Comparison of different methods to detect genetic barriers in a small mammal population

Habitat fragmentation and the consequently the loss of connectivity between populations can reduce the individuals interchange and gene flow, increasing the chances of inbreeding, and the increase the risk of local extinction. Landscape genetics is providing more and better tools to identify genetic barriers.. To our knowledge, no comparison of methods in terms of consistency has been made with observed data and species with low dispersal ability. The aim of this study is to examine the consistency of the results of five methods to detect barriers to gene flow in a Mediterranean pine vole population Microtus duodecimcostatus: F-statistics estimations, Non-Bayesian clustering, Bayesian clustering, Boundary detection and Simple/Partial Mantel tests. All methods were consistent in detecting the stream as a non-genetic barrier. However, no consistency in results among the methods were found regarding the role of the highway as a genetic barrier. Fst, Bayesian clustering assignment test and Partial Mantel test identifyed the highway as a filter to individual interchange. The Mantel tests were the most sensitive method. Boundary detection method (Monmonier’s Algorithm) and Non-Bayesian approaches did not detect any genetic differentiation of the pine vole due to the highway. Based on our findings we recommend that the genetic barrier detection in low dispersal ability populations should be analyzed with multiple methods such as Mantel tests, Bayesian clustering approaches because they show more sensibility in those scenarios and with boundary detection methods by having the aim of detect drastic changes in a variable of interest between the closest individuals. Although simulation studies highlight the weaknesses and the strengths of each method and the factors that promote some results, tests with real data are needed to increase the effectiveness of genetic barrier detection.

Characterization of water-soluble organic compounds in bioaerosols by liquid chromatography and fluorescence spectroscopy

In the last decades, the effects of the air pollution have been increasing, especially in the case of the human health diseases. In order to overcome this problem, scientists have been studying the components of the air. As a part of water-soluble organic compounds, amino acids are present in the atmospheric environment as components of diverse living organisms which can be responsible for spreading diseases through the air. Liquid chromatography is one technique capable of distinguish the different amino acids from each other. In this work, aiming at separating the amino acids found in the aerosols samples collected in Aveiro, the ability of four columns (Mixed-Mode WAX-1, Mixed-Mode HILIC-1, Luna HILIC and Luna C18) to separate four amino acids (aspartic acid, lysine, glycine and tryptophan) and the way the interaction of the stationary phases of the columns with the analytes is influenced by organic solvent concentration and presence/concentration of the buffer, are being assessed. In the Mixed-Mode WAX-1 column, the chromatograms of the distinct amino acids revealed the separation was not efficient, since the retention times were very similar. In the case of lysine, in the elution with 80% (V/V) MeOH, the peaks appeared during the volume void. In the Mixed-Mode HILIC-1 column, the variation of the organic solvent concentration did not affect the elution of the four studied amino acids. Considering the Luna HILIC column, the retention times of the amino acids were too close to each other to ensure a separation among each other. Lastly, the Luna C18 column revealed to be useful to separate amino acids in a gradient mode, being the variation of the mobile phase composition in the organic solvent concentration (ACN). Luna C18 was the column used to separate the amino acids in the real samples and the mobile phase had acidified water and ACN. The gradient consisted in the following program: 0 – 2 min: 5% (V/V) ACN, 2 – 8 min: 5 – 2 % (V/V) ACN, 8 – 16 min: 2% (V/V) ACN, 16 – 20 min: 2 – 20 % (V/V) ACN, 20 – 35 min: 20 – 35 % (V/V) ACN. The aerosols samples were collected by using three passive samplers placed in two different locations in Aveiro and each sampler had two filters - one faced up and the other faced down. After the sampling, the water-soluble organic compounds was extracted by dissolution in ultra-pure water, sonication bath and filtration. The resulting filtered solutions were diluted in acidified water for the chromatographic separation. The results from liquid chromatography revealed the presence of the amino acids, although it was not possible to identify each one of them individually. The chromatograms and the fluorescence spectra showed the existence of some patterns: the samples that correspond to the up filters had more intense peaks and signals, revealing that the up filters collected more organic matter.