scholarly journals Distribution and Rate of Microbial Processes in an Ammonia-Loaded Air Filter Biofilm

2009 ◽  
Vol 75 (11) ◽  
pp. 3705-3713 ◽  
Author(s):  
Susanne Juhler ◽  
Niels Peter Revsbech ◽  
Andreas Schramm ◽  
Martina Herrmann ◽  
Lars D. M. Ottosen ◽  
...  
Keyword(s):  
Heterotrophic Bacteria ◽  
Experimental Period ◽  
Protonated Form ◽  
Product Inhibition ◽  
Nitrifying Bacteria ◽  
Outlet Section ◽  
Ammonia Oxidizing Bacteria ◽  
Biological Degradation ◽  
Activity Measurements

ABSTRACT The in situ activity and distribution of heterotrophic and nitrifying bacteria and their potential interactions were investigated in a full-scale, two-section, trickling filter designed for biological degradation of volatile organics and NH3 in ventilation air from pig farms. The filter biofilm was investigated by microsensor analysis, fluorescence in situ hybridization, quantitative PCR, and batch incubation activity measurements. In situ aerobic activity showed a significant decrease through the filter, while the distribution of ammonia-oxidizing bacteria (AOB) was highly skewed toward the filter outlet. Nitrite oxidation was not detected during most of the experimental period, and the AOB activity therefore resulted in NO2 −, accumulation, with concentrations often exceeding 100 mM at the filter inlet. The restriction of AOB to the outlet section of the filter was explained by both competition with heterotrophic bacteria for O2 and inhibition by the protonated form of NO2 −, HNO2. Product inhibition of AOB growth could explain why this type of filter tends to emit air with a rather constant NH3 concentration irrespective of variations in inlet concentration and airflow.

Significance of substrate C/N ratio on structure and activity of nitrifying biofilms determined by in situ hybridization and the use of microelectrodes

2000 ◽  
Vol 41 (4-5) ◽  
pp. 317-321 ◽  
Author(s):  
H. Satoh ◽  
S. Okabe ◽  
N. Norimatsu ◽  
Y. Watanabe
Keyword(s):  
In Situ Hybridization ◽  
Heterotrophic Bacteria ◽  
Outer Part ◽  
Experimental Result ◽  
Nitrifying Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Spatial Distributions ◽  
Interspecies Competition ◽  
Oxidation Rates

The effect of substrate C/N ratio on the spatial distributions of ammonia-oxidizing bacteria and their activity was investigated by using microelectrodes with high spatial resolution and fluorescent in situ hybridization (FISH) technique. In this study, an interspecies competition for O2 between ammonia-oxidizing bacteria and heterotrophic bacteria was experimentally evaluated. An autotrophic nitrifying biofilm originally cultured at C/N=0 was used as a model biofilm to study changes in specific NH4+ oxidation rate profiles in the biofilm when the substrate C/N ratio was varied. As C/N ratio increased, specific NH4+ oxidation rates decreased in the outer part of the biofilm due to interspecies competition, while they were unchanged in the inner part. The increase in substrate C/N ratio (i.e., addition of acetate) immediately induced the interspecies competition for O2 between ammonia-oxidizing bacteria and heterotrophic bacteria at the outer part of the biofilm. As a result of the interspecies competition, NH4plus; oxidation was restrained, resulting in a decrease in the ammonia-oxidizing bacterial populations. This experimental result clearly explains the stratified spatial distributions of ammonia-oxidizing bacteria within the biofilms at higher substrate C/N ratios. The combined application of microelectrodes and FISH techniques provides new insights into microbial ecology and population dynamics of nitrifying bacteria within multi-species biofilms.

