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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SpringerOpen
2025-07-01
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| 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 |
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| 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. |
| format | Article |
| id | doaj-art-ace767a263c44b45af3ec879a2c1b474 |
| institution | Kabale University |
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| language | English |
| publishDate | 2025-07-01 |
| publisher | SpringerOpen |
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| series | AMB Express |
| 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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