91% 0% 0.65% Fusarium spp. 0% 0% 0.65%
Data from a Five-year review of Fungal Isolates at UPM-CPH ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Data from a Five-year review of Fungal Isolates at UPM-CPH ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Data from a Five-year review of Fungal Isolates at UPM-CPH ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Data from a Five-year review of Fungal Isolates at UPM-CPH ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Data from a Five-year review of Fungal Isolates at UPM-CPH ,[object Object],[object Object],[object Object],[object Object],[object Object]
Laboratory Methods for Diagnosis of Mycoses ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Laboratory Methods for Diagnosis of Mycoses ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Laboratory Methods for Diagnosis of Mycoses ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Mycotic Infections ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Superficial mycoses ,[object Object],[object Object],[object Object],[object Object],[object Object]
Superficial mycoses ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Superficial mycoses ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Pityriasis versicolor ,[object Object],[object Object],[object Object],Superficial mycoses www.ethnomed.org
Pityriasis versicolor ,[object Object],[object Object],[object Object],Superficial mycoses Modified from www.columbia.edu
Pityriasis versicolor ,[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Pityriasis versicolor ,[object Object],[object Object],[object Object],Superficial mycoses
Pityriasis versicolor ,[object Object],[object Object],Superficial mycoses
Pityriasis versicolor ,[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Pityriasis versicolor ,[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Pityriasis versicolor ,[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Tinea nigra ,[object Object],[object Object],[object Object],[object Object],Superficial mycoses 11th.blogspot.com
Tinea nigra ,[object Object],[object Object],[object Object],Superficial mycoses
Tinea nigra ,[object Object],[object Object],Superficial mycoses
Tinea nigra ,[object Object],[object Object],[object Object],Superficial mycoses
Tinea nigra ,[object Object],[object Object],[object Object],Superficial mycoses
Tinea nigra ,[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Tinea nigra ,[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Tinea nigra ,[object Object],[object Object],Superficial mycoses
Piedra ,[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Piedra ,[object Object],[object Object],[object Object],Superficial mycoses
Black piedra ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses www.doctorfungus.org
Black piedra - distribution Superficial mycoses
Black piedra ,[object Object],[object Object],[object Object],Superficial mycoses
Black piedra - lab diagnosis ,[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Black piedra - lab diagnosis ,[object Object],[object Object],Superficial mycoses
Black piedra - lab diagnosis ,[object Object],[object Object],[object Object],Superficial mycoses
Black piedra - lab diagnosis ,[object Object],[object Object],[object Object],[object Object],Superficial mycoses Heaped center Flat periphery
White piedra ,[object Object],[object Object],[object Object],Superficial mycoses
White piedra ,[object Object],[object Object],Superficial mycoses scalp eyebrow eyelashes
White piedra ,[object Object],[object Object],[object Object],[object Object],Superficial mycoses
White piedra ,[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
White piedra ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
White piedra - lab diagnosis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
White piedra - lab diagnosis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
White piedra - lab diagnosis ,[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
White piedra - lab diagnosis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Piedra - Treatment ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Superficial mycoses
Cutaneous mycoses ,[object Object],[object Object],skin hair nails keratinase
Cutaneous mycoses ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],ringworm
Cutaneous mycoses ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object]
Geophilic ,[object Object],[object Object]
Zoophilic ,[object Object],www.saanendoah.com   www.kolumbus.fi
