PROGRESSIVE BREWERY WASTEWATER MANAGEMENT STRATEGIES©

A Thesis

Presented to

The Faculty of Graduate Studies

of

The University of Guelph

by

SCOTT JAMES MASSEN

In partial fulfilment of requirements

For the degree of

Master of Science

June 2011

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While these forms may be included Bien que ces formulaires aient inclus dans in the document page count, their la pagination, il n'y aura aucun contenu removal does not represent any loss manquant. of content from the thesis. Canada ABSTRACT

PROGRESSIVE BREWERY WASTEWATER MANAGEMENT STRATEGIES

Scott James Massen Advisors:

University of Guelph, 2011 Professor Hamidreza Salsali

Professor Ed McBean

Brewery wastewater effluents exhibit high concentrations of nutrients and suspended solids. Treatment is further complicated by the batch-discharge nature of brewery wastes, and a widely variable pH.

Effluents discharged directly to municipal sanitary systems are often subject to a high strength sewage surcharge (HSS) and can be a major expense to the brewer. The proper identification of contributory waste streams, by-product diversion and disposal techniques, and end of pipe treatment are essential to the development of an effective wastewater management strategy. This thesis contains a compliment of initiatives directed at reducing HSS fees at a local brewery, including a wastewater characterization study, value-added through anaerobic digestion of brewery by-product study, and investigation into a biological packed tower for end-of-pipe wastewater treatment. The major conclusions of this study will contribute to the formation of a stronger brewery wastewater management program. ACKNOWLEDGEMENTS

The author of this report wishes to extend a special thanks to the following contributors:

University of Gueloh

Dr. Edward McBean: Academic Advisor

Dr. Hamid Salsali: Academic Advisor

Joanne Ryks: Laboratory Coordinator

Laura Wright Head Technican Elora Dairy Cattle Research Station

Joel Citulski: PhD Candidate

Victoria Hillbom: MASc. Graduate

Allison Chan: MASc. Graduate

Victoria Sharpe: Co-op Student

Labatt Breweries London. ON

Charlotte Armstrong: Brewery Support Manager - Environment Health, Safety

Debbie Cassel: Environment and Safety Specialist

Emily Hahn-Trnka: Environment and Safety Specialist

Jordan Meunier: Environment and Safety Specialist

Dan Swystun: Co-op Student

Alex Demianiuk: Co-op Student

Steve Isaacson: Brewery Operations Specialist

John McAllister: Brewery Operations Specialist

Scott Durnin: Capital & Utilities Specialist

Jay Cooke: Brewhouse Floor Manager

Terry Mine: Brewmaster

Patrick Hoofnaggles: Maintenance - Plumber

Jim Fragily: Maintenance - Electrician

Steve Eastendon: Maintenance - Electrician

Ashley Farrington: Quality Control Laboratory Technician TABLE OF CONTENTS

Chapter 1: Proposal arid Objectives 1

Chapter 2: A Literature Review on Brewery Water and Wastewater Management 4

2.1 Introduction to Brewing and Brewery Wastewater 4

2.2 A Background on the Brewery Industry 5

2.2.1 Labatt London, ON 5

2.2.2 Product Ownership 6

2.2.3 ABInBev Environmental Policy 6

2.3 The Brewing Process 7

2.3.1 Malt Preparation 9

2.3.2 Milling and Mashing 10

2.3.3 Wort Cooling and Fermentation 13

2.3.4 Maturation and Clarification 17

2.3.5 Bottling and Pasteurization 18

2.3.6 Powerhouse 23

2.3.7 Clean In Place 24

2.4 Wastewater Legislation for Breweries 25

2.5 Brewery Wastewater Constituents 31

2.5.1 Oxygen Demand 31

2.5.2 Total Suspended Solids 34

2.5.3 pH 34

2.5.4 Inorganic Pollutants 35

Chapter 3: Labatt London Wastewater Characterization 37

3.1 Wastewater Characterization Project Introduction 37

3.2 Wastewater Characterization Methodology 39

3.3 Wastewater Characterization Results 39

i 3.3 Wastewater Characterization Discussion and Recommendations 43

3.3.1 Brewhouse 43

3.3.2 Fermentation and Aging 43

3.3.3 Filtration 46

3.3.4 Bright Beer 49

3.3.5 Packaging 49

3.3.6 Powerhouse 52

3.3.7 Clean In Place 53

3.3.8 Main Effluent 53

Chapter 4: Added Value from Rebate Beer in Anaerobic Digestion 58

4.1 Introduction to Anaerobic Digestion 60

4.1.1 Background on Anaerobic Digestion 60

4.1.2 Biogas Facilities 63

4.1.3 Rebate Beer as a Co-Substrate 64

4.1.4 Digester Objectives 65

4.2 Bench Scale Digester Project Methodology 66

4.2.1 Source of Waste 66

4.2.2 Preliminary Biochemical Methane Potential Test 67

4.2.3 Digester Design 69

4.2.4 Start up Procedure and Operation 71

4.2.5 Digester Sampling Plan 75

4.2.6 Physical and Chemical Analyses of Digestate 76

4.3 Digester Results and Discussion 81

4.3.1 Feed Characteristics 81

4.3.2 Steady-State Digester Performance 84

4.4 Recommendations for Anaerobic Digestion of Brewery Wastes 91

ii 4.5 Anaerobic Digestion Project Conclusions 94

Chapter 5: Biological Packed Tower Retrofit Pilot 96

5.1 Introduction to BioTower Pilot 96

5.1.1 Problem Definition 96

5.1.2 Retrofit Opportunity 97

5.1.3 Background on Attached Growth Processes 98

5.2 BioTower Pilot Methodology 102

5.2.1 Experimental Apparatus 102

5.2.2 Daily Maintenance 106

5.2.3 Sampling Analysis 106

5.3 Results and Discussion 107

5.3.1 Operational Issues 108

5.4 Recommendations for End-of Pipe Treatment 113

5.5 Conclusions 114

Works Cited 116

Glossary of Terms 120

iii LIST OF FIGURES

Figure 1: Basic Overview of Labatt Brewing Process (Labatt, 2007) 8

Figure 2: Clean-ln-Place Flow Diagram 24

Figure 3: Comparative Sewer System Charge Overview - Industrial (City of London 2008) 26

Figure 4: 2000-2009 High Strength Sewage Surcharge for London, ON 30

Figure 5: Average Wastewater Profile of Manual Clean Process for Krausen Tank 45

Figure 6: Diatomaceous Earth Slurry Particle Size Distribution 48

Figure 7: COD and BOD5 Loading of Brewery Effluent May 2009 to February 2010 54

Figure 8: TSS and VSS Loading of Brewery Effluent from May 2009 to February 2010 55

Figure 9: Labatt Brewery Main Effluent Particle Size Distribution 55

Figure 10: TP Loading and pH of Brewery Effluent May 2009 to February 2010 56

Figure 11: The carbon flow in the methane production process 61

Figure 12: BMP of dairy manure and rebate beer 68

Figure 13: Schematic drawing of lab digester 70

Figure 14: Methane production normalized by volatile solids destruction 87

Figure 15: Comparison of the oxygen demand reduction from digested feed stocks with varying dairy manure to beer ratios during steady state operation of bench scale anaerobic digester 89

Figure 16: A comparison of the solids destruction from digested feed stocks with varying dairy manure to beer ratios during steady state operation of bench scale anaerobic digester 91

Figure 17: Biological Packed Tower Schematic 103

Figure 18: Equalization Tank Arrangement 105

Figure 19: Results of COD reduction 24 hr. time series in brewery wastewater with ammonia nitrate supplements for investigation into the possibility if a nitrogen deficiency affecting biological wastewater treatment Ill

iv Figure 20: Digester operation on trial 1of the 50% DM/50% RB feedstock 178

Figure 21: Digester operation for trial 2 of the 50%RB/50%DM feedstock 179

Figure 22: Solids analysis for the 50% DM/50% RB 180

Figure 23: Solids destruction for the 50% DM/50% RB 180

Figure 24: Solids analysis for the 75% DM/25% RB 180

Figure 25: Solids destruction for the 75% DM/25% RB 180

Figure 26: Solids analysis for the 100% DM 180

Figure 27: Solids destruction for the 100% DM 180

Figure 28: Digester Performance on the 75% DM/25% RB blend 181

Figure 29: Digester Performance on the 100% DM blend 182

Figure 30: Effluent VFA and Bicarbonate Concentrations of the 2nd 50% DM/50% RB trial 183

Figure 31: Effluent VFA and Bicarbonate Concentrations of 75% DM/25% RB trial 183

Figure 32: Effluent VFA and Bicarbonate Concentrations of 100% DM trial 184

LIST OF TABLES

Table 1: The composition of barleys grown in Sweden and Montana (% (w/w) d.b.) (Briggs 1997) 10

Table 2: City of London 2007 Limitations on Discharges to Sanitary Section 4.8 29

Table 3: Approximate COD Content of Various Brewery Waste Streams (United Nations 1999) 33

Table 4: Loading of BOD and Suspended Solids from Various Brewery Waste Streams 34

Table 5: Results From Brewery Wastwater Characterization 41

Table 6: Dairy Cow Feeding Regiment 67

Table 7: Amount of substrates required for BMP test of rebate beer and dairy manure 68

Table 8: Primary Digester Components List 71

Table 9: Digester Feedstock Blends 71

v Table 10: Frequency of analytical parameters 75

Table 11: Average Characterization of Undiluted Feedstock 82

Table 12: Summary of Digester Operations 85

Table 13: Summary of Chemical Parameters in Digester Effluent 88

Table 14: Summary of Digester Performance through Reduction of Physical and Chemical Parameters..90

vi LIST OF APPENDICES

Appendix A: Brewery Wastewater Characterization Sample Detail 127

Appendix B: labatt Wastewater Process Flow Diagram 150

Appendix C: Standard Methods for Laboratory Analysis 152

Appendix D: Supplementary Bench-scale Anaerobic Digester Performance Data 177

Appendix E: BioTower Drawings 213

Appendix F: BioTower Maintenance and Performance Summary 216

Appendix G: US Centrifuge Results. 224

vii LIST OF SYMBOLS

ABInBev Anheuser-Busch InBev

AGP Attached Growth Process

AS Aerobic Sludge

0 Temperature activity coefficient

BA Bicarbonate

BODs Five-day Biochemical Oxygen Demand

CDAF Centrifuged Dissolved Air Flotation Sludge

CIP Cleaning In Place

COD Chemical Oxygen Demand

CSTR Continuously Stirred Tank Reactor

Cvs Concentration of volatile solids in feedstock cw City Water

DAF Dissolved Air Floatation

DCW De-aerated Carbon-Filtered Water

DE Diatomaceous Earth

DM Dairy Manure

F Actual to allowable wastewater strength ratio

FOG Fats, Oils and Greases

GC Gas Chromatographer

H2S Hydrogen Sulfide

HDPE High Density Polyethylene

HRT Hydraulic Retention Time (h)

HSS High-strength Sewage Surcharge kd Microbial decay coefficient

Ks Saturation constant (mg/L)

KPI Key Performance Indicator

KW Kitchen Waste

viii L Unit of Length

M Unit of Mass

MW Molecular Weight (g/mol)

NH3 Ammonia

OLR Organic Loading Rate

PM Pig Manure

Q Flow rate rsu Substrate Utilization Rate

R High Strength Sewage Service Rate

S Concentration of growth limiting substrate

SRT Sludge Retention Time t Time (hr)

T Unit of Time tCOD Total Chemical Oxygen Demand sCOD Soluble Chemical oxygen Demand

TKN Total Kjeldahl Nitrogen

TP Total Phosphorous

TS Total Solids

TSS Total Suspended Solids

1 |jm Maximum Specific Growth (h )

V Volume

V0 Working Digester Volume

Vfeed Volume of digester influent

VFA Volatile Fatty Acids

VS Volatile Solids

VSS Volatile Suspended Solids

X Initial Microbial Concentration (mg/L)

Y Maximum Biomass Yield Coefficient (gbiomass/gsubstrate)

ix Chapter 1: Proposal and Objectives

Labatt Brewery of London Ontario has actively sought methodologies and practices to effectively reduce their wastewater generation and loading. Labatt's ultimate ambition for wastewater management is to satisfy increasingly stringent regional wastewater Bylaw objectives. This would alleviate the $1 million plus per annum High-strength Sewer Surcharge (HSS) expenditures incurred by discharge of brewery effluent to sanitary. The University of Guelph (U of G) was requested to submit a proposal in January

2009 to evaluate optimization strategies to improve the effluent quality of the Labatt brewing facility located in London, Ontario.

