Comparación de técnicas para contar procariotas en muestras de biofilm y plancton marino

Autores/as

  • Vanessa Ochi Agostini Laboratory of Zooplankton, Universidade Federal do Rio Grande (FURG), Institute of Oceanography (IO) - Laboratory of Biofilms and Microbial Diversity, Universidade Federal do Rio Grande do Sul (UFRGS) - Programa de Pós-Doutorado Empresarial do Conselho Nacional de Desenvolvimento Científico e Tecnológico (PDI-CNPq), Regenera Moléculas do Mar https://orcid.org/0000-0002-8325-254X
  • Letícia Terres Rodrigues Laboratory of Cell Culture, Universidade Federal do Rio Grande do Sul (UFRGS) https://orcid.org/0000-0002-9402-0306
  • Alexandre José Macedo Laboratory of Biofilms and Microbial Diversity, Universidade Federal do Rio Grande do Sul (UFRGS), Pharmacy Faculty and Biotechnology Centre https://orcid.org/0000-0002-8951-4029
  • Erik Muxagata Laboratory of Zooplankton, Universidade Federal do Rio Grande (FURG), Institute of Oceanography (IO) https://orcid.org/0000-0002-4210-5252

DOI:

https://doi.org/10.3989/scimar.05117.019

Palabras clave:

ecología, enumeración de bacteria, citometría de flujo, métodos microbianos, microscopía

Resumen


A pesar de la gran cantidad de técnicas disponibles para el estudio de bacterias acuáticas, el objetivo de este estudio fue aplicar y proponer una técnica para el análisis de células procariotas de vida libre y asociado a biofilms en ensayos de laboratorio. En particular, deseamos analizar la eficiencia de de la aplicación de ultrasonidos para separar y romper el biofilm, obtener un tratamiento de tinción eficiente para cuantificar procariotas de vida libre y de biofilm por citometría de flujo (CF), y comparar microscopía de epifluorescencia (MEP), microscopía electrónica de barrido (MEB) y CF para cuantificar los procariotas de vida libre y de biofilm. Los sustratos de madera contrachapada de grado marino se sumergieron en agua marina natural que se acondicionó durante 12 días. A los 6 y 12 días, se retiraron alícuotas de agua y sustratos para estimar la densidad de procariotas de vida libre y asociados a los biofilms. Los ultrasonidos eliminaron de manera eficiente el biofilm marino de los sustratos (hasta 94%) sin dañar las células. El análisis por CF (sin marcador) cuantificó de manera fiable las células del plancton marino y los procariotas de biofilms jóvenes o maduros en comparación con otras técnicas de marcación (naranja de acridina, 4’, 6-diamidino-2-fenilindol, yoduro de propidio, ácido nucleico fluorescente verde), MEP o MEB. CF y MEB lograron resultados similares, mientras que se observó una alta variabilidad en la técnica EFM. Cuando se compara, CF es más rápido y más preciso que MED, ya que el recuento no depende del observador.

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Citas

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Publicado

2021-09-02

Cómo citar

1.
Ochi Agostini V, Terres Rodrigues L, Macedo AJ, Muxagata E. Comparación de técnicas para contar procariotas en muestras de biofilm y plancton marino. Sci. mar. [Internet]. 2 de septiembre de 2021 [citado 27 de julio de 2024];85(3):211-20. Disponible en: https://scientiamarina.revistas.csic.es/index.php/scientiamarina/article/view/1908

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Conselho Nacional de Desenvolvimento Científico e Tecnológico
Números de la subvención 11/2014 (6/2551-000244-4)