High-strength nitrogen removal of opto-electronic industrial wastewater in membrane bioreactor - a pilot study

2003 ◽  
Vol 48 (1) ◽  
pp. 191-198 ◽  
Author(s):  
T.K. Chen ◽  
C.H. Ni ◽  
J.N. Chen ◽  
J. Lin
Keyword(s):  
Wastewater Treatment ◽  
Industrial Wastewater ◽  
Organic Nitrogen ◽  
Membrane Bioreactor ◽  
High Strength ◽  
Experimental Period ◽  
Ammonia Nitrogen ◽  
Nitrifying Bacteria ◽  
Biological Degradation ◽  
The Past

The membrane bioreactor (MBR) system has become more and more attractive in the field of wastewater treatment. It is particularly attractive in situations where long solids retention times are required, such as nitrifying bacteria, and physical retention critical to achieving more efficiency for biological degradation of pollutant. Although it is a new technology, the MBR process has been applied for industrial wastewater treatment for only the past decade. The opto-electronic industry, developed very fast over the past decade in the world, is high technology manufacturing. The treatment of the opto-electronic industrial wastewater containing a significant quantity of organic nitrogen compounds with a ratio over 95% in organic nitrogen (Org-N) to total nitrogen (T-N) is very difficult to meet the discharge limits. This research is mainly to discuss the treatment capacity of high-strength organic nitrogen wastewater, and to investigate the capabilities of the MBR process. A 5 m3/day capacity of MBR pilot plant consisted of anoxic, aerobic and membrane bioreactor was installed for evaluation. The operation was continued for 150 days. Over the whole experimental period, a satisfactory organic removal performance was achieved. The COD could be removed with an average of over 94.5%. For TOC and BOD5 items, the average removal efficiencies were 96.3 and 97.6%, respectively. The nitrification and denitrification was also successfully achieved. Furthermore, the effluent did not contain any suspended solids. Only a small concentration of ammonia nitrogen was found in the effluent. The stable effluent quality and satisfactory removal performance mentioned above were ensured by the efficient interception performance of the membrane device incorporated within the biological reactor. The MBR system shows promise as a means of treating very high organic nitrogen wastewater without dilution. The effluent of TKN, NOx-N and COD can fall below 20 mg/L, 30 mg/L and 50 mg/L.

Combining fluorescent in situ hybridization (fish) with cultivation and mathematical modeling to study population structure and function of ammonia-oxidizing bacteria in activated sludge

1998 ◽  
Vol 37 (4-5) ◽  
pp. 441-449 ◽  
Author(s):  
Michael Wagner ◽  
Daniel R. Noguera ◽  
Stefan Juretschko ◽  
Gabriele Rath ◽  
Hans-Peter Koops ◽  
...  
Keyword(s):  
Mathematical Modeling ◽  
In Situ Hybridization ◽  
Activated Sludge ◽  
Heterotrophic Bacteria ◽  
Sewage Treatment Plant ◽  
Laser Scanning Microscopy ◽  
Industrial Plant ◽  
Ammonia Oxidizing Bacteria ◽  
Ammonia Oxidizers

16S rRNA-targeted oligonucleotide probes for phylogenetically defined groups of autotrophic ammonia-oxidizing bacteria were used for analyzing the natural diversity of nitrifiers in an industrial sewage treatment plant receiving sewage with high ammonia concentrations. In this facility discontinuous aeration is used to allow for complete nitrification and denitrification. In situ hybridization revealed a yet undescribed diversity of ammonia oxidizers occurring in the plant. Surprisingly, the majority of the ammonia oxidizers were detected with probe combinations which indicate a close affiliation of these cells with Nitrosococcus mobilis. In addition, low numbers of ammonia-oxidizers related to the Nitrosomonas europaea - Nitrosomonas eutropha cluster were present. Interestingly, we also observed hybridization patterns which suggested the occurrence of a novel population of ammonia oxidizers. Confocal laser scanning microscopy revealed that all specifically stained ammonia oxidizers were clustered in microcolonies formed by rod-shaped bacteria. Combination of FISH and mathematical modeling was used to investigate diffusion limitation of ammonia and O2 within these aggregates. Model simulations suggest that mass transfer limitations inside the clusters are not as significant as the substrate limitations due to the activity of surrounding heterotrophic bacteria. To learn more about the ammonia-oxidizers of the industrial plant, we enriched and isolated ammonia-oxidizing bacteria from the activated sludge by combining classical cultivation techniques and FISH. Monitoring the isolates with the nested probe set allowed us to specifically identify those ammonia oxidizers which were found in situ to be numerically dominant. The phylogenetic relationship of these isolates determined by comparative 16S rDNA sequence analysis confirmed the affiliation suggested by FISH.

