Protocolos de extracción de material genético de excretas de palomas (Columba livia) para la detección de hongos patógenos mediante ensayos LAMP

Abstract

Pathogenic fungi are the cause of infectious mycoses in humans and animals. The domestic pigeon (Columba livia) is one of the main reservoirs of these fungi that can be aspirated producing a series of respiratory and systemic disorders. The objective of this research was to evaluate protocols for the extraction of pigeon genetic material and pathogenic fungi in pigeon excreta samples through 4 protocols (A, B, C and D) with the use of physical, chemical and enzymatic methods to later determine the quantity, quality, integrity and viability of the genetic material obtained with these protocols using LAMP assays. Pathogenic fungi are diagnosed with culture techniques, microscopy, and serology, so implementing a molecular biology technique offers an effective diagnostic tool for these fungi in pigeons and their relationship with public health. For this study, samples of dry excreta from nest boxes and wet excreta from the cloaca of birds were taken at strategic points of high density of pigeons. In the statistical analysis of the quantity and quality of DNA obtained, the estimation of means was performed using the Ky.Plot 5.0 program and Duncan`s test in which the quantity measured by DNA concentration was higher in the protocols of the dry excreta extracts than in the wet ones and the quality for the C protocol of dry and wet excreta was found in the ideal parameter of ratio 260/280; in relation of the integrity analysis, the contingency table of relative frequencies was made using the Infostat program. The results obtained for the amplification by LAMP assays were negative for each one of the protocols in the two types of extracts, whether dry or wet excreta.