Brewery wastewater from various process contributories is collected and directed to a central effluent pit for pH adjustment before discharge to the municipal sanitary sewer. There is presently no other treatment applied to the main effluent. The total average wastewater flow rate is 5,000 and 6,000

m3/day. The biological oxygen demand (BODs) and total suspended solid (TSS) concentrations for the final effluent range from 1,000 to 1,400 mg/L and 400 to 700 mg/L, respectively. Wastewater strength and volume is subject to seasonal variability based on production. The facilities water-use to beer- produced ratio of 5.9 to 1 is average relative to industrial brewery standards.

Industrial wastewater concentrations in Canada are regulated under the municipality. The City of

London's Waste Discharge Bylaw (WDB) is tailored to the unique capabilities of its wastewater treatment plant. Under the Bylaw, BOD, TSS, and TP contributions that exceed 300 mg/L, 350 mg/L and

10 mg/L respectively are subject to a High-strength Sewage Surcharge (HSS) (City of London 2007). The current HSS Rate of 40.8C/m3 (2009) has been subject to an average annual increase of 9% over the past several years.

Preliminary wastewater testing of the various process contributories were previously completed by

Labatt. Wastewater volumes, concentrations, and discharge characteristics were identified for several

1 primary unit processes. Labatt has estimated that approximately 40 percent of the wastewater is generated from packaging, 50 percent from brewing processes and an additional 10 percent from miscellaneous sources. Limited data sets and omission of certain processes resulted in a recommendation for further wastewater characterization. An independent wastewater characterization project was formulated between Labatt and the University of Guelph to verify and expand upon the initial findings.

Labatt's specific objectives for this project were:

1. To conduct a wastewater characterization of the various brewing process effluent streams;

2. To investigate alternatives for waste byproduct disposal, particularly whether waste byproducts

can be sold to generate additional revenue; and

3. To identify and assess biological packed tower for reducing BOD5 and TSS in the discharge

stream that are applicable to the Labatt facility.

Criteria

1. HSS savings are to be maximized;

2. Proposed new equipment to have minimal area footprint;

3. Emphasis on solutions that can be implemented with minimal impact on production.

Over the course of the project, the terms of the initial proposal began to evolve as new opportunities presented themselves. The final work resulted in the completion of four technical memoranda that have become chapters within this thesis:

1. A critical review and analysis of the technical literature in regard to the impact of brewing waste

on organic loading of the local municipal sewage was completed (see Chapter 2).

2. Wastewater characterization study of brewery unit process streams (see Chapter 3).

2 3. The application of rebate beer as a co-substrate for the anaerobic digester of dairy manure was

investigated as an avenue for brewery byproduct diversion (see Chapter 4).

4. A pilot scale biological packed tower for end-of-pipe treatment to test the feasibility of

converting existing wastewater treatment infrastructure to have a biological wastewater

treatment component (see Chapter 5).

3 Chapter 2: A Literature Review on Brewery Water and Wastewater

Management

2.1 Introduction to Brewing and Brewery Wastewater

The brewing of beer is an art form steeped with cultural heritage and significance. As the fifth most widely consumed beverage in the world behind tea, milk, carbonates, and coffee, beer is a lucrative economic commodity. In 2002, over 1.34 billion hectoliters of beer were brewed (Fillaudeau. L, 2006).

Brewers largely remain faithful to ancestral practices to preserve the quality and traditional flavours of their product. Brewing is one of the most water-intensive food processing industries in practice, and generates a correspondingly large quantity of high strength wastewater. Excessive concentrations of biochemical oxygen demand (BOD), total suspended solids (TSS), and total phosphorous are naturally present in brewery effluents. Increasingly stringent environmental regulations are challenging breweries to drastically reduce water consumption and wastewater generation to remain economically formidable as an industry. Modern technologies and innovations to streamline the process are being implemented to create a higher quality product while reducing the environmental impacts of the industry.

The following is a literature review on the brewing process that will provide the reader insight into the development of a brewery wastewater characterization. The challenges described herein are shared by breweries across the world; however, the examples contained within the body of this report are most relevant to Labatt London. This chapter aims to identify brewing process improvement opportunities and alternatives in the context of resource management for the benefit of the brewery, the community, and the environment. An examination of the brewing process with an environmentally mindful perspective is necessary to understand the distribution and fate of resources. With the brewing process in mind water usage, solid waste generation, and wastewater can be better understood. To situate the

4 challenges for breweries within legislative context, a discussion of the municipal guidelines is provided.

This is intended to exemplify the current state of understanding and concern for wastewater management. Strategies will then be suggested for brewery process improvements to achieve environmental policy obligations while epitomizing sustainability.

2.2 A Background on the Brewery Industry

2.2.1 Labatt London, ON

John Kinder Labatt built his first brewery in London Ontario in 1847. The first brewery made close to one thousand bottles of beer a year. Full production began in 1852. Presently the London Brewery has an annual production capacity of 5 million hectoliters, and operates 6 days a week, 24 hrs a day. Labatt produces more than 60 brands. Labatt Breweries currently produce 15 National Domestic Brands, 20

Regional Domestic Brands, 13 International Premium Brands, as well as a diverse portfolio of low alcohol beers. Labatt products have been well recognized for their quality, and have been the recipient of numerous awards. The London division produces many notable national brands including Labatt Blue,

Budweiser, and Bud Light, specialty beers including Bass Ale (British), and Brahma (Brazil), and local flavour beers such as Labatt Crystal (Ontario).

As the Labatt brewery marked its success, the City of London thrived. Municipal development continued to intensify around the brewery until present day. Currently Labatt is nestled at the heart of London on

150 Simcoe St. The underlining implication of this location is in the procurement of new real estate for future expansions. The economics of future development are constrained to the optimization of existing manufacturing processes and spaces in an effort to reduce the facility's environmental footprint.

5 2.2.2 Product Ownership

In 1945 Labatt became a publically-traded company issuing 900,000 shares. In 1946 marked the purchase of the Copland Brewery in Toronto, marking the company's first step to becoming a national brewer. This triggered a period of rapid national expansion, acquiring ownership of Lucky Lager in British

Columbia (1958) to Bavarian Brewing Limited in Newfoundland (1962), and with multiple breweries in between. Currently eight Labatt breweries exist in Canada.

Labatt was acquired by Interbrew in 1995 which merged with Brazilian company AmBev in 2004 to create the existing parent company InBev. Labatt beverages are now brewed globally. In 2008 InBev merged with Anheuser-Busch. The new conglomerate adopted the title Anheuser-Busch InBev or

ABInBev for short (Anheuser-Busch InBev 2009). There are presently 124 beverage plants operating globally under the ownership ABInBev, including Labatt London (Anheuser-Busch InBev 2009)

2.2.3 ABInBev Environmental Policy

ABInBev has a number of progressive accomplishments concerning environmental stewardship. They utilize Voyager Plant Optimization (VPO) and Key Performance Indicator (KPI) programs as an internal auditing system to monitor progress. They actively work with legislative authorizes and non-government organizations.

U.S. brewery water usage was reduced by almost 7% over the past five years, resulting in a saving in excess of 4.7 billion litres of water. ABInBev breweries and soft drink plants report a water use ratio of

5.29 hIwater/hIbeverage for 2007. The best plant is in Hannover with a ratio of 3.19 hl/hl.

ABInBev energy conservation programs achieved a 22.2% reduction in usage since 2005. 20 years ago a

Bio-Energy Recovery Systems (BERS) technology was introduced. BERS reclaim brewery wastewater to create renewable energy (biogas) that now provides up to 15 percent of the fuel for nine of its U.S. breweries, as well as its brewery in Wuhan, China.

6 ABInBev reduced C02 emissions per hectoliter of production by 5.5% for a total reduction of 13.6% since

2005 (InBev 2008). However with 32 additional plants reporting data for the first time in 2007 an overall increase of 16.8% compared to 2006 was experienced. In 2007, ABInBev reused or recycled 97.2% of their total solid waste and byproducts (Anheuser-Busch InBev 2009).

Wastewater management, however, remains an area ripe for improvement in many jurisdictions of the beverage industry.

2.3 The Brewing Process

Intimate knowledge of all employed materials, manufacturing process, and techniques are required to accurately assess the environmental impacts of a given industry. The beer industry is highly guarded about the unique brewing techniques to create their beverages of distinction; however the overall brewing process is relatively the same. Figure 1 depicts the generic sequence of brewing operations.

Labatt brews approximately 54 batches a week, and produces over a dozen brands. One batch of beer can take upward of a month to complete. Each brand has a distinct material composition, tailored brewing process, and final presentation. Tweaks to the brewing process from ingredients composition, process temperatures, to aging, carbonation, filtering, and packaging methods are what define the qualities of a given beer. The following sections present a brief description of major brewing operations.

This section will detail the general materials and practices involved in brewing beer on an industrial scale to help the reader identify key areas of water consumption, waste production, and wastewater generation.

7 The Brewing Process 10 *> HH

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Figure 1: Basic Overview of Labatt Brewing Process (Labatt, 2007)

8 2.3.1 Malt Preparation

Malt is derived from cereal grains. Typical malt beverages include beer, ale, porter, and stout (Madigan and Martino 2003). Malted grain is principally barley, occasionally amended with corn, rice, or wheat at the brewer's discretion. The nutritional characteristics of a given grain are dependent on the climate and growing season. A dry season will produce a grain with a higher percentage of proteins, whereas a wet season will produce a grain with a higher starch count. As a result, the necessary amount of water used to make malt from cereals may vary slightly from batch to batch.

To create malt, the grain is germinated for a limited period then dried. During the malting process, naturally occurring enzymes in the cereals convert starches to sugars. These sugars will be metabolized by brewing yeast to produce alcohol during fermentation. In addition, some proteins and amino acids are hydrolyzed which increases their accessibility to yeast. Malt preparation from barley involves the following steps (Nemerow and Agardy 1998); (United Nations 1999).

i. Screening: kernels are separated according to size, the smallest usually being sold as fodder;

ii. Steeping: bleaches out colour and increases barley water content to about 45%. Up to three

water changes may be required;

iii. Germination: steeped grain is transferred into boxes with perforated steel plate bases that are

agitated and aerated for 120-190 hours. Temperature and moisture content are controlled by

the aeration. (Note: barley grain may start germinating before being moved to the germination

boxes following steeping in this process);

iv. Drying: the germinated grain is dried in a kiln to reduce its moisture content to around 4%. This

halts the germination process, develops flavour characteristics, and prolongs the malts shelf life.

