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Phylogenetic characterization and quantification of ammonia-oxidizing archaea and bacteria from Lake Kivu in a long-term microcosm incubation

A microcosm cultivation-based method was set up to investigate the growth of ammonia-oxidizing archaea (AOA), isolated from a water sample acquired at a depth of 50 m from the northern basin of Lake Kivu. For this purpose, both CARD-FISH and qPCR targeting of archaeal 16S rRNA and amoA genes were used. Archaeal cell growth at the end of the 246-day microcosm experiment accounted for 35 % of the SybrGold-stained cells, which corresponded to 6.61 × 106 cells/ml and 1.76 ± 0.09 × 106 archaeal 16S rRNA gene copies/ml. Clone libraries and DGGE fingerprinting confirmed the dominance of AOA phylotypes in the archaeal community microcosm. The majority of the identified archaeal 16S rRNA gene sequences in the clone libraries were affiliated with Thaumarchaeota Marine Group 1.1a. Subsequent cultivation of the AOA community on deep-well microtiter plates in medium containing different carbon sources to stimulate archaeal growth failed to show significant differences in archaeal abundance (ANOVA t14 = -1.058, P = 0.308 and ANOVA t14 = 1.584, P = 0.135 for yeast extract and simple organic acids, respectively). The lack of growth stimulation by organic compounds is in concordance with the oligotrophic status of Lake Kivu. Finally, the addition of antibiotics to the growth medium resulted in archaeal cell counts that were significantly lower than those obtained from cultures in antibiotic-free medium

This work was founded by the Spanish Government through projects CRENYC (CGL2006-12058-C02-01), ARKI (CGL2007-29823-E) and ARCANOX (CGL2009-13318-C02-02). A.P.C. and M.F.H. are recipients of pre-doctoral fellowships from the Catalan (2006FI-109) and theSpanish (FPI BES-2010-035225) Governments, respectively

© International Microbiology, 2013, vol. 16, núm. 3, p. 177-189

Spanish Society for Microbiology (SEM)

Manager: Ministerio de Ciencia y Tecnología (Espanya)
Ministerio de Educación y Ciencia (Espanya)
Ministerio de Ciencia e Innovación (Espanya)
Author: Plasencia, Anna
Gich Batlle, Frederic
Fillol Homs, Mireia
Borrego i Moré, Carles
Date: 2013 December 24
Abstract: A microcosm cultivation-based method was set up to investigate the growth of ammonia-oxidizing archaea (AOA), isolated from a water sample acquired at a depth of 50 m from the northern basin of Lake Kivu. For this purpose, both CARD-FISH and qPCR targeting of archaeal 16S rRNA and amoA genes were used. Archaeal cell growth at the end of the 246-day microcosm experiment accounted for 35 % of the SybrGold-stained cells, which corresponded to 6.61 × 106 cells/ml and 1.76 ± 0.09 × 106 archaeal 16S rRNA gene copies/ml. Clone libraries and DGGE fingerprinting confirmed the dominance of AOA phylotypes in the archaeal community microcosm. The majority of the identified archaeal 16S rRNA gene sequences in the clone libraries were affiliated with Thaumarchaeota Marine Group 1.1a. Subsequent cultivation of the AOA community on deep-well microtiter plates in medium containing different carbon sources to stimulate archaeal growth failed to show significant differences in archaeal abundance (ANOVA t14 = -1.058, P = 0.308 and ANOVA t14 = 1.584, P = 0.135 for yeast extract and simple organic acids, respectively). The lack of growth stimulation by organic compounds is in concordance with the oligotrophic status of Lake Kivu. Finally, the addition of antibiotics to the growth medium resulted in archaeal cell counts that were significantly lower than those obtained from cultures in antibiotic-free medium
This work was founded by the Spanish Government through projects CRENYC (CGL2006-12058-C02-01), ARKI (CGL2007-29823-E) and ARCANOX (CGL2009-13318-C02-02). A.P.C. and M.F.H. are recipients of pre-doctoral fellowships from the Catalan (2006FI-109) and theSpanish (FPI BES-2010-035225) Governments, respectively
Format: application/pdf
Citation: 022977
ISSN: 1139-6709 (versió paper)
1618-1905
Document access: http://hdl.handle.net/10256/12659
Language: eng
Publisher: Spanish Society for Microbiology (SEM)
Collection: MICYT/PN 2006-2009/CGL2006-12058-C02-01
MEC/PN 2008-2009/CGL2007-29823-E
MICINN/PN 2010-2012/CGL2009-13318-C02-02
Reproducció digital del document publicat a: http://dx.doi.org/10.2436/20.1501.01.192
Articles publicats (D-B)
Is part of: © International Microbiology, 2013, vol. 16, núm. 3, p. 177-189
Rights: Attribution-NonCommercial-ShareAlike 3.0 Spain
Rights URI: http://creativecommons.org/licenses/by-nc-sa/3.0/es/
Subject: Microbiologia d’aigua dolça
Freshwater microbiology
Microbiologia -- Cultius i medis de cultiu
Microbiology -- Cultures and culture media
Title: Phylogenetic characterization and quantification of ammonia-oxidizing archaea and bacteria from Lake Kivu in a long-term microcosm incubation
Type: info:eu-repo/semantics/article
Repository: DUGiDocs

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