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Previous issue date: 2016-02-26 / CAPES / The study on parabasalids in companion animals is gaining more attention for its association with diarrhea. The flagellate Pentatrichomonas hominis has been reported in association with domestic cats since the early 20th century. As a eurixenic parasite has been described in several hosts, including humans, dogs, bovines, rats, and a variety of wild animals. Another parabasalid with great importance in livestock, Tritrichomonas foetus has been also described in cats, pigs, and humans. These observations raise questions about the zoonotic potential of both species and their host specificity. This study aimed to diagnose parabasalid species found in cats using morphological and molecular analysis. Therefore, this study was divided into two parts to assess two different cat populations. In the first part, 41 animals in a trial cattery were evaluated. Twenty-six percent of the animals (11) were positive for P. hominis, at both techniques as fresh examination and culture. The DNA was extracted from the samples in culture and rRNA genes were amplified by polymerase chain reaction (PCR) using universal primers (TFR1 and 2) and other two specific species for T. foetus (TFR3 and 4) and P. hominis (TH3 and 5). Morphological analysis of trophozoites revealed the presence of five previous plagues core round and axostyle tapering uniform, characteristic way of P. hominis. The results of morphology were confirmed by molecular study. The sequencing of the isolates revealed a sequence with 100% similarity to P. hominis isolated from cats and dogs deposited in Genbank. This is the first study in Brazil with pointed out the presence of Parabasalids in cats by using morphological and molecular data and it is the one in the literature where P. hominis was isolated. In the second part of this study, 77 samples of feces from cats from the clinical care of HVPA-UFRRJ were examined. Only four of 77 samples tested were positive. Morphological analysis showed predominantly pear-shaped protozoa with three previous scourges, elongated nucleus and ax?stilo abruptly ending in characteristic fillet in T. fetus. In scanning electron microscopy and transmission, were visualized the identifying characters were similar to those previously reported for T. foetus. Molecular analysis confirmed the morphological diagnosis in the organism from four samples showed a sequence with 99.7 to 100% of similarity. It was deposited in Genbank as T. foetus. Despite the morphological analysis have recognized only T. foetus in the four samples examined, three of them were also positive for P. hominis in molecular analysis used as a differential diagnosis using species-specific primers (TH3 and TH5). The molecular analysis was used as a confirmatory tool for the presence of only one species present in evaluated feces. This demonstrated that not only T. foetus was identified in this study, but a co-infection by P. hominis cats could be considered. This indication was only confirmed as a diagnostic techniques when the morphological analysis and molecular biology were used to confirm both species. / O estudo sobre parabasal?deos em animais de companhia vem ganhando cada vez mais aten??o por sua associa??o a quadros de diarreia. O flagelado Pentatrichomonas hominis foi relatado em associa??o com gatos dom?sticos desde o in?cio do s?culo 20. Por ser um parasito eurix?nico, j? foi descrito em diversos hospedeiros, incluindo os seres humanos, c?es, bovinos, ratos e uma variedade de animais selvagens. Assim tamb?m outra esp?cie de parabasal?deo com grande import?ncia na pecu?ria, Tritrichomonas foetus j? foi descrito em gatos, su?nos, e em humanos tamb?m. Tais observa??es levantam d?vidas sobre o potencial zoon?tico de ambas as esp?cies e sua inespecificidade hospedeira. Este estudo teve por objetivos diagnosticar esp?cies de parabasal?deos encontrados em gatos utilizando de an?lise morfol?gica e molecular. Para tanto, este estudo foi dividido em duas partes para avaliar duas popula??es felinas distintas. Na primeira parte, 41 animais de um gatil de experimenta??o foram avaliados. Vinte e seis por cento dos animais (11) foram positivos para P. hominis, tanto no exame a fresco quanto na cultura. O DNA foi extra?do das amostras em cultura e os genes de rRNA foram amplificados por rea??o em cadeia da polimerase (PCR), utilizando iniciadores universais (TFR1 e 2) e outros dois esp?cies espec?ficos para T. foetus (TFR3 e 4) e P. hominis (TH3 e 5). A an?lise morfol?gica dos trofozo?tos revelou a presen?a de cinco flagelos anteriores, n?cleo redondo e ax?stilo afunilando de maneira uniforme, caracter?stico de P. hominis. Os resultados da morfologia foram confirmados pelo estudo molecular. O sequenciamento dos isolados revelou 100% de similaridade de sequ?ncia com P. hominis isolado de gato e de c?o depositados no Genbank. Este ? o primeiro estudo realizado no Brasil sobre a presen?a de parabasal?deos em gatos utilizando dados morfol?gicos e moleculares e o ?nico na literatura onde somente P. hominis foi isolado. Na segunda parte deste estudo, 77 amostras de fezes de gatos provenientes do atendimento cl?nico do HVPA-UFRRJ foram examinadas. Somente quatro amostras apresentaram positivas. A an?lise morfol?gica demonstrou protozo?rios predominantemente piriformes com tres flagelos anteriores, n?cleo alongado e ax?stilo terminando bruscamente em filete caracter?stico de T. foetus. Na microscopia eletr?nica de varredura e de transmiss?o, foram visualizados caracteres de identifica??o semelhantes aos descritos na literatura para T. foetus. ? an?lise molecular, confirmou o diagn?stico morfol?gico nas quatro amostras, e no sequenciamento apresentaram 99,7-100% de similaridade com sequencias de T. foetus depositadas no Genbank. Apesar da an?lise morfol?gica ter reconhecido somente T. foetus nas quatro amostras, tr?s delas foram positivas tamb?m para P. hominis na an?lise molecular utilizada como diagn?stico diferencial utilizando iniciadores esp?cie-espec?fica (TH3 e TH5). O estudo molecular foi utilizado como ferramenta confirmat?ria da presen?a de somente uma esp?cie presente nas fezes avaliadas. Isso demonstra que n?o s? T. foetus foi identificado pelo presente estudo, como tamb?m a coinfec??o por P. hominis em felinos. Estas informa??es s? foram confirmadas quando se utilizou as t?cnicas de diagn?stico em conjunto como an?lise morfol?gica simples e biologia molecular.
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:jspui/1821 |
| Date | 26 February 2016 |
| Creators | SANTOS, Caroline Spitz dos |
| Contributors | Lopes, Carlos Wilson Gomes, Jesus, Vera L?cia Teixeira de, Berto, Bruno Pereira, Cardozo, Sergian Vianna, Almeida, Elan Cardozo Paes de, Leal, Paulo Daniel Sant'Ana, Jesus, Vera L?cia Teixeira de |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Programa de P?s-Gradua??o em Ci?ncias Veterin?rias, UFRRJ, Brasil, Instituto de Veterin?ria |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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