Made available in DSpace on 2014-06-12T23:01:24Z (GMT). No. of bitstreams: 2 arquivo8552_1.pdf: 533604 bytes, checksum: a260b9f76c8b82b2713fa5d4b3ad9d38 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2006 / A água doce é um recurso natural finito, cuja qualidade vem decaindo devido ao aumento da população e à deficiência de políticas públicas voltadas para a sua preservação. A rota de transmissão ambiental para protozoários e helmintos é particularmente significativa e envolve a água, o solo e o alimento. A partir da década de 1980, acrescentou-se a preocupação quanto à ocorrência de protozoários patogênicos tais como Cryptosporidium spp. e Giardia spp., em resposta ao crescente número de surtos envolvendo um grande número de pessoas, estando relacionados ao consumo de água e à ineficiência do tratamento aplicado na remoção de cistos e oocistos nas Estações de Tratamento de Água (ETA). No Brasil foi instituído a partir de 2003 o Sistema de Informação de Vigilância da Qualidade da Água de Consumo Humano (SISAGUA), instrumento de coleta de análises de dados a ser desenvolvido em âmbito nacional pelas áreas de vigilância ambiental em saúde, visando à garantia da qualidade da água distribuída à população. O presente estudo teve como objetivo detectar a presença de enteroparasitas, com ênfase em Cryptosporidium spp. em amostras de água consumida por parte da população do Distrito Sanitário VI, na cidade do Recife. As amostras foram coletadas em pontos considerados como início, meio e fim da rede de distribuição. A metodologia de sedimentação espontânea foi utilizada para pesquisa de enteroparasitas e outros organismos. Para a detecção de Cryptosporidium spp., utilizouse o método de concentração de oocistos através de membrana filtrante, seguido da identificação e contagem de estruturas álcool-ácido resistentes através da coloração histoquímica de Kinyoun. Como ensaios confirmatórios para Cryptosporidium spp., foram utilizadas as metodologias de Ensaio Imunoenzimático e Imunofluorescência Direta. Os parâmetros físico-químicos e microbiológicos foram avaliados de acordo com os padrões da Portaria nº 518/04/MS. Em 95,56% (86/90) das amostras de água, analisadas neste estudo foram encontrados organismos como: oocistos Cryptosporidium spp. (15,55%), cistos de Giardia spp. (3,85%), além de ovos de helmintos (47,78%), larvas de nematóides (28,78%) e organismos do zooplâncton (84,44%). Conclui-se que existe vulnerabilidade nos processos de tratamento da água para consumo humano, principalmente na etapa da filtração, que deveria ser eficiente na remoção física de partículas, com especial atenção aos organismos patogênicos e que a qualidade da água está comprometida desde a saída da ETA até o ponto de consumo. Torna-se necessário intensificar a vigilância em saúde ambiental relacionada à qualidade da água para consumo humano, garantindo que a água não ofereça riscos a saúde da população

No description available.

We describe a simple, sensitive, nonradioactive, relatively rapid and relatively inexpensive protocol to measure DNA synthesis in cultured cells by a chemiluminescent bromodeoxyuridine (BrdU) enzyme-linked immunosorbent assay (ELISA). We show that it exhibits similar sensitivity and activity as traditional 3H-thymidine incorporation assays and a commercial chemiluminescent BrdU ELISA kit when tested in commonly used cell lines, such as NIH 3T3 cells, mink lung epithelial cells (Mv1LU), and baby hamster kidney (BHK-21) fibroblasts. This assay also exhibits a wider dynamic range than colorimetric BrdU ELISA methods. Besides being a viable, nonradioactive alternative to 3H-thymidine incorporation assays, our BrdU ELISA is less expensive than a commercial chemiluminescent BrdU ELISA kit.

No description available.

