Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá)

Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá)

The amilolytic thermophilic anaerobic microorganisms of a mineral hot spring in the Andean region (5° 45' 69’’ N, 73° 6'61’’W, 2500 m) were quantified through the Most Probable Number (MPN) technique. The microbial recounts of the populations presented values between 1.4*102cell/100 mL and 5.8*102 cell/100 mL in the presence of starch and thiosulphate as electron acceptors and 3.4*102 cel/100 mL in the presence of starch alone. Microbial isolations were conducted starting with the last positive dilutions of the MPN and 8 bacterial strains were isolated and denominated P4-6, P4-7, P4-8, P4-9, P4-10, P4-11, P4-12, and P4-13. These strains grew at optimum temperatures between 60 and 65 °C and exhibited fermentative metabolism. The main product of fermentation was ethanol followed by acetate together with hydrogen and CO2. Thiosulphate was used as external acceptor of electrons by all the isolated strains, but not sulphate or ferric iron. The phylogenetic diversity of these 8 strains was evaluated through denaturing gel gradient electrophoresis (DGGE) analysis. The sequence of the gene encoding for the 16S rRNA of two of the isolated strains (P4-6 and P4-9) was analyzed. These analyses showed that they belong to the family Thermoanaerobiaceae, domain Bacteria. From their phenotypical and genotypical characteristics, it is inferred that these organisms belong to the genus Thermoanaerobacter with similarity of 98% of the 16S rDNA with Thermoanaerobic italicus and Thermoanaerobacter mathranii. Key words: thermophile, hot springs, anaerobiosis, Thermoanaerobacter, DGGE.