Sulfur dioxide may be used to bleach the kernels and lower the pH of the malt during this

operation. During kilning, some non-enzymatic reactions occur between sugars and amino acids

9 which contribute to the colour of beer. Darker beers tend to be kilned at higher temperatures to

promote browning reactions (Moss, 2007);

v. Polishing: sprouted rootlets from germination are removed;

vi. Storage: dried and polished grain is finally stored in elevators for no less than four weeks until

the next operation.

2.3.2 Milling and Mashing

Malted barley is ground in a manner that leaves husks intact while pulverizing the grain interior into a coarse starch and enzyme-rich powder. The method of malting may alter the constituent distribution of barley. Labatt considers approximately 75-77% of their malt as available sugar, which may be a slightly high approximation. Table 1 shows the average composition of barely.

Table 1: The composition of barleys grown in Sweden and Montana (% (w/w) d.b.) (Briggs 1997)

Two-rowed barlevs Six-rowed barlevs Sweden Montana Sweden Montana Mean Range Mean Range Mean Range Mean Range Number of varieties analyzed 81 16 11 7

Glucose 0.3 0.1-0.8 0.2 0.06-0.3 0.6 0.2-1.4 0.2 0.06-0.3

Fructose 0.1 tr.-0.4 0.1 0.05-0.3 0.2 0.1-0.5 0.1 0.09-0.2

Sucrose 1.6 0.6-3.1 1.9 1.4-2.3 1.9 1.1-3.9 2.0 1.6-2.3

Fructan 0.4 tr.-0.8 0.5 0.4-0.8 0.3 tr.-l.O 0.5 0.4-0.7

Starch 62.2 55.9-66.6 57.2 53.3-59.8 58.9 52.9-64.1 57.2 53.0-60.6

Crude protein 10.7 8.6-13.4 14.4 12.2-16.8 11.5 8.9-14.0 13.8 12.2-15.8

Crude fat 3.0 2.7-3.3 2.8 2.3-2.7 3.3 2.8-3.7 2.7 2.1-3.3

Ash 2.4 1.8-2.9 2.9 2.3-4.0 2.4 2.2-2.7 3.1 2.3-3.8

Total Fibre 19.3 14.0-24.7 19.9 17.6-22.9 20.9 17.8-23.8 20.5 17.6-23.0

*tr. Trace amount

Starches and fructans degrade into simple sugars by way of enzymatic action the instant ground malt comes in contact with brewing water in a vessel called a mash tun. It can be assumed that enzyme

10 activity is sufficient to completely mobilize these materials into solution. The source and treatment of the brewing water is essential to the quality of the beer. Brewing water is often acidified to prevent the excessive dissolution of chill haze forming proteins. Budweiser brands include hulled rice with malted grains in the lauter tun. Rice is primarily starches. Brewmasters at Labatt suggest rice has an 81% available sugar content.

The mash is heated to 75°C -78"C and transferred to the lauter tun to separate the husks from the sweet liquid, termed wort (United Nations 1999). The lauter tun has a false, screened bottom that captures solids and allows the liquid to seep through. Sweet wort is filtered as it percolates through the bottom of the established grain bed, where it is then sent on to the brew kettle. At Labatt, the lauter tun diameter is 9.75 m. Malt bed depth is generally 0.36 to 0.41 m. The initial glucose concentration of the

wort is generally around an upper limit of 22°Plato, or 239 800 mg giUCose/l- Plato and specific gravity are related by the following empirical equation at a reference temperature of 15.5°C (Manning 2009):

259 ^ SG = 259 -°P where: SG = Specific gravity

°P = Degrees plato

Applying the above equation, the density of Labatt and Budweiser wort is 1070 kg/m3 and 1061 kg/m3 respectively. Once the lauter tun runs dry, the spent grain bed are further sprayed with water in a process called sparging to assist in mobilizing additional sugars. Washings are monitored for sucrose content, and extraction is stopped when the sucrose strength reaches the 1-2° level on the Plato scale.

Weak wort is diverted to the post runoff (PRO) tank instead of the brew kettle once percolate has

reached a lower bound of l'Plato for Labatt Blue (10 000 mgg|UCOse /L) and 2°Plato for Budweiser (or 20

100 mg glucose/L)' This weak wort is termed PRO. The PRO Tank at Labatt has a 25 m3 capacity. Labatt brands reuse PRO, whereas Budweiser brands do not.

11 The residual solid material in the lauter tun after sparging is termed 'spent grains'. One hundred kilograms of malt produces 125-170 kg of spent grains with 80-85% moisture content. Spent grains contain 20-23% dry matter, 19-28% crude protein, 3-5% ether extract, 18-20% crude fibre, 8% fat, and a good profile of amino acids (T. Goldammer 2008); (Saxena 2004).

The wort is boiled in the brew kettle for about 1.5 hours. This results in:

• Halting enzymatic action;

• Wort sterilization and concentration;

• Coagulation and precipitation of undesirable proteinaceous material.

Adjuncts, such as maltose syrup (81% maltose, 19% water)1, hops and hop extracts, gruit, minerals, and/or coagulants, are added during wort boiling. The adjuncts act as flavouring/bittering agents. Hops are the most common bitter agents. They are related to the cannabis family, and have beneficial antibiotic properties that increase the shelf life of the finished product. It is estimated that the production of one hectoliter of normal lager beer requires about 15 kg of solid materials, including malt.

When brewing is complete, the wort is then clarified in a hydro-cyclone or whirlpool, or strained to remove precipitates that may impart undesirable odours and flavours on the wort. This material is called trub, and consists of spent hops and/or other adjunct. Mature unfermented Labatt wort is 17.5°Plato

(188 900 mg glucose/1-)- Mature unfermented Budweiser wort is 15°Plato (161 700 mg giUcose/t-).

The principal polluting load in the brewhouse is the stillage or spent grain, the residual grain mash from in the lauter tun. As much of this as possible is recovered by the industry as a by-product for manufacturing animal feed or for conversion to chemical products. Trub is often combined with the spent grain for animal food; however it may impart a bitterness that makes it less desirable to livestock.

Trub is very high in nitrogen. When combined with sawdust, it makes a good fertilizer or soil amendment for use on mushroom farms. Without such recovery, the population equivalent of distillery

1 Labatt Blue only

12 wastes, based on BOD, would be about 50 000 for each 1000 bushels of grain mash. Screening out the dried grains reduces the population equivalent to 30 000. Complete stillage recovery makes possible a population equivalent of only about 2500 and a large volume, but weak waste (Hardwick 1995).

2.3.3 Wort Cooling and Fermentation

Clarified wort is cooled to about 10°C before entering fermentation. Further precipitation of proteins and tannin components occurs at this stage. The wort is then aerated with pure oxygen to 15-20 ppm before transfer. The solids in the product at this stage are almost entirely dissolved and colloidal; the suspended-solids content is seldom above 200 ppm (Hardwick 1995).

The fermentation process converts sugar wort into alcohol by the action of yeast on the wort. Yeast is pitched into the wort in large steel tanks known as fermenters. 550 kg of yeast are pitched in a 700 hL

3 brew (7860 gyeast/mwort )- Yeast is a facultative anaerobe. During the 5-14 day residency time in the

Fermenters, the yeast population roughly doubles. For aerobic metabolism, oxygen is limiting substrate, whereas for anaerobic metabolism, glucose is the limiting substrate.

There are two major of categories of brewery yeast: top fermenting (Saccharomyces Cerevisiae) and bottom fermenting (Saccharomyces Carlsbergensis). Top fermenting yeasts have a uniform distribution

within the fermenting wort due to re-suspension by the C02 gas generated during fermenting; bottom yeast settle out. Top yeasts are used to brew ales, whereas bottom yeasts are used in lager beers.

Fermentation by top yeast usually occurs at higher temperatures (14-23'C) than bottom yeast (6-12°C) and is accomplished in a shorter number of days (5-7 for top yeast versus 8-14 days for bottom yeast)

(Madigan and Martino 2003).

Low fermentation beer is produced through two fermentation steps, the primary fermentation being when 90% of the fermentable matter is consumed. A rapid cooling of the tank halts this fermentation and causes the flocculation of insoluble particles and the sedimentation of yeast. The tank bottom

13 becomes full of yeast and "green beer". At present, the fermentation tank bottom generates a beer loss of around 1-2% of production (Fillaudeau. L 2006).

A unit of yeast biomass can be estimated as having a molecular composition of C5H7NO2 (Metcalf and

Eddy, Wastewater Engineering: Treatment and Reuse: Fourth Edition 2003). Therefore:

glu cos e yeast 3 C6Hn06+802 + 2NH3 2 C5H7N02+8C02 +UH 20 3(180) 8(32) 2(17) 2(113)

12]

A(CSH7NQ2) 2(113 glmok) SmD A(Ct«,A as BOD) 3(l80g/mOfeXl.07s,OT/gs„„,J '

where Y = maximum biomass yield coefficient (gbiomass/gsubstrate)

The unbalanced stoichiometric equation for anaerobic metabolism of glucose by yeast is:

glucose veast ethanol glycerol succinate C6Hl206+ NH3 -> C5H7N02+C2H50H+ C3H5(OH)3+ C4H404+ C02 + H20 [3] (180) (17) (113)

Aerobic metabolism is much quicker than anaerobic metabolism. The wort is aerated to quickly promoted cell growth; however alcohol (ethanol) is not produced until the majority of oxygen is used up.

The final glucose concentration of the fermented wort termed 'green beer' is 2"P (21000 g/m3) to 2.5°P

(25 400 g/m3) for Labatt Blue and Budweiser. Dry yeast contains 50-60 percent proteins, 15-35 percent carbohydrates and 2-12 percent fats (T. Goldammer 2008).

Yeast growth can be modelled by applying the Monod equation:

dS) _ ju.XS rsu ~ [4] dtju Y(Ks+S)

where rsu= substrate utilization rate (mg/L-h)

14 S = the concentration of limiting substrate (mg/L)

t = time (h)

1 (jm = maximum specific growth (h )

X = initial yeast concentration (mg/L)

Ks = saturation constant (mg/L)

Y = maximum biomass yield coefficient (gbi0mass/gsubstrate)

The maximum biomass yield coefficient may be expressed as in equation [5].

Substitution of [5] into [4] yields an expression for the concentration of yeast at given time.

3»"jc.+s ' as 161

Growth kinetics for a variety of yeasts is available in literature. For example, aerobic glucose-limited

Yeast CBS 0866 kinetics at 30°C coefficients for |Jm, Ks, and Y are 0.5 h"\ 40 mg/L, and 0.39 gyeast/ggiucose

respectively (deKock 2000). A kd of 0.1 at 20°C may be assumed using the average ranges given by

Metcalfe & Eddy 2003. Laboratory analysis using the IWA Protocol's respirometric method would be required in order to determine the actual growth kinetics of Labatt brewing yeast for use in equation 10.

The initial conditions for the aerobic metabolism are:

3 S(0) = 18,7go2/m

3 X(0) = 7857 gyeast/m

15 Both kd and |Jm must be corrected for temperature using the Arrhenius equation:

.. .. actual ^rcf') ' mTactual ' mTrcf

[7]

Jf _ b djacmal~Tref) d Tactual ~ K,dTrefU

where: 8 = temperature activity coefficient (1.024)

Tactual = actual recorded temperature

Tref = reference temperature for the empirical nm or kd value.

