taller de microscopia ii
TRANSCRIPT
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TALLER DE TALLER DE DIAGNOSTICO MICOLOGICO Y DIAGNOSTICO MICOLOGICO Y APLICACIÓN CLÍNICAAPLICACIÓN CLÍNICA
Dr. Gustavo E. Giusiano
Facultad de MedicinaInstituto de Medicina Regional Universidad Nacional del [email protected]
Guayaquil Guayaquil -- 20112011
Módulo II
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Rhinosporidium seeberi
Scientific classification
Kingdom: ProtistaPhylum: Choanozoa
Class: Ichthyosporea
Order: Dermocystida
Genus: Rhinosporidium
Species: R. seeberiSpecies: R. seeberi
Rhinosporidium
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Micosis SubcutáneasMicosis Subcutáneas
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EsporotricosisEsporotricosis
Agente etiológico: Agente etiológico: Sporothrix schenckiiSporothrix schenckii
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DiagnósticoDiagnóstico
�� Examen directo en frescoExamen directo en fresco
Cuerpos asteroides
�� ElEl agenteagente raramenteraramente eses observadoobservado enen elel examenexamen microscópicomicroscópico directodirecto..
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Coloraciones
Giemsa, Gram, Grocott, PASPAS
Giemsa
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ESPOROTRICOSISESPOROTRICOSISPAS
MICOLOGIAMICOLOGIA::
�� BIOPSIABIOPSIA:: HiperplasiaHiperplasia seudoepiteliomatosa,seudoepiteliomatosa, microabscesosmicroabscesos intradérmicos,intradérmicos, infiltracióninfiltración porpor
leucocitosleucocitos yy célulascélulas plasmáticas,plasmáticas, eosinófilos,eosinófilos, granuloma,granuloma, cuerposcuerpos asteroidalesasteroidales..
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CultivoCultivo
Sabouraud con ATB
Cultivo a 28ºC
�Hongo dimórfico
Cultivo a 37ºC
�Hongo dimórfico
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�CULTIVO: 37ºC: Colonia levaduriforme
� Células brotantes ovales, fusiformes.
�Forma parasítica o cultivo a 37ºC de Sporothrix schenckii
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�CULTIVO: 25-28ºC: Hifas finas (1 a 2 micras de diam.), septadas, conidios ovales a piriformes (2-3 a 6µ) producidos en bouquet en el extremo del conidióforo. La acumulación de estas microconidias da aspecto de flor o “margarita”.
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MicrocultivoMicrocultivo
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Micetomas
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Grano con hifas hialinas o dematiaceas
�Sospechar grano micótico
Eumicetoma
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GRANOS NEGROS
Corte histológico
MO 10X
MO 100X
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Principales Agentes Etiológicos de Principales Agentes Etiológicos de
EumicetomasEumicetomas
GRANOS NEGROS GRANOS BLANCOS
�� Madurella griseaMadurella grisea
�� Madurella mycetomatisMadurella mycetomatis
�� Pyrenochaeta romeroiPyrenochaeta romeroi
�� Leptosphaeria senegalensisLeptosphaeria senegalensis
�� Exophiala jeanselmeiExophiala jeanselmei
�� Exophiala Exophiala wernekii wernekii
�� Scedosporium apiospermumScedosporium apiospermum
�� Acremonium kilienseAcremonium kiliense
�� Acremonium recifeiAcremonium recifei
�� Acremonium falciformeAcremonium falciforme
�� Fusarium moniliforme Fusarium moniliforme
�� Fusarium solaniFusarium solani
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por Madurella mycetomatis
GRANOS NEGROS
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Conidióforos cilíndricos pigmentados con anélide en la porción apical dando aspecto de espinaPuede estar ramificado
Exophiala spinifera
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Scedosporium apiospermumScedosporium apiospermum
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Acremonium sp.Acremonium sp.
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Fusarium sp.Fusarium sp.
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CromomicosisCromomicosisCromomicosisCromomicosis
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Examen directo en fresco
Cuerpos escleróticos (células fumagoides)Esclerotes de Medlar
elementos esféricos, dematiáceos, tabicadosy agrupadosde 2, 4, 6 o más. tabicadosy agrupadosde 2, 4, 6 o más.
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Examen en fresco
Cuerpos escleróticos (células fumagoides)Esclerotes de Medlar
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Principales Agentes Etiógicos de Principales Agentes Etiógicos de CromomicosisCromomicosis
►► Phialopora verrucosaPhialopora verrucosa
►►Cladosporium carrioniiCladosporium carrionii
►► Fonsecaea pedrosoiFonsecaea pedrosoi
►► Fonsecaea compactaFonsecaea compacta
►►Rhinocladiella aquaspersaRhinocladiella aquaspersa
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Phialophora verrucosaPhialophora verrucosa
Fialides con forma de botella con collarete conico
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Cladosporium sp.Cladosporium sp.
