DOCSLIB.ORG

Detection of Anammox Activity and Diversity of Anammox Bacteria- Related 16S Rrna Genes in Coastal Marine Sediment in Japan

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Detection of Anammox Activity and Diversity of Anammox Bacteria- Related 16S Rrna Genes in Coastal Marine Sediment in Japan Microbes Environ. Vol. 22, No. 3, 232–242, 2007 http://wwwsoc.nii.ac.jp/jsme2/ Detection of Anammox Activity and Diversity of Anammox Bacteria- Related 16S rRNA Genes in Coastal Marine Sediment in Japan TERUKI AMANO1, IKUO YOSHINAGA1*, KENTARO OKADA1, TAKAO YAMAGISHI2, SHINGO UEDA3, AKIRA OBUCHI2, YOSHIHIKO SAKO1, and YUICHI SUWA2 1 Laboratory of Marine Microbiology, Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Oiwake-cho, Kitashirakawa, Sakyo-ku, Kyoto 606–8502, Japan 2 Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology, 16–1, Onogawa, Tsukuba, Ibaraki 305–8569, Japan 3 College of Bioresource Sciences, Nihon University, 1866, Kameino, Fujisawa, Kanagawa 252–8510, Japan (Received April 16, 2007—Accepted May 10, 2007) A first assessment of anammox activity, which is a unique N2 emission process, was conducted in samples of coastal marine sediment from Japan with a 15N tracer. The occurrence and diversity of bacteria possibly responsi- ble for the anammox process were also evaluated by selective PCR-amplification of the 16S rRNA gene for known anammox bacteria. Anammox activity, detected by measuring 14N15N gas production, was only found in samples collected at the intertidal sand bank located at the Yodo River estuary. In the Yodo River samples, 16S rRNA gene fragments affiliated with the known anammox bacterial lineage were also recovered, and the two major phylotypes were both “Candidatus Scalindua wagneri” relatives with 95% and 98% sequence similarity. Even from the other samples in which no recognizable anammox activity was detected, 16S rRNA gene frag- ments related to known anammox bacteria, but not to “Ca. S. wagneri”, were detected. This is the first report of anammox-mediated N2 emission in coastal marine environments in Japan. Notably, the PCR-based analysis allowed us to discover unexpected phylogenetic diversity of anammox bacteria-related 16S rRNA gene sequences. The selective PCR primer set developed in this study could be a powerful tool to unveil the ecology of anammox bacteria in natural environments. Key words: anammox, 16S rRNA gene, nitrogen cycle, marine sediment, 15N tracer Coastal and enclosed sea areas, into which excess fication. A specific group of bacteria belonging to the order amounts of terrestrial nitrogen flow, can easily become Planctomycetales in the phylum Planctomycetes is consid- eutrophic and suffer from harmful algal blooms. For the ered to mediate the anammox reaction under anoxic sustainable management of such marine environments, a conditions28). Since anammox has a different mechanism better understanding of the nitrogen cycle, especially the from denitrification in N2 formation, the presence of anam- emission of N2 gas as a mechanism of removing nitrogen mox will cause one to overestimate true N2 production when 15 from the fixed nitrogen pool, is essential. N2 gas emission in the nitrogen isotope ( N) pairing technique (IPT), which marine environments may also be a key regulator of ecosys- has been widely used for measuring N2 production by deni- tem function and global biogeochemistry2). trification in natural samples, is employed19). Therefore, Recently, it was demonstrated that anaerobic ammonium investigating the distribution of anammox activity and oxidation (anammox) was also involved in the emission of microorganisms responsible in marine environments is 12,30) N2 gas from marine environments in addition to denitri- required for a precise estimation of N2 gas emission. Most of the information on anammox has been obtained * Corresponding author. E-mail address: [email protected]. from enriched cultures with high concentrations of inor- jp; Tel.: +81–75–753–6219; Fax: +81–75–753–6226. ganic nitrogen substrates. In the anammox reaction, the two Anammox in Coastal Marine Sediment 233 + − nitrogen atoms in N2 are derived from NH4 and NO2 Sea, Osaka Bay, Ise Bay and the intertidal sand bank at the 36) respectively , while denitrification forms N2 gas from two Yodo River estuary in 2005 and 2006, using a grab sampler − − 37) molecules of NO2 (NO3 ) through a stepwise reduction. (Rigo Co. Ltd., Tokyo, Japan) (for