Characterization of bacterial biofilm communities in tertiary treatment processes for wastewater reclamation and reuse

2008 ◽  
Vol 58 (5) ◽  
pp. 1023-1030 ◽  
Author(s):  
Tadashi Shoji ◽  
Shuichi Ochi ◽  
Masaaki Ozaki
Keyword(s):  
Urban Areas ◽  
Heterotrophic Bacteria ◽  
Bacterial Biofilm ◽  
Nitrifying Bacteria ◽  
Water Supply Systems ◽  
Ammonia Oxidizing Bacteria ◽  
Tertiary Treatment ◽  
Treatment Processes ◽  
Related Group ◽  
Biofilm Development

The concern with wastewater reuse as a sustainable water resource in urban areas has been growing. For the reclamation and distribution of wastewater, biofilm development deserves careful attention from the point of view of its promotion (e.g. biofiltration) and inhibition (e.g. clogging and hygiene problems). As the first step to control biofilm development, bacterial biofilm communities in tertiary treatment processes were characterized by using molecular biological methods. The result of clone library analysis showed that Nitrospirae-related (nitrite-oxydizing bacteria) and Acidobacteria-related (probably oligotrophic bacteria) groups were dominant. The ratio of the Nitrospirae-related group to the Acidobacteria-related group was associated with ammonia load, whereas other operational conditions (process, media, temperature, salt) did not clearly affect the phylum-level community or the dominant sequence of nitrifying bacteria. The result of real-time PCR also indicated that high ammonia load promotes the proliferation of nitrite- and ammonia-oxidizing bacteria. Regarding water supply systems, some researchers also have suggested the dominance of Nitrospirae- and Acidobacteria-related groups in biofilm formed on water distribution pipes. In tertiary wastewater treatment, therefore, it is concluded that oligotrophic and autotrophic bacteria are the dominant groups in biofilm samples because assimilable organic carbon is too poor to proliferate various heterotrophic bacteria.

Transition of bacterial spatial organization in a biofilm monitored by FISH and subsequent image analysis

2004 ◽  
Vol 49 (11-12) ◽  
pp. 365-370 ◽  
Author(s):  
Y. Aoi ◽  
S. Tsuneda ◽  
A. Hirata
Keyword(s):  
Image Analysis ◽  
Outer Layer ◽  
Spatial Organization ◽  
Heterotrophic Bacteria ◽  
Great Influence ◽  
Ammonia Oxidizing Bacteria ◽  
Nitrite Oxidizing Bacteria ◽  
Growth Activity ◽  
Do Concentration

The dynamic transition of bacterial community structure in a biofilm was monitored by the fluorescence in situ hybridization (FISH) technique and subsequent image analysis. Heterotrophic bacteria that had occupied the outer layer were gradually decreased whereas ammonia-oxidizing bacteria (AOB) gradually increased their growth activity and extended their existence area to the outer layer of the biofilm through the gradual reduction of the C/N ratio. The spatial organization of AOB in the biofilm dynamically changed responding to the environmental conditions such as pH fluctuation and lack of dissolved oxygen (DO) and had great influence on the nitrification activity. The accumulation of nitrite was observed at lower DO concentration, which might be due to the property that nitrite-oxidizing bacteria (NOB) possess of higher Km values for oxygen than AOB.

