Technology

AMX - Anammox

 

Anaerobic ammonium oxidation (anammox) has been discovered in 1990's from one of the side-stream treatment systems. Unlike conventional biological nitrogen removal processes involving complete ammonia oxidation followed by chemoorganoheterotrophic denitrification, anammox involves autotrophic conversion of a partially nitrified stream containing nearly equimolar mixtures of ammonia and nitrite to dinitrogen gas.

Anammox process is an attractive sustainable nitrogen control solution as no external carbon is required and can reduce energy costs by 60% while removing nitrogen from the liquid stream (i.e., the transformation of inorganic nitrogen species to dinitrogen gas). Additionally, a 90% less biomass yield can be achieved by anammox processes. Thus, this process is an energy and cost-effective alternative for conventional nitrogen removal. 

  • Mainstream Annamox

  • Sidestream Annamox

  • Industrial Annamox

 
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Technology

AMX - Mainstream

 

Mainstream Anammox Process for Nutrient Removal and Recovery

Municipal Tertiary Treatment

  • 2-stage Partial Nitritation + Anammox (PN+A) Upflow Biofiltration System

  1. Low Temperature

  2. High C/N ratio

  • Stable operation via separate enrichment of AOB & AMX

Technical Information

Stable nitritation and AOB granule generation in the sidestream

  • Achieved nitritation by supplying AOB from sidestream to the mainstream

  • BBF-AMX :

    • Maintain high concentration anammox microorganisms on pores and surfaces of EPP media

    • Physical filtration ensures treatment quality (<2 NTU)

    • Separate solid-liquid separation (sedimentation, filtration) is unnecessary

LED Nitritation Process

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Combination of blue LED and reactor operating conditions for effective NOB suppression

  • Simultaneously achieve organic removal and nitritation

  • BBF-AMX:

    • Maintain high concentration anammox microorganisms on pores and surfaces of EPP media

    • Physical filtration ensures treatment quality (<2 NTU)

    • Separate solid-liquid separation (sedimentation, filtration) is unnecessary

Photo-Granule Nitritation Process

 

Simultaneous organic removal and nitritation using algae granule

  • Easily separate solid-liquid by granule formation composed of algae, heterotrophs, and AOB

  • Utilizes air generated from photosynthesis and does not require a separate air supply

  • BBF-AMX :

    • Maintain high concentration anammox microorganisms on pores and surfaces of EPP media

    • Physical filtration ensures treatment quality (<2 NTU)

    • Separate solid-liquid separation (sedimentation, filtration) is unnecessary

 

Dr. Sandeep Sathyamoorthy, PhD, PE, Black and Veatch presenting at 2018 CWEA on Tomorrow Water’s Sidestream & Mainstream Nutrient Workshop

 
 

It’s time to take Anammox mainstream

 

Technology

AMX - Sidestream

 

Sidestream Anammox Process for Nutrient Removal 

  • Municipal Side-stream Treatment

  • 2-stage Partial Nitritation + Anammox (PN+A) System

  1. Wide Range of Influent Temperature (Mesophilic to Thermophilic Digester)

  2. Low C/N ratio

  3. Low alkalinity

  • Stable operation via separate enrichment of AOB & AMX

Streams such as filtrates, centrates, and digester supernatants make up only 1-2% of the total flow at WWTPs, but add approximately 15-40% of the nitrogen load to the mainstream treatment line.

With our Anammox Sidestream process, plants can decrease their operating costs by 15-40% and avoid costly violation fees.

Compared to conventional nitrification/denitrification (NDN):

  •  Reduces carbon requirements by 90% (for complete removal of inorganic nitrogen)

  •  Reduces oxygen requirements by approximately 60%

  •  Reduces biomass production by approximately 90%

 

Our versatile Anammox solution is a highly effective method of removing nitrogen from sidestream before returning it to the headworks.