samples S1, S2, S3, IB1, This biochemical difference allows us to distinguish and IB2), a KK core sampler (Hashimoto Kagaku Co. Ltd., 15 between N2 production by the two processes by using N as Kyoto, Japan) (for samples OB1 and OB2), or a plastic core a tracer. Regarding microorganisms responsible for the sampler (for samples YR-M and YR-J) (Fig. 1 and Table 1). anammox reaction, four genera of Planctomycetales (“Bro- The surface layer of each sediment sample was collected cadia”, “Kuenenia”, “Scalindua” and “Anammoxoglobus”) into plastic containers, tightly sealed, and stored at 4°C in are proposed as anammox bacteria10,24,25,28), although no iso- the dark until experiments. The measurement of anammox late has been established so far. activity was performed within a week at longest after sam- However, there has been limited information available pling except for the samples S1, S2, and S3 which were about the contribution of anammox to the nitrogen cycling stored for one and a half months at 4°C in the dark. and diversity of anammox-related bacteria in natural envi- ronments. At present, the activity and related bacteria at Measurement of anammox and denitrification activity several sites has been investigated with a 15N tracer and The occurrence of the anammox process was examined 12,13,20,26,27,29) 14 15 29 molecular techniques . A PCR amplification of by quantifying N N ( N2) emissions using gas chroma- the 16S rRNA gene related to known anammox bacteria tography-quadrupole mass spectrometry (GC-MS) in a was carried out in these investigations, however, sequences strictly anaerobic incubation of sediment slurries amended 15 affiliated with the anammox bacterial lineage were recov- with N-labeled or non-labeled NH4Cl and NaNO2 (Suwa et ered at a relatively low efficiency. For instance, when a al. in preparation). Each sediment sample was suspended at Planctomycetales-specific primer was used for PCR ampli- a concentration of 8–25% (wt/vol) in seawater that was pre- fication, only 4 out of 50 clones were recovered as anam- liminarily filtrated through a 0.22 µm cellulose acetate filter mox bacteria-related sequences in a library constructed system (Corning Inc., NY, USA), and the suspension was from the sediment sample showing anammox activity29). thoroughly purged with Ar to remove oxygen. The salinity This could be explained by the inadequacy of the primer in the slurry was adjusted to in situ levels with an oxygen- sets previously available, including nucleotide sequences free inorganic medium developed previously35) except that targeting a broader range of phyla as bacterial universal the medium used in this study contained 7.15 mM NaHCO3 primers or a primer for all Planctomycetales. The low instead of 5 mM KHCO3. Portions (40 ml) of the slurry recovery of anammox bacteria-related 16S rRNA gene frag- were transferred to 67-ml glass vials, and capped tightly ments limits information on the diversity and community with a butyl rubber stopper and an aluminum seal. These structure of anammox bacteria in natural environments, and procedures were carried out inside a glove box under an prevents us from understanding the ecology of the anam- Ar+H2 (anoxic) atmosphere. Subsequently, the headspace mox bacterial community. gas of a vial was replaced with He by repeatedly vacuuming The ultimate goal of our project is to elucidate the eco- and purging with high-purity He gas. The final pressure of logical significance of the anammox process in marine envi- ronments. As a first step to achieving this, here we demon- strate the occurrence of anammox activity and anammox bacteria-related 16S rRNA gene fragments in coastal marine sediment in Japan. Our research areas, the Seto Inland Sea, Osaka Bay and Ise Bay are representative enclosed sea areas in Japan; however, no report has been available on anammox activity there up to now. We also propose effec- tive PCR primer sets to detect anammox bacteria-related 16S rRNA genes with a relatively high recovery efficiency. Materials and Methods Sampling Sediment samples were collected from the Seto Inland Fig. 1. Sampling sites in western Japan. 