1 1 Application of equation [7] yields a of 0.31 h' (or 7.44 day" ) and a kd of 0.0789 gyeast/gyeast day at

10°C. The Monod equation coefficients are assumed constant. Ideally these equations would be solved using two separate sets of Monod kinetics; one for aerobic oxygen limited metabolism and one for anaerobic glucose limited metabolism. It is assumed that anaerobic metabolism will dominate once

anoxic conditions are achieved in the fermenter (02 < 2 mg/L).

Excess yeast, carbon dioxide, and heat are the main by-products of fermentation. Excess yeast, along with residual trub that escaped the whirlpool, deposits in thick slurry at the bottoms of fermentation and aging vessels. Yeast wastes consist primarily of the spent nutrient (although only 20% of the wastes by volume, they account for 75-80% of the total BOD). They are yellowy brown, have the typical odour of yeast, and are highly hygroscopic. Part of this mixture is used to seed the following batch of wort with yeast; the rest is sold as a by-product. One application of surplus yeast is for animal feed formulations

(United Nations 1999). Approximately 3% of the beer in the fermentation vessels is lost during the

removal of surplus yeast and trub (Van der Merwe and Friend 2002). C02 is continuously removed from the tops of the fermentation vessels. Water scrubbers followed by an activated carbon filter to remove

16 organic impurities are often employed as C02 recovery systems to perform this process. Collected C02 is reused in carbonation. The heat generated by yeast metabolism is continually removed at Labatt by a central refrigerated glycol loop that maintains process temperature.

2.3.4 Maturation and Clarification

Green beer then enters the lagering process where it stabilizes, self carbonates, and further develops flavour. The beer is siphoned off the top the fermentation vats, and transferred into storage or maturation vessels in a process known as racking. Aging tanks are an oxygen free environment that

often prepared by injecting a blanket of C02 in beforehand (Moss and Adams 2007). Here the beer ages for several weeks at a cool temperature (-1°C). Green beer flavours are removed by the enzymatic reduction of yeast (Van der Merwe and Friend 2002). Further fermentation and precipitation of yeast takes place during storage while the product matures, stabilizes, and naturally carbonates (Madigan and

Martino 2003). Product transfer from fermentation to aging may utilize de-aerated carbon-filtered water as a pushing interface between brews.

Wood chips are utilized in some brews as flavoring agents in aging processes. The wood chips contribute additional TSS to the tank bottoms over time as they degrade. This material is mixed with the surplus yeast (by far the larger contributor) in the Krausen tanks. Beech wood chips are coated in NaMS.

Next, the green beer is filtered to improve its aesthetic qualities and remove undesirable, lingering by­ products such as yeast and chill haze. It is estimated that 1.5% by volume of processed beer is lost during filtration (Van der Merwe and Friend 2002). Diatomaceous earth (DE) is the most commonly employed filter media in brewing, but membranes and resins are becoming more prevalent in the industry. A candle filter is the most common vessel to support DE. The filter is prepared by circulating DE slurry into the filter at a high flow rate to deposit a uniform layer or filter cake over surface of the filter leaves (Hardwick 1995). This process is called pre-coating. Additional DE is also injected during operation to prevent early break through and channelling. The filtered beer is continuously monitored for

17 turbidity. When effluent turbidity values are high, the filter is dislodged or dropped using high pressure water jets. The candle filter is CIP'd, and in the case of DE, pre-coated once more. Filter slurry produced by DE is a fine, dense material and the source of waste management problems (United Nations 1999).

After filtration, the beer is centrifuged and cooled down to between -1 ± 0.5°C to remove further

dissolved and suspended solid material. Sterile de-aerated water and C02 may be added to adjust the concentration of high-gravity beer and its gaseous content. Stabilizers, colourants, additional sugar and foam improvement agents may also be added following filtration (United Nations 1999).

Finally it is placed in bright beer tanks until ready for packaging. Top fermented ales are stored for shorter periods at higher temperatures (4-8°C) before packaging, which assists in the development of the characteristic ale flavour (Madigan and Martino 2003).

Spent charcoal and Kieselguhr sludge from filtering operations are a major source of solid waste. Dead­ end filtering with diatomaceous earth and filter-aids has been the standard industrial practice for the past 100 years. Approximately two thirds of DE is used by the beverage industry (wine, beer, fruit juices, and liqueurs). DE use will be increasingly scrutinized from economic, environmental and technical standpoints in the coming century. DE is a finite, mineral resource. The conventional filter practice consumes approximately 1-2 g of DE per litre of clarified beer. The resulting sludge triples in weight through the aggregation of organics and water. DE is a fine particulate considered a hazardous waste by world health organization that requires safe handling. It is a compound of crystalline silica; chronic inhalation can contribute to lung disease. Disposal routes are agriculture and recycling with an average cost of $269.89 (US)/ton (Fillaudeau. L 2006).

2.3.5 Bottling and Pasteurization

Bright beer is transferred into bottles using an automated system of pipes, pumps, fillers and bottle conveyors. It is common practice to spray line lubricants continuously on the conveyors, even when the line is not in production. Returned bottles (from the Beer Stores) are thoroughly washed in a bottle

18 washer with a caustic sanitizer to remove ambient dust and residues from previous usage. Bottles are of the 340 mL and 341 mL variety. The bottle washer removes labels from the exterior of the bottle, and cigarette butts, and organics from the interior of the bottle. Large volumes of water are used during cleaning.

Once the bottles are cleaned, they are split and conveyed to the respective filling areas. Bottles are inspected by computer controlled electronic systems and manually by operators. An OMNI check is an automated process that uses eight points of inspection to determine the cleanliness of a given bottle.

The parameters tested are bottle weight, integrity, transparency, height or volume, and glass type.

Chipped mouths, the presence of liquid, incorrect size, or fogged glass are all reasons to reject bottles.

Out of spec bottles are discarded accordingly. Manual tests are conducted hourly to ensure the OMNI check system is functioning optimally. Clean bottles are then forwarded to the filler bowl to be filled by product.

Each packaging line handles one brand at a time. There is one filler bowl for each packaging line. Beer is conveyed through a piping network from the Bright Beer tanks to the filler. The fillers are outfitted with vacuum pumps that have an inherent water dependency. Each filler bowl is outfitted with 120 to 140 valves. Bottles are mechanically loaded up into the filler bowl positioned under a valve outfitted with a tube that is inserted into the mouth of the bottle. A tight seal is created. Air is first sucked out of the bottle to remove oxygen and create a vacuum to allow a smooth, quick, gravity driven fill. Bottles are filled one at a time in quick succession as they move in a carousel fashion around the filler bowl. Foam intentionally overflows in every bottle in a process called Fobbing to blast air out of bottle. Fobbing minimizes oxygen transfer into the bottle between filling and capping to maintain quality assurance. A very fine stream of de-aerated carbon filtered water is jetted across the top of the mouth of the bottle when it exits the filler bowl carousel to enhance the "fobbing" effect. This nozzle is referred to as the

'Jetter\ After fobbing, the bottles are then Crowned or capped. Bottles are rejected if they are miss-

19 capped. There are a number of faults that may occur in the filler bowl that result in down time and the loss of beer. Bottle break fault at the in feed of the filler, missing filler tubes faults (the tube inserted into the mouth of the bottle to deliver the product), a presence fault (no bottle detected) and a centering cup fault (bottle is improperly aligned) will halt line production until an operator can correct the problem. During a bottle break, five rounds (filler bowl rotations) of seven bottles (three on either side of the bottle in question) are rejected to ensure that there is no glass in the finished product. These bottles are diverted to Rebate Beer. Malfunctioning valves may result in low fills. Low fills are monitored as an indicator of which values require maintenance. If low fills continue to increase on a given valve, a mechanic is called in. All valves are inspected and cleaned weekly. Percent losses are recorded hourly while the line is in operation. A filler bowl operating at full capacity will produce 1020 bottles/min.

Pasteurization of the final product is carried out either by passage through a pasteurization tunnel following bottling or using flash pasteurization prior to bottling. The tunnel is separated into a series of

11 chambers that hold a water bath of a given temperature. The pasteurizers use carbon filtered water to reduce scaling deposits on the bottles. The beer is then packaged in boxes and sent to a large warehouse termed staging. The capped bottles enter the first chamber 21°C. The proceeding chambers incrementally increase in temperature until the sixth chamber at 64.1C. This temperature thermally inactivates residual yeast to inhibit further fermentation.

After pasteurization, the bottles are then labelled, visually inspected, and boxed. Labeller rejects go to

Rebate Beer. The boxed bottles or cans of beer are stored in Staging (a large warehouse) where they await transport to various markets and venues.

Labatt London also has keg and canning lines. The keg line aims to produce 330 - 58.6L kegs/hr. The keg line has a unique labelling system. Domestic product is packaged in 58L silver kegs. US product is packaged in 58L keg with a blue strip. Bass Ale is packaged in 50L Orange kegs. There are also 15L, 20L,

20 and 30L kegs, but each has a separate labelling system. Foreign kegs are pulled off - many are identified with a white stripe.

The main wastewater constituents produced from Packaging operation include caustic sanitizer rinses, wasted beer (from some brand changes), beer spillage (fobbing), breakages, and residues in returned bottles. The packaging and repackaging streams generate a wide range of solid wastes. The beer bottles are moved through the packaging lines by a series of conveyor belts. The conveyance systems are rarely faultless; jams and broken glass are common occurrences. The conveyors are continuously sprayed with a line lube than helps prevent the mechanism from snagging and/or jerking that may impart enough force to upset a bottle.

Bottle-cleaning water generally constitutes a significant proportion of the total pollution from breweries. This pollution is attributed to different gluing methods, glue types and paper types, packaging, and the standing times of the lye. Bottles obtained from collection sites are thoroughly cleaned for reuse. Old brand labels and foreign materials cigarette butts are removed using high pressure sprays of caustic solutions that create a pulpy material that is often filtered off using coarse screens. Waste label disposal is related to product decoration and design and the waste label mass fluctuate greatly. On average, a weight of 282 kg/1000 hi of produced beer has been calculated. Waste labels should be avoided or at least limited since they are not simple papers but wet-strength paper impregnated with caustic solution. The average disposal cost in the United Kingdom is equivalent to

$60.67/ton (min, $0; max, $146.90 /ton) in US dollars (Fillaudeau. L, 2006). Luxurious print work of the labels and the other accoutrements of the beer bottle (metal foil) can leach heavy metals into the wastewater. Heavy metals can be controlled by selecting printing colours for labels free of all heavy metals and eliminating the use metal foils (Murauer Bier, 2004).

21 2.3.5.1 Brand Line Change

Near the end of run, a Bright Beer tank change is initiated at the Brewing side. When 350 to 450 dozen bottles of a given brew (341 ml each) remain in the piping network between the bright beer tank and the filler (this translates to approximately 14 hL), a diverter valve is activated. A new product is then introduced into the same piping network. This is termed a Brand Change. Mixing occurs between the old and new batches which is unmarketable and must be sent to beer pack or the Butts Tank. Beer in the

Butts Tank is sent back to the carbonating room in filtration for reprocessing. Due to the difference in distances between a given Bright Beer tank and the filler in question, the volume of beer that remains in the line varies between successive brand changes. Each individual brand change pushes a fixed the amount of waste beer due to mixing. Some beers, such as Lakeport Ale and Bud with Lime must be sent directly to drain.

Brand changes utilize two full Filler Bowl pushes, and series partial fills to fully dispel the past brand residuals. The new brand pushes out (or chases) the current brand in the conveyance pipes to the filler.

