Thesis (MSc (Veterinary Science))--University of Pretoria, 2002. / Includes bibliographical references.

Thesis (Ph. D.)--Ohio State University, 2006. / Title from first page of PDF file. Includes bibliographical references (p. 180-198).

Thesis (doctoral)--Universität Konstanz, 2005.

Thesis (Ph. D.)--Ohio State University, 2005. / Title from first page of PDF file. Document formatted into pages; contains xvi, 216 p.; also includes graphics (some col.). Includes bibliographical references (p. 180-216). Available online via OhioLINK's ETD Center

Tierärztl. Hochsch., Diss., 2004--Hannover.

Tierärztl. Hochsch., Diss., 2004--Hannover.

Freie Universiẗat, Diss., 2005--Berlin. / Dateiformat: zip, Dateien im PDF-Format. Erscheinungsjahr an der Haupttitelstelle: 2005.

Made available in DSpace on 2014-08-13T14:50:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-11-14Bitstream added on 2014-08-13T18:00:47Z : No. of bitstreams: 1 000753373.pdf: 543622 bytes, checksum: 568807a679f23679d4c54335db59a4d0 (MD5) / O reaparecimento de manifestações clínicas após tratamento eficaz, neste texto identificado como recaída, tem sido pouco avaliado. Assim, foram estudados os casos de recaída observados em 400 pacientes, 93 com a forma aguda/subaguda (FA) e 307 com a crônica (FC), que já tinham apresentado cura aparente, isto é, com cura clínica, normalização da velocidade de hemossedimentação e cura sorológica – caracterizada pela presença de teste negativo à imunodifusão dupla em gel de agar por dois anos. Vinte e um (5,2%) desses pacientes apresentaram recaídas da doença. Três (14,3%) eram do sexo feminino e 18 (85,7%) do masculino, com razão de masculinidade de 6:1. Dos 21 pacientes com recaída, 15 (4,8%) apresentavam a FC e 6 (6,4%) a FA (p>0,05). As reativações ocorreram de 46 a 296 meses após introdução do tratamento (Md=96) e de 4 a 267 meses (Md= 60) após sua suspensão. As formas clínicas não diferiram quanto aos tempos decorridos até a reativação. O diagnóstico sorológico de recaída pela IDD foi feito em apenas 45% dos casos, o que levou à avaliação de outros testes para esse fim. Assim, foi realizado o enzimaimunoensaio (ELISA) e o immunoblotting (IB). A sensibilidade da IDD e do ELISA / 0,710 foi 76,1% em amostras de soro obtidos no pré-tratamento (p=0,25). No diagnóstico de recaída, a sensibilidade da IDD foi menor que no pré-tratamento (80% vs 45%; p=0,017), enquanto o ELISA / 0,710 foi igual (80% vs 65%;p=0,125). A sensibilidade do IB para diagnóstico de recaída foi de 12,5% na identificação da gp70 e 43,8% na da gp43 (p<0,05). A avaliação da acurácia do teste ELISA revelou sensibilidade de 96%, especificidade de 95%, valor preditivo positivo de 95%, valor preditivo negativo de 96% e acurácia de 95,5% quando o cut-off utilizado foi a densidade óptica de 0,710, obtido em função da construção da receiver operator characteristc – ROC, para um intervalo de confiança ... / The reappearance of clinical manifestations after efficacious treatment, here identified as relapse, has been rarely evaluated. Thus, the cases of relapse observed in a cohort of 400 patients, 93 with acute/subacute form (AF) and 307 with chronic form (CF) were studied. They had already reached apparent cure, characterized as clinical cure, normalization of the erythrocyte sedimentation rate and serological cure, with a negative double agar gel immunodiffusion test (DID) for two years after antifungal discontinuation. Twenty-one (5.2%) of these patients had relapses. Three (14.3%) were female and 18 (85.7%) were male, with a male:female ratio of 6:1. Of the 21 relapsed patients, 15 (4.8%) presented the CF and 6 (6.4%) the AF (p>0.05). The relapse occurred 46-296 months after introduction of the treatment (Md=8 yrs) and from 4 to 267 months (Md=5 yrs) after withdrawal. Clinical forms did not differ regarding to the time elapsed until relapse. DID was positive in only 45% of the relapsed cases, which led to the evaluation of other tests to diagnose this condition. Thus, the enzyme-linked immunosorbent assay (ELISA) was standardized and the cut off was determined using the curve receiver operator characteristic – ROC and confidence intervals of 95% and 99%, showing optical densities of 0.710 and 0.850, respectively. Then, serological evaluation was performed using ELISA/0.710 and ELISA/0.850, and immunoblotting identifying gp43 (IBgp43) and gp70 (IBgp70). ELISA 0.710 and DID showed the same sensitivity, 76.1%, in serum samples obtained before treatment (p=0.25). DID sensitivity was lower at relapse than before the initial treatment (45% vs 80%; p=0.017), whereas ELISA/0,710 was the same (65% vs 80%; p=0.125). IBgp70 showed a 12.5% and IBgp43 a 43.8% sensitivity for diagnosing relapse (p<0.05). ELISA/0.710 showed a 96% sensitivity, 95% specificity, 95% positive predictive value, 96% negative ...

