Aislamiento e identificación de microorganismos del género Methanococcus y Methanobacterium de cuatro fuentes de Bogotá D.C.

Abstract

<p>Las bacterias metanogénicas obtienen su energía mediante la producción metabólica de gas metano, utilizando sustratos como dióxido de carbono, acetato y sustratos de metilo a través de procesos de hidrólisis y acetogenesis y son esenciales en la degradación anaerobia de la materia orgánica en la naturaleza. El propósito de esta investigación fue aislar bacterias metanogénicas para conservarlas en la colección de cultivos de la Universidad Colegio Mayor de Cundinamarca. El muestreo se realizó por duplicado de cuatro fuentes ubicadas en Bogotá D.C, Colombia, las cuales ofrecían las características ambientales para su desarrollo. El procedimiento incluyó la toma de la muestra en ambiente anaerobio, aislamiento en medios selectivos e identificación por observación de las características microscópicas con coloración de Gram y de características macroscópicas en los medios selectivos y verificación de producción de metano mediante prueba piloto.</p><p>Los resultados permitieron evidenciar la presencia de bacterias de los géneros <em>Methanococcus </em>y <em>Methanobacterium </em>a partir de las fuentes seleccionadas para el estudio. Se concluyó que el mejor método para la conservación de estos géneros es la congelación con la adición de agentes reductores y glicerol como criopreservante.</p>

Genetic Characterization of Anaerobic Micro-Organisms Applied to Wastewater Treatment: An Alternative in Areas of Height

“…The sampling of effluents discharges from stables was performed in triplicate with sterile 60 ml syringes, which were introduced approximately 40 cm under the water mirror [17], according to the anaerobic treatment to be carried out, the sampling was separated into three areas: (1) wetland of Tarqui river sub-basin; (2) stagnant water near the experimental farms; and (3) sewage running of the investigated farms. The samples were kept under anaerobic conditions and below 10°C for transportation to the laboratory [18].…”

“…For the inoculation of the sample, 100 μL of each liquid means was taken and incubated for 15 days at 37°C in an anaerobic atmosphere (AnaeroGen OXOID), afterwards the bacterial growth was verified by the presence of turbidity in the liquid means. In solid means was planted MC and MB pre-reduced, they were incubated with the same conditions for liquid means previously described, and finally the presence of bacilli and cocci growth obtained in the liquid and solid means was confirmed by Gram staining [17], [21].…”

“…The phenotypic identification was carried out based on traditional schemes such as the observable characteristics in morphology, development, biochemistry and metabolism [22] In addition to the Gram staining, the metabolic capacity of the bacteria to produce methane gas was evaluated, by means of: 16 x 150 mm with lateral detachment, rubber stoppers, latex hoses of 20 cm and tweezers; the system was assembled in a biosafety cabinet, 6 ml of MC and MB liquid culture means were supplied, providing the space for gas storage that bacteria will generate; the colonies obtained from the solid media of MC and MB were inoculated before sealing the system subsequently, the qualitative measurement was made by the water displacement technique, demonstrated by the presence of bubbles that were generated by the gas production and the increasing of water volume [17], [21].…”