M?todos de diagn?stico molecular e metagen?mico de agentes anaplasmataceae em felinos dom?sticos (domiciliados e errantes) e gamb?s da Regi?o Metropolitana do Rio de Janeiro

• Autor(a) principal: Guimar?es, Andresa
• Data de Publicação: 2017
• Tipo de documento: Tese
• Idioma: por
• Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRRJ
• Texto Completo: https://tede.ufrrj.br/jspui/handle/jspui/2633
• topic: gatos; hemoparasitoses; diagn?stico; cats; hemoparasitosis; diagnosis; Medicina Veterin?ria

Anaplasmataceae agents are gram-negative bacteria that are found in leukocytes, platelets or erythrocytes, forming morulae, characteristic of their infection. The transmission of these agents to man and animals occurs mainly by arthropod vectors. The participation of domestic felines and wild reservoirs has not yet been defined, with few studies on hemoparasitosis affecting these animals. The present study had as objective to investigate the presence of infection by Anaplasmataceae family agents in domestic felines and possums (Didelphis sp.) of the state of Rio de Janeiro, using molecular methods. We sought to sequence the PCR positive samples for validation of the same and to compare them with those deposited in Genbank, as well as to develop new diagnostic methods aiming at fast and high yield results, such as "deep sequencing", used in metagenomic assays. The aim of this study was to determine which hematological changes (hemogram, hematological morphology and hemoparasite inclusions) are more frequent and which associated factors are related to these in two distinct cat populations in the Metropolitan Region of Rio de Janeiro. Two populations of domestic cats, wandering cats (n = 105) and domiciled cats (n = 178) from the Metropolitan region of Rio de Janeiro were studied. Also, were studied 37 opossums collected from nature for rehabilitation, these were tested for the same agents as cats. The diagnosis of Ehrlichia canis, E. chaffeensis and Anaplasma spp. was performed by Polymerase Chain Reaction (PCR). In the comparison between the two populations of cats, there were significant differences between them (p <0.05), revealing that the wandering population had non-standard haematological values for the species, being thrombocytopenia, eosinophilia and hyperproteinemia the most relevant findings. In contrast, the population of domiciled cats presented, on average, within the reference parameters for the species. Only one animal (0.56%) of the domiciled cats population was positive for Ehrlichia chaffeensis based on the 16S rRNA gene, the other cats were negative for all the agents studied. Only one opossum (2.7%) was positive in the PCR for E. canis and E. chaffeensis simultaneously, after sequencing demonstrated 100% identity with E. canis. Part of the populations of cats was submitted to the diagnosis by deep sequencing, where it was observed that the genera of pathogenic bacteria present in these populations are: Mycoplasma and Bartonella, several bacteria were also present in the domestic feline microbiota. It can be concluded that environmental and management conditions interfere in the hematological parameters of domestic cats, so the difference between domiciled and errant. This is the first report of E. chaffeensis in the domestic cat population of Rio de Janeiro. A new strain of E. canis circulates in peridomicile opossum population of Rio de Janeiro. New generation sequencing and metagenomic studies are diagnostic forms and tools for studying the microbiota of domestic animals and should be more widespread in Veterinary Medicine