Orientador: Wilma Aparecida Starke Buzetti / Banca: Edson Guilherme Vieira / Banca: Willian Marinho Dourado Coelho / Resumo: A leishmaniose visceral (LV), doença causada pelo protozoário Leishmania infantum e transmitida pelo flebotomíneo Lutzomyia longipalpis, é considerada um problema de saúde pública no Brasil. O objetivo deste trabalho foi realizar um estudo epidemiológico sobre a ocorrência da LV em cães e gatos e de flebotomíneos em dois abrigos de animais, durante o período de um ano, no município de Ilha Solteira, SP. Amostras de material biológico foram coletadas de 192 cães e de 197 gatos, para realização de exames sorológicos por meio do ensaio imunoenzimático indireto (ELISA), da reação de imunofluorescência indireta (RIFI) para ambas espécies animais e da reação intradérmica de Montenegro para cães. Dos cães examinados, 17,2% (33/192) estavam sorologicamente positivos para LV pelo método ELISA, 19,8% (38/192) pela RIFI e 45,4% (15/33) pela reação intradérmica. Para cães, a concordância entre as técnicas sorológicas foi classificada de razoável a boa, mas quando comparadas à reação de Montenegro foi considerada ruim. No inquérito felino das 197 amostras, a soroprevalência foi de 32,4% (64/197) e 31,9% (63/197) no ELISA e RIFI, respectivamente. Embora a maioria dos cães e dos gatos entregue nos abrigos já estivesse infectada, seis cães e cinco gatos infectaram-se no interior desses abrigos, após algum tempo de permanência nesses recintos. Um total de 131 flebotomíneos foram capturados por meio das armadilhas luminosas, colocadas no interior dos dois abrigos durante o período de dois anos. Os fatores de riscos associados à LV foram determinados estatisticamente pela análise univariada, com valor significativo (p ≤ 0,05), onde o porte pequeno dos cães, o uso de coleira de deltametrina e o diagnóstico precoce foram variáveis determinantes que influenciaram a positividade dos animais, ou seja, as duas últimas variáveis determinaram significativamente na diminuição de casos positivos da doença nos... / Abstract: Visceral leishmaniasis (VL), a disease caused by Leishmania infantum and transmitted by the sand fly Lutzomyia longipalpis, is considered a public health problem in Brazil. The study aimed to carry out an epidemiological study on the occurrence of VL in dogs and cats, and sand flies during the period of one year in two animal shelters from Ilha Solteira, SP. A total of 197 and 192 biological material samples were collected, respectively from cats and dogs for serological survey by an indirect enzyme-linked immunosorbent assay (ELISA) and an indirect fluorescent antibody test (IFAT) for both species of animals, and the intradermal reaction of Montenegro (only for dogs). During the study with dogs, 17,2% (33/192) were serologically positive for LV by ELISA, 19,8% (38/192) by IFAT and 45,4% (15/33) by the intradermal method. The correlation analysis between the serological techniques was ranged from reasonable to good for dogs, but was bad when compared to the Montenegro reaction. The feline seroprevalence was 32,4% (64/197) and 31,9% (63/197) for ELISA and IFAT, respectively. Although the most of dogs and cats in shelters were infected, six dogs and five cats infected inside the shelters. A total of 131 sand flies were captured by the light traps in both shelters, during two years. The risk factors, statistically determined by univariate analysis (p ≤ 0.05), demonstrated that the small size of the dogs, the use of deltamethrin collar and the precocious diagnosis, significantly influenced the animals positivity for VL. It was concluded that these animal shelters were vulnerable to this parasitic infection because of the presence of sand flies and positive animals for LV / Mestre

Two hybridoma clones, labelled 4D10 C1 and 2H4 H12, produced monoclonal antibodies which recognized the outer membrane of an enteropathogenic Escherichia coli (EPEC) 0142:K86:H6 in an enzyme-linked immunosorbent assay (ELISA) and the whole cell in an immunofluorescence assay. Large scale production of the monoclonal antibodies was accomplished through ascites production in balb/c mice. Purification of the ascites fluid was achieved by gel filtration and ion exchange chromatography. Isotyping of the purified fractions showed 4D10 C1 to be an IgG2 and 2H4 H12 an IgM. These monoclonal antibodies were screened by immunofluorescence assay against several pathogenic and nonpathogenic strains of E.coli in addition to other Enterobacteriaciae. Results of the screening showed these antibodies to be specific for the E.coli serotype to which they were raised. Minimal cross-reactivity with other Enterobacteriaceae was observed. In a separate and concurrent project, the use of an ELISA capable of detecting ß-N-acetylglucosaminidase (NAGase) was examined. White Leghorn hens were injected with commercially prepared bovine NAGase. Eggs were collected and the immunoglobulin fraction separated from the egg yolk by polyethylene glycol precipitation followed by ion exchange on a DEAE-Sephacel column. The use of the purified immunoglobulins was examined in a sandwich, double-sandwich and a competitive ELISA. A statistically significant standard curve for the detection of NAGase was successfully derived using a double-sandwich ELISA when rabbit immunoglobulin was used to coat the microwell plates. This assay was used to measure the NAGase concentration in press juice and fish extract of fresh and frozen salmon muscle samples. The ratio of the NAGase concentration in the press juice to the total NAGase concentration was compared. No significant difference was found between the calculated concentration ratios of the fresh muscle samples and samples frozen for 1 week at -20°C. / Land and Food Systems, Faculty of / Graduate