234 AMANO et al. Table 1. Sample characteristics Sample Location Water depth (m) Date Sediment layer (cm) Seto Inland Sea S1 34°04'N, 133°17'E 28 10 Aug. 05 0–5 S2 34°25'N, 134°30'E 40 9 Aug. 05 0–5 S3 34°30'N, 134°31'E 42 8 Aug. 05 0–5 Osaka Bay OB1 34°39'N, 135°22'E 10.5 18 Apr. 06 0–5 OB2 34°38'N, 135°23'E 11 18 Apr. 06 0–5 Ise Bay IB1 34°43'N, 135°44'E 36 20 Jul. 06 0–2 IB2 34°39'N, 135°55'E 21 20 Jul. 06 0–2 Yodo River estuary YR-M 34°41'N, 135°27'E intertidal 12 May. 06 0–1 YR-J 13 Jun. 06 0–2 29 15 + − the headspace gas with He was 0.15 MPa. The vials con- of N2 gas production in vials amended with NH4 +NO2 . 30 + 15 − taining anoxic sediment slurries were pre-incubated for 1–4 Increases in N2 accumulated in vials with NH4 + NO2 hours in the dark at 20°C before substrates were added, in were expressed as potential denitrification activity. A time order to eliminate the remaining nitrite and nitrate as well as course was used for determining rate constants of both reac- 29 30 dissolved oxygen. tions. Abundances of N2 and N2 gas in vials for a nega- 15 15 Oxygen-free stock solutions of N-labeled NH4Cl tive control experiment with NH4Cl but without nitrite (Shoko-Co., Ltd., Tokyo, Japan, 15N atom%: >99) and were monitored to see if nitrification and denitrification 15 29 NaNO2 (ISOTEC Inc, OH, USA, N atom%: >99) or non- coupled to produce N2.
Load more

Recommended publications
  • Deammonification
    Deammonification MARCH 2019 The Deammonification Compendium was prepared in 2012 and later updated in 2014 as part of the Nutrient Removal Challenge. The Deammonification Compendium serves as a succinct primer on the deammonification process and applications in water resource recovery facilities. This 2019 compendium updates prior versions based on reports and documents generated by the Nutrient Removal Challenge. As such, updates are limited in scope and not intended to capture all recent industry and research activity related to deammonification. ACKNOWLEDGMENTS We acknowledge the following contributors and reviewers (alphabetically). The findings from the Nutrient Removal Challenge were incorporated into this 2019 edition by: Bryce Figdore JB Neethling David Stensel HDR Inc. HDR Inc. University of Washington The following individuals contributed to earlier versions of this compendium: Charles Bott Sudhir Murthy Julian Sandino Hampton Roads Sanitation DC Water CH2M Hill District JB Neethling David Stensel Gregory Bowden HDR Inc. University of Washington AECOM Amit Pramanik Ryujiro Tsuchihashi Ramesh Goel The Water Research AECOM University of Utah Foundation Please use the following citation for this document: WRF (The Water Research Foundation). 2019. “Deammonification” from the Nutrient Removal Challenge. 1199 North Fairfax Street, Suite 900 6666 West Quincy Avenue Alexandria, VA 22314-1445 Denver, CO 80235-3098 [email protected] www.waterrf.org TABLE OF CONTENTS DEAMMONIFICATION | 1 1 | What is deammonification? DEAMMONIFICATION PROCESS
    [Show full text]
  • Spatiotemporal Distributions and Environmental Drivers Of
    Journal of Oceanology and Limnology Vol. 37 No. 4, P. 1211-1228, 2019 https://doi.org/10.1007/s00343-019-8200-3 Spatiotemporal distributions and environmental drivers of diversity and community structure of nosZ -type denitrifi ers and anammox bacteria in sediments of the Bohai Sea and North Yellow Sea, China* CAI Youjun 1 , ZHANG Xiaoli 2 , LI Guihao 2, 3 , DONG Jun 3, 4 , YANG Anjing 2, 3 , WANG Guangyu 5 , ** , ZHOU Xiaojian 1 , ** 1 College of Environmental Sciences and Engineering, Yangzhou University, Yangzhou 225000, China 2 Key Laboratory of Coastal Environmental Processes and Ecological Remendation, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China 3 University of Chinese Academy of Sciences, Beijing 100049, China 4 Shenzhen Lightsun Technology Co. Ltd., Shenzhen 518000, China 5 Department of Bioengineering, School of Marine Science and Technology, Harbin Institute of Technology, Weihai 264209, China Received Jul. 26, 2018; accepted in principle Sep. 5, 2018; accepted for publication Oct. 23, 2018 © Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2019 Abstract Denitrifi cation and anammox processes are major nitrogen removal processes in coastal ecosystems. However, the spatiotemporal dynamics and driving factors of the diversity and community structure of involved functional bacteria have not been well illustrated in coastal environments, especially in human-dominated ecosystems. In this study, we investigated the distributions of denitrifi ers and anammox bacteria in the eutrophic Bohai Sea and the northern Yellow Sea of China in May and November of 2012 by constructing clone libraries employing nosZ and 16S rRNA gene biomarkers.
