Continuous succinic acid production by Actinobacillus succinogenes ...
ACTINOBACILLUS PLEUROPNEUMONIAEhistolab.com.co/images/2017/enfermedades-bacterianas/APP.pdf ·...
Transcript of ACTINOBACILLUS PLEUROPNEUMONIAEhistolab.com.co/images/2017/enfermedades-bacterianas/APP.pdf ·...
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ACTINOBACILLUS PLEUROPNEUMONIAE
Jersson Avila Coy MV, Esp Member of Latin Comparative Pathology Group
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Etiología
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Etiología � Cocobacilo; Gram- � 2 biovares/biotipos
� 1: NAD-depend � 2:NAD-independ
� Normal/ no invasiva � No sobrevive mucho en el amb
Immuno-gold electron photomicrograph of A. pleuropneumoniae. A. pleuropneumoniae is an encapsulated gram-negative coccobacilli that in some culture conditions can have a pleomorphic phenotype and express pili.
Síntesis en presencia de piridinas o sus precursores
NAD-NADP
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Etiología � 15 serotipos/serovares (según sus Ag’s polisacáridos capsulares)
� 1, 5, 7: Norteamer � 2, 9: Europa � 1, 7, 12: Australia � 1, 5, 9, 11: secrec tox Apx I y II: + severidad � Protección cruzada parcial
� Subdiferenciación � 1: 1a y 1b � 5: 5a y 5b
Según ≠ menores en la estructura del polisacárido
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Etiología � Polisacáridos Grupos capsulares
� I: Oligosacáridos repetidos unidos por enlaces fosfato � II: Oligosacáridos repetidos � III: Polímeros de ác teicoico unidos por enlace fosfato diéster
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Epidemiología
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
Edad
6 sem-6 meses
Fatalidad
20-80% en brotes agudos
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Epidemiología
Colombia: Endémica
Países de Cría Intensiva
SubCx
Difícil Cultivo –aún en medios selectivos
Limitados serovares x región
De baja virulencia pueden ser pasados x alto
Cepas/serovares ≠ virulencia x país
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Epidemiología
Lugar Serovar Asia 1, 2, 5 Australia 1, 2, 5, 15 Gran Bretaña y Europa 2, 9 Canadá, USA, México 1, 5 Japón 15 China 1, 3, 4, 5
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Epidemiología
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Epidemiología
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
Hampshire: Enf - severa
Landrace alemán y Piétrain: Enf + severa
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Factores Virulencia � Apx
� Citotox de la fam RTX � Apx I, II, III � Apx IV: significancia desconocida? � Citólisis: N0, M0 alveolares, GR, cél epiteliales � Bajas []: estallido resp en N0
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Factores Virulencia � LPS
� Activación M0s � Secrec de quemoatrayentes de N0s � Procoagulante � Activ complemento
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Factores Virulencia � Cápsula
� Altera fagocitosis x M0s � Previene activ complemento
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Factores Virulencia � Otros
� Superóxido dismutasa � Catalasa � Hidroperóxido reductasa � Adhesinas fímbricas � Prots de memb ext � Prots fijadoras de Fe � Metaloproteinasa � Ureasa
Protección muerte oxidativa x N0 y M0
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Transmisión
Contacto directo Gotas aerosol
Distancias cortas
OJO: Portadores asintomáticos Nasofaringe Ppal forma
ingreso a granja
1. Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654. 2. MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
Sec nasales Exudados pulmonares
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Factores predisponentes Aumento severidad
� No necesarios � Sobrepoblación � Pobre ventilación � Fluctuaciones temp
� Mycoplasma � Pseudorrabia � Éstasis ciliar
1. Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654. 2. MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Fisiopatología
Inhalación
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Unión bact-epitelio Respuesta Macrófagos
Liberación quemoatrayentes de N0
Infiltración N0 y exudado alveolar
Necrosis leucocitos mediada x Apx
90 min No invasión epitelio
1. Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654. 2. MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
Fagocitosis x N0 y M0
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Etapas Básicas de la Patogénesis
COLONIZACIÓN
EVASIÓN DE MEC DE LIMPIEZA
DAÑO TISULAR
Adhesinas Adquisición
de nutrientes
Limpieza específica
Limpieza No específica
PERSISTENCIA
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COLONIZACIÓN
Adhesinas
LPS Fimbria tipo 4
OMP 55-kDa
OMP 60-kDa
ComE1 Productos locus tad Aasp
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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LPS
Centro interno
Centro externo
Conservado 2 tipos O-
oligosacárido
Interacción moléculas hosp Ej: fosfatidiletanolamina
Débil Fuerte
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Receptores para App y LPS Glicoesfingolípidos
� Glucosilceramida (Glc β 1Cer) � Galactosilceramida (Gal β 1Cer) con ác grasos hidroxi y no
hidroxi � Sulfaturo (SO(3) - 3Gal β 1Cer) � Lactosilceramida (Gal β 1 -‐ 4Glc β 1Cer) � Gangliotriaosilceramida GgO3 (GalNAc β 1 -‐ 4Gal β 1 -‐ 4Glc β 1Cer)
� Gangliotetraosilceramida GgO4 (Gal β 1 -‐ 3GalNAc β 1 - 4Gal β 1 - 4Glc β 1Cer)
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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OMP 60-kDa
Interacción con Matriz EC
Interacción con colágenos Tipo I Tipo III Tipo IV Tipo V
Ca2+ depend
ComE1
Unión a Fibronectina DNA
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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COLONIZACIÓN
Adquisición de Nutrientes
Escasos nutrientes en T resp
Funciones esenciales
Prod ATP Resp Replic DNA Reparación
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Adquisición de Nutrientes
Utilización compuestos heme
Heme libre Hemina Hematina Hb Transferrina
Sideróforos
Ferricromo Bis-catecol
Prot Fur Ferric uptake regulator
Regulación de captación
TbpB (TfbA) TbpA (TfbB)
Otros
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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TbpB (TfbA) TbpA (TfbB)
Remoción Fe de la transferrina
Transporte a través de memb ext
Unión a prot de unión periplásmicas
TbpA TbpB
TbpA
Prot Fur Ferric uptake regulator
Prot HbgA regulada x Fur
Unión a Hb y hemina
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Evasión Limpieza No específica
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
Mucociliar Complemento
Fagocitos
Mucociliar
No efecto
Afección previa: ñ severidad
Complemento Polisacárdio capsular
(CPS) LPS
Limitación deposición C9
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Fagocitosis x N0 y M0 N0
M0
Muerte APP
Superviv APP
Lib Apx
Lisis fagocitos
Alteración función fagocítica
Apx I, II, III
Bajas [] Altas []
êquemotaxis M0
Daño M0 y PMN
Cápsula Apx IV estudio
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Supervivencia APP
LPS Limpieza R libres
Prot de rta a estrés: DnaK y Factor disparador
Amoniaco prod x hidrólisis urea
Inhibre fusión fagosoma-lisosoma
ñpH intralisosomal: êactividad hidrolasa
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Evasión Limpieza específica
� APP patógeno extracel � Rta humoral-Ac específicos: Muy imp � IgG1-Anti Apx: + imp
� Rta celular (CMIR) � CD4+
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Evasión Limpieza específica
� APP patógeno extracel � Rta humoral-Ac específicos: Muy imp � IgG1-Anti Apx: + imp
� Rta celular (CMIR) � CD4+
Proteasas (47 y 101 kDa)
Interferencia Opsonofagocitosis?
