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Biological Treatment of Ammonia-Rich Wastewater by

PartialNitrification/ANAMMOX in the BioCAST Reactor

Conference Paper · May 2017

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Biological Treatment of Ammonia-Rich Wastewater by Partial
Nitrification/ANAMMOX in BioCAST Reactor

N. Saborimanesh*, E. Castillo Arriagada**, D. Walsh*, L. Yerushalmi*, C.N. Mulligan*1

* Concordia University
1
corresponding author, Dept. Building, Civil and Environmental Engineering, Concordia University-1455 de
Maisonneuve Blvd. West, Montreal, Quebec, Canada H3G 1M8, mulligan@civil.concordia.ca, (514) 848-2424
ext. 7925
** Institute of Parasitology, McGill University, Montreal, Canada

Abstract:

Nitrogenous pollution has deleteriously impacted human health and aquatic environments. The objective of this
study was to investigate the potential application of a biological reactor (BioCAST) for the treatment of an
ammonia-rich synthetic wastewater by providing a balance in the environmental conditions for aerobic nitrifying
bacteria and anaerobic ammonium oxidizers ("Anammox") in a single reactor. The bioreactor was inoculated
with Anammox sludge and fed with an ammonia-rich wastewater (NH 3 -N: 350 mg/L) at a hydraulic retention
time of 2 days and temperature of 35°C. The DO levels in the aerobic and anoxic zones were maintained at 1.5
mg/L and 0.1 mg/L, respectively. Despite nitrite limited feeding conditions (NO 2 : 0 mg-N/L), microbial
communities contributed to nearly 80% and 91% of TN and NH 3 -N removal, respectively. This suggests that the
favorable conditions for the activity of nitrifying and Anammox communities were provided in this system under
the tested conditions.

Keywords: Partial Nitrification/Anammox; Ammonia; Wastewater Treatment

Introduction
Nitrogenous pollution as a result of the discharge of untreated or improperly treated
wastewater has deleteriously impacted human health and aquatic environments (Environment
Canada & Health Canada, 2001). Traditional and innovative biological treatment technologies
have proven to effectively remove nitrogenous pollutants to non-polluting forms (e.g., N 2 )
using nitrifying and anaerobic ammonium oxidizers (“Anammox”) (Figure 1) (Mulder et al.,
1995; Dijkmall and Strous, 2002). However, obtaining successful removal in a one-vessel
treatment system that can simultaneously provide a balance in the environmental conditions
(e.g., dissolved oxygen, DO) for aerobic nitrifying and anoxic Anammox communities is
extremely challenging.

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 Figure 1. Biological Treatment of Ammonia Through Nitrification (a) and Partial
Nitrification and Anammox (b) Processes (adapted from (Mulder et al., 1995; Dijkmall and
Strous, 2002).
The BioCAST technology is a biological wastewater treatment system and was developed to
maximize the growth of aerobic and anaerobic microbial communities in a single multi-zone
reactor. It was hypothesized that this reactor can be efficient in the treatment of rich-ammonia
wastewater if the optimal environmental conditions (e.g., dissolved oxygen and temperature)
were maintained at the optimal levels for the nitrifying microorganisms in the aerobic zone
(which produce the nitrite required for the Anammox microorganisms) and for the Anammox
microorganisms in the anoxic zone (which can convert the remaining ammonia and the
produced nitrite to nitrogen gas). The aim of this study is to investigate the feasibility of
treatment of ammonia-rich wastewater with partial nitrification/Anammox processes in this
reactor. This technology can be beneficial in several ways including minimum sludge
production and low oxygen consumption and simplicity of operation.

Material and Methods

BioCAST Reactor Start-Up and Operation
The BioCAST is a lab-scale continuous biological treatment reactor containing aerobic,
microaerophilic, anoxic and clarification zones (Figure 2). The aerobic zone is in the middle
of the bioreactor and contains a microbial support covered by a nonwoven geotextile and
stripes of geotextile to support attached bacteria growth (Figure 3). The bioreactor was
wrapped with a black plastic sheet/aluminum foil to prevent the microorganisms from
exposure to light. The biological sludge was wasted from the bottom of the anoxic zone under
gravity flow using a clear vinyl tubing. Aeration provided both the dissolved oxygen (DO)
necessary for the bacterial growth and mixing. The air was introduced upward from the
bottom of the bioreactor to the aerobic zone (final concentration of <2 mg/L) using a timer-
controlled aerator. The heat was maintained at 35°C using a 1000 W titanium heating system
with a digital controller (JBJ TRUE TEMP) and pH (7.8 ± 0.5) was monitored by a portable
pH meter (OAKTON PD-650).
 Figure 2. Schematic diagram of the BioCAST reactor.