Anthropophilic fungi ,[object Object],[object Object],[object Object]
Anthropophilic fungi ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Classification of  Dermatophytes
Microsporum Macroconidia Rough walled Microconida present
Trichophyton Macroconidia Smooth walled Microconidia present
Epidermophyton Chlamydoconidia Macroconidia Smooth walled Microconidia none
[object Object]
 
Tinea capitis www.emedicinehealth.com   ,[object Object],[object Object]
Tinea capitis ,[object Object],[object Object]
Tinea favosa ,[object Object],[object Object],[object Object],www.mf.uni-lj.si
Tinea corporis ,[object Object],[object Object],[object Object],www.cut.ee/
Tinea corporis ,[object Object],[object Object],[object Object]
Tinea imbricata Cutaneous ,[object Object],[object Object]
Tinea barbae ,[object Object],Cutaneous www.merck.com   www.emedicine.com
Tinea cruris Cutaneous www.dermnetnz.org   ,[object Object],[object Object],[object Object]
Tinea pedis Cutaneous www.doctorfungus.org   dermatologie.free.fr   ,[object Object],[object Object],[object Object]
Tinea manuum ,[object Object],Cutaneous www.dermnetnz.org
Tinea unguium Cutaneous www.dermnetnz.org   ,[object Object],[object Object],[object Object],[object Object]
Laboratory diagnosis
Skin scraping specimen Wipe with water www.doctorfungus.org   scalpel Paper / envelope active edge
Direct Examination ,[object Object],[object Object],[object Object],[object Object],[object Object],KOH parker
KOH of skin scrapings Cutaneous Septate hypha
KOH of skin scrapings Cutaneous arthrospores septate hypha
Ectothrix   invasion of hair ,[object Object],[object Object],[object Object],Cutaneous
Ectothrix   invasion of hair ,[object Object],[object Object],[object Object],[object Object],[object Object],Cutaneous
Ectothrix   invasion of hair ,[object Object],[object Object],[object Object],Cutaneous
Endothrix  invasion of hair ,[object Object],[object Object],[object Object],Cutaneous
Endothrix  invasion of hair ,[object Object],[object Object],[object Object],Cutaneous
Culture: ,[object Object],[object Object],[object Object],[object Object],[object Object]
Culture: ,[object Object],[object Object],[object Object]
Identification ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Mycology
Microsporum canis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Cutaneous netti.nic.fi   www.vet.ohio-state.edu
Microsporum canis ,[object Object],[object Object],Cutaneous ,[object Object],www2.provlab.ab.ca
Microsporum canis ,[object Object],[object Object],[object Object],[object Object],Cutaneous www.doctorfungus.org
Microsporum gypseum ,[object Object],[object Object],[object Object],[object Object],Cutaneous
Microsporum gypseum ,[object Object],[object Object],[object Object],[object Object],Cutaneous www.ukneqasmicro.org.uk
Microsporum gypseum ,[object Object],[object Object],[object Object],[object Object],[object Object],Cutaneous vtpb-www.cvm.tamu.edu   www.medmicro.wisc.edu
Trichophyton mentagrophytes ,[object Object],[object Object],[object Object],[object Object],[object Object],Cutaneous
Trichophyton mentagrophytes ,[object Object],[object Object],Cutaneous danival.org
Trichophyton mentagrophytes Cutaneous Microscopic ,[object Object],[object Object],vtpb-www.cvm.tamu.edu
Trichophyton mentagrophytes ,[object Object],Cutaneous ,[object Object],Microscopic www.vet.ohio-state.edu   vtpb-www.cvm.tamu.edu
Trichophyton mentagrophytes ,[object Object],Cutaneous www2.provlab.ab.ca   ,[object Object],[object Object],www2.provlab.ab.ca
Trichophyton rubrum ,[object Object],[object Object],[object Object],[object Object],[object Object],Cutaneous
Trichophyton rubrum ,[object Object],[object Object],[object Object],Cutaneous www.pfizer.ch   www4.medfak.lu.se
Trichophyton rubrum ,[object Object],[object Object],[object Object],Cutaneous www2.provlab.ab.ca
Trichophyton concentricum Cutaneous ,[object Object],[object Object],[object Object],[object Object]
Trichophyton concentricum Cutaneous ,[object Object],[object Object],[object Object],[object Object]
Trichophyton concentricum Cutaneous ,[object Object],[object Object],[object Object],[object Object]
Trichophyton concentricum Cutaneous ,[object Object],[object Object],[object Object]
Trichophyton schoenleinii Cutaneous ,[object Object],[object Object],[object Object],[object Object],[object Object]
Trichophyton schoenleinii Cutaneous ,[object Object],[object Object],[object Object],[object Object]
Trichophyton schoenleinii Cutaneous ,[object Object],[object Object],[object Object],[object Object],[object Object]