Eco-physiology of autotrophic nitrifying biofilms

2005 ◽  
Vol 52 (7) ◽  
pp. 225-232 ◽  
Author(s):  
S. Okabe ◽  
T. Kindaichi ◽  
Y. Nakamura ◽  
T. Ito
Keyword(s):  
Organic Matter ◽  
Fluorescent In Situ Hybridization ◽  
Direct Evidence ◽  
Heterotrophic Bacteria ◽  
Microbial Interactions ◽  
Nitrifying Bacteria ◽  
Low Molecular Weight ◽  
Sulfur Bacteria ◽  
First Time

Microautoradiography combined with fluorescent in situ hybridization (MAR-FISH), a powerful tool for linking physiology with identification of individual cells, was applied to investigate microbial interactions between nitrifying bacteria and coexisting heterotrophic bacteria in an autotrophic nitrifying biofilm community fed with only ammonia as the sole energy source and bicarbonate as the sole carbon source. First, nitrifying bacteria were radiolabeled by culturing the biofilm samples with [14C]bicarbonate for 6 h, and then the transfer of radioactivity from nitrifying bacteria to heterotrophic bacteria was monitored by using MAR-FISH. MAR-FISH revealed that the heterotrophic bacterial community was composed of bacteria that were phylogenetically and metabolically diverse. We could obtain direct evidence that organic matter derived from nitrifiers was subsequently utilized by mainly filamentous bacteria belonging to the Chloroflexi (green non-sulfur bacteria) group or CFB group in the biofilm, which was clearly visualized by MAR-FISH at single cell resolution for the first time. On the other hand, the members of the α- and γ-Proteobacteria were specialized to utilize low-molecular-weight organic matter. This community represents functionally integrated units that assure maximum access to and utilization of metabolites of nitrifiers.

Effect of dissolved oxygen concentration on the biofilm and in situ analysis by fluorescence in situ hybridization (FISH) and microelectrodes

2003 ◽  
Vol 47 (1) ◽  
pp. 49-57 ◽  
Author(s):  
A. Jang ◽  
P.L. Bishop ◽  
S. Okabe ◽  
S.G. Lee ◽  
I.S. Kim
Keyword(s):  
In Situ Hybridization ◽  
Dissolved Oxygen ◽  
Nitrifying Bacteria ◽  
Dry Density ◽  
Ammonia Oxidizing Bacteria ◽  
Biofilm Thickness ◽  
Experimental Findings ◽  
And Control ◽  
Microelectrode Measurements

A better understanding of microbiology and ecology of nitrifying bacteria in inner biofilms is an important part of improving process performance and control. Microelectrodes and fluorescent in situ hybridization (FISH) in biofilm research have been used to investigate the spatial distributions of various microbial activities in biofilms and have led to new experimental findings as well as modifications of the homogeneous assumptions in the biofilm kinetic models. The objective of this study is to try the combination of two methods, both FISH and microelectrode measurements, and to provide reliable and in situ information on nitrifying bacterial activity in biofilms. The characteristics of biofilm developed on tygon slides were different according to the change of dissolved oxygen (DO). When the DO increased from 2 to 10 μg DO/L, the rate of the biofilm thickness increased and its dry density changed from 50-70 to 25-90 mg/cm3. Ammonia oxidizing bacteria were not uniformly distributed in biofilm, and were found at the deeper layer where oxygen is depleted, they were detected primarily in the upper and middle layers of the biofilm.

Fluorescence in situ hybridization analysis of nitrifiers in piggery wastewater treatment reactors

2004 ◽  
Vol 49 (5-6) ◽  
pp. 333-340 ◽  
Author(s):  
D.J. Kim ◽  
T.K. Kim ◽  
E.J. Choi ◽  
W.C. Park ◽  
T.H. Kim ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Ammonia Concentration ◽  
Biofilm Reactor ◽  
Nitrifying Bacteria ◽  
Fish Analysis ◽  
Ammonia Oxidizing Bacteria ◽  
Piggery Wastewater

