El objetivo del presente estudio fue determinar la presencia de anticuerpos contra Encephalitozoon cuniculi en conejos destinados como mascotas. Se evaluaron las muestras serológicas de 76 conejos clínicamente sanos provenientes de 7 granjas de la provincia de Lima. Las muestras se procesaron utilizando un kit de ELISA. Anticuerpos contra el Encephalitozoon cuniculi fueron encontrados en el 43/76 (56.6%) de la muestras analizadas. El análisis de regresión logística de los resultados mostro que la edad en meses (9 meses) se asociaba al riesgo que un conejo sea positivo a la prueba serológica (P<0.05). Los resultados demostraron la presencia de anticuerpos contra Encephalitozoon cuniculi con una alta prevalencia en conejos destinados como mascota en la provincia de Lima.

Thesis--Copenhagen. / Summary in Danish. Bibliography: p. 84-89.

Thesis (Ph. D.)--Rutgers University, 2009. / "Graduate Program in Biology." Includes bibliographical references (p. 150-164).

Thesis--Copenhagen. / Summary in Danish. Bibliography: p. 84-89.

No description available.

Conventional methods such as serology and microscopy are unreliable for diagnosis of encephalitozoonosis in domestic rabbits. Previous studies have reported PCR to be insensitive but it is unclear whether this is because of inherent limitations or the lack of assay optimization and validation. The studies described in this thesis assess DNA quality and quantity for combinations of six DNA extractions kits and four spore disruption methods. The resulting DNA underwent PCR using a published primer set. The optimal method had a detection threshold of 100 spores/ml in saline. However, when repeated in urine, the detection threshold was much higher (10,000 spores/ml) and non-target DNA amplification was present. Various methods were used to improve analytical sensitivity and eliminate non-target amplification. One method involving PEG 8000 treatment produced a detection threshold of 1,000 spores/ml and decreased non-target DNA amplification. Ultimately, new primers were designed and when the optimized method was tested with these primers, a detection threshold of 100 spores/ml with no non-target DNA amplification was achieved. The optimal method and new primers were tested using clinical samples of rabbit urine and 32.4% were found to be positive for E. cuniculi. The final assay was shown to be both analytically sensitive and specific; however further clinical investigation is warranted to determine clinical utility. / OVC Pet Trust

Thesis (Ph. D.)--Cornell University, April, 2008. / Vita. Includes bibliographical references (leaves 151-166).

Memoria para optar al Título Profesional de Médico Veterinario Departamento de Medicina Preventiva Animal / Los microsporidios son protozoos, parásitos intracelulares obligados de distintas especies animales. Pertenecen al Phylum Microspora, caracterizada por producir esporas muy pequeñas y por carecer de mitocondrias. Entre los microsporidios, está el Encephalitozoon cuniculi, el cual es un parásito oportunista emergente. El reservorio, en estudio, que pone en peligro la salud humana es el conejo (Oryctolagus cuniculus), el cual elimina al parásito principalmente por la orina, contaminando el medio ambiente. En la prevalencia de E. cuniculi en conejos se reportan rangos de 15 al 76 % a nivel mundial (Ansbacher et al, 1988). Con la finalidad de detectar la presencia de este parásito en nuestro país, se realizó la pesquisa de éste, en el laboratorio de parasitología del Instituto de Salud Pública de Chile (ISP). La muestra estuvo constituida por 100 conejos normales sin distinción de sexo, pertenecientes a: Bioterio de ISP, Criaderos familiares de la localidad de Pirque, “Mundo Granja” perteneciente a la Universidad de Chile y a un Criadero comercial. Todos los conejos fueron previamente evaluados mediante un examen clínico. Luego de procesar la muestra, de deposición y de orina, se procedió a teñirlas con colorantes tricrómicos: Cromotropo 2R y Gram Cromotropo. Se obtuvo una positividad en un 55% de los individuos, donde el mayor porcentaje se presenta en conejos inmunosuprimidos y de crianza convencional. El objetivo de este estudio es contribuir a la salud, en especial la del ser humano como la de los animales determinando la presencia de E. cuniculi en conejos de distintos orígenes / Servicio Agrícola y Ganadero SAG) y Proyecto FAVET /04-35.

In this study was described the role of the inflammation in activation of latent microsporidiosis in immunocompetent BALB/c and MMF knock out mice and immunodeficient SCID mice and the correlation between inflammation induction and microsporidia occurrence. The number of spores in selected organs was located by using molecular and histology methods. Moreover, the occurrence of spores in faecal samples was monitored during the whole time of experiment.

Encephalitozoon cuniculi is a zoonotic protozoan parasite in the phylum Microspora that has been shown to naturally infect several host species, including humans, rabbits and dogs. Currently, serological diagnosis of infection is made using the immunofluorescense assay (IFA) or enzyme-linked immunosorbent assay (ELISA). Although these methods are sensitive and reliable, there are several drawbacks to both tests. Cross-reactivity between other Encephalitozoon species is common, and specialized equipment is required for IFA and ELISA. Most wildlife species are unable to be tested using these methods, because species-specific antibodies are required. One goal of this work was to develop a new serological test for diagnosing E. cuniculi infection that would be more practical for use in small veterinary and medical clinics. The effectiveness of the agglutination test was examined in CD-1 and C3H/He mice infected with E. cuniculi or one of 2 other Encephalitozoon species. The results indicate that the agglutination test is 86% sensitive and 98% specific for E. cuniculi, with limited cross-reactivity to E. intestinalis. The test is fast and easy to conduct, and requires no specialized equipment or species-specific antibodies. Recent reports of microsporidial DNA in crop irrigation waters suggest that unpasteurized juice products may be contaminated with E. cuniculi. High pressure processing (HPP) is an effective means of eliminating bacteria and extending the shelf life of products while maintaining the sensory features of food and beverages. The effect of HPP on the in vitro infectivity of E. cuniculi spores was examined. Spores were exposed to between 140 and 550 MPa for 1 min, and then spores were loaded onto cell culture flasks or were kept for examination by transmission electron microscopy (TEM). Spores treated with between 200 and 275 MPa showed reduction in infectivity. Following treatment of 345 MPa or more, spores were unable to infect host cells. No morphologic changes were observed in pressure-treated spores using TEM. The effect of disinfectants on in vitro infectivity of E. cuniculi spores was also examined. Spores of E. cuniculi were exposed to several dilutions of commercial bleach, HiTor and Roccal, and 70% ethanol for 10 minutes and then loaded onto Hs68 cells. The results of this study showed that all concentrations of disinfectants tested were lethal to E. cuniculi spores. Encephalitozoon cuniculi spores are more sensitive to disinfectants than are coccidian oocysts and other parasite cysts. / Master of Science