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Chapter 16 : Nitrification in Wastewater Treatment

Authors: Satoshi Okabe1, Yoshiteru Aoi2, Hisashi Satoh3, Yuichi Suwa4
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Affiliations: 1: Department of Urban and Environmental Engineering, Graduate School of Engineering, Hokkaido University, Sapporo 060-8628, Japan; 2: Waseda Institute for Advanced Study, Tokyo 169-8050, Japan; 3: Department of Urban and Environmental Engineering, Graduate School of Engineering, Hokkaido University, Sapporo 060-8628, Japan; 4: Faculty of Science and Engineering, Chuo University, Tokyo 112-8551, Japan
Content Type: Monograph
Publication Year: 2011

Category: Environmental Microbiology

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Abstract:

Microbial nitrification is a necessary step in removing nitrogen from wastewaters via biological denitrification and is becoming more important due to strict regulations on nitrogen discharge. However, microbial nitrification is recognized as being difficult to maintain in practical wastewater treatment plants (WWTPs) owing to the lower kinetics, yields, and sensitivity of nitrifying bacteria to physical, chemical, and environmental disturbances as mentioned, even though nitrification has been studied more than any other specific biochemical reactions occurring in wastewater treatment to date. Influent NO , chromium, and nickel influenced the AOB community structure, while correlations between other metals analyzed in this study and the AOB community structure were insignificant. As an oxidation process, nitrification significantly consumes oxygen, and dissolved oxygen (DO) concentration is a key factor for maintaining nitrification stably as well as pH. In an activated sludge process, 3% salt inhibited both the maximum utilization rate and the saturation constant, suggesting uncompetitive inhibition. Nitrification in wastewater treatment systems has been studied extensively. Despite their importance, knowledge about the identity and ecology of nitrifying bacteria carrying out nitrification in WWTPs has been scarce. Thus, biological nitrogen removal processes have been regarded as “a black box” in practice because the lack of fundamental microbiological understanding hampers knowledge-driven process design and operation.

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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Nitrification in Wastewater Treatment
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Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
/content/10.1128/9781555817145.ch16.ch16fig01
FIGURE 1
Typical process flow sheets for biological nitrogen removal. (A) A portion of the wastewater can be bypassed to the anoxic tank (denitrifying tank). (B) Bardenpho process.
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Image of FIGURE 1
FIGURE 1

Typical process flow sheets for biological nitrogen removal. (A) A portion of the wastewater can be bypassed to the anoxic tank (denitrifying tank). (B) Bardenpho process.

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 2
(A) Photomicrograph of an activated sludge floc. (B and C) Confocal laser-scanning microscope images of an activated sludge floc showing the in situ spatial organization of bacteria and AOB. FISH was performed using a fluorescein isothiocyanate-labeled EUB338-mixed probe and a tetramethylrhodamine 5-isothiocyanate-labeled Nso190 probe. The probe Nso190-stained AOB appear to be yellow because of binding of both probes, and bacterial cells are green.
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Image of FIGURE 2
FIGURE 2

(A) Photomicrograph of an activated sludge floc. (B and C) Confocal laser-scanning microscope images of an activated sludge floc showing the in situ spatial organization of bacteria and AOB. FISH was performed using a fluorescein isothiocyanate-labeled EUB338-mixed probe and a tetramethylrhodamine 5-isothiocyanate-labeled Nso190 probe. The probe Nso190-stained AOB appear to be yellow because of binding of both probes, and bacterial cells are green.

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 3
Typical concentration profiles of O, NH , and NO in an activated sludge floc at 45 µM O. The shaded area indicates the floc. The center of the floc is at a depth of 0 µm. (From ], with permission from .)
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Image of FIGURE 3
FIGURE 3

Typical concentration profiles of O, NH , and NO in an activated sludge floc at 45 µM O. The shaded area indicates the floc. The center of the floc is at a depth of 0 µm. (From ], with permission from .)

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 4
The consumption rates of NH and inorganic nitrogen (Ni) defined as the sum of NH , NO , and NO as determined by the batch experiments at various O concentrations in the bulk liquid. Rates shown are mean values, and error bars indicate standard deviations. (From ], with permission from .)
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Image of FIGURE 4
FIGURE 4

The consumption rates of NH and inorganic nitrogen (Ni) defined as the sum of NH , NO , and NO as determined by the batch experiments at various O concentrations in the bulk liquid. Rates shown are mean values, and error bars indicate standard deviations. (From ], with permission from .)

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 5
Steady-state concentration profiles of O and NH in the autotrophic nitrifying biofilm incubated in the media at C/N = 0 (A), C/N = 1 (B), and C/N = 3.4 (C), respectively. The modeled profiles are indicated by solid lines. The spatial distributions of NH oxidation rates are indicated by stippled area. Surface is at a depth of 0 µm. (From Okabe et al. [2004b], with permission from .)
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Image of FIGURE 5
FIGURE 5

Steady-state concentration profiles of O and NH in the autotrophic nitrifying biofilm incubated in the media at C/N = 0 (A), C/N = 1 (B), and C/N = 3.4 (C), respectively. The modeled profiles are indicated by solid lines. The spatial distributions of NH oxidation rates are indicated by stippled area. Surface is at a depth of 0 µm. (From Okabe et al. [2004b], with permission from .)

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 6
Spatial distributions of surface fractions of AOB in biofilms cultured in the media at C/N = 0, 1, and 2, respectively. The biofilm surfaces are indicated by the dotted lines.
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Image of FIGURE 6
FIGURE 6

Spatial distributions of surface fractions of AOB in biofilms cultured in the media at C/N = 0, 1, and 2, respectively. The biofilm surfaces are indicated by the dotted lines.