    [Show full text]
  • 16S Rrna and Hydrazine Gene-Based Profiling of the Candidatus Scalindua Community from the Arabian Sea Hypoxic Sediments
    RESEARCH ARTICLES 16S rRNA and hydrazine gene-based profiling of the Candidatus Scalindua community from the Arabian Sea hypoxic sediments Jovitha Lincy* and Cathrine Sumathi Manohar Biological Oceanography Division, CSIR-National Institute of Oceanography, Goa 403 004, India and Academy of Scientific and Innovative Research, Ghaziabad 201 002 India The nitrogen loss contribution of anammox to the total Anammox bacterial diversity and abundance were denitrification in marine sediments can range from near studied from the organic-rich hypoxic sediments of 7 the Arabian Sea utilizing the partial 16S rRNA, and 0% to 80%, especially in deposits underlying ODZ . In hydrazine synthase, hzsA and hydrazine oxidoreduc- the southeastern Arabian Sea, benthic nitrogen metabo- 8 tase, hzo genes. Among all the clades obtained, phylo- lism is driven by sinking organic matter which is excep- typic diversity was high within the Candidatus genus tionally high during the southwest monsoon period9. The Scalindua with an abundance of ≤7 × 104 copies/g dry prevailing conditions are assumed to favour heterotrophic wt. As such, Scalindua is known to play a significant denitrifying communities that rely on organic sources10. role in fixed nitrogen removal through anaerobic Hence the possible occurrence of a chemolithoautotroph ammonium oxidation (anammox) pathway. From these like anammox microbes is not studied thoroughly. Be- analyses, it is inferred that searching for hzo gene sides ammonium ion available in situ11, dissimilatory yields robust evidence for detecting anammox com- nitrate reduction to ammonia (DNRA) is reported1. In the munity than the widely used 16S rRNA gene marker. Oman coast of the Arabian Sea, a coupling of DNRA to anammox resulted in intense nitrogen loss12, suggesting Keywords: Anammox, community gene-based profil- that anammox occurrence is controlled by the availability ing, hydrazine, hypoxic sediments, Scalindua.
    [Show full text]
  • 1 Single Cell Genomic and Transcriptomic Evidence for the Use of Alternative Nitrogen
    1 Single cell genomic and transcriptomic evidence for the use of alternative nitrogen 2 substrates by anammox bacteria 3 4 Contributors: Sangita Ganesh1,2*, Anthony D. Bertagnolli1*, Laura A. Bristow3, Cory C. 5 Padilla1, Nigel Blackwood4, Montserrat Aldunate5,6, Annie Bourbonnais7,8, Mark A. Altabet8, 6 Rex R. Malmstrom9, Tanja Woyke9, Osvaldo Ulloa6, Konstantinos T. Konstantinidis10, Bo 7 Thamdrup11, Frank J. Stewart1# 8 9 Affiliations and footnotes: 10 11 * These authors contributed equally to this work. 12 # Corresponding author. Mailing address: School of Biological Sciences, Georgia Institute of 13 Technology, Ford ES&T Building, Rm 1242, 311 Ferst Drive, Atlanta GA 30332 14 Email: [email protected] 15 16 1 School of Biological Sciences, Georgia Institute of Technology, Atlanta GA, USA, 30332 17 2 Radiant Genomics, Emeryville CA, USA, 94608 18 3 Biogeochemistry Group, Max Planck Institute for Marine Microbiology, Bremen, Germany 19 4 Department of Biology, University of Pennsylvania, Philadelphia PA, USA, 19104 20 5 Graduate Program in Oceanography, Department of Oceanography, Faculty of Natural Sciences 21 and Oceanography, University of Concepción, Casilla 160-C, Concepción, Chile 22 6 Departamento de Oceanografía, Universidad de Concepción, Casilla 160-C, 4070386, 23 Concepción, Chile 24 7 Marine Chemistry & Geochemistry, Woods Hole Oceanographic Institution, 266 Woods Hole 25 Road, Woods Hole MA, USA, 02543 26 8 School for Marine Science and Technology, University of Massachusetts Dartmouth, 706 27 Rodney French