Epidermophyton floccosum ,[object Object],[object Object],[object Object],[object Object],Cutaneous
Epidermophyton floccosum ,[object Object],[object Object],[object Object],[object Object],[object Object],Cutaneous
Epidermophyton floccosum ,[object Object],[object Object],[object Object],[object Object],Cutaneous botit.botany.wisc.edu www.fns.uniba.sk

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41. laboratory diagnosis of common fungal diseases

  • 1. LABORATORY DIAGNOSIS OF COMMON FUNGAL DISEASES Prof. Louella A. Dancel, RMT, MSc. University of Perpetual Help System DALTA 42 nd PAMET ANNUAL CONVENTION Manila Hotel – December 2,2006
  • 2. Clinical specimens submitted for Fungal Isolation 2004-2006* Clinical specimen 2004 2005 2006 Total Respiratory 29 35 34 98 Body fluids 37 17 34 88 Tissues 9 0 23 32 Skin 1 0 0 1 Nails 29 31 20 80 Hair 0 1 0 1 CSF 8 5 8 21 Others 5 20 24 49 TOTAL 118 109 153 380
  • 3. Fungal Isolates 2004-2006 * Isolate 2004 2005 2006 C.albicans 5.1% 13.8% 11.8% C.tropicalis 1.8% 3.7% 5.2% C.parapsilosis 5.1% 6.4% 3.9% C.glabrata 0.91% 0.91% 3.9% C.famata 1.8% 4.6% 2.0% Aspergillus spp. 0.91% 0% 0.65% Fusarium spp. 0% 0% 0.65%
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  • 35. Black piedra - distribution Superficial mycoses
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  • 59. Classification of Dermatophytes
  • 60. Microsporum Macroconidia Rough walled Microconida present
  • 61. Trichophyton Macroconidia Smooth walled Microconidia present
  • 62. Epidermophyton Chlamydoconidia Macroconidia Smooth walled Microconidia none
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  • 77. Skin scraping specimen Wipe with water www.doctorfungus.org scalpel Paper / envelope active edge
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  • 79. KOH of skin scrapings Cutaneous Septate hypha
  • 80. KOH of skin scrapings Cutaneous arthrospores septate hypha
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Editor's Notes

  1. Cutaneous mycoses are fungal infections of the integument and its appendages. They are superficial infections of the skin, hair & nails The fungus localizes only in the cornified layer, stratum corneum, No living tissue is invaded Pathological changes occur in the host because of her reaction to the fungus and its products of metabolism or life. A llergic & inflammatory eczematous host response
  2. Dermatophytosis are caused by a closely related group of organisms, called dermatophytes Which are keratinophilic Very similar & closely related group of fungi that cause a wide variety of clinical conditions A single species may be involved in several disease types, each with its distinctive pathology Most commonest infectious agents of man, the infections are distributed worldwide (geographically & racially) Common term is ringworm
  3. Dermatomycosis are skin infections that are caused neither by dermatophytes nor by the genus candida. They may be caused by soil inhabiting fungi
  4. Tinea capitis is a dermatophyte infection of the scalp, eyebrows and eyelashes caused by species of Microsporum and Trichophyton. type & severity of response is related to the species & strain of dermatophyte causing infection
  5. Tinea capitis is a dermatophyte infection of the scalp, eyebrows and eyelashes caused by species of Microsporum and Trichophyton. type & severity of response is related to the species & strain of dermatophyte causing infection
  6. Favus refers to the similarity of appearance of scutula and honeycombs. Favus is characterized by the occurrence of dense masses of mycelium & epithelial debris which form yellowish, cup-shaped crusts called SCUTULA. The scutulum develops in a hair follicle, with the hair shaft in the center of the raised lesion. Removal of these crusts reveals an oozing, moist red base. After a period of years, atrophy of the skin occurs, leaving a cicatricial alopecia & scarring. Scutula may be formed on the scalp or the glabrous skin.
  7. Tinea corporis is a dermatophyte infection of the glabrous skin most commonly caused by species of the gnera Trichophyton and Microsporum . The infection is generally restricted to the stratum corneum of the epidermis. The clinical symptoms are a result of the fungal metabolites acting as toxins & allergens. Lesions vary from simple scaling, scaling with erythema & vesicles, to deep granulomata. Villous hair in the involved area may be invaded, & the follicle often acts as a reservoir for recrudescence of the disease.