A Bottle Stop (i.e. no more bottle enter the filler) is initiated at end of the final dozen bottles packaged before the mixed beer is anticipated to reach the filler. The remaining beer in the line between the primary valve and the Filler Bowl is pushed to the Butts Tank. The filler bowl is the vessel bottles are loaded onto as they spin around the Filler Enclosure. The Butts Tank collects beer from the filler bowl and conveyance pipes that may be reprocessed when sent back to the carbonation room in Filtration.

However, brands that have flavourings and other additives are discharged directly to the drain (i.e

Budweiser with Lime).

The filler bowl is then allowed to fill up with the mixed beer volume from the packing line. This beer is once again pushed to the Butts Tank. The actual volume wasted in the Beer Line Pack is fairly low because the new brand is pushing the old brand of liquid to the filler (chasing it) and only enough beer is diverted to the Butts tank to ensure any mixed product is packed past the inlet header at the filler.

22 The filler bowl is then allowed to fill once more, and it diverted to the filler. As the filler bowl is never

100% empty, there is likely lingering mixed beverage. A series of Partial Bottle Fills with the new brand are run and delivered to the Rebate Beer Tank until the desired quality is met. Nothing is lost to drain on the Partial Fills. Only the volume of beer required to flush out the filler valves is used. The amount rejected is one full round (approximately 35 dozen bottles worth).

The volume of beer rejected varies by the filler volume, and the number of valves on the particular filler.

The fillers for Packaging Lines 1 and 4 are outfitted with 120 and 140 valves respectively; hence Line 4 will require more beer to fill and dump than the Line 1 filler. The waste volume will vary from 2 to 4 hL.

Actual beer loss at a Brand Change is at the start of new brand line pack (4-6 hL), the filling of the first bowl and the push to Butts tank (3-5 hL), and then rejection of first round of partial fills from filler (1-2 hL) for a net loss around 8-13 hL.

2.3.6 Powerhouse

The powerhouse is operation hub for several supportive brewery processes. Electrical distribution is managed by the powerhouse. Many large breweries are outfitted with emergency backup generators in case of power outages. Thermal regulation is one of the staples to brewing as the product goes through multiple heating and cooling processes. Many of these control systems are located in the powerhouse.

Labatt uses a glycol cooling loop with heat exchangers for thermal regulation of fermentation vessels.

The powerhouse also manages a large centralized boiler for hot water production, and water conditioning systems including softeners and deaerators. The powerhouse operates wet scrubber for

C02 reclamation from the off gases of primary fermenters and aging tank. C02 is a valuable resource used in many brewery operations that is also a byproduct of the brewing process. The offgases are vacuumed off the top of the brewing vessels and directed to the wet scrubber. De-aerated carbon

filtered water (DCW) is used as the scrubbing agent. DCW prevents 02 addition to the C02. Once the fermentation gases have passed through the scrubber, the gas is then carbon filtered, compress to 60

23 psi, and dewatered for immediate use. Surplus C02 may be further compressed to 240 psi, dried, and

cooled to -24°C for liquid C02 storage.

2.3.7 Clean In Place

All vessels and pipes are routinely cleaned in a brewery 'NawVeptl Pre Rinse (To Drain) using a clean in place (CIP) process to ensure the product is X AcWCaustlc DCW Wish _ . _ _ .. WasH free of undesirable tastes and odours. These processes use (Reclaimed} CLEAN (Reclaimed) IN a variety of cleaning agents and a substantial amount of PLACE

Sanltlzer Acld/Caustlc water. A CIP cycle may include any combination of acid, (CIO,) Rinse Post Rinse (To Drain) (To Drain) caustic, chlorinated, and/or pure water rinses. Acid washes Figure 2: Clean-ln-Place Flow Diagram are either phosphoric (H3P04) or nitric (HN03) acid. These acids do not introduce any unnatural elements into the system; malt is already latten with phosphorous

and nitrogen. HN03 is more abrasive on metals than H3P04. Labatt currently utilizes a 1.13% warm

Divosheen solution at 31°C for the vertical and horizontal fermenters. Divosheen is 60% H3P04. A caustic cleanser of 3.6% Sodrox BW at 53°C is used in Krausen CIPs, Aging Tank CIPs, and bottle washing. Caustic

3 soda in particular is used in amounts ranging between 5 and 10 kg30%Na0H/m (United Nations 1999).

According to Labatt's RSView, significant heat loss in the wash water occurs during transmission between the central CIP pit and the vessels. Acid and caustic washes are generally reclaimed and stored in a central tank for reuse. The acid/caustic washes are followed by a post rinse to remove residual

cleaning agent. A single use chlorinated sanitizing wash, generally chlorine dioxide (CI02), follows. It is discharged directly to drain. To ensure that there are no lingering chemicals that may impart into the new product, a final post rinse of de-aerated carbon-filtered water (DCW) is applied. The post rinse is reclaimed to be reused as the pre-rinse in future CIPs. There are two central CIP pits at the Labatt

London Brewery. One services the brewing side of operations, while the other services the packaging

24 side. The direct discharge of either caustic or acids may cause dramatic fluctuations in the pH of the resulting wastewater.

Fermentation uses an Acid wash CIP cycle. From September '09 Vertical Fermenter 83 data:

• 45.7°C Pre-rinse water is pumped at 36.5 m3/hr for 30 minutes over a 70 minute runtime;

• 29.5°C Acid wash water is pumped at 34.2 m3/hr for 30 minutes over a total of 40 minutes;

• 12.1°C Post Rinse 1is pumped at 35.5 m3/hr for 21.5 minutes over a 28 minute runtime;

• The sanitizer rinse for 10 min over an 18 minute runtime; and

• The second post rinse for 10 minutes over an 18 minute runtime.

Krausen and Aging Tanks use a Caustic wash CIP cycle. From June '09 from Krausen Tank 1402 data:

• 45.7°C Pre-rinse water is pumped at 36.5 m3/hr for 10 minutes over a 16 minute runtime;

• 53.1°C caustic wash water is pumped at 34.2 m3/hr for 30 minutes over a total of 41minutes;

• 12.1°C Post Rinse 1is pumped at 35.5 m3/hr for 22 minutes over a 30 minute runtime;

• The sanitizer rinse for 5 min over a 10 minute runtime; and

• The second post rinse for 30 minutes over a 36 minute runtime.

2.4 Wastewater Legislation for Breweries

To protect the health and safety of the public and the environment, wastewater discharges into sanitary systems and water ways are subject to strict limitations on concentrations of a variety of chemical constituents, particularly in organic load, suspended solids, pH, temperature, and chlorine. Discharge into water ways is even stricter. The maximum allowable BOD limit for discharge of effluents into public waters near densely populated areas can be as low as 10 ppm.

The City of London categorizes breweries as an industrial consumer of water. From Figure 3, it is evident that industry in the City of London pays less in sanitary charges than other southern Ontario cities.

25 Comparative Sewer System Charge Overview - Industrial Outlined Mow Is a comparison of th« proposed 2009 City of London sewer system charge compared to 2008 charges for other munietpalittfti In Ontario.

2009 charges for London compared to 2008 charges for other municipalities

Jo0,000

$50,000

03 #40,000

$30,000

$20,000

$10,000

— Hftfcoe MMM** Si. lotxtoa KiWhantr Cambritfge St Thorn.** KiftgttOft Ottawa Sarota Rtg&n CattMurii-KMi •oe 08 *oa •&s Hlh 06 *08 **e 08 08 a Sanitary W3.074 $5X3*2 541,590 545356 S39.561 S4Q.S94 530.729 SA2.2W *45,151 MW1 $41,758 •Stotm $3/58 MUNICIPALITY

Note Anno usarcitMpniisMa at 38 800 m3watwconsum«

Figure 3: Comparative Sewer System Charge Overview - Industrial (City of London 2008)

Wastewater from a brewery may be managed in any combinations of several methods:

a. Directly discharge into a municipal sewer system;

b. Treated on site with a private wastewater facilities;

c. Discharge into a waterway after pretreatment; and/or

d. Diversion to an alternate party by means of trucking.

Wastes streams discharge into municipal systems in excess of BOD, TSS, total phosphorous (TP) and

ammonia (NH3) may be allowed assuming a high-strength sewage surcharge is paid. The fee is based

upon average volumetric load, average load of BOD, TSS, TP and NH3, and the peak discharge rates

(Hardwick 1995). Box 1 provides an example of billing practices.

26 Box 1: The City of London, ON 2007 Waste Discharge Bylaw: High Strength Sewage Service Charge Billing Program

3.11 Discharge - prohibited waste - permission - conditions

(1) The City Engineer may permit the discharge of waste which has a five-day B.O.D. exceeding

300 milligrams per litre and/or contains more than 350 milligrams per litre of suspended

solids and/or contains more than 50 milligrams per litre of ammonia and/or contains more

than 10 milligrams per litre of phosphorous, into a sanitary sewer, notwithstanding any other

section of this by-law:

(a) if the land from which the wastes are to be discharged is equipped with the

following inspection facilities and apparatus, to the satisfaction of the City Engineer:

(i) a flow measuring, sampling and recording device within a compartment

which may be kept locked by the City and to which no other person shall

have access, at a point between the public sewage works and the outlet of

all plumbing systems serving the land; and

(ii) an observation manhole, having a diameter of not less than 1.2 metres

located so as to permit inspection of all wastes being discharged into the

6.11 Measurement

(1) High Strength Waste

(a) The volume of high strength waste shall be as measured by the flow measuring

device located upon the land served or as estimated by the City Engineer should

such device malfunction or fail to provide complete data.

(b) The concentration of ammonia, B.O.D., suspended solids and phosphorous in excess

27 6.12 Billing

(1) The High Strength Sewage Service Charge shall be based upon:

(a) the strength of ammonia, B.O.D., suspended solids and phosphorous which have

been treated and shall be calculated as follows:

F x Q x R S= 2

Where:

S represents the High Strength Sewage Service Charge in dollars

F means the ratio of:

actual strength - allowable strength allowable strength

Table 2 lists the wastewater limits for the City of London ON discharge to sanitary sewers. Only BOD5(

TSS, and TP are considered for the HSS billing. All other parameters MUST fall within the outlined criteria.

