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®

OWEA 2013 Technical Conference and Exposition June 18 – 21, 2013

Michael H. Gerardi ArchaeaSolutions, Inc. www.archaeasolutions.com [email protected]

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Nitrogenous Wastes in Domestic Wastewater

• Urine ▫ Urea as major component of urine

▫ Chemical formula: NH2COH2N ▫ Urea contains two (2) amine groups (-NH2) ▫ Urea is hydrolyzed in the sewer system ▫ Hydrolysis results in the release of reduced + nitrogen (NH4 /NH3)

3 Dominant Form of Reduced Nitrogen as Determined by pH 100 0

% % + NH3 NH4

50 50

0 100

pH 7 pH 9 pH 11 4 Nitrogenous Wastes in Domestic Wastewater  Feces  Proteins (organic-nitrogen) as a major component  Proteins made of amino acids

 Amino acids contain amine groups (-NH2)  Deamination of proteins and degradation of amino acids in sewer system and aeration tank results in the release of amine groups and the release of + 3 reduced nitrogen (NH4 /NH )  Therefore, organic-nitrogen represents bonded nitrogen (amine groups) that may/must be released in aeration tank

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Amino acid, cysteine

HSCH2CHNH2COOH

Amine group + - - NH3/NH4 /NO2 /NO3 Thiol group

- 2- H2S/H2S/HS /SO4 H2S (1 to 3 mg/L) toxic to nitrifying microbes 6

Free and Organic-Nitrogen

• Free reduced nitrogen + ▫ NH4 /NH3 in solution ▫ Determined by ammonia testing • Organic-nitrogen ▫ Bonded nitrogen that may/must be released ▫ Determined by total Kjeldahl nitrogen (TKN)

and ammonia testing: TKN – NH3 7

Archaeal and Bacterial Roles in Nitrification and Denitrification

• Remove soluble cBOD • Provide maximum time for nitrification • Provide acceptable nitrification and denitrification • Perform cold weather nitrification 8

Archaea

What are Archaea? 9

What are Archaea?  Separate domain of life

 Formerly known as “ancient” bacteria

 Significant differences  Some Archaea degrade wastes that bacteria cannot  Some Archaea out-compete bacteria for substrate and nutrients  Some have a higher metabolic rate than bacteria  Provides for more rapid degradation of cBOD  Provide for more HRT for nitrification  Removes soluble, short chain, inhibitory cBOD  Some have a syntrophic relationship with bacteria 10

Inhibitory Soluble cBOD

Methanol CH3OH 1

Methylamine CH3NH2 1

Ethanol CH3CH2OH 2 n-propanol CH3CH2CH2OH 3

Ethyl acetate CH3CO2C2H5 4 11

What is Arkea®? 12

What is Arkea®?

• ArchaeaSolutions, Inc. produces Arkea®, a proprietary blend of Archaea organisms and other microbes. • Several species of Archaeal methanogens have

been identified in Arkea® through gene sequencing analysis.

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Nitrogen Removal 14

Nitrogen Removal from Wastewater

• Air stripping of ammonia (NH3) + • Assimilation of ammonium (NH4 ) • Nitrification/denitrification + - ▫ Nitrification: NH4 ----- > NO3 ▫ Denitrification NO3------>

 Insoluble molecular nitrogen (N2)  Insoluble nitrous oxide (N2O) ▫ Simultaneous nitrification/denitrification (SND) ▫ Anammox 15

+ Conversion of NH4 to NH3 with increasing pH; Air stripping of NH3 to atmosphere

NH3

Influent Effluent + NH4 16

+ Assimilation of NH4 as nitrogen nutrient for microbial growth (MLVSS production)

Influent Effluent + NH4 MLVSS

Approximately 12 – 15% of microbial dry weight is nitrogen. + NH4 is the preferred nitrogen nutrient. NH3 is toxic! 17

Nitrification*/Denitrification

*Nitrification does not remove nitrogen from the wastewater! 18

What is Nitrification?