 

Technology

AMX - Industrial

 

Low Cost Nutrient Removal

Indiana’s new law has enacted stringent nitrogen regulations for dairy farms across the state. To mitigate the harmful effects of nitrogen in the environment, Indiana farmers are now required to remove significant amounts of nitrogen from manure or from the effluent in their digesters. Otherwise farmers need to purchase 10 times more land for their cows (the new law requires 10 acres of land per one head of cow). 

HIGH STRENGTH NITROGEN REMOVAL FOR LIVESTOCK MANURE OR DIGESTATE

With decades of experience in livestock wastewater treatment, Tomorrow Water is developing an innovative nitrogen removal process using our Anammox bacteria. The Anammox process can remove nitrogen from either from the source or after anaerobic digestion.

Tomorrow Water has many experiences with the problems farmers face themselves such as unstable dewatering system or expensive chemical dosing for coagulation.

Using Anammox Process, Tomorrow Water can offer stable and economical nitrogen removal solution with these benefits: 

  •  Reduced footprint

  •  Stable operation tactics

  • Economical solution

Compared to conventional nitrification/denitrification (NDN):

  •  Reduces carbon requirements by 90% (for complete removal of inorganic nitrogen)

  •  Reduces oxygen requirements by approximately 60%

  •  Reduces biomass production by approximately 90%

 

Our versatile Anammox solution is a highly effective method of removing nitrogen from sidestream before returning it to the headworks.

High Strength Nitrogen Water

  • Livestock wastewater treatment

  • Single-stage Partial Nitritation/Anammox (PN/A) System

  1. High NH4-N & NO2-N inhibition

  2. Effective inhibition prevention via intermittent aeration & NO2 monitoring

Technology

AMX Species 

 

THE MOST EFFICIENT ANAMMOX SPECIES- "BROCADIA SINICA" JPN1

  • BKT’s unique Anammox Solution: Candidatus “Brocadia sinica” JPN1
     

  •  Original enrichment at Hokkaido University by Professor Satoshi Okabe

  • High growth rate & nitrogen removal rate
    Typical range of full-scale sidestream deammonification (Lackner et al., 2014): 60 - 250 mg N/g VSS/d
    Maximum achieved by highly enriched BKT Anammox species (Okabe et al.): 370 mg N/g VSS/d (33 kg N/m3/d)
     

  • Outperforms other species in mainstream application

 

Interactive Chart

The link on the image provides a Krona chart to demonstrate a progressive increase in Candidatus Brocadia over the pilot operation.

 
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Technology

Overcoming Anammox Challenges

 
 

Using CRISPR to Establish Transgenic Derivatives of Anammox Bacteria

RESEARCH OUTLINE

Anammox (anaerobic ammonium oxidation) bacteria is one of the novel microorganisms used for the removal of nitrogen contained in wastewater. We intend to induce genetic editing by introducing CRISPR gene scissors to anammox bacteria. Ultimately, we want to modify wild-type species with improved treatment efficiency (i.e. have a high growth rate or the ability to grow in low temperature) compared to the standard anammox bacteria. 

RESEARCH GOALS

  • Explore the pure culture conditions of the industrial-level anammox bacterial species

  • Secure functional gene sequences via high-throughput sequencing

  • Develop CRISPR gene editing platform to specifically target anammox functional genes

  • Establish a way to transfer CRISPR gene scissors into the anammox cell

  • Ultimately, establish a way to edit desired functionality through introducing CRISPR gene scissors to the anammox genome strain

RESEARCH BACKGROUND 

Internationally, there is a growing interest in water treatment technologies due to strict regulations on water quality.  More specifically, there is a growing demand on the environmentally friendly technologies for significantly reducing energy consumed by biological treatment of nitrogen in sewage and wastewater. Anammox bacteria has attracted much attention in the field of wastewater treatment due to its ability to reduce nitrogen contents to dinitrogen gas in an anaerobic setting while maintaining a low energy usage and low operating cost.Despite having many advantages, anammox technology is often limited by cold temperature and slow rate of cell division. Because of these limitations, there needs to be technological differentiations to make anammox technology a viable treatment solution.