Fluorescence in situ hybridization (FISH) was performed to analyze the nitrifying microbial communities in an activated sludge reactor (ASR) and a fixed biofilm reactor (FBR) for piggery wastewater treatment. Heterotrophic oxidation and nitrification were occurring simultaneously in the ASR and the COD and nitrification efficiencies depend on the loads. In the FBR nitrification efficiency also depends on ammonium load to the reactor and nitrite was accumulated when free ammonia concentration was higher than 0.2 mg NH3-N/L. FISH analysis showed that ammonia-oxidizing bacteria (NSO1225) and denitrifying bacteria (RRP1088) were less abundant than other bacteria (EUB338) in ASR. Further analysis on nitrifying bacteria in the FBR showed that Nitrosomonas species (NSM156) and Nitrospira species (NSR1156) were the dominant ammonia-oxidizing and nitrite-oxidizing bacteria, respectively, in the piggery wastewater nitrification system.

In situ analysis of nitrifying bacteria in sewage treatment plants

1996 ◽  
Vol 34 (1-2) ◽  
pp. 237-244 ◽  
Author(s):  
Michael Wagner ◽  
Gabriele Rath ◽  
Hans-Peter Koops ◽  
Janine Flood ◽  
Rudolf Amann
Keyword(s):  
Activated Sludge ◽  
Sewage Treatment ◽  
Nitrifying Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Microbiological Methods ◽  
Nitrite Oxidizing Bacteria ◽  
Identification Technique ◽  
Engineered Systems ◽  
Simultaneous Identification

Autotrophic microbial nitrification is the key process in the removal of ammonia from wastewater. To avoid the limitations of traditional microbiological methods an in situ identification technique for ammonia- and nitrite-oxidizing bacteria was developed. Based on comparative sequence analyses we designed a collection of 16S ribosomal RNA-targeted oligonucleotide probes for all validly described members of the genusNitrobacter . Whole cell hybridizations of target and reference cells with fluorescent probe derivatives were used to determine the optimal hybridization stringency for each of the probes. These probes were applied together with a recently developed probe for important members of the genus Nitrosomonas for simultaneous identification of ammonia- and nitrite-oxidizing bacteria in natural and engineered systems. Ammonia-oxidizing bacteria were identified in situ in river water, epiphytic biofilms from eutrophic wetlands, oligotrophic biofilms, a nitrifying trickling filter biofilm as well as in all analyzed nitrifying activated sludge samples. In none of these samples could Nitrobacter cells be detected in situ. However, all hitherto describedNitrobacter species and a strain of Nitrobacter sp. isolated from one of the analyzed nitrifying activated sludge samples showed bright hybridization signals with all Nitrobacter specific probes. Possible reasons for the absence of in situ detectable Nitrobacter cells are discussed.

Membrane bioreactor versus conventional activated sludge system: population dynamics of nitrifiers

2005 ◽  
Vol 52 (10-11) ◽  
pp. 417-425 ◽  
Author(s):  
R. Manser ◽  
W. Gujer ◽  
H. Siegrist
Keyword(s):  
Activated Sludge ◽  
Community Composition ◽  
Membrane Bioreactor ◽  
Membrane Separation ◽  
Heterotrophic Bacteria ◽  
Microbial Community Composition ◽  
Laser Scanning Microscopy ◽  
Nitrifying Bacteria ◽  
Ammonia Oxidizing Bacteria ◽  
Conventional Activated Sludge

Although membrane bioreactors have attracted increasing attention in recent years, little research has been undertaken on the influence of the membrane separation on the microbial community composition. This paper compares the startup behaviour and the performance of the subsequent eight months of a membrane bioreactor with a conventional activated sludge pilot plant. Both plants were operated in parallel at the same sludge age and treated the same domestic wastewater. The identification of the nitrifying community composition using fluorescent in situ hybridization revealed only minor differences between the two reactors for both ammonia-oxidizing bacteria and nitrite-oxidizing bacteria. Accordingly, both systems exhibited the same maximum nitrification rates. Confocal laser scanning microscopy showed that the aggregates formed by nitrifying bacteria were located mostly in the inner part of the flocs and were overgrown by heterotrophic bacteria. It is concluded that the membrane separation itself does affect neither the nitrifying community composition nor the nitrification performance. However, impacts on kinetic parameters are emphasized.

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