Monitoring complete ammonia oxidizers bacteria: relevant players for nitrogen removal from wastewater

Abstract COMAMMOX (Complete Ammonia Oxidizers) bacteria seem to play an important role in nitrification, being able to directly transform ammonia into nitrates. Due to the relevance of this process in WWTPs, different approaches were used in this work to monitor COMAMMOX bacteria, as well as other m...

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Main Authors: Javier Duque, Leire Besga-Oyanarte, Miguel De Celis, Susana Serrano, José Luis Alonso, Antonio Santos, Lucía Arregui
Format: Article
Language:English
Published: SpringerOpen 2025-07-01
Series:AMB Express
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Online Access:https://doi.org/10.1186/s13568-025-01878-6
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author Javier Duque
Leire Besga-Oyanarte
Miguel De Celis
Susana Serrano
José Luis Alonso
Antonio Santos
Lucía Arregui
author_facet Javier Duque
Leire Besga-Oyanarte
Miguel De Celis
Susana Serrano
José Luis Alonso
Antonio Santos
Lucía Arregui
author_sort Javier Duque
collection DOAJ
description Abstract COMAMMOX (Complete Ammonia Oxidizers) bacteria seem to play an important role in nitrification, being able to directly transform ammonia into nitrates. Due to the relevance of this process in WWTPs, different approaches were used in this work to monitor COMAMMOX bacteria, as well as other microorganisms involved in the biological removal of nitrogen occurring in the bioreactor of a full-scale WWTP. This facility operates with low dissolved oxygen concentrations and prolonged sludge retention times. Fluorescence in situ hybridization confirmed the presence of COMAMMOX bacteria showing their distribution within the activated sludge flocs. Primers targeting the amoA gene, which encodes the A subunit of the ammonia monooxygenase enzyme, were employed as genetic markers in PCR techniques to simultaneously detect and amplify both COMAMMOX clades A and B, as well as specifically clade A, clade B, or various COMAMMOX Nitrospira species (Ca. N. nitrosa, Ca. N. inopinata, and Ca. N. nitrificans). Using digital PCR, a set of these primers enabled the estimation of the proportion of amoA COMAMMOX genes versus other nitrifiers (AOB, AOA and NOB). The data obtained not only confirmed the presence of COMAMMOX bacteria, but also their abundance, comparable to that of NOB and AOB. The 16S rRNA gene-based meta-taxonomy analysis gave complementary data about the microbial diversity and community composition within the samples. The results also highlight the biases of each technology used as monitoring tools, which must be accurate for the effective management of wastewater treatment plants (WWTPs) and for exploiting the potential of COMAMMOX bacteria.
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spelling doaj-art-ace767a263c44b45af3ec879a2c1b4742025-08-20T03:45:32ZengSpringerOpenAMB Express2191-08552025-07-0115111210.1186/s13568-025-01878-6Monitoring complete ammonia oxidizers bacteria: relevant players for nitrogen removal from wastewaterJavier Duque0Leire Besga-Oyanarte1Miguel De Celis2Susana Serrano3José Luis Alonso4Antonio Santos5Lucía Arregui6Unit of Microbiology, Department of Genetics, Physiology and Microbiology, Complutense University of MadridUnit of Microbiology, Department of Genetics, Physiology and Microbiology, Complutense University of MadridUnit of Microbiology, Department of Genetics, Physiology and Microbiology, Complutense University of MadridUnit of Microbiology, Department of Genetics, Physiology and Microbiology, Complutense University of MadridResearch Institute of Water and Environmental Engineering (IIAMA), Universitat Politècnica de ValènciaUnit of Microbiology, Department of Genetics, Physiology and Microbiology, Complutense University of MadridUnit of Microbiology, Department of Genetics, Physiology and Microbiology, Complutense University of MadridAbstract COMAMMOX (Complete Ammonia Oxidizers) bacteria seem to play an important role in nitrification, being able to directly transform ammonia into nitrates. Due to the relevance of this process in WWTPs, different approaches were used in this work to monitor COMAMMOX bacteria, as well as other microorganisms involved in the biological removal of nitrogen occurring in the bioreactor of a full-scale WWTP. This facility operates with low dissolved oxygen concentrations and prolonged sludge retention times. Fluorescence in situ hybridization confirmed the presence of COMAMMOX bacteria showing their distribution within the activated sludge flocs. Primers targeting the amoA gene, which encodes the A subunit of the ammonia monooxygenase enzyme, were employed as genetic markers in PCR techniques to simultaneously detect and amplify both COMAMMOX clades A and B, as well as specifically clade A, clade B, or various COMAMMOX Nitrospira species (Ca. N. nitrosa, Ca. N. inopinata, and Ca. N. nitrificans). Using digital PCR, a set of these primers enabled the estimation of the proportion of amoA COMAMMOX genes versus other nitrifiers (AOB, AOA and NOB). The data obtained not only confirmed the presence of COMAMMOX bacteria, but also their abundance, comparable to that of NOB and AOB. The 16S rRNA gene-based meta-taxonomy analysis gave complementary data about the microbial diversity and community composition within the samples. The results also highlight the biases of each technology used as monitoring tools, which must be accurate for the effective management of wastewater treatment plants (WWTPs) and for exploiting the potential of COMAMMOX bacteria.https://doi.org/10.1186/s13568-025-01878-6Wastewater treatment plantsNitrificationCOMAMMOX bacteriaFluorescent in situ hybridization (FISH)Digital PCR (dPCR)Next generation sequencing (NGS)
spellingShingle Javier Duque
Leire Besga-Oyanarte
Miguel De Celis
Susana Serrano
José Luis Alonso
Antonio Santos
Lucía Arregui
Monitoring complete ammonia oxidizers bacteria: relevant players for nitrogen removal from wastewater
AMB Express
Wastewater treatment plants
Nitrification
COMAMMOX bacteria
Fluorescent in situ hybridization (FISH)
Digital PCR (dPCR)
Next generation sequencing (NGS)
title Monitoring complete ammonia oxidizers bacteria: relevant players for nitrogen removal from wastewater
title_full Monitoring complete ammonia oxidizers bacteria: relevant players for nitrogen removal from wastewater
title_fullStr Monitoring complete ammonia oxidizers bacteria: relevant players for nitrogen removal from wastewater
title_full_unstemmed Monitoring complete ammonia oxidizers bacteria: relevant players for nitrogen removal from wastewater
title_short Monitoring complete ammonia oxidizers bacteria: relevant players for nitrogen removal from wastewater
title_sort monitoring complete ammonia oxidizers bacteria relevant players for nitrogen removal from wastewater
topic Wastewater treatment plants
Nitrification
COMAMMOX bacteria
Fluorescent in situ hybridization (FISH)
Digital PCR (dPCR)
Next generation sequencing (NGS)
url https://doi.org/10.1186/s13568-025-01878-6
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