Since its successful development in early 2013, genome editing using CRISPR / Cas system gene scissors has advanced dramatically in recent years with applications in various fields. CRISPR gene scissors technology is a revolutionary tool for editing specific genes, and is expected to induce the desired functional transformation when applied to anammox bacteria. NGS (next generation sequencing) for genome analysis will be performed after making and applying the CRISPR gene editing platform to the anammox bacteria.  There is a worldwide competition for improving functions in bacterial strains for wastewater treatment technology. By introducing CRISPR gene editing tool to anammox genome, we expect to make a highly versatile transformant that is not limited to industrial use. 

RESEARCH MATERIALS AND METHOD

Strategy 1.
Establishment of optimized culture condition for Anammox bacteria

  • Anammox strain has become largely used in industrial sector, but it must be cultured as a highly enriched strain for genome editing.

  • Establish optimal culture conditions for pure culture strain

Strategy 2.
Acquire functional gene sequences to target in Anammox bacteria

  • When a gene is mutated in a demonstration case, target easily observable genes when they are expressed (i.e., ones that either change colors or react to specific chemicals)

  • Investigate target gene’s nucleotide sequences through databases such as literature and NCBI

  • Obtain the genome sequences in the present strains through next-gen DNA sequencing

Strategy 3.
Making gene scissors for target genes & transforming the scissors to cells

  • Produce a CRISPR gene scissors that can target gene sequences obtained in strategy 2

  • Create at least four CRISPR gene scissors for each gene to determine the most efficient gene scissors

  • Establish a way to transform the produced CRISPR gene scissors most efficiently into cells (make various attempts such as electroporation and lipofectamine on the carrier)

  • Anamox strains which gene scissors have been introduced to are screened for transformants through NGS sequencing

  • Obtain transformants that have been knocked out by CRISPR

EXPECTATIONS

  • Using the CRISPR gene scissors to produce transformants in Anammox is expected to be revolutionary in the process of environment-friendly water treatment

  • In the future, apply CRISPR gene scissors to not only a single gene but also several gene families related to bacterial growth

  • Through screening experiments, produce highly efficient anammox transformant.

 

Bio-Algae Nutrient Removal (BANR)

The Bio-Algae Nutrient Removal (BANR) process is a GRI patented technology that allows nitrification and algae growth to occur in a single reactor. The dissolved oxygen necessary for wastewater treatment using the BBF is supplied through the byproducts of accelerated algal growth, for which the BANR process has overcome the limitation of sunlight dependency by employing LED light in the absence of sunlight.

BANR is a part of several new emerging technologies. Different species of algae are utilized in nitrigen and phosphorus removal as well as for energy neutralization in wastewater treatment plants. This is considered an effective organic pretreatment (BBF). Results of using BANR include CO2 reduction and resource recovery through algae production and reuse.

BANR integration

  • Zero Energy & Zero Chemical consumption

  • Energy production through carbon diversion and biomass processing

  • By removing not only solid organics but also soluble organics, BBF ensures that new energy saving processes such as OBA and BANR, can work properly alongside WWF

 

HOLLOW FIBER MEETS ANAMMOX

AOBs, AMXs, and denitrifiers coexist and organic removal, conventional denitrification, and Anammox reaction are simultaneously carried out in one reactor. 

Using a membrane diffuser, supply oxygen needed only for nitritation

  • Denitrifiers (XHD) denitrify NO2 and NO3 using organic matter in influent

  • AMXs can be used to remove nitrogen through single-nitrogen reaction with NO2 generated by AOB from NH4 in the influent.

  • Maximize oxygen transfer rate (OTE 90-100%) to save energy

CONTROL THE SUPPLIED OXYGEN, CARBON DIOXIDE, AND NITROGEN CONCENTRATION

  • Supply only the amount of oxygen required for nitritation

  • Supply inorganic carbon required for nitrification and anammox reaction

  • Supply gas volume required for reactor agitation and biofilm thickness control

 
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