28 Table 2: City of London 2007 Limitations on Discharges to Sanitary Section 4.8

pH Between 6 and 10.5 1.5-12 (avg. 7)

Temperature < 55°C 18-40"C (Khan, 1984)

BOD, < 300 mg/L 650-1300 mg/L (Hardwick, 1995)

Suspended Solids < 350 mg/L 105 -772 mg/L (Hardwick, 1995)

Natural FOG (Animal, Vegetable) < 100 mg/L 8-55 mg/L (Labatt New Westminister, 1988)

Synthetic Oils, and Greases < 15 mg/L N/A

Aluminum <50 mg/L 1.21 mg/L (Labatt London Effluent, 1999)

Ammonia (NH3) < 50 mg/L 30-57 mg/L (Gu, 1996)

Arsenic (As) < 1.0 mg/L <0.01 mg/L (Labatt London Effluent, 1999)

Barium (Ba) < 5 mg/L <0.04 mg/L (Labatt London Effluent, 1999)

Beryllium (Be) <5 mg/L <0.01 mg/L (Labatt London Effluent, 1999)

Cadmium (Cd) < 3 mg/L <0.02 mg/L (Gu, 1996) (Labatt London Effluent, 1999)

Chloride (CI) < 1500 mg/L 22-82.0 mg/L (Khan, 1984)

Chromium (Cr) <5 mg/L 0.02 mg/L (Labatt London Effluent, 1999)

Copper (Cu) <5 mg/L 0.03 mg/L (Labatt London Effluent, 1999)

Cyanide (CN) < 2 mg/L 0.038 mg/L (Labatt London Effluent, 1999)

Fluoride (F) < 10 mg/L 0.8 mg/L (Labatt London Effluent, 1999)

Iron (Fe) < 50 mg/L 3.0 mg/L (Khan, 1984)

Lead (Pb) <5 mg/L <0.03 mg/L (Labatt London Effluent, 1999)

Manganese (Mn) < 5 mg/L <0.02 mg/L (Labatt London Effluent, 1999)

Mercury (Hg) < 0.1 mg/L <0.001 mg/L (Gu, 1996)

Molybdenum (Mo) < 5 mg/L 0.06 mg/L (Labatt London Effluent, 1999)

Nickel (Ni) <5 mg/L <0.05 mg/L (Labatt London Effluent, 1999)

Phenolic compounds < 1.0 mg/L 0.007 mg/L 4-AAP (Labatt London Effluent, 1999)

Phosphorous (P) < 10 mg/L 3.2 -56 mg/L (Shu-Guang Wang, 2007)

Selenium (Se) < 5 mg/L <0.02 mg/L (Labatt London Effluent, 1999)

Sliver (Ag) <2 mg/L <0.1 mg/L (Labatt London Effluent, 1998)

Sulphates (S04) < 1 500 mg/L 75.0 mg/L (Khan, 1984)

Sulphides (S) <2 mg/L 0.09 mg/L (Labatt London Effluent, 1999)

Tin (Sn) < 5 mg/L <0.07 mg/L (Labatt London Effluent, 1999)

Zinc (Zn) <5 mg/L 0.49 mg/L (Labatt London Effluent, 1999)

Nonylphenol < 0.02 mg/L N/A

Nonylphenol Ethoxylate < 0.2 mg/L N/A

Max. Solid Size < 6.7 mm2 N/A

29 In recent years, the high strength sewage service charges have been subject to substantial increases as environmental regulations continue to tighten. As of 2004, the high strength service charge fee has increased annually by roughly 9% (see Figure 4). In terms of water management, strict legislation favours a reduction of water consumption and wastewater production in order to reduce the volume to treat (Fillaudeau. L 2006).

High-strength Sewage Surcharge (HSS) Rate for London, ON. from 2000 to 2010

0.45

0.40

SoiI > g> d> 0.35 5 E

0.30

Trendlirte: 0.25 y = 0.G293x- 58.517 RJ = 0.971 0.20 \ %, % % %, %s \ %, \ \ Time (Year)

Figure 4: 2000-2009 High Strength Sewage Surcharge for London, ON.

Occasionally breweries are required to contribute to construction costs for municipal wastewater treatment facilities because of the strengths of their wastes. It is becoming common for large breweries to construct their own complete wastewater treatment facility or pretreat their effluent. The high costs that are often required for wastewater treatment off brewers an additional incentive to eliminate unnecessary wastes and to optimize the reuse of effluents (Hardwick 1995).

30 2.5 Brewery Wastewater Constituents

The brewing industry is water intensive with high ratios of water used to beer produced. In the developed world, 4-11 m3 of water are used per m3 of beer produced (Fillaudeau. L 2006); (United

Nations 1999); (Van der Merwe and Friend 2002). In developing countries, such as China, breweries may generate 20-30m3 of effluent per m3 beer (Wang, et al. 2007). The primary water consumptive areas in the average brewery are the brewhouse, cellars, packaging and general water use. Water demand attributed to these areas includes all water used in the product, vessel washing, general washing and cleaning in place (CIP). Each water-consuming application is of particular importance both in terms of water intake and effluent produced (Van der Merwe and Friend 2002).

The brewing process consists of batch operations that result in intermittent and substantial wastewater discharges. Wastewater volumes, pH, and concentrations vary constantly. Brewery wastewater usually has temperatures ranging from 25°C to 38°C (T. Goldammer 2008). A good characterization of wastewater requires simultaneous measurement of flow rates and concentrations over an extended period of time. The flow-proportional sampling method is most suited for this purpose (Hardwick 1995).

The quantity and quality of brewery wastewater depends on the production and specific water usage.

Effluents from breweries and fermentation operations are generally of special consideration to municipal wastewater systems as contaminant concentrations of several items are commonly in excess of guidelines. The most troublesome regulated wastewater parameters for breweries include biochemical oxygen demand (BOD), total suspended solids (TSS), pH and inorganics such as phosphorous and nitrogen compounds. This section will detail the nature of each of these compounds in brewery wastewater.

2.5.1 Oxygen Demand

BOD is an indicator of the amount of oxygen required by microorganisms to decomposed readily available organics in wastewater. Brewery effluents are generally characterized as having a high BOD in

31 the neighbourhood of 1500 mg/L. This results from all the organics that enter the waste stream during the brewing process; primarily in the form of maltose, dextrose, soluble starches, ethanol, and volatile fatty acids. The concentration of organic substances in brewery wastewater discharges is directly dependent upon the amount of beer and organic auxiliary materials (i.e. hops, yeast, wort, trub, spent grain filter slurry and intermediate and final fermentation products, etc.) that are permitted to enter the effluent stream. For example, if trub and spent hops are discharged, they can contribute up to 20 per cent of the total daily organic loading. Increased pollution in wastewater often results from a non-

optimal solid/liquid phase separation. Fluctuations in BOD5 (and pH) values result from the various cleaning processes and CIP systems (Murauer Bier 2004). Product at various stages in development enters the effluent stream as a result of leakage and breakage. Great care should be taken to minimize or divert these spills from wastewater as beer has a very high BOD.

Chemical Oxygen Demand (COD) represents the amount of dissolved oxygen required oxidize all organic materials present, and may be empirically correlated to BOD. BOD represents a fraction of COD. The average brewery main effluent is generally related by an empirical factor of approximately 0.51; this does not represent the ratio of unique contributing waste streams.

Wastewater from the malting process has a COD of 800-1,200 mg/L. A study carried out in the mid-

1980s estimated that approximately 0.5-1.5 per cent (by weight) of the barley ends up in the wastewater as organics (e.g., pentose, sucrose, glucose, cellulose, protein and minerals).

Effluent from the brewhouse may have a COD value in the 3,000-5,000 mg/L range, high suspended solids content, relatively high temperatures and high pH values. Concentrated COD loading principally emanates from the brew kettle and the fermenter, as well as from cleaning operations. Residues such as trub, spent grain, kieselguhr and yeast are major waste items at this stage. Rinsing and cleaning of process and bottling equipment, as well as bottle washing, imposes further waste loading. Bottle washing produces a wastewater with moderate COD values of 2,000-3,000 mg/L and temperatures of

32 25°C to 30°C. Non-alcoholic beer production further increases the proportion of organic substances in the corresponding wastewater by adding condensed alcohol enters the waste stream.

Table 3: Approximate COD Content of Various Brewery Waste Streams (United Nations 1999)

Source effluent

Trub from Hot wort 3200 Last runnings: - Fermentation vessel to storage vessel 2700 - Lauter tun 2500 Fermentation vessels 1400 Spent filter slurry 1400

High concentrations of both BOD and COD have an adverse effect on aquatic organisms by removing available oxygen from the water. High organic loadings may generate noxious gases with offensive

odours (i.e. H2S), and contribute to corrosion of sewer infrastructure. The amount of BOD in brewery wastewaters yields a high of degree biodegradability that lends credence to biological treatment technologies as a highly suitable remediation option.

Malt wastes were shown to contain an average of 72 ppm of suspended solids and 390 ppm of BOD, when 6996 bushels of barely were processed per day. The volume of waste generated averaged 75 gallons per bushel, or 524 700 gallons per day. The solids were mainly organic and high in nitrogen, indicating considerable protein material. A major portion of the solids were in solution, as indicated by the low suspended-solids content (Nemerow and Agardy 1998).

The process water inputs must be considered separately from the brewing water. Process water is used for the automated CIP system for vessel cleaning. Each brewhouse tank will have a small amount of residual wort once it is transferred to fermentation. The residual wort will be swept away by the cleaning water which is discharged directly to drain. Based on Labatt RSView2 data, it is assumed that

the hot wort tank residual of 0.9hL of wort is the only serious BOD5 contributor.

2 RSView is Labatt's operational control software, and is similar to SCADA (system computing and data acquisition).

33 2.5.2 Total Suspended Solids

Brewery effluents are notably high in total suspended solids (TSS, >300 mg/L). A high proportion of TSS originates from DE slurry, spent grain, trub, surplus yeast, and label pulp - all of which are primarily organic in nature (T. Goldammer 2008); (Hardwick 1995). The organic portion of the TSS can significantly contribute to the biochemical oxygen demand (BOD) and chemical oxygen demand (COD) loading in municipal sewage as they deteriorate. Dissolved solids are mainly from beer, wort, and cleaning/sanitizing solutions (Hardwick 1995).

Table 4: Loading of BOD and Suspended Solids from Various Brewery Waste Streams

Lauter tun rinse and drain 1.00 0.39 Trub and wort losses 0.43 0.23 Other brewhouse losses, rinse and CIP 0.31 0.19 Press liquor 1.784 0.85" Surplus yeast handling waste 0.31 0.19 Spent filter materials 0.62 2.01b Fermenting and finishing waste 1.70 0.39 Packaging waste 2.78 0.31 Total 8.93 4.56 a Can be avoided when brewer's grain is sold wet, or when liquor is used as a by-product b The majority of this can be disposed of as solid waste, if special equipment is available

(Table Adapted from Hardwick, 1995)

2.5.3 pH

The considerable fluxes in effluent pH are attributed to the caustic and acidic effluents from regular clean in place (CIP) operations, caustic-laden water from sanitizing rinses in the bottling area, and discharges of beer at various stages of fermentation. Neutralization of these agents to within prescribed specification requires a robust pH adjustment system. Strong caustics and acids are commonly utilized in cleaning procedures. Rinse water represents 45 percent of the total water use in a brewery (United

Nations 1999). Beer has a pH of around 4, and inevitably gets discharged to drain during regular

34 operations (i.e., brand changes), and accidental spills (i.e., equipment failure). Residual beer lost during production stages constitutes 1 to 5% of total production. Most residual beer can be collected and, depending infrastructure and quality criteria, either reclaimed or repurposed (i.e. sold as by-product).

Large volumes and concentrations of these materials may shorten the lifespan of sewer infrastructure.

Some cleaning chemicals can also be toxic to aquatic organisms (CRD 2002). This naturally has a decided influence upon waste treatment methods. If any sort of biological treatment technology were to be employed, equalization tanks would be necessary.

Regeneration of ion exchange units (i.e. water softeners) in water treatment plants associated with beer production is also responsible for further discharges of strong acids and caustic materials. Waste from such operations has a wide range of pH values, from 2-12. Therefore it is essential to collect and neutralize regeneration waste. Neutralization should occur as close to the source as possible to protect sanitary infrastructure. It should be noted that the pH of neutralized brewery wastewater tends to drop

due to ongoing bacterial action which releases C02 into the water. This is common where sewage conveyance time between the brewery outlet and the treatment plant is in the order +10 minutes

(Hardwick 1995).

2.5.4 Inorganic Pollutants

Phosphorous, ammonia, and sulfide compounds respectively constitute the troublesome inorganic portion of brewery wastewater; phosphorous of which is the only one consistently over municipal guidelines. These ions are principally derived from malt, nutrient additives, the yeast, and acid-based CIP procedures. A study conducted by Labatt London in 2007 suggests that trub may contain up to 170 mg/L of total phosphorous. Ammonium salts are commonly used to promote yeast growth. Vessels that require a more thorough cleaning such as fermenters often employ acid CIP processes over caustic rinses. Nitric and phosphoric acids are favoured as these elements are already present in the beer.