    [Show full text]
  • Effects of Recirculating Aquaculture System Wastewater on Anammox Performance and Community Structure
    processes Article Effects of Recirculating Aquaculture System Wastewater on Anammox Performance and Community Structure Jonathan A. C. Roques 1,2,† , Federico Micolucci 1,2,† , Suguru Hosokawa 3, Kristina Sundell 1,2 and Tomonori Kindaichi 3,* 1 Department of Biological and Environmental Sciences, University of Gothenburg, P.O. Box 463, SE-405 30 Gothenburg, Sweden; [email protected] (J.A.C.R.); [email protected] (F.M.); [email protected] (K.S.) 2 Swedish Mariculture Research Center (SWEMARC), University of Gothenburg, P.O. Box 463, SE-405 30 Gothenburg, Sweden 3 Department of Civil and Environmental Engineering, Graduate School of Advanced Science and Engineering, Hiroshima University, 1-4-1 Kagamiyama, Higashihiroshima 739-8527, Japan; [email protected] * Correspondence: [email protected]; Tel.: +81-82-424-5718 † Contributed equally to the study. Abstract: Recirculating aquaculture systems (RAS) are good candidates for the sustainable devel- opment of the aquaculture sector. A current limitation of RAS is the production and accumulation of nitrogenous waste, which could affect fish health. We investigated the potential of the anaerobic ammonia oxidation (anammox) process to treat marine wastewater from a cold-water RAS. We show that the marine anammox bacteria Candidatus Scalindua is a promising candidate. However, its activity was affected by unknown compounds in the RAS wastewater and/or the sub-optimum + Citation: Roques, J.A.C.; Micolucci, content of essential trace elements (TEs). Anammox activity dropped to 2% and 13% in NH4 and F.; Hosokawa, S.; Sundell, K.; − NO2 removal, respectively, when NO3-rich RAS wastewater was used as a medium in the absence Kindaichi, T.
    [Show full text]
  • Changes in Benthic Denitrification, Nitrate Ammonification, And
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Digital.CSIC APPLIED AND ENVIRONMENTAL MICROBIOLOGY, May 2009, p. 3171–3179 Vol. 75, No. 10 0099-2240/09/$08.00ϩ0 doi:10.1128/AEM.02511-08 Copyright © 2009, American Society for Microbiology. All Rights Reserved. Changes in Benthic Denitrification, Nitrate Ammonification, and Anammox Process Rates and Nitrate and Nitrite Reductase Gene Abundances along an Estuarine Nutrient Gradient (the Colne Estuary, United Kingdom)ᰔ† Liang F. Dong,1‡ Cindy J. Smith,1,2‡ Sokratis Papaspyrou,1§ Andrew Stott,3 A. Mark Osborn,1,2 and David B. Nedwell1* Department of Biological Sciences, University of Essex, Colchester CO4 3SQ, United Kingdom1; Department of Animal and Plant Sciences, University of Sheffield, Western Bank, Sheffield S10 2TN, United Kingdom2; and NERC Life Sciences Mass Spectrometer Facility, Centre for Ecology & Hydrology, Lancaster Environment Centre, Lancaster LA1 4AP, United Kingdom3 Received 3 November 2008/Accepted 11 March 2009 Estuarine sediments are the location for significant bacterial removal of anthropogenically derived inorganic nitrogen, in particular nitrate, from the aquatic environment. In this study, rates of benthic denitrification (DN), dissimilatory nitrate reduction to ammonium (DNRA), and anammox (AN) at three sites along a nitrate concentration gradient in the Colne estuary, United Kingdom, were determined, and the numbers of functional genes (narG, napA, nirS, and nrfA) and corresponding transcripts encoding enzymes mediating nitrate reduc- tion were determined by reverse transcription-quantitative PCR. In situ rates of DN and DNRA decreased toward the estuary mouth, with the findings from slurry experiments suggesting that the potential for DNRA increased while the DN potential decreased as nitrate concentrations declined.