  8. Tinea corporis is a dermatophyte infection of the glabrous skin most commonly caused by species of the gnera Trichophyton and Microsporum . The infection is generally restricted to the stratum corneum of the epidermis. The clinical symptoms are a result of the fungal metabolites acting as toxins & allergens. Lesions vary from simple scaling, scaling with erythema & vesicles, to deep granulomata. Villous hair in the involved area may be invaded, & the follicle often acts as a reservoir for recrudescence of the disease.
  9. Tinea imbricata is a geographically restricted form of tinea corporis caused by Trichophyton concentricum. It is characterized by polycyclic, concentrically arranged rings of papulosquamous patches of scales scattered over and often covering most of the body
  10. Tinea barbae is a dermatophyte infection of the bearded areas of the face and neck, and therefore is restricted to adult males.
  11. Tinea cruris is a dermatophyte infection of the groin, perineum, & perianal region, which is acute or chronic and generally pruritic. The lesion is sharply demarcated, with a raised, erythematous margin & thin, dry epidermal scaling. The disease is found in all parts of the world but is more prevalent in the tropics. It tends to occur when conditions of high humidity lead to maceration of the crural region. A similar condition may involve the axilla or other intertriginous areas. The disease is more common in men, but frequently involves women, usually when it is transmitted by intimate contact or fomite. It may reach epidemic proportions in athletic teams, troops, ship crews, & inmates of institutions. In such cases, it is probably most commonly transmitted by towels, linens, & clothing. E. Floccosum has been isolated from blankets & sheets as well as rugs & other fomites where it can survive for years as infective arthroconidia.
  12. Tinea pedis is a dermatophyte infection of the feet involving particularly the toe webs and soles. The lesions are of several types, varying from mild, chronic, and scaling to acute, exfoliative, pustular, & bullous disease.
  13. Most dermatophyte infections of the hand, particularly of the dorsal aspect, are similar to tinea corporis. Tinea manuum refers to those infections in which the interdigital areas & the palmar surfaces are involved & show characteristic pathologic features. Along with tinea pedis, tinea manuum is one of the commoner types of chronic dermatophytosis in the adult. It has been postulated that this is related to the lack of sebaceous glands & their fungistatic lipids in these two areas.
  14. Tinea unguium is an invasion of the nail plates by a dermatophyte. Onychomycosis is an infection of the nails caused by nondermatophytic fungi and yeasts. Tinea unguium is of at least 2 types: Superficial white onychomycosis – invasion is restricted to patches or pits on the surface of the nail invasive subungual dermatophytosis (ringworm of the nail), in which the lateral or distal edges of the nail are first involved, followed by establishment of the infection beneath the nail plate.
  15. For skin specimen, first clean lesion & periphery with 70% alcohol. Use sterile scalpel or edge of microscope slide, scrape perpendicular to the skin. Scrape around the active edge where the fungus is actively growing. The center of lesion heals first, so the laboratory results are negative using this sample. If the lesion is inflamed or with fissures, clean it with sterile distilled water. Collect skin scrapings in paper envelope or petri dish, or place between 2 slides. Store at room temperature. If patients are young children and are scared of the scalpel, use can use scotch tape to collect specimen for microscopy. Collect moist exudate for candida
  16. These Fungi exhibit characteristic structures in clinical specimens that can be seen microscopically using a brightfield or phase contrast microscope. You can prepare a wet mount, a specimen plus sterile water or NSS, or specimen alone, like exudates. We can use potassium hydroxide, from 10 to 30% depending upon the type of specimen, for skin use 10%, for nails use a stronger concentration, 30%. Potassium hydroxide digests or dissolves proteins, fats & carbohydrates, the tissue clears Making the fungal cell wall, which is resistant to alkali, visible. Addition of Parker blue-black ink, will stain the fungal structures, facilitating microscopic examination, fungi appear bluish green. The preparation is passed 2 or 3 times over an alcohol lamp or bunsen burner, to hasten the reaction. If the result is negative, especially in nail scrapings, you can leave the preparation overnight on the lab table, to give time for digestion to occur. View the preparation again the next day. Question: Can you use Parker permanent blue or black ink? YES
  17. Microscopic examination
  18. Use several media