Conidios ovoidales dispuestos en largas cadenasuni o bicelulares
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Cladosporium carrioni
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RhinocladiellaRhinocladiella
Conidios ovoidales - fusiformes, laterales, unicelulares
que se forman en la extremidad de
largos conidióforoserectos cilíndricos
((AcrothecaAcrotheca))
erectos cilíndricos
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Fonsecaea pedrosoiFonsecaea pedrosoiTres tipos de fructificaciónTres tipos de fructificación
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Fonsecaea pedrosoiFonsecaea pedrosoiTres tipos de fructificaciónTres tipos de fructificación
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Micosis Sistémicas EndémicasMicosis Sistémicas Endémicas
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Agente: Agente: Paracoccidioides brasiliensisParacoccidioides brasiliensis
ParacoccidioidomicosisParacoccidioidomicosis
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Examen directo
FRESCO
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400X
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TODAS LAS MUESTRAS
Examen directo
GiemsaCOLORACIONES
GroccotPass
Z-N (TBC)Kinyoun (Nocardia)Gram Weigert (P. jiroveci)Azul de toluidina (P. jiroveci)
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Coloración de Giemsa
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Grocott
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Z -N * importante por la dificultad en algunos casos de diferenciar con TBC por radiología o clínica
* Asociaciones
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Tº : 25-28ºC y 35-37ºC
Forma saprofítica Forma parasitaria
TERMODIMORFOTERMODIMORFO
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35-37ºC – Forma parasitaria
Cultivo
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25-28ºC – Forma saprofítica
microconidias
Cultivo
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Histoplasmosis
Agente: Histoplasma capsulatumvar capsulatum (Argentina)
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Histoplasmosis
Diagnóstico
Fase parasitaria
Examen directo
intracelular1-5 micras
Coloración de Giemsa(100X)
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Citoplasma en casquete Citoplasma en casquete (forma semilunar, desplazado hacia hacia uno de los polos)(forma semilunar, desplazado hacia hacia uno de los polos)
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Grocott – metenamina plata
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Histoplasmosis
HEMOcultivo
Método de elección – Lisis centrifugación
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Tº : 25-28ºC y 35-37ºC
Forma saprofítica Forma parasitaria
TERMODIMORFOTERMODIMORFO
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Forma parasitaria
Fase Levaduriforme Fase Levaduriforme
unicelular, unicelular, in vitroin vitro 3434--3737°°CC
HHistoplasma capsulatum istoplasma capsulatum
varvar. capsulatum. capsulatum
Elementos levaduriformes(unicelulares) ovoidescon un solo brote -Tamaño 1-5 micras
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Micelio hialino tabicadoMicroconidiasMicroconidias pequeñas, piriformes ( < 5 pequeñas, piriformes ( < 5 µµµµµµµµm)m)
Fase MicelialFase Micelial
multicelular en la naturaleza e multicelular en la naturaleza e in vitroin vitro 2525--28 28 °°CC
MacroconidiasGlobosas (8-15micras)Pared gruesa tuberculada
HHistoplasma capsulatum istoplasma capsulatum
varvar. capsulatum. capsulatum
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HHistoplasma capsulatum istoplasma capsulatum
varvar. capsulatum. capsulatum
Fase Fase saprofíticasaprofítica
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CoccidioidomicosisCoccidioidomicosis
Agente: Coccidioides immitis (Valle de San Joaquín – California)
Coccidioides posadasiiCoccidioides posadasii
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TODAS LAS MUESTRAS
Examen directo
FRESCO
Fase parasitaria
esférulas de 20 a 80 µ de diámetro con endosporos
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Examen directo
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Examen directo
Coccidioides immitis Coccidioides posadasii
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Examen directo
Coccidioides immitis Coccidioides posadasii
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Calcofluor
Coccidioides immitis Coccidioides posadasii
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Coloraciones• Giemsa• Gram• Grocott• PAS
Z-N (TBC)Kinyoun (Nocardia)Kinyoun (Nocardia)Gram Weigert (Pneumocistis)Azul de toluidina (Pneumocistis)
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CultivoTº : 25-28ºC y 35-37ºC
Forma saprofítica
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113Forma saprofítica
micelio hialino tabicado fragmentado en clamido-artroconidias Coccidioides immitis
Coccidioides posadasii
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112
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Micosis oportunistas IIMicosis oportunistas IIMicosis oportunistas IIMicosis oportunistas II
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Examen microscópico de colonias levaduriformes
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Cryptococcus sp Cryptococcus sp
Saccharomyces cerevisiaeSaccharomyces cerevisiae
Pichia membranifaciensPichia membranifaciens Candida globosaCandida globosa
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Examen microscópico de colonias levaduriformes
Saccharomyces cerevisiaeSaccharomyces cerevisiae
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Candida kruseiCandida krusei
Examen microscópico de colonias levaduriformes
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Trichosporon sppTrichosporon cutaneum
Blasto-artroconidios
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25ºC por 25ºC por 2 a 7 días2 a 7 días
� en agar harina de maíz - Tween 80
� agar arroz -Tween 80
MicrocultivoMicrocultivo ( Dalmau, 1929 ) Estudio micromorfológico de levaduras
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Candida albicans – Candida dubliniensis
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CryptococcusCryptococcus
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Cryptococcus en LCR Test de tinta china
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Pneumocystis sp.
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Trofozoitos (formas troficas): formas libres de pneumocystis, pequeñas, aparece frecuentemente en acúmulos
Formas quísticas : redondos o en media luna, de 5-8 micras de diametro. En su interior se encuentran los esporozoitos(cada quiste puede tener hasta 8 esporozoitos).
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Giemsa
5-8 µm
Formas quísticas
Lavado broncoalveolar (BAL)
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4-7 µm
Contienen 1 o más núcleos
Formas tróficas(pre-quiste)