Figure 3. Microbial support structure made of stainless steel covered by nonwoven

geotextiles and strips of a nonwoven geotextile in the aerobic zone.
A sludge seed containing nitrifying and Anammox communities was obtained from Hampton
Roads Sanitation District (USA) to start up the system. A synthetic wastewater was prepared
with the addition of main chemicals, (NH 3 ) 2 CO 3 (final concentration of 50-350 mg/L NH 3 -
N), and trace elements to the feeding tank containing tap water (100 L) according to Graaf et
al. (1996) and Strous et al. (1997) with modifications. The bioreactor operation was started
with an NH 3 -N concentration of 50 mg/L and increased stepwise to 350 mg/L at a hydraulic
retention time (HRT) of 4 days. When the nitrogen removal efficiency reached above 80% the
HRT was reduced to 2 days and operation was continued at this HRT using a peristaltic pump
to maintain a flow rate of 2.1 L/h.
Sample Collection & Analysis
Two sets of samples including liquid and solid/biomass were collected. The liquid sampling
was conducted daily or at least three times a week. The liquid samples were taken from
influent, aerobic, microaerophilic, and anoxic zones for determining the total alkalinity, NH 3 -
N, NO 2 -N, and NO 3 -N concentrations using colorimetric methods (TNT 870, 832, 840, 836;
HACH Canada). The solid/biomass sampling was conducted biweekly or monthly for the
settling time, TSS, and VSS tests (Standard Methods 2540 D/E).

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 Results and Conclusions
A maximum removal of 92% and 80% for NH 3 and total nitrogen were achieved in this
system at HRT 2 days and 35°C (Table 1). Following the startup phase, nitrite and nitrate
were produced as the main intermediate products and the concentration of nitrate was 42 ±19
mg/L. In the operating phase when no excess sludge was withdrawn, the biomass
concentration in the anoxic zone was increased rapidly following three months of startup.
These results suggests that the favorable environmental conditions for the activity of
nitrifying and Anammox communities were provided in this system under the tested
conditions and the removal of NH 4 + in an anoxic zone can be due to the Anammox activity.

Table 1. Ammonium removal from synthetic wastewater with BioCAST technology.

Parameter BioCAST Technology

Duration of experiment (days) 30
Influent NH 4 + concentration (mg NH 4 +-N/L) 344.69 ± 22.7
Effluent NH 4 + concentration (mg NH 4 +-N/L) 19.23± 18.4
Influent NO 2 - concentration (mg NO 2 - -N/L) 0.00
Ammonium removal (%) 90.8 ± 9.2
Total nitrogen removal (%) 80.1 ± 5.8

Figure 3. Total nitrogen (a) and ammonium (b) removals (%)

References
Dijkmall, H., Strous, M., 2002. (12) United States Patent. Patent N0.: US 6,485,646 B1.
Environment Canada & Health Canada, 2001. Ammonia in the aquatic environment. Priority substances list
assessment report., Canadian Environmental Protection Act, 1999. Minister of Public Works and
Government Services, Cat. No.: En 40-215/55E.
Graaf, A.A. Van De, Bruijn, P. De, Robertson, L.A., Jetten, M.S.M., Kuenen, J.G., 1996. Autotrophic growth of
anaerobic in a fluidized bed reactor. Microbiology 142, 2187–2196.
Mulder, A., van de Graaf, A.A. Robertson, L.A., Kuenen J.G.K., 1995. Anaerobic ammonium oxidation
discovered in a denitrifying fluidized bed reactor. FEMS Microbiology Ecology 16, 177-183.
doi:http://dx.doi.org/10.1111/j.1574-6941.1995.tb00281.x
Strous, M., Van Gerven, E., Zheng, P., Kuenen, J.G., Jetten, M.S.M., 1997. Ammonium removal from
concentrated waste streams with the anaerobic ammonium oxidation (anammox) process in different
reactor configurations. Water Research 31, 1955–1962. doi:10.1016/S0043-1354(97)00055-9

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