Concentrations of inorganic phosphorus in brewery wastewater may range from 30-100 mg/L. Such

35 concentrations are toxic to some forms of aquatic life; contribute to groundwater pollution and eutrophication of surface water (United Nations 1999).

Other inorganic chemicals introduced into brewery wastewater include potassium and calcium salts as well as silicates derived from DE (United Nations 1999). The frequent problem of scale or deposits of calcium salts and magnesium salts in bottle washing machines may be prevented by adding chelating agents and softeners to the rinsing solutions (Murauer Bier 2004).

Not all wastewater requires treatment. Noncontact cooling water and rinse water for nonreturnable bottles and cans, for example, is relatively clean and may be discharged directly into a river or storm sewer depending on the temperature and chlorine limitations (Hardwick 1995).

36 Chapter 3: Labatt London Wastewater Characterization

3.1 Wastewater Characterization Project Introduction

The University of Guelph (UoG) was requested by Labatt's Brewery of London ON to conduct an extensive investigation on their wastewater effluent and various process contributories over the course of two years. The purpose of the investigation was to develop data for a process evaluation on water-use reduction and/or waste minimization opportunities. A wastewater sampling program was formed by Labatt personnel in cooperation with Dr. Hamidreza Salsali of

UoG and in February 2009.

A wastewater characterization study was conducted at a commercial brewery over the course of eight months to assist in the development of wastewater reduction program. Specifically, the objective was to find opportunities to save on the quarterly High-strength Sewer Surcharge

(HSS) fee imposed by the municipality. Process wastewater streams across the brewery from

Brewing, Fermentation, Aging, Filtration, Racking, Powerhouse operations, Central Clean-ln-

Place, to Packaging, which includes Bottling, Canning, and Kegging, were targeted and sampled for various chemical and physical parameters. Wastewater analysis was conducted in-house

using Standard Methods procedures to determine Biochemical Oxygen Demand (BOD5), Total

Suspended Solids (TSS), and Total Phosphorous (TP) concentrations, the parameters monitored by the local municipal wastewater treatment plant. Chemical Oxygen Demand (COD), Volatile

Suspended Solids (VSS) were also analyzed to provide further insights into the nature of the waste. A brewery process flow diagram was created to conceptualize unit process inputs. Some of the major findings were 1) Aging tank bottoms were the most concentrated waste with BOD5

and TSS in excess of 100 000 mg/L 2) C02 Scrubber water was identified as a previously unknown contributor had a BOD over 1000 mg/L. 3) The rotary drum filter which was used to

37 remove diatomaceous earth (DE) from Filtration wastewater was under-performing and resulted in higher overall solids loading than expected. The data suggested that the avoidance and/or reduction of beer, and beer byproduct (yeast, DE) discharges to drain would result in HSS fee savings. However, minor product wastewater inputs are integral to brewing operations, such as in vessel washing (CIP) and thus unavoidable. End-of-pipe biological treatment was recommended.

The chapter summarizes the wastewater sample locations and results conducted from April to

November 2009 at the London Ontario Labatt Brewery. The lengthy duration was intended to capture seasonal variability in production on the final effluent.

The approximate location and unique sampling details are described in Appendix A. The sample locations were selected to quantify nutrient loading in wastewater streams generated across the entire brewing process. Average wastewater parameter concentrations for each sample point are provided. The standard deviation for each value is indicated by a '±'. Each sample location is identified by a unique sample number. A reference to the relevant Labatt Brewery AutoCAD drawings that illustrate the sampling locations is provided where applicable.

A process flow diagram (PFD) has been generated to situate the contributories (Appendix B).

Each sample location has been identified by a reference number to the more detail summaries provided in this Document. The PFD consists of a series of box models around each process in the format of a mass balance. Box model inputs consist of processing materials (i.e. ingredients, beer at various stages of development), and cleaning water (i.e. clean in place (CIP) water plus manual pre-rinse water where applicable). Outputs are processed beer, by-products (i.e. spent grains, surplus yeast), wastewater, evaporative losses and recaptured volatile organic compounds (VOCs). The productive outputs from one box model (i.e. beer) are fed into the

38 receiving end of the succeeding process. The majority of the BOD5 in effluents is from residual or waste beer, surplus yeast, and VOCs.

3.2 Wastewater Characterization Methodology

Thirteen locations were initially selected to be tested for a variety of wastewater parameters.

The sampling program was later refined and broadened in July 2009 after the reviewal of preliminary results. Four additional areas were incorporated for a grand total of 18 unique locations. Over 100 samples grab were obtained. Several sub processes such as centrifuging have been neglected as insufficient data is available at this time.

Each sample was collected and stored in a clean 500 mL glass mason jars samples maintained at

4°C. No preservatives were used. Testing completed using University of Guelph School of

Engineering Resources in the Biohazard Lab, Rm 1196. All tests were completed in duplicate. The

pH was determined immediately. BOD5 (Std. M. 5210 B) tests were initiated within 24 hours

after sampling. Three dilutions were used for each BOD5 sample. COD (Std. M. 5220 D) was completed within 2 days of sampling, TP (Std. M. 4500 P.E.) within 3-4 days, and TSS/VSS (Std.

M. 2540 D) within 3-4 days. Retests were completed thereafter with the exception of BOD.

Copies of the Standard Methods employed are available in Appendix C.

3.3 Wastewater Characterization Results

The results for the wastewater characterization are displayed in Table 5. The samples have been organized in accordance to the relative progression of the brewing process, from Post Runoff

(PRO) generated at the Brewhouse waste streams, all the way to the final effluent. A more detailed description of each sample point is available in Appendix A. The detailed summary provides complete details about each sample point, including a unique process description, discharge frequency, flow rates and the number of samples taken. Each sample also has a

39 number preceding its description in Table 5; this number corresponds to both the detailed summary document, and a Wastewater Process Flow Diagram (PFD). The PFD traces the brewing process to shows where each sample point enters the wastewater stream (see Appendix B).

40 Table 5: Results From Brewery Wastwater Charaaerization • POWERHOUSE 4: COt Scrubber Water 2710 ±233 1760 ±259 0.1 ±0.1 1.6 ±0.1 29.2 ±1.4 0.3 ±0.4 0 ±0 0 ±0 5.28 ±0.25 5: Boiler Blow Down Water 93 ±84 28 ±6 1.8 ±0.5 5.1 ±2.5 7.4 ±0.4 0.3 2 ±114 0 ±0 11.90 ±0.28

BREWHOUSE

18: PRO 17500 - 12400 -- 19.0 - 150.0 ------420 -- 411 - 6.06 -- 2: Brewhouse & Bottle Line 1 Sewer 1890 ±627 1100 ±361 2.5 ±0.8 21.5 5.7 31.7 2.8 0.6 ±0.2 69 ±17 34 ±9 10.34 ±0.25

FERMENTING

7: Vertical Fermenter Bottoms 115000 - 25.3 ------3500 - -- - 4.21 -

7: VF CIP - Initial Rinse from Reclaim 1520 ±879 904 ±609 4.8 ±1.3 27.2 - - - - 197 -- 223 ±140 9.52 ±2.01 „ 7: VF CIP - Acid Cycle Post Rinse 790 ±864 39 ±68 32.0 ±13.3 11.5 - - -- 346 ±567 19 ±20 3.50 ±2.13 .. 7: VF CIP - Sanitizer Rinse (City Water) 37 ±38 4 ±5 4.0 ±0.3 ~ - -- - 65 ±83 7 - 5.90 ±0.82

39 ±19 0 ±0 1.3 ±1.7 - - - -- 201 ±281 3 -- 6.27 ±0.34 7: VF CIP - CI02 Rinse " 7: Recycled Rinse Water for initial Rinse 102 ±77 44 ±29 7.4 ±3.1 2.6 -- -- 18 ±25 11 ±15 8.05 ±5.35

AGING

8: Aging Tank Bottoms 218000 - 100000 -- 40.7 -- 432 -- 469 - 3.0 1990 -- 1990 - 4.03 --

11: Krausen Tank Bottoms 226000 ±27400 109000 ±30100 322 ±227 699 -- 102 -- 2.1 91800 ±21300 82700 ±25700 5.42 ±0.35

11: Krausen - Manual - 1st Rinse 17600 ±141 10800 ±193 39.4 ±12.8 112 ±9.7 24.3 -- 0.2 10600 ±52 10000 13 5.54 ±0.10

11: Krausen - Manual - 2nd Rinse 10200 ±4350 4980 ±1330 24.5 ±4.1 76.2 ±22.2 34.6 -- 0.0 5261 5060 - 5.82 --

11: Krausen - Manual - 3rd Rinse 11400 ±3980 6500 ±3800 22.0 ±2.6 89.4 ±37.6 36.0 -- 0.3 6442 ±3460 6170 ±3120 5.78 ±0.22

11: Krausen - Manual - 4th Rinse 3740 ±530 1770 ±207 8.6 ±4.4 34.8 ±9.5 - •- -- 1704 ±501 1660 ±476 6.54 ±0.38

11: Krausen • Manual • Chips Split 56900 - 40000 -- 21.2 -- - -- 73.1 -- 1.0 57000 -- 54000 - 5.28 --

11: Krausen - Manual - 5th Rinse 744 -- 614 - 3.8 - 11.3 - 28.4 -- 0.0 392 -- 389 - 6.88 -- 11: Krausen - Manual - 6th Rinse 1100 ±11 858 ±133 3.5 ±1.3 13.9 ±2.9 28.4 -- 0.0 415 ±71 395 ±50 7.44 ±0.79 11: Krausen Pre-CIP Rinse (Chips 994 ±185 940 ±124 3.6 ±2.0 16.0 ±8.2 28.7 0.1 565 ±350 531 ±320 6.70 ±0.01 removed) - 11: Krausen Tank CIP - Initial Rinse 8570 ±1480 3950 ±911 17.7 ±0.9 80.3 -- -- - .. 4420 ±5280 890 ±291 7.52 ±2.84 11: Krausen CIP - Caustic Cycle Post 1280 ±171 789 -- 11.6 ±7.2 31.6 -- -- 363 ±105 254 ±8 13.29 ±0.02 Rinse •• - .. 11: Krausen Tank CIP - CI02 Rinse 103 ------1.5 ------17 - 16 -- 11.52 - 6: Aging, Fermenting, &Powerhouse 762 ±334 782 ±396 9.4 ±6.3 25.6 ±26.8 38.0 ±7.6 0.7 ±0.6 44 ±28 32 ±21 6.30 ±1.02 Sewer flfll FILTRATION 9: DE Filtration Pit 3930 ±2560 2960 ±1950 3.4 ±3.4 29.5 ±12.7 40.5 ±6.7 0.2 ±0.0 7410 ±7420 3781 ±6408 6.13 ±0.72 9: DE Slurry Tank 8680 ±720 5950 ±1520 11.0 ±2.9 41.4 ±3.2 37.0 -- 0.5 - 19800 ±2880 2633 ±445 5.03 ±0.58

9: Beer Filter DE Washout 11600 9410 ~ 13.3 67.6 - 45.5 - 0.4 - 42900 - 5490 -- 5.59 --

BRIGHT BEER 10: Bright Beer/Fermenting 5180 ±4700 2330 ±1420 4.5 ±0.9 29.5 ±22.1 27.4 ±2.7 0.6 ±0.2 589 ±726 169 ±237 6.49 ±0.83