Degradación IgG, IgA
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Daño Tisular
Apxs Citotóxicas
Hemolíticas
Endotelios Fagocitos Leucocitos PMN, M0
Mediadores Infl:(IL) -1α, IL-1β, IL-6, IL-8, IL10, IL12p35, TNF-α, y INFα
Fibrinolisis
Sist kininas
Coagulación
Activ Plaq
Microtrombos
Isquemia/Necrosis
LPS
FXII
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Daño Tisular
Apxs LPS
Activ M0s alveol e intravasc
Metabolitos O2: Anión superóxido,
peróxido de H, radical hidroxilo
Enzimas proteolícas
Citokinas
Injuria celular
Interacción
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Apxs
ApxI, ApxII, ApxIII, ApxIV Familia de toxinas RTX (Repeats in ToXin)
Región altamente conservada con nonapéptidos de ác aspártico y glicina repetidos en el tercio carboxiterminal
Involucradas en la unión a Ca y la inducción de toxicidad a leucocitos Posee sitio de reconocimiento para el transporte de las toxinas a través de las membranas biológicas por el sist sec tipo 3 dependiente de tolC
Singh K, et al, 2011. Mannheimia haemoly,ca: Bacterial−Host Interac2ons in Bovine Pneumonia. Vet Pathol, 48: 338-‐348.
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Persistencia prolonagada en tej pulmonar necrótico: Baja disp O2
Persistencia
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Dimetil sulfóxido reductasa
Persistencia
Liasa amonio aspartato Resp anaeróbica
Flagelos y motiidad ?
Sintasa acetohidroxiácido Biosíntesis aa cad ramif
ArcA Resistencia a A-B
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Presencia de APP en fluido alveolar e
interlobulillar
Invasión pleural
Bacteremia (rara)
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Supervivencia
Secuestros necróticos
Abscesos encapsulados
Permanencia en criptas tonsilares
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Inmunidad � IgA e IgG � Contra
� Apx: Ppal/ � Polisacárido capsular
� Aumenta fagocitosis x N0 y M0 � Muerte efectiva de Bact opsonizadas x N0 (no x M0)
� Granjas endémicas: 12 sem caen Ac’s maternos
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Inmunidad
Ac´s
Tiempo (sem)
Desafío
Niveles detectables
Pto máx 5
2
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Inmunidad
Ac´s maternos
Tiempo
Ac´s
Desafío
Granjas endémicas
12 sem
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Sx Cx
Peraguda Aguda Crónica
Según: serovar, estado inmune, # bact
Tos Fiebre
Taquipnea Estornudos
Disnea Anorexia
Sec nasal sang
Letargia Ataxia Vómito Diarrea Distres Cianosis
Fiebre: leve o ausente
Tos: Espontánea o intermitente Inapetencia
Falta desarrollo Intol ejercicio
1. Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654. 2. MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Macro � Neumonía/pleuroneumonía lobar fibrinosupurativa,
hemorrágica y necrotizante � Lóbulos medios y caudales � Uni o bilateral � Lesiones rojas, profundas, firmes-duras, protruyen, al
corte son quebradizas � En superficie de corte focos de necrosis de coagulación
bien demarcados de 1-10cm, friables y pálidos
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Macro � Casos peragudos-agudos:
� Expansión de septos interlobulillares por fibrina y edema � Exudado fibrinoso en pleura � Cav torácica llena de fluido sang y coágulos de fibrina � Pericarditis y Pleuritis fibrinosa difusa
� Bronquios con fluido espumoso o sang � NL mediastinales y bronquiales: grandes, edematosos y
congestionados � Pericardio y peritoneo: escaso fluido serosang
1. Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654. 2. MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Macro � Crónico:
� Adherencias pleurales fibrosas � Fibrosis pulmonar o abscedación local/ extensas � Secuestros (a partir de grandes focos de necrosis de coagulación) � Abscesos encapsulados
1. Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654. 2. MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Parte cranial de lóbulo caudal es negra y dura. Pleura cubierta x fibrina
Áreas bien demarcadas de 3-10 cm, irregulares, rojas, firmes, dispersas en
todos los lóbulos Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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A. Foco neumónico consolidado (flecha) en lóbulo cardiaco B. Focos neumónicos en lóbulo apical, cardiaco y parte dorsal del caudal del pulmón izq Además intenso edema intersticial y pleuritis severa (flecha)
