En embalses, la eutrofización es consecuencia de procesos naturales y de actividades humanas, lo cual puede facilitar la aparición de afloramientos de cianobacterias potencialmente tóxicas. En este trabajo, se utilizaron dos técnicas moleculares en la detección de genes presentes en cepas potencialmente tóxicas de cianobacterias y evaluaciones de diversidad molecular de cianobacterias en los embalses para potabilización de agua, Riogrande II y La Fe, Colombia. Entre 2010-2011, se tomaron 12 muestras de agua en ambos embalses y se realizaron extracciones de ADN para un análisis de marcadores moleculares mediante PCR y DGGE. Se amplificaron secuencias entre 250-300pb de los genes mcyA y mcyE, implicados en la toxicidad de cepas de cianobacterias. Asimismo, se amplificaron secuencias de la región 16S del ARN ribosomal (422pb), para la técnica DGGE. Se corrieron geles de poliacrilamida en gradientes de desnaturalización, se realizó agrupamiento genético (UPGMA), y se separaron por patrones de bandeo las muestras de cada embalse evaluado. Se demuestra la utilidad de las técnicas moleculares en estudios relacionados con la búsqueda de genes asociados con toxicidad y diversidad molecular de cianobacterias en muestras de agua provenientes de embalses de agua con fines de potabilización para centros urbanos.

In lentic water bodies as reservoirs occur eutrophication processes, originated mainly from human activities (i.e. agriculture, animal exploitation). This influx of nutrients in aquatic ecosystems could promote blooms of potentially toxic cyanobacteria. The purpose of this work is to detect the presence of cyanobacteria strains in water samples, using molecular techniques to help in preventive management of reservoirs dedicated to water purification. We used two molecular techniques to detect genes implied with the synthesis of hepatotoxic microcystins from potentially toxic cyanobacteria strains, and to evaluate the molecular diversity of cyanobacteria in water samples from two high-mountain reservoirs used for purification of drinking water for the metropolitan area of Medellin, Colombia. Between 2010-2011 collections of 12 water samples were taken and DNA extraction together with PCR and DGGE analyses where carried out. We amplified 22 sequences between 250-300bp of the genes mcyA and mcyE, and these sequences were related with several strains and cyanobacteria genera accessions from NCBI-GenBank databases. Moreover, sequence amplifications of the 16S small ribosomal RNA subunit - 16S rRNA- between 400-800bp were also performed in order to use them for the DGGE technique. The amplification products of DGGE were set in polyacrilamide gel with posterior denaturing electrophoresis, and the scanned images of the gel bands were analysed with the software GelCompar II. For Riogrande II and La Fe reservoirs we found 35 and 30 different DGGE bands, respectively, as a measurement of molecular diversity in these artificial ecosystems. Here, we demonstrated the utility of two molecular techniques for the detection of genes associated with toxicity and molecular diversity of cyanobacteria in reservoirs destined for drinking water in urban centers. We recommend strongly following with periodically molecular biology studies in these ecosystems combined with limnological and ecological data, as new tools for management of plants of water purification and for the prevention of potentially public health concerns. Rev. Biol. Trop. 62 (1): 403-419. Epub 2014 March 01.

Brycon henni es una especie nativa de la cuenca del río Magdalena, Colombia, donde es fuertemente explotada, por lo cual en este trabajo se estudió la variabilidad genética y estructura poblacional en individuos provenientes de la cuenca media de los ríos Nare y Guatapé, Antioquia. Se recolectaron 195 ejemplares y se les extrajo ADN genómico total del tejido muscular y/o sanguíneo. Se evaluaron 14 cebadores para la técnica RAPD (Random Amplifyed Polimorphic DNA), siendo cuatro de ellos polimórficos, los cuales generaron 66 fragmentos diferentes (63% polimorfismo). Por medio de un análisis de varianza molecular (AMOVA), se determinó la estructuración poblacional para todos los sitios evaluados y la cuenca del río Nare ( ΦST =0.297 y ΦST =0.163, respectivamente). Las distancias genéticas de Nei mostraron que todas las poblaciones diferían entre sí y que la mayor diferenciación genética se dio para las poblaciones del río Guatapé y la quebrada El Cardal. Un test de Mantel mostró correlación entre las distancias genéticas y geográficas (ambas cuencas, r=0.431; cuenca del río Nare, r=0.377). Esto sugiere aislamiento por distancia y separación de grupos genéticos producidos por los embalses sobre los ríos Nare y Guatapé, lo cual puede tener implicaciones relevantes en la conservación de la variabilidad genética de las poblaciones naturales de esta especie en el oriente antioqueño.

Genetic variability of Brycon henni (Characiformes: Characidae) in the middle basin of Nare and Guatapé rivers, Magdalena River system, Colombia. Brycon henni is a native species in Magdalena´s River basin, and because of its cultural and economic importance, is strongly overexploited. This study aimed to describe the genetic variability and population structure of this species from Nare and Guatapé rivers basins. A total of 195 individuals were collected and DNA extractions were obtained from muscle and blood tissue. Fourteen primers were evaluated with the RAPD technique, being four of them polymorphic, and produced 66 different fragments (63% polymorphism). Besides, using the molecular variance (AMOVA) analysis, the population structure was described for all sites ( ΦST=0.297, ΦST=0.163; Nare and Guatapé river basins, respectively), and suggested the importance of the migratory behaviour of the species in the genetic differentiation. Genetic distances among sampled sites showed that most of the genetic differentiation occurred between sites Guatapé and El Cardal with respect to the others. A Mantel Test demonstrated a correlation between genetic and geographic distances (r=0.431 both of the basins evaluated; r=0.377, Nare river middle basin), and suggested isolation by distance. The outcomes obtained in this study have valuable implications in species conservation and the genetic variability of natural populations of B. henni, and should be complemented with morphological analyses. Rev. Biol. Trop. 59 (1): 269-282. Epub 2011 March 01.