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 7
(A) Spatial distribution of average microbial cluster sizes of AOB hybridized with probe Nso190 in different biofilms cultured at C/N = 0 and 1. (B) Cross-section of biofilm cultured at C/N = 0.
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Image of FIGURE 7
FIGURE 7

(A) Spatial distribution of average microbial cluster sizes of AOB hybridized with probe Nso190 in different biofilms cultured at C/N = 0 and 1. (B) Cross-section of biofilm cultured at C/N = 0.

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 8
(A) Steady-state concentration profiles of O, NH , NO , and NO in a biofilm cultured at C/N = 2. (B) Spatial distribution of the estimated volumetric consumption and production rates of NH , NO , and NO . The biofilm surface is at a depth of 0 µm.
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Image of FIGURE 8
FIGURE 8

(A) Steady-state concentration profiles of O, NH , NO , and NO in a biofilm cultured at C/N = 2. (B) Spatial distribution of the estimated volumetric consumption and production rates of NH , NO , and NO . The biofilm surface is at a depth of 0 µm.

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 9
Proposed ecophysiological interactions between nitrifiers and heterotrophic bacteria in a carbon-limited autotrophic nitrifying biofilm fed with only NH as energy source.
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Image of FIGURE 9
FIGURE 9

Proposed ecophysiological interactions between nitrifiers and heterotrophic bacteria in a carbon-limited autotrophic nitrifying biofilm fed with only NH as energy source.

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 10
(A) Two-dimensional (2-d) IbM model description of actions occurring at the individual scale. (B) Spatial scales in the 2-d model of nitrifying granules. (a) Representative biomass granule comprised in a square computational domain; (b) the square grid elements discretizing the space, each containing several biomass particles; (c) individual biomass particles, of different possible biomass types. All biomass particles within a single grid element experience the same substrate concentrations. The biomass concentrations within a grid element are calculated from the mass of all individual biomass particles within the element volume ( ).
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Image of FIGURE 10
FIGURE 10

(A) Two-dimensional (2-d) IbM model description of actions occurring at the individual scale. (B) Spatial scales in the 2-d model of nitrifying granules. (a) Representative biomass granule comprised in a square computational domain; (b) the square grid elements discretizing the space, each containing several biomass particles; (c) individual biomass particles, of different possible biomass types. All biomass particles within a single grid element experience the same substrate concentrations. The biomass concentrations within a grid element are calculated from the mass of all individual biomass particles within the element volume ( ).

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 11
Detailed insight into the spatial localization of HetU (a), HetB (b), and HetO (c) provided by a two-dimensional IbM simulation, at day 100. White dotted line shows the granule surface ( ).
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Image of FIGURE 11
FIGURE 11

Detailed insight into the spatial localization of HetU (a), HetB (b), and HetO (c) provided by a two-dimensional IbM simulation, at day 100. White dotted line shows the granule surface ( ).

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 12
Spatial distributions of bacteria along the radius of a nitrifying granule as detected by FISH. The abundance ratio of each bacterium was quantified in 50-µm-thick shells starting at the granule surface (left). Abundance data presented are averages of six replicate measurements. The total bacterial volumetric occupancy in the granule is derived from fluorescence of EUB338 mix-tagged cells (open circles along the -axis). The α- constitute the bacterial group that hybridized with probe ALF1b, excluding the genus that hybridized with probe NIT3. (The figure was reconstructed from the data of ].)
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Image of FIGURE 12
FIGURE 12

Spatial distributions of bacteria along the radius of a nitrifying granule as detected by FISH. The abundance ratio of each bacterium was quantified in 50-µm-thick shells starting at the granule surface (left). Abundance data presented are averages of six replicate measurements. The total bacterial volumetric occupancy in the granule is derived from fluorescence of EUB338 mix-tagged cells (open circles along the -axis). The α- constitute the bacterial group that hybridized with probe ALF1b, excluding the genus that hybridized with probe NIT3. (The figure was reconstructed from the data of ].)

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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FIGURE 13
Comparison of the steady-state solute concentrations in the biofilm calculated in two-dimensional (2-d) IbM model simulations, with the experimental microelectrode data (open circles, triangles and squares for O, NH , NO , NO ). The model results are at day 100. To obtain the comparable profiles along the radius, the 2-d concentration distributions were averaged in concentric shells with different radii. (The figure was reconstructed from the data of ].)
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Image of FIGURE 13
FIGURE 13

Comparison of the steady-state solute concentrations in the biofilm calculated in two-dimensional (2-d) IbM model simulations, with the experimental microelectrode data (open circles, triangles and squares for O, NH , NO , NO ). The model results are at day 100. To obtain the comparable profiles along the radius, the 2-d concentration distributions were averaged in concentric shells with different radii. (The figure was reconstructed from the data of ].)

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
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Tables

Nitrification in Wastewater Treatment
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Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16
/content/10.1128/9781555817145.ch16.ch16tab01
TABLE 1
Stoichiometric parameters for microbial reactions
Generic image for table
TABLE 1

Stoichiometric parameters for microbial reactions

Citation: Okabe S, Aoi Y, Satoh H, Suwa Y. 2011. Nitrification in Wastewater Treatment, p 405-433. In Ward B, Arp D, Klotz M (ed), Nitrification. ASM Press, Washington, DC. doi: 10.1128/9781555817145.ch16

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