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Micro � Álveolos y bronquiolos terminales llenos con
� Fibrina, PMN, pocos M0 � Muchos GB necróticos probl/ N0 � Proliferación de M0
� Focos necrótico centrados en septos alveolares, delineados por bandas de GB (PMN) degenerados
� Exudado � Edema proteináceo-fibrina-GB
Crónico
Agudo
Crónico
1. Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654. 2. MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Micro
� En algunas áreas los GB contienen detritos de cromatina dispersos homogéneos ligera/ basofílicos, similiar a las cél avena, similar a pasteurelosis neumónica
� Mucosa bronquios y bronquiolos: invadida x N0 y pocos M0 � Epitelio bronquiolar: necrótico o desprendido
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Micro � Septos:
� VS: Trombos en vénulas y capilares, vasculitis fibrinoide � Vasculitis: Hemorragias
� Linfáticos: Distendidos por exudado serofibrinoso con N0
� Lesiones extrapulmonares � Trombosis glomerular renal, vasculitis renal, osteomielitis
1. Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654. 2. MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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Líneas distintivas de respuesta inflamatoria
Extensa zona de necrosis de coagulación rodeando un bronquiolo parcialmente
colapsado y delineada por una banda basofílica oscura
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A. Infiltrado alveolar compuesto x M0s activados y PMNs (flecha) y edema alveolar leve B. Intenso infiltrado leucocitario alveolar rodeado por un halo inflamatorio y la
neoformación de tej fibroso en la periferia (secuestro) (flechas)
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Forma Granulomatosa-Pulmón � 14818 cerdos-25
con pleuropneumonía porcina-20 con App- 8 con granulomas y cuerpos asteroides
Ohba T, et al, 2009. Prevalence of Granulomatous Pleuropneumonia Associated with Ac,nobacillus pleuropneumoniae Serotype 2 in Slaughter Pigs. J Vet Med Sci. 71: 1089-‐1092.
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Múltiples focos blanco amarillentos Bastones radiantes eosinofílicos en el centro de una lesión granulomatosa con
numerosos N0
Ohba T, et al, 2009. Prevalence of Granulomatous Pleuropneumonia Associated with Ac,nobacillus pleuropneumoniae Serotype 2 in Slaughter Pigs. J Vet Med Sci. 71: 1089-‐1092.
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Hepatitis granulomatosa � 66894 cerdos-11 hepatitis granulomatosa multifocal x App 2
Múltiples focos blanco amarillentos difusos
Varios bastones radiantes en las lesiones granulomatosas
Ohba T, et al, 2008. Mul2focal Granulomatous Hepa22s Caused by Ac2nobacillus pleuropneumoniae Serotype 2 in Slaughter Pigs. J Comp Pathol. 139: 61-‐66.
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Linfadenitis granulomatosa � 151653 cerdos-6 linfadenitis granulomatosa multifocal x App 2
Múltiples focos blanco amarillentos difusos, en la superficie de corte, NL hiliares pulmonares y
mediastinales
NL mandibular. Granuloma multifocal rodeado x TC denso
Ohba T, et al, 2010. Granulomatous lymphadeni2s associated with Ac,nobacillus pleuropneumoniae serotype 2 in slaughter barrows. CVJ. 51: 733-‐737.
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Dx � HxPx � Aislamiento
� Agar sangre
� ELISA/Aglutinación en látex: frotis pulmón/cultivos 1rios � PCR: cruce con otros Actinobacillus, No uso común � Serología
� Fijación comp: baja sens � ELISA: Pobre repetibilidad
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Dx ≠
� Actinobacillus suis + � Salmonella cholerasuis -
Patrón macro
Neumonía fibrinosa y necrotizante local o localmente extensa
Caswell JL, Williams KJ, 2007. Respiratory System, en: Maxie G. Jubb, Kennedy and Palmer´s Pathology of Domes2c Animals, 523-‐654.
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Prevención y control � Buenas Prácticas de
manejo � Aires � Temp � Sobrepoblaciones � Todo dentro/fuera � Limpieza
� Penicilinas � Tetraciclinas � Gentamicina � Tiamulina
MacInnes JI, et al, 2010. Ac2nobacillus, en: Gyles CL, et al. Pathogenesis of Bacterial Infec2ons in Animals, 363-‐386.
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?