    [Show full text]
  • Phylogenetic and Metabolic Diversity of Planctomycetes from Anaerobic, Sulfide- and Sulfur-Rich Zodletone Spring, Oklahomaᰔ Mostafa S
    APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Aug. 2007, p. 4707–4716 Vol. 73, No. 15 0099-2240/07/$08.00ϩ0 doi:10.1128/AEM.00591-07 Copyright © 2007, American Society for Microbiology. All Rights Reserved. Phylogenetic and Metabolic Diversity of Planctomycetes from Anaerobic, Sulfide- and Sulfur-Rich Zodletone Spring, Oklahomaᰔ Mostafa S. Elshahed,1* Noha H. Youssef,1 Qingwei Luo,2 Fares Z. Najar,3 Bruce A. Roe,3 Tracy M. Sisk,2 Solveig I. Bu¨hring,4 Kai-Uwe Hinrichs,4 and Lee R. Krumholz2 Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma 740781; Department of Botany and Microbiology and Institute for Energy and the Environment2 and Department of Chemistry and Biochemistry and the Advanced Center for Genome Technology,3 University of Oklahoma, Norman, Oklahoma; and Deutsche Forschungsgemeinschaft-Research Center Ocean Margins and Department of Geosciences, University of Bremen, Bremen, Germany4 Received 14 March 2007/Accepted 27 May 2007 We investigated the phylogenetic diversity and metabolic capabilities of members of the phylum Planctomy- cetes in the anaerobic, sulfide-saturated sediments of a mesophilic spring (Zodletone Spring) in southwestern Oklahoma. Culture-independent analyses of 16S rRNA gene sequences generated using Planctomycetes-biased primer pairs suggested that an extremely diverse community of Planctomycetes is present at the spring. Although sequences that are phylogenetically affiliated with cultured heterotrophic Planctomycetes were iden- tified, the majority of the sequences belonged to several globally distributed, as-yet-uncultured Planctomycetes lineages. Using complex organic media (aqueous extracts of the spring sediments and rumen fluid), we isolated two novel strains that belonged to the Pirellula-Rhodopirellula-Blastopirellula clade within the Planctomycetes.
    [Show full text]
  • Enrichment Using an Up-Flow Column Reactor and Community Structure of Marine Anammox Bacteria from Coastal Sediment
    Microbes Environ. Vol. 26, No. 1, 67–73, 2011 http://wwwsoc.nii.ac.jp/jsme2/ doi:10.1264/jsme2.ME10158 Enrichment Using an Up-flow Column Reactor and Community Structure of Marine Anammox Bacteria from Coastal Sediment TOMONORI KINDAICHI1*, TAKANORI AWATA1, YUJI SUZUKI2, KATSUICHIRO TANABE3, MASASHI HATAMOTO4, NORIATSU OZAKI1, and AKIYOSHI OHASHI1 1Department of Civil and Environmental Engineering, Graduate School of Engineering, Hiroshima University, 1–4–1 Kagamiyama, Higashihiroshima 739–8527, Japan; 2Wakayama Prefectural Bureau, 1–1 Komatsubaradori, Wakayama 640–8585, Japan; 3Matsuo Corporation, 1–10 Hachimankoji, Saga 840–8666, Japan; and 4Department of Environmental Systems Engineering, Nagaoka University of Technology, 1603–1 Kamitomioka, Nagaoka 940–2188, Japan (Received August 18, 2010—Accepted November 8, 2010—Published online December 11, 2010) We established an enrichment culture of marine anaerobic ammonium oxidation (anammox) bacteria using an up- flow column reactor fed with artificial sea water supplemented with nitrogen and minerals and inoculated with coastal + surface sediment collected from Hiroshima Bay. After 2 months of reactor operation, simultaneous removal of NH4 − and NO2 was observed, suggesting that an anammox reaction was proceeding. A total nitrogen removal rate of 2.17 g-N L−1 day−1 was attained on day 594 while the nitrogen loading rate was 3.33 g-N L−1 day−1. Phylogenetic analysis revealed that at least two dominant “Candidatus Scalindua” species were present in this reactor. Moreover, many uncultured bacteria and archaea, including candidate division or ammonia-oxidizing archaea, were present. Fluores- cence in situ hybridization (FISH) revealed that anammox bacteria accounted for 85.5 ± 4.5% of the total bacteria at day 393.