Ornithobacterium rhinotracheale (ORT) is a Gram-negative bacterium that causes respiratory disease in poultry characterized by rhinitis, tracheitis, and pneumonia with mortality averaging 2-3%. In the Shenandoah Valley of Virginia, the seroprevalence for ORT among turkey flocks as determined by enzyme-linked immunosorbent assay (ELISA) was found to be 70.9% (n=175). Additionally, the seroprevalence for hemorrhagic enteritis virus (vaccine induced), Bordetella avium, and paramyxovirus-1 was 100%, 74.8%, and 6.3% respectively. No significant interactions were detected. The type strain of ORT is characteristically non-hemolytic at least for 96 hours at 37°C on Columbia Blood Agar. In recent years, atypical isolates that rapidly produce hemolysis have been isolated with increasing frequency. A variety of in vitro tests were used to determine differences between representative isolates of the hemolytic (H) and non-hemolytic (NH) phenotypes. Findings suggest that the H isolate contains a 4 kb plasmid similar to that found in Reimerella anatipestifer. No plasmid was found in the NH isolate. Differences in growth characteristics and resistance to tetracyclines were also noted. No differences in proteins, biochemical characteristics or 16S rRNA sequences were found, the latter serving as confirmation that the isolates were both ORT. Embryo inoculation was used to assess virulence. No significant differences were observed and most embryos survived through to the day of hatch (pip) despite the fact that ORT could be re-isolated. In turkey poults however, the H phenotype did appear less virulent. A significant depression in weight gain was noted for birds inoculated intratracheally with the NH isolate at 7 days post-inoculation (dpi). NH inoculates also had significantly higher antibody levels on ELISA at 14 and 21 dpi and histopathological lesion scores for lung at 7, 14, and 21 dpi. The NH isolate could be re-isolated from NH-inoculated poults through 21 dpi; whereas the H isolate could only be re-isolated through 14 dpi. In conclusion, there are numerous differences between the NH and H isolates found in the field with the H isolate appearing less virulent and as such, making it a potential vaccine candidate. The phenotypic difference appears to correlate with this, but may not suffice to explain it. / Ph. D.

Background: Hemophilia A is X-linked recessive congenital bleeding disorder. Exogenous infusion of FVIII is the treatment of choice in hemophilia A patients. However, inhibitor development remains the major problem in management of Hemophilia A. It has been showed that IVIG has immunomodulatory effects and it has been being used in the treatment of several autoimmune and inflammatory disorders. Here, we investigated the effect of co-administration of FVIII with IVIG on the development of inhibitor in naive and previously immunized hemophilia A mice. Methods: Initially, hemophiliac mice were immunized by weekly intraperitoneal injection of human recombinant FVIII (rFVIII). The mice then were treated, either by rFVIII/IVIG co-injection or rFVIII alone. In the other experimental group, naive hemophiliac mice were treated with rFVIII/IVIG co-injection for four weeks followed by injection of either rFVIII or rFVIII/IVIG. Plasma's anti-FVIII Ab titer was measured using ELISA. Results: Weekly injection of rFVIII led to the development of anti-FVIII Ab in all previously untreated mice. Treatment of those immunized mice with rFVIII/IVIG co-injection did not reduce the level of pre-existing Ab. On the other hand, naive mice treated with rFVIII/IVIG co-injection showed significantly less Ab titer compared to the mice received rFVIII alone after 4 weeks (mean Ab titre of 1 compared to 39, in rFVIII/IVIG co-injection and rFVIII groups respectively). Although the rFVIII/IVIG-treated mice developed immune response following the injection of rFVIII alone, Ab titer in those that kept receiving rFVIII/IVIG co-injection remained lower compared to other groups during the whole twelve weeks of the experiment. Conclusions: Co-injection of rFVIII with IVIG decreased the anti-FVIII immune response in previously untreated hemophilia A mice. These findings suggest that IVIG co-administration can be effective in management of hemophilia A patients at risk of inhibitor development. / Thesis / Master of Applied Science (MASc)

