[논문]아나목스 농후배양에서 암모니아 산화균의 자생 특성

배효관; 정진영

doi:10.15681/kswe.2020.36.3.220

아나목스 농후배양에서 암모니아 산화균의 자생 특성
Presence and Growth of Ammonia-oxidizing Bacteria in Anaerobic Ammonium Oxidation Enrichment 원문보기

한국물환경학회지 = Journal of Korean Society on Water Environment, v.36 no.3, 2020년, pp.220 - 228

배효관 (부산대학교 사회환경시스템공학과) , , 정진영 (영남대학교 환경공학과)

Abstract ▼ AI-Helper

Anaerobic ammonium oxidation (AMX) is a cost-efficient biological nitrogen removal process. The coexistence of ammonia-oxidizing bacteria (AOB) in an AMX reactor is an interesting research topic as a nitrogen-related bacterial consortium. In this study, a sequencing batch reactor for AMX (AMX-SBR) was operated with a conventional activated sludge. The AOB in an AMX bioreactor were identified and quantified using terminal restriction fragment length polymorphism (T-RFLP) and real-time qPCR. A T-RFLP assay based on the ammonia monooxygenase subunit A (amoA) gene sequences showed the presence of Nitrosomonas europaea-like AOB in the AMX-SBR. A phylogenetic tree based on the sequenced amoA gene showed that AOB were affiliated with the Nitrosomonas europaea/mobilis cluster. Throughout the enrichment period, the AOB population was stable with predominant Nitrosomonas europaea-like AOB. Two OTUs of amoA_SBR_JJY_20 (FJ577843) and amoA_SBR_JJY_9 (FJ577849) are similar to the clones from AMX-related environments. Real-time qPCR was used to quantify AOB populations over time. Interestingly, the exponential growth of AOB populations was observed during the substrate inhibition of the AMX bacteria. The specific growth rate of AOB under anaerobic conditions was only 0.111 d-1. The growth property of Nitrosomonas europaea-like AOB may provide fundamental information about the metabolic relationship between the AMX bacteria and AOB.

주제어

표/그림 (6)

그림 Fig. 1. The profile of nitrogen and TN removal efficiency of the anaerobic ammonium oxidation in the sequencing batch reactor.
그림 Fig. 2. Nitrogen loading and removal rates of the anaerobic ammonium oxidation in a sequencing batch reactor.
그림 Fig. 3. The T-RFLP pattern for AOB, digested by TaqI using amoA-1F and amoA-2R primer set.
그림 Fig. 4. Neighbor-joining phylogenetic tree based on amoA gene of β-proteobacterial AOB.
표 Table 1. Affiliation of the amoA gene clones analyzed in this study
그림 Fig. 5. The growth of AOB during the enrichment of anaerobic ammonium oxidation bacteria.

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문제 정의

However, the relationship between AMX activity and the quantity of AOB is not known. In this research, the objectives were to identify and quantify AOB in an AMX enrichment bioreactor during the start-up and substrate inhibition periods using molecular biological techniques. The qualitative and quantitative approaches revealed the close relationship between AOB and AMX bacteria in the dynamic conditions of the common substrate, i.

제안 방법

5 µL of template DNA. All manipulations were performed in laminar airflow and in low light to prevent light-activated degradation of the fluorescently labeled oligonucleotide probes. The PCR cycling condition was as follows: 1 cycle of 2 min at 50℃, 1 cycle of 10 min at 95℃, 30 cycles of 15 sec at 95℃ and 60 s at 60℃.
In this study, An AMX-SBR with a conventional activated sludge was operated to reveal the relationship of AOB growth and AMX activity during the AMX enrichment for 159 days. The AMX-SBR showed efficient TN removal efficiency of 99.
All PCR runs included control reactions without template DNA to test for possible non-specific amplification. The standard curves for AOB were constructed using a series of DNA concentrations prepared from the plasmid vector carrying the 16S rDNA gene of a Nitrosomonas europaea-like AOB related clone, which was obtained from a clone library constructed during this study.
Nine samples were collected for Days 28, 56, 77, 101, 108, 115, 124, 127 and 153. To investigate the microbial community structure of AOB in the AMX-SBR, real-time qPCR, T-RFLP, and sequencing were conducted. To isolate DNA, samples of homogenous activated-sludge were obtained from the SBR.

대상 데이터

The PCR cycles were as follows: 1 cycle of 2 min at 95℃, 30 cycles of 20 s at 95℃, 40 s at 57℃, 40 s at 72℃ then 1 cycle of 5 min at 72℃. Cycles were performed using a MyCycler^TMThermal Cycler (BIO-RAD, USA). The amplified amoA gene was cloned in a pGEM^®-T Easy Vector System (Promega, USA).

이론/모형

Clones with more than 97% sequence similarity were grouped into the same operational taxonomic unit (OTU). The phylogenetic tree was constructed using the neighbor-joining (NJ) method. The reliability of internal branches was assessed using 1,000 bootstrap replicates.