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Main Authors: Posada, Yully, Agudelo, Alejandra, Álvarez, Elsa, Díaz, Carolina, Joulian, Catherine, Ollivier, Bernard, Baena, Sandra
Format: Digital revista
Language:spa
Published: Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología 2004
Online Access:https://revistas.unal.edu.co/index.php/biotecnologia/article/view/530
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country Colombia
countrycode CO
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access En linea
databasecode rev-biotecnologia-co
tag revista
region America del Sur
libraryname Sistema Nacional de Bibliotecas de la UNAL
language spa
format Digital
author Posada, Yully
Agudelo, Alejandra
Álvarez, Elsa
Díaz, Carolina
Joulian, Catherine
Ollivier, Bernard
Baena, Sandra
spellingShingle Posada, Yully
Agudelo, Alejandra
Álvarez, Elsa
Díaz, Carolina
Joulian, Catherine
Ollivier, Bernard
Baena, Sandra
Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá)
author_facet Posada, Yully
Agudelo, Alejandra
Álvarez, Elsa
Díaz, Carolina
Joulian, Catherine
Ollivier, Bernard
Baena, Sandra
author_sort Posada, Yully
title Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá)
title_short Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá)
title_full Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá)
title_fullStr Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá)
title_full_unstemmed Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá)
title_sort counts, isolation and identification of amilolytic anaerobic communities from a hot spring in paipa (boyacá)
description The amilolytic thermophilic anaerobic microorganisms of a mineral hot spring in the Andean region (5° 45' 69’’ N, 73° 6'61’’W, 2500 m) were quantified through the Most Probable Number (MPN) technique. The microbial recounts of the populations presented values between 1.4*102cell/100 mL and 5.8*102 cell/100 mL in the presence of starch and thiosulphate as electron acceptors and 3.4*102 cel/100 mL in the presence of starch alone. Microbial isolations were conducted starting with the last positive dilutions of the MPN and 8 bacterial strains were isolated and denominated P4-6, P4-7, P4-8, P4-9, P4-10, P4-11, P4-12, and P4-13. These strains grew at optimum temperatures between 60 and 65 °C and exhibited fermentative metabolism. The main product of fermentation was ethanol followed by acetate together with hydrogen and CO2. Thiosulphate was used as external acceptor of electrons by all the isolated strains, but not sulphate or ferric iron. The phylogenetic diversity of these 8 strains was evaluated through denaturing gel gradient electrophoresis (DGGE) analysis. The sequence of the gene encoding for the 16S rRNA of two of the isolated strains (P4-6 and P4-9) was analyzed. These analyses showed that they belong to the family Thermoanaerobiaceae, domain Bacteria. From their phenotypical and genotypical characteristics, it is inferred that these organisms belong to the genus Thermoanaerobacter with similarity of 98% of the 16S rDNA with Thermoanaerobic italicus and Thermoanaerobacter mathranii. Key words: thermophile, hot springs, anaerobiosis, Thermoanaerobacter, DGGE.
publisher Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología
publishDate 2004
url https://revistas.unal.edu.co/index.php/biotecnologia/article/view/530
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spelling oai:www.revistas.unal.edu.co:article-5302017-06-12T21:54:35Z Counts, isolation and identification of amilolytic anaerobic communities from a hot spring in Paipa (Boyacá) Cuantificación, aislamiento e identificaciónde comunidades anaerobias amilolíticas de un manantial termomineral de Paipa, Boyacá Posada, Yully Agudelo, Alejandra Álvarez, Elsa Díaz, Carolina Joulian, Catherine Ollivier, Bernard Baena, Sandra termofilia manantiales termominerales anaerobiosis Thermoanaerobacter DGGE thermophile hot springs DGGE. The amilolytic thermophilic anaerobic microorganisms of a mineral hot spring in the Andean region (5° 45' 69’’ N, 73° 6'61’’W, 2500 m) were quantified through the Most Probable Number (MPN) technique. The microbial recounts of the populations presented values between 1.4*102cell/100 mL and 5.8*102 cell/100 mL in the presence of starch and thiosulphate as electron acceptors and 3.4*102 cel/100 mL in the presence of starch alone. Microbial isolations were conducted starting with the last positive dilutions of the MPN and 8 bacterial strains were isolated and denominated P4-6, P4-7, P4-8, P4-9, P4-10, P4-11, P4-12, and P4-13. These strains grew at optimum temperatures between 60 and 65 °C and exhibited fermentative metabolism. The main product of fermentation was ethanol followed by acetate together with hydrogen and CO2. Thiosulphate was used as external acceptor of electrons by all the isolated strains, but not sulphate or ferric iron. The phylogenetic diversity of these 8 strains was evaluated through denaturing gel gradient electrophoresis (DGGE) analysis. The sequence of the gene encoding for the 16S rRNA of two of the isolated strains (P4-6 and P4-9) was analyzed. These analyses showed that they belong to the family Thermoanaerobiaceae, domain Bacteria. From their phenotypical and genotypical characteristics, it is inferred that these organisms belong to the genus Thermoanaerobacter with similarity of 98% of the 16S rDNA with Thermoanaerobic italicus and Thermoanaerobacter mathranii. Key words: thermophile, hot springs, anaerobiosis, Thermoanaerobacter, DGGE. Se cuantif icaron microorganismos anaerobios termofílicos amilolíticos de un manantial termomineral en la región andina (5° 45' 69’’ N, 73° 6' 61’’ W, 2500 msnm) a través del Número Más Probable (NMP). Los recuentos microbianos de las poblaciones presentaron valores entre 1,9*102 células/100 mL y 5.8*102 células/100 mL en presencia de almidón y tiosulfato como aceptor de electrones y 1,4*102células/100 mL y 3,4*102 células/100 mL en presencia solamente de almidón. Se realizaron aislamientos microbianos a partir de las últimas diluciones positivas del NMP y se aislaron 8 cepas bacterianas denominadas P4-6, P4-7, P4-8, P4-9, P4-10, P4-11, P4-12 y P4-13. Estas cepas crecieron a temperaturas óptimas entre 60 y 65 °C, y exhibieron un metabolismo fermentativo. El principal producto de fermentación fue etanol seguido de acetato, CO2 e hidrógeno. El tiosulfato fue utilizado como aceptor externo de electrones, pero el sulfato o el hierro férrico no fue reducido. La diversidad filogenética de estas 8 cepas fue evaluada por medio de geles de electroforesis de gradiente denaturalizante (DGGE). Se analizó la secuencia del gen 16S rRNA de dos de las cepas aisladas (P4-6 y P4-9) y el análisis indicó que éstas pertenecen a la familia Thermoanaerobiaceae del dominio Bacteria. Del análisis fenotípico y genotípico se deduce que estos organismos pertenecen al género Thermoanaerobacter, y con base en el análisis de las secuencias del 16S rDNA se observa una similitud del 98% con Thermoanaerobacter italicus y Thermoanaerobacter mathranii. Palabras clave: termofilia, manantiales termominerales, anaerobiosis, Thermoanaerobacter, DGGE. Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología 2004-07-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Artículo revisado por pares text/html application/pdf application/msword https://revistas.unal.edu.co/index.php/biotecnologia/article/view/530 Revista Colombiana de Biotecnología; Vol. 6 No. 2 (2004); 90-99 Revista Colombiana de Biotecnología; Vol. 6 Núm. 2 (2004); 90-99 1909-8758 0123-3475 spa https://revistas.unal.edu.co/index.php/biotecnologia/article/view/530/980 https://revistas.unal.edu.co/index.php/biotecnologia/article/view/530/1023 https://revistas.unal.edu.co/index.php/biotecnologia/article/view/530/1024 Derechos de autor 2004 Revista Colombiana de Biotecnología https://creativecommons.org/licenses/by/4.0

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