    [Show full text]
  • Anaerobic Ammonium Oxidation in an Estuarine Sediment
    AQUATIC MICROBIAL ECOLOGY Vol. 36: 293–304, 2004 Published September 14 Aquat Microb Ecol Anaerobic ammonium oxidation in an estuarine sediment Nils Risgaard-Petersen1,*, Rikke Louise Meyer2, Markus Schmid3, Mike S. M. Jetten4, Alex Enrich-Prast5, Søren Rysgaard1, Niels Peter Revsbech2 1National Environmental Research Institute, Department of Marine Ecology, Vejlsøvej 25, 8600 Silkeborg, Denmark 2University of Aarhus, Institute of Biological Sciences, Department of Microbial Ecology, Ny Munkegade Building 540, 8000 Århus C, Denmark 3Delft University of Technology, Department of Biotechnology, Delft, The Netherlands 4KU Nijmegen, Department of Microbiology, Nijmegen, The Netherlands 5Depto. Ecologia, CCS, Ilha do Fundao, Cidade Universitaria, Universidade Federal do Rio de Janeiro, Rio de Janeiro, 21951-590, Brasil ABSTRACT: The occurrence and significance of the anammox (anaerobic ammonium oxidation) process relative to denitrification was studied in photosynthetically active sediment from 2 shallow-water estuaries: Randers Fjord and Norsminde Fjord, Denmark. Anammox accounted for 5 to 24% of N2 pro- duction in Randers Fjord sediment, whereas no indication was seen of the process in sediment from Nors- minde Fjord. It is suggested that the presence of anammox in Randers Fjord and its absence from Nors- – – – minde Fjord is associated with differences in the availability of NO3 + NO2 (NOx ) in the suboxic zone – – of the sediment. In Randers Fjord, NOx is present in the water column throughout the year and NOx – porewater profiles showed that NOx penetrates into the suboxic zone of the sediment. In Norsminde – Fjord, NOx is absent from the water column during the summer months and, via assimilation, benthic mi- – croalgae may prevent penetration of NOx into the suboxic zone of the sediment.
    [Show full text]
  • Anaerobic Ammonium-Oxidizing Bacteria Rapid and Simple
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Ghent University Academic Bibliography Rapid and Simple Cryopreservation of Anaerobic Ammonium-Oxidizing Bacteria Kim Heylen, Katharina Ettwig, Ziye Hu, Mike Jetten and Boran Kartal Appl. Environ. Microbiol. 2012, 78(8):3010. DOI: 10.1128/AEM.07501-11. Downloaded from Published Ahead of Print 3 February 2012. Updated information and services can be found at: http://aem.asm.org/content/78/8/3010 http://aem.asm.org/ These include: REFERENCES This article cites 25 articles, 5 of which can be accessed free at: http://aem.asm.org/content/78/8/3010#ref-list-1 CONTENT ALERTS Receive: RSS Feeds, eTOCs, free email alerts (when new articles cite this article), more» on March 25, 2012 by UNIVERSITEIT GENT/UZGENT Information about commercial reprint orders: http://aem.asm.org/site/misc/reprints.xhtml To subscribe to to another ASM Journal go to: http://journals.asm.org/site/subscriptions/ Rapid and Simple Cryopreservation of Anaerobic Ammonium-Oxidizing Bacteria Kim Heylen,a,b Katharina Ettwig,b Ziye Hu,b Mike Jetten,b,c and Boran Kartalb Ghent University, Department of Biochemistry and Microbiology, Laboratory of Microbiology, Ghent, Belgiuma; Radboud University Nijmegen, IWWR, Department of b c Microbiology, Nijmegen, The Netherlands ; and Delft University of Technology, Department of Biotechnology, Delft, The Netherlands Downloaded from A quick and simple protocol for long-term cryopreservation of anaerobic ammonium-oxidizing bacteria (anammox bacteria) was developed. After 29 weeks of preservation at ؊80°C, activity recovery for all tested cultures under at least one of the applied sets of preservation conditions was observed.