Vaccine Brucella abortus strain RB51, unlike the wild strain 2308 and another vaccine strain (strain 19) does not induce anti-O-chain antibodies. An efficacious vaccine strain that fails to produce an O-chain and thus a lack of an anti-O-chain humoral response greatly simplifies identification of vaccinated versus field strain infected animals. The three primary objectives of this research were the following: 1) to develop a serological assay to detect anti-RB51 antibodies in vaccinated elk (Cervus elaphus nelsoni), 2) to identify potential antigenic alterations in RB51 after vaccination of elk and BALB/c mice, and 3) to confirm the general stability of RB51. Elk were divided into four groups based upon gender and the route of inoculation (subcutaneous or ballistic) of RB51 bacteria. This study developed a highly reliable ELISA (using a monoclonal anti-bovine IgG 1 antibody and acetone killed whole RB51 bacteria) which can identify RB51-vaccinated elk. Also, isolates recovered from RB51-vaccinated elk were inoculated into female BALB/c mice whose spleens were then cultured. All elk and mice isolates were bacteriologically, biochemically, and serologically evaluated. This study showed that RB51 is a highly stable strain, which does not revert to smooth morphology or initiate synthesis of LPS-O-chain, maintains it biochemical characteristics, does not undergo detectable antigenic variations, and remains attenuated even after successive passages in elk and mice. Overall, this research indicates that RB51 is a vaccine candidate for the prevention of brucellosis in elk. Further studies are needed to determine the protective capabilities of RB51 in elk. / Master of Science

La producción ovina representa un rubro importante dentro del sistema agropecuario de la Argentina y la brucelosis causada por Brucella ovis es considerada el motivo principal de los problemas reproductivos en esta especie. Si bien a la fecha no ha sido reportada como causa de enfermedad en el humano, hay trabajos que informan sobre la presencia de anticuerpos en sangre de personal expuesto. La enfermedad se encuentra en todas las regiones del país donde se crían ovinos, con prevalencias que varían de 3 a 50%. Hasta la fecha no existe un programa nacional o provincial para el control, siendo pocos los laboratorios de diagnóstico que ofrezcan un servicio con técnicas de alta sensibilidad y especificidad. El diagnóstico clínico de la infección por B. ovis, mediante palpación de los epidídimos y testículos, no es suficientemente sensible y debe considerarse como presuntivo. El único diagnóstico certero es el aislamiento e identificación de la bacteria, pero es un método poco práctico para realizarlo en gran número de animales y un resultado negativo no asegura que el animal no esté enfermo. Para el diagnóstico de rutina el uso de pruebas serológicas está muy difundido, y las recomendadas como más eficientes son: inmunodifusión en gel de agar (IDGA), fijación del complemento (FC) e IELISA. El primer objetivo del presente trabajo fue identificar pruebas serológicas que además de ser sencillas y prácticas, presenten alta sensibilidad y especificidad. B. ovis y B. canis comparten componentes antigénicos, por lo cual ambas podrían ser utilizadas como antígeno con resultados similares. Sin embargo el uso de la cepa de B. canis (M-) permite desarrollar un antígeno estable, útil para pruebas de aglutinación. Se estudiaron las pruebas de IDGA e IELISA, utilizando antígeno B. ovis, en el suero de 225 animales. En las mismas muestras se realizó una prueba rápida de microaglutinación (RSAT), 2-mercapto-etanol RSAT (2ME-RSAT) e IELISA utilizando antígeno B.canis. Los valo / López, GA. (2008). Estudio de la brucelosis causada por brucella ovis en ovinos [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/1996

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Background: The monitoring of infectious diseases like the porcine reproductive and respiratory syndrome (PRRS) using pen-wise oral fluid samples becomes more and more established. The collection of individual oral fluid, which would be useful in the monitoring of PRRSV negative boar studs, is rather difficult. The aim of the study was to test two methods for individual oral fluid collection from pigs and to evaluate the specificity of a commercial ELISA for detection of PRRSV antibodies in these sample matrices. For this reason, 334 serum samples from PRRSV negative pigs (group 1) and 71 serum samples from PRRSV positive pigs (group 2) were tested for PRRSV antibodies with a commercial ELISA. Individual oral fluid was collected with a cotton gauze swab from 311 pigs from group 1 and 39 pigs from group 2. Furthermore, 312 oral fluid samples from group 1 and 67 oral fluid samples from group 2 were taken with a self-drying foam swab (GenoTube). The recollected oral fluid was then analysed twice with a commercial ELISA for detection of PRRSV antibodies in oral fluid.