Bacterias y Actinobacterias aisladas del PNN los Nevados para estudios de actividad antimicriobana

Study Extent

Soil samples from the National Natural Park los Nevados Suelos en el Parque Nacional Natural los Nevados

Sampling

Soils (approximately 100 - 200g) were sampled in sterile containers, transported to the labs for processing. In general, several sub-samples were taken per site, and these were consolidated into a single sample for processing and analysis. Se tomaron muestras de diferentes suelos (aproximadamente 100 - 200g) que se transportaron a los laboratorios para ser procesados. Por lo general se tomaron varias muestras por punto de muestreo que luego se consolidaron en una sola muestra para procesamiento y análisis.

Method steps

1. Serial dilutions of soil samples in saline solution were plated on diverse culture media in the lab, like Oatmeal Agar or Actino medium (Difco), among others. Plates were incubated at different temperatures for several days until growth was observed. Colonies were purified by streaking on the same media and strains were stored at -80oC in medium with 20% glycerol. Strains were characterized based on colony morphologies on various media, macro and microscopic observations, and the production of metabolites with antimicrobial activity. Taxonomy was determined by sequence analysis of the 16S rRNA gene. Se hicieron diluciones seriadas de las muestras de suelo en solución salina que luego se sembraron sobre diferentes medios de cultivo como agar avena o medio actino (Difco), entre otros. Las muestras se incubaron a diferentes temperaturas por varios días hasta ver crecimiento. Las colonias se purificaron re-estriando en los mismos medios y se guardaron a -80oC en medio con 20% glicerol. Se determinaron morfologías en estos medios, se hicieron observaciones macro y microscópicas y se analizó la capacidad de producción de moléculas con actividad antimicrobiana. Se determinó la taxonomía haciendo amplificación, secuenciación y análisis del gen ribosomal 16S rRNA.

All specimens are Bacteria, most are classified to the phylum level as Actinobacteria. There are also some Proteobacteria and some Firmicutes. All isolates are classified to the Class level, only a few were identified at the species level.

1. Actinobacteria
   rank: phylum
2. Proteobacteria
   rank: phylum
3. Firmicutes
   rank: phylum

The strains were collected from soils located in the National Natural Park Los Nevados in Colombia located on the Cordillera Central in Colombia. The Park reaches altitudes between 2,600 and 5,321 m.a.s.l. with an area of approximately 58,300 hectares and diverse ecosystems (Van Wyngaarden, W., & Fandiño, M. 2002). It is located within the Departments of Caldas (Municipio de Villamaría), Risaralda (Municipios de Santa Rosa de Cabal y Pereira), Quindío (Municipio de Salento) and Tolima (Municipios de Ibagué, Anzoátegui, Santa Isabel, Murillo, Villahermosa, Casabianca y Herveo)(www.parquesnacionales.gov.co/).