성능/효과

The predominant AOB were confirmed by cloning and sequencing based on the amoA gene. Four OTUs of amoA_ SBR_JJY_20 (FJ577843), amoA_SBR_JJY_9 (FJ577849), amoA_SBR_JJY_63 (FJ577881) and amoA_SBR_JJY_71 (FJ577885) were defined among 47 clones at the cutoff of 3%. OTUs were closely related to the Nitrosomonas europaea/mobilis cluster (Fig.
0 mg/L resulted in the serious inhibition of AMX activity. Overall the enrichment period, T-RFLP and sequencing based on the amoA gene revealed that the AOB population was stable with predominant Nitrosomonas europaea-like AOB. In particular, the OTUs of amoA_ SBR_JJY_20 (FJ577843) and amoA_SBR_JJY_9 (FJ577849) are similar to the clones from AMX-related environments.
In this research, the objectives were to identify and quantify AOB in an AMX enrichment bioreactor during the start-up and substrate inhibition periods using molecular biological techniques. The qualitative and quantitative approaches revealed the close relationship between AOB and AMX bacteria in the dynamic conditions of the common substrate, i.e., NH₄⁺, in an AMX reactor.

참고문헌 (31)

Abeliovich, A. and Vonshak, A. (1992). Anaerobic metabolism of nitrosomonas europaea, Archives of Microbiology, 158, 267-270.

상세보기
Abzazou, T., Salvado, H., Cardenas-Youngs, Y., Becerril-Rodriguez, A., Cebiran, E. M. C., Huguet, A., and Araujo, R. M. (2018). Characterization of nutrient-removing microbial communities in two full-scale WWTP systems using a new qPCR approach, Science of the Total Environment, 618, 858-865.

상세보기
Anjali, G. and Sabumon, P. C. (2017). Development of simultaneous partial nitrification, anammox and denitrification (SNAD) in a non-aerated SBR, International Biodeterioration & Biodegradation, 119, 43-55.

상세보기
Bae, H., Park, J. H., Jun, K. S., and Jung, J. Y. (2011). The community analysis of ammonia-oxidizing bacteria in wastewater treatment plants revealed by the combination of double labeled T-RFLP and sequencing, Journal of Environmental Science and Health Part A, 46(4), 345-354.
Bae, H., Park, K. S., Chung, Y. C., and Jung, J. Y. (2010). Distribution of anammox bacteria in domestic WWTPs and their enrichments evaluated by real-time quantitative PCR, Process Biochemistry, 45(3), 323-334.

상세보기
Dalsgaard, T., Thamdrup, B., and Canfield, D. E. (2005). Anaerobic ammonium oxidation (anammox) in the marine environment, Research in microbiology, 156, 457-464.

상세보기
Hagopian, D. S. and Riley, J. G. (1998). A closer look at the bacteriology of nitrification, Aquacultural Engineering, 18, 223-244.

상세보기
Hermansson, A. and Lindgren, P. E. (2001). Quantification of ammonia-oxidizing bacteria in arable soil by real-time PCR, Applied and Environmental Microbiology, 67, 972-976.

상세보기
Jetten, M. S. M., Strous, M., van de Pas-Schoonen, K. T., Schalk, J., van Dongen, L., van de Graaf, A. A., Logemann, S., Muyzer, G., van Loosdrecht, M. C. M., and Kuenen, J. G. (1998). The anaerobic oxidation of ammonium, FEMS Microbiology Review, 22, 421-437.
Jin, R. C., Hu, B. L., Zheng, P., Qaisar, M., Hu, A. H., and Islam, E. (2008). Quantitative comparison of stability of ANAMMOX process in different reactor configurations, Bioresource Technology, 99(6), 1603-1609.

상세보기
Kowalchuk, G. A., Stephen, J. R., De Boer, W., Prosser, J. I., Embley, T. M., and Woldendorp, J. W. (1997). Analysis of ammonia-oxidizing bacteria of the ${\beta}$ -subdivision of the class Proteobacteria in coastal sand dunes by denaturing gradient gel electrophoresis and sequencing of PCR-amplified 16S ribosomal DNA fragments, Applied and Environmental Microbiology, 63, 1489-1497.
Lackner, S., Gilbert, E. M., Vlaeminck, S. E., Joss, A., Horn, H., and van Loosdrecht, M. C. (2014). Full-scale partial nitritation/anammox experiences-an application survey, Water Research, 55, 292-303.

상세보기
Li, J., Huang, B., Wang, Q., Li, Y., Fang, W., Yan, D., Guo, M., and Cao, A. (2017). Effect of fumigation with chloropicrin on soil bacterial communities and genes encoding key enzymes involved in nitrogen cycling, Environmental Pollution, 227, 534-542.

상세보기
Lim, J., Lee S., and Hwang S. (2008). Use of quantitative real-time PCR to monitor population dynamics of ammonia-oxidizing bacteria in batch process, Journal of Industrial Microbiology and Biotechnology, 35, 1339-1344.

상세보기
Limpiyakorn, T., Shinohara, Y., Kurisu and F., and Yagi, O. (2005). Communities of ammonia-oxidizing bacteria in activated sludge of various sewage treatment plants in Tokyo, FEMS Microbiology Ecology, 54, 205-217.