+  Biological oxidation of ammonium (NH4 ) to - nitrite (NO2 ) and/or the biological oxidation of - nitrite to nitrate (NO3 )

+ + - NH4 + 1.5 O2 --- > 2 H + NO2 + H2O

- - NO2 + 0.5 O2 --- > NO3 19

Nitrification

Influent Effluent + - NH4 NO2 NO3- 20

Nitrifying Systems

• One-stage system • Two-stage system 21

One-stage Nitrification System

Aeration Tank Removes Clarifier cBOD and Nitrifies 22

Two-stage Nitrification System

Aeration Tank Aeration Tank Clarifier Removes cBOD Nitrifies 23

Microbes That Perform Nitrification?

• Ammonium-oxidizing bacteria + - ▫ NH4 ----- > NO2 ▫ Nitrosomonas and Nitrosospira • Nitrite-oxidizing bacteria - - ▫ NO2 ----- > NO3 ▫ Nitrobacter and Nitrospira • Ammonium-oxidizing Archaea (Crenarachaeota) ▫ AOA more competitive than AOB ▫ AOA more competitive than NOB • Some protozoa ▫ Epistylis ▫ Vorticella 24

Stalk ciliates, Epistylis 25

Stalk ciliate, Vorticella

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Monitoring Nitrification

• Alkalinity/pH

• Temperature

• cBOD including slug discharges

• Inhibitory/toxic wastes + • NH3/NH4 concentration in aeration tank • Dissolved oxygen • Phosphorus • Form of nitrification (complete, incomplete #1, 2, 3, 4) 27

Forms of Nitrification*

Complete < 1 mg/L < 1 mg/L > 1 mg/L Incomplete 1 < 1 mg/L > 1 mg/L < 1 mg/L Incomplete 2 > 1 mg/L < 1 mg/L > 1 mg/L Incomplete 3 < 1 mg/L > 1 mg/L > 1 mg/L Incomplete 4 > 1 mg/L > 1 mg/L > 1 mg/L

*Based upon mixed liquor effluent filtrate from an on-line aeration tank. 28

What is Denitrification?

- • The biological use of typically nitrate (NO3 ), - atypically nitrite (NO2 ) by denitrifying microbes to degrade soluble cBOD. • Denitrification occurs in the absence of free

molecular oxygen (O2). 29

Denitrification Tank

1-stage or 2-stage Denitrification Clarifier nitrification system Tank Denitrification tank: Slow, surface mixing Carbon Source 30

Denitrification

- Soluble cBOD (methanol) + NO3 --- >

cells (sludge) + H2O +

   - CO2 + N2 + N2O + OH

3 gases produced return of alkalinity 31

Monitoring Denitrification

• Soluble cBOD or “carbon” source • Residual dissolved oxygen 32

Simultaneous Nitrification/Denitrifica tion (SND) 33 Oxygen Gradient

- X-sectional view of floc particle with oxic (O2 use) and anoxic (NO3 use) zones

> 100 μm Soluble cBOD OXIC ZONE

Dissolved oxygen

Nitrate ANOXIC ZONE

Perimeter: DO < 1.0 mg/L; cBOD removal via O2; nitrification occurs; destroy alkalinity; grow low dissolved oxygen filamentous organisms.

- Core; DO is “0;” cBOD removal via NO3 ; denitrification occurs; return alkalinity; destroy low dissolved oxygen filamentous organisms. 34

Anaerobic Ammonia Oxidation

(Anammox) 35

+ Organic-nitrogen  ammonification  NH4

AOB + O2

↑ - N2 NO2

Denitrifying bacteria NOB + O2 + carbon source - NO3

Nitrification/Denitrification 36

+ Organic-nitrogen  ammonification  NH4 Anammox Archaea Anammox bacteria

+ O2

↑ - N2 NO2 Anammox Archaea Anammox bacteria

Denitrifying bacteria NOB + O2 + carbon source - NO3 Anammox Reduced dissolved oxygen requirement; “carbon” addition not required. 37

Benefits of Anammox

• Use represents 10% of current cost for nitrification and denitrification

• Reduces greenhouse gas carbon dioxide (CO2) addition 38

ArchaeaSolutions, Inc. Consulting.Bioaugmentation.Results.

Scientists, Engineers, and Wastewater Professionals Specializing in Process Consulting, Bioaugmentation Strategies, Plant Optimization, and Troubleshooting Support

100 Lloyd Ave., Suite D, Tyrone, GA 30290 770-487-5303 www.archaeasolutions.com [email protected]

Low Cost “Performance-Based” Arkea® Trials Available Call us today to schedule a consultation and site visit!