    [Show full text]
  • Investigation of Aerobic and Anaerobic Ammonium-Oxidizing Bacteria
    Investigation of Aerobic and Anaerobic Ammonium-oxidizing bacteria presence in a small full-scale wastewater treatment system comprised by UASB reactor and three polishing ponds J. C. Araujo*, M. M. S. Correa**, E. C. Silva**, V. M. Godinho**, M. Von Sperling** and C.A.L. Chernicharo** *Centro Federal de Educação Tecnológica de Minas Gerais (CEFET/MG), Belo Horizonte, Brazil – (E-mail: [email protected] ) ** Universidade Federal de Minas Gerais, De partment of Sanitary and Environmental Engineering. Belo Horizonte, Brazil Abstract This work applied PCR amplification method and Fluorescence in situ hybridization (FISH) with primers and probes specific for the aerobic ammonia-oxidizing β-Proteobacteria and anammox organisms in order to detect these groups in different samples from a wastewater treatment system comprised by UASB reactor and three polishing ponds. Seven primer pairs were used in order to detect Anammox bacteria, such as: the forward primer Pla46rc (specific for the Planctomycetales ) together with either the reverse primer 1392r (E ubacteria) , Amx368 (specific for “ Brocadia ”, “ Kuenenia ” and “ Scalindua ”); or Amx820 (specific for “ Brocadia ” and “Kuenenia ”), or Amx1240 (specific for “ Brocadia anammoxidans ”), or Amx667R. Primer pairs Brod541f-Brod1260r (specific for Scalindua brodae and Scalindua wagneri ) and An7f-An1388r (specific for all anammox organisms Genera “ Ca. Brocadia”, “Ca. Kuenenia ”, and “Ca. Scalindua” ) were also tested. Results with primers Pla46rc/Amx368r and Pla46rc/Amx667r suggest the presence of Anammox bacteria in the UASB reactor and also in the three polishing ponds. However, FISH results indicated the absence of Anammox and aerobic ammonia-oxidizing bacteria in these samples, suggesting that nitrification and anammox were not the major reactions responsible for total nitrogen removal in this wastewater treatment system.
    [Show full text]
  • JMB024-07-01 FDOC 1.Pdf
    J. Microbiol. Biotechnol. (2014), 24(7), 879–887 http://dx.doi.org/10.4014/jmb.1401.01016 Research Article Review jmb Successful Enrichment of Rarely Found Candidatus Anammoxoglobus propionicus from Leachate Sludge Shu-Chuan Hsu, Yen-Chun Lai, Ping-Heng Hsieh, Pun-Jen Cheng, Suen–Shin Wong, and Chun-Hsiung Hung* Department of Environmental Engineering, National Chung Hsing University, Taichung 402, Taiwan Received: January 9, 2014 Revised: March 4, 2014 Bacteria that mediate the anaerobic oxidation of ammonium (anammox) have been detected in Accepted: April 3, 2014 natural ecosystems, as well as various wastewater treatment systems. In this study, sludge from a particular landfill leachate anaerobic treatment system was selected as the incubation seed for anammox microorganism enrichment owing to its possible anammox activity. First published online Transmission electron microscopy observation, denaturing gradient gel electrophoresis April 7, 2014 analysis, and cloning/sequencing techniques were applied to identify the diversity of *Corresponding author anammox microorganisms throughout the incubation. During the early stage of operation, the Phone: +886-4-22840441; diversity of anammox microorganisms was similar to the original complex microbes in the Fax: +886-4-22862589; seed sludge. However, as incubation time increased, the anammox microorganism diversity E-mail: [email protected] within the system that was originally dominated by Candidatus (Ca.) Brocadia sp. was replaced by Ca. Anammoxoglobus propionicus. The domination of Ca. Anammoxoglobus propionicus produced a stable removal of ammonia (70 mg-N/l) and nitrite (90 mg-N/l), and the total nitrogen removal efficiency was maintained at nearly 95%. The fluorescence in situ hybridization results showed that Ca.
    [Show full text]