상세보기
Lotti, T., Van der Star, W. R. L., Kleerebezem, R., Lubello, C., and Van Loosdrecht, M. C. M. (2012). The effect of nitrite inhibition on the anammox process, Water research, 46(8), 2559-2569.

상세보기
Mannucci, A., Caretti, C., Ducci, I., Lubello, C., Gori, R., Coppini, E., and Munz, G. (2020). Ammonia Oxidizing Bacteria (AOB) kinetic parameters estimated using the actual maximum ammonia oxidation rate, International Biodeterioration & Biodegradation, 147, 104876.
Nitahara, S., Kato, S., Usui, A., Urabe, T., Suzuki, K., and Yamagishi, A. (2017). Archaeal and bacterial communities in deep-sea hydrogenetic ferromanganese crusts on old seamounts of the northwestern Pacific, PloS one, 12(2), e0173071.

상세보기
Park, H. D. and Noguera, D. R. (2004). Evaluating the effect of dissolved oxygen on ammonia-oxidizing bacterial communities in activated sludge, Water Research, 38, 3275-3286.

상세보기
Purkhold, U., Pommerening-Roser, A., Juretschko, S., Schmid, M. C., Koops, H. P., and Wagner, M. (2000). Phylogeny of all recognized species of ammonia oxidisers based on comparative 16S rRNA and amoA sequence analysis: implications for molecular diversity surveys, Applied and Environmetal Microbiology, 66, 5368-5382.
Qiao, S., Kawakubo, Y., Cheng, Y., Nishiyama, T., Fujii, T., and Furukawa, K. (2009). Identification of bacteria coexisting with anammox bacteria in an upflow column type reactor, Biodegradation, 20, 117-124.

상세보기
Quan, Z. X., Rhee, S. K., Zuo, J. E., Yang, Y., Bae, J. W., Park, J. R., Lee, S. T., and Park, Y. H. (2008). Diversity of ammonium-oxidizing bacteria in a granular sludge anaerobic ammonium-oxidizing (anammox) reactor, Environmental Microbiology, 10, 3130-3139.

상세보기
Rowan, A. K., Snape, J. R., Fearnside, D., Barer, M. R., Curtis, T. P., and Head, I. M. (2003). Composition and diversity of ammonia-oxidising bacterial communities in wastewater treatment reactors of different design treating identical wastewater, FEMS Microbiology Ecology, 43, 195-206.

상세보기
Schmid, M., Twachtmann, U., Klein, M., Strous, M., Juretschko, S., Jetten, M., Metzger, J. W., Schleifer, K. H., and Wagner, M. (2000). Molecular evidence for genus level diversity of bacteria capable of catalyzing anaerobic ammonium oxidation, Systematic and Applied Microbiology, 23(1), 93-106.

상세보기
Schmidt, I., Sliekers, O., Schmid, M., Cirpus, I., Strous, M., Bock, E., Kuenen, J. G., and Jetten, M. S. M. (2002). Aerobic and anaerobic ammonium oxidizing bacteria-competitors or natural partners?, FEMS Microbiology Ecology, 39, 175-181.
Schramm, A., De Beer, D., Wagner, M., and Amann, R. (1998). Identification and activities in situ of Nitrosospira and Nitrospira spp. as dominant populations in a nitrifying fluidized bed reactor, Applied and Environmental Microbiology, 64, 3480-3485.

상세보기
Sedlacek, C. J., Giguere, A. T., Dobie, M. D., Mellbye, B. L., Ferrell, R. V., Woebken, D., Sayavedra-Soto, L. A., Bottomley, P. J., Daims, H., Wagner, M., and Pjevac, P. (2020). Transcriptomic response of Nitrosomonas europaea transitioned from ammonia-to oxygen-limited steady-state growth, mSystems, 5(1), e00562-19.
Sliekers, A. O., Derwort, N., Gomez, J. L. C., Strous, M., Kuenen, J. G., and Jetten, M. S. M. (2002). Completely autotrophic nitrogen removal over nitrite in one single reactor, Water Research, 36, 2475-2482.

상세보기
Strous, M., Heijnen, J. J., Kuenen, J. G., and Jetten, M. S. M. (1998). The sequencing batch reactor as a powerful tool for the study of slowly growing anaerobic ammonium-oxidizing microorganisms, Applied Microbiology and Biotechnology, 50, 589-596.

상세보기
Third, K. A., Sliekers, A. O., Kuenen, J. G., and Jetten, M. S. M. (2001). The CANON system (completely autotrophic nitrogen-removal over nitrite) under ammonium limitation: interaction and competition between three groups of bacteria, Systematic and Applied Microbiology, 24, 588-596.

상세보기
Wett, B., Hell, M., Nyhuis, G., Puempel, T., Takacs, I., and Murthy, S. (2010). Syntrophy of aerobic and anaerobic ammonia oxidisers, Water Science and Technology, 61(8), 1915-1922.

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표제어: PCR

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