PACKAGING 1: Packaging Lines 1-5 Main Sewer Hub 3380 ±2050 2370 ±1250 2.4 ±0.6 35.8 ±16.3 66.6 ±33.2 0.5 ±0.5 141 ±100 132 ±108 9.41 ±1.81 3: Bottle Line 1 Sewer (Normal Operation) 1060 ±505 931 ±637 1.6 ±0.6 16.1 ±5.9 32.3 ±4.9 0.4 ±0.2 62 ±31 33 ±8 10.70 ±0.67

15: Bottle Soaker Packaging Line 1 4230 ±4360 1570 ±827 3.2 ±1.1 45.7 ±3.6 ------179 ±31 145 ±18 12.34 ±0.08 „ 16: Vacuum Filler Packaging Line 1 1330 ±476 592 ±270 0-6 ±0.1 2.8 10.2 - - 5 ±2 4 2 6.41 ±0.88

17: Pastuerizer Packaging Line 1 54 ±94 46 ±41 0.2 ±0.1 2.0 ±0.2 ------0 ±0 0 ±0 8.04 ±1.32 13: Keg Line Sump Pit (Online) 7370 ±3180 5870 ±2120 4.5 ±0.5 91.3 ±48.8 135 ±171 1.7 ±0.3 117 ±44 81 ±56 10.17 ±0.91 „ 13: Keg Line Sump Pit (Offline) 91 ±87 111 ±99 0.8 ±0.4 31.4 ±35.2 124 - 0.5 35 ±17 35 ±17 8.09 ±2.42

BREWERY FINAL EFFLUENT

12: Main Sewer Line (SAN 34) 2860 ±1070 1700 ±348 4.8 ±3.6 22.7 ±16.2 28.3 ±8.5 0.1 ±0.0 1020 ±1050 1270 ±1170 7.95 ±1.74 14: Reactor 4 Effluent 2370 ±1270 1770 ±699 11.3 ±3.2 14.4 ±4.9 38.5 ±8.0 0.4 ±0.0 1010 ±408 546 ±219 6.43 ±0.90 •• m •I

Notes: The number (#:) beside each description under the Sample Identification column corresponds to the detailed sample point summary

tables and process flow diagram located in Appendix A and B respectively. 3.3 Wastewater Characterization Discussion and Recommendations

3.3.1 Brewhouse

Wastewater is well managed within the Brewhouse at Labatt London. The nutrient loading in the shared sewer is predominantly from packaging. Post Run Off (PRO) and Trub are the only identified major contributor for BOD within the Brewhouse. PRO is a very concentrated waste stream with a BOD in excess of 12g/L, however, it is discharged infrequently and in relatively low quantities. Each batch of beer produces about 3 hL of PRO. PRO is only discharged once the 18 hL PRO tank reaches capacity during Budweiser production. This only occurs a few times a week.

The amount of PRO discharged depends upon the amount of Budweiser brands brewed that weak. Emphasis for process improvements to minimize wastewater loading should be placed on other departments within the Brewery that contain more concentrate waste streams that are discharged to drain in larger quantities, such as Fermentation and Aging.

3.3.2 Fermentation and Aging

Krausen tank bottoms are the most concentrated source of BOD5 and TSS sent to sewer. The residual volume of Krausen yeast remaining in the tank before the manual clean (5 to 10 hL) costs approximately $63K to $126K /year to discharge to drain, assuming the April 2009 sewage

3 surcharge of 40.70/m . Aging tank bottoms are also a significant source of BOD5. Aging Tanks would cost approximately $65K to $130K /year to sewer using the same methodology. Both of these sources have identified in previous studies as major contributors to the high strength sewage service charge. The Krausen tank bottoms separate into three distinct layers; beer, yeast, and fine wood chip sediments. Settleability and centrifuging tests have been conducted on Krausen Yeast, but require further refinement. The conventional Imhoff cone test to determine settleable solids test is somewhat impractical as each layer has its own settling rates;

43 an alternate settleable solids test should be devised. Various speeds and times could be tested to optimize the centrifuging of these materials. When left undisturbed at 20°C, the wood chip materials generally settle by gravity in 45 minutes, whereas yeast requires at least 4 hours.

Ideally the tank bottoms would be source separated from the Krausen and aging tanks instead of direct discharge to drain. There is presently no existing infrastructure to divert this waste stream from sewer.

Chemical oxidation of the aging tank bottoms with hydrogen peroxide (H202) and ferric sulphate

(FeS04) before the pre-CIP rinse may be an effective way to reduce BOD5. This method has been applied for COD removal in high strength, low volume wastewater in the oil refining industry where biological treatment was impractical due to toxicity of the wastestream (Dincer, et al.

2008). Brewery wastewater is nontoxic to the microorganism associated with biological treatment. However, as Labatt London does not currently operate biological wastewater treatment technology, chemical oxidation of select wastewater streams may be a viable option for the interim. Extensive laboratory-scale testing should be completed to determine the feasibility and suitability of this pre-treatment option. The chemical reaction is very exothermic and significantly alters the character of the wastewater. If adopted, the pretreatment method could potentially result in the savings of tens of thousands of dollars a year on the HSS, but would likely necessitate additional changes to the CIP process.

Increased agitation of the Krausen tank bottoms during a manual clean may facilitate nutrient removal and yield additional water savings. Splitting the chips demonstrates the effectiveness of agitation. The yeast and wood chip sediments are relatively heavy and sticky compared to the beer, and become lodged within the networks of wood chips. A simple increase in agitation may be observed by 1) providing a sweeping motion of the hose over the tank bottoms or 2) increasing the water pressure. Effort should be made to keep the drains clear from clogs to

44 prevent back up and pooling of wastes. Removal of the chips from the tanks bottoms before the manual clean may would likely require a tank retrofit, but would likely assist in water reductions.

Labatt Krausen Tank Manual Clean Wastewater Prof ile 110000 100000 _ 90000 "a 80000 E 70000 c o 60000 '£3 (0 50000 40000 • BOD o 30000 w • TSS 20000 10000 0 Initial 1st Rinse 2nd Rinse 3rd Rinse Chips Split 4th Rinse 5th Rinse Pre-CIP Burst Rinse

Process

Figure 5: Average Wastewater Profile of Manual Clean Process for Krausen Tank

Assuming that a viable strategy could be implemented to divert tanks bottoms from the floor drains into a separate holding vessel, sedimentation may be an effective way to reduce solids loading and reclaim beer. Sedimentation of Krausen tank bottoms was briefly examined in

February '10 (see Figure 11C and 11D in Appendix A). After approximately 4 hours at 4°C, well defined stratification occurred in a 1L sample. Three separate layers are observable. The upper layer is thought to be beer, the middle - yeast, and the bottom - beech chip sediments. The beer supernatant is easily decanted, and contains only 3.9% of the TSS concentration from the homogenized sample. Further sedimentation tests should be conducted to confirm the validity of this technique. Centrifuging may also be an option.

45 3.3.3 Filtration

In Filtration, drop beer filter cake or diatomaceous earth (DE) slurry results in a direct and highly concentrated total suspended solid discharge to sewer if not adequately pretreated. A Komline

Rotary Drum Vacuum Filter (RDVF) has been employed for this purpose since 1982. The RDVF generally runs for 1 hr, and is idle for 6 hr. The poor performance of the RDVF was noted, especially during a dramatic decline in August 2009. A similar decrease in performance was noted in 2007. The Komline RDVF should consistently achieve 90% solids removal. During the sampling program, only 40% TSS removal was observed. In RDVF operation, a filter cake accumulates on the surface of a fabric coated drum via vacuum pressure as it rotates through a vat of diatomaceous earth slurry. The drum is generally 37.5% submerged in the vat. The vacuum has a dual purpose. It picks up solid in the vat to form the filter cake, and dries the filter cake as it emerges from the vat. Optimum operation requires the filter cake be dry, even, and relatively thick upon cake discharge via the scraper blade. The scraper blade is generally set to clear the cloth surface by approximately 1/8". Cake thickness should be greater than %" for this unit. Solids removed by the scraper are collected in a bin at the bottom of the unit for disposal.

Komline-Sanderson Engineering Corp. was contacted by Labatt to assess the RDVF on Tues.,

February 2nd, 2010. Inspection revealed the RDVF operational setup resulted in an inferior cake formation that was both thin and moist. There are several possible explanations for poor cake formation. The applied vacuum pressure may not be sufficient. This would result in a thin cake formation in the vat that would not air dry well before cake discharge. The vacuum pump, and all the pipes and fittings that maintain vacuum may need to be examined and repaired. The speed of rotation may not allow sufficient contact time for cake formation. The switch to a finer grade of DE, or a difference in its composition could account for variations in the RDVF performance. The possibility of an alteration in the type of DE used for filter beer has not been

46 investigated. As a result, the filter material may now be inadequate for the application. Too fine a mesh would blind the filter material, resulting in a thin, dense filter cake difficult to dry. Too coarse a mesh will cause solids break through. The filter mesh should be cleaned routinely to prevent fouling. Filter cloth life should be recorded, and the location, type of wear, rip or combination of the two should be noted (Komline Sanderson 1982).

Labatt London submitted a DE Slurry Sample to LEX Scientific Inc. for particle size analysis on

June 23, 2010 (see Figure 6). The mean particle diameter is 5.2 |jm, and less than 0.1% of the DE

Slurry has a diameter greater than 50 |Jm. The filter cloth pore size during sampling was 150 pm, suggesting that at least 99.9% of the particles pass through the filter cloth upon initial start up.

This explained the significant lag observed before a filter cake began to accumulate on the RDVF filter cloth. There are three options to address this issue:

1) Test a new filter material with a finer mesh size using a leaf test kit and install the optimum

candidate. The vacuum pressure and speed of rotation would also require adjustment

whenever the properties of the filter cake are significantly altered. A finer filter material

would incur a higher resistance to air flow, thus the vacuum pressure would likely have to

be increased. The filter cake would also accumulate faster. A balance must be struck

between filter cake depth and its porosity to ensure the cake can still be air dried once it

leaves the vat. Filter cloth installation is detailed in Section 4 of the Komline RDVF Manual,

1982. Filter leaf tests must reproduce operational parameters of the RDVF;

2) A course pre-coat of DE on the RDVF with a particle diameter greater than the pore size of

the fabric could stimulate filter cake development; or

3) The particle size must be increased with polymer addition. UoG has been cooperating with

Ocean Chemicals in exploring the possibility on RDVF performance enhancement with the

addition of polyacrylamide to improve cake formation. The polymer acts as a flocculating

47 agent to increase the size of suspended materials, thus assisting in their removal. Flocculant

removal using the RDVF should be investigated to determine the maximum applicable

vacuum pressure before floe complexes breakdown. Strong turbulent forces could tear the

larger coagulated particles apart into their smaller constituents. It is recommended that the

RDVF influent and effluent be tested regularly for TSS to ensure performance is as expected.

Ml* t«nM Cumulative rtrt«nug«

18 8-22.9 37 20 l«.t-12.» 97.7 229-270 21 1.1 is.t - iT.a r 98 3 27 8 33.8 12 0 6 17.t - »3.t I 99.5 :«H-4i i K U3 33.3-411 1 41 1 -53 2 G.I 411 -50 99 9 V O <.*» 2 0.1 » 99 100 0

Total Count: 1864 Mean: 5.2 microns Standard Deviation: 6 microns

Figure 6: Diatomaceous Earth Slurry Particle Size Distribution

A Labatt filtration operator estimated that DE slurry is sent directly to drain three to four times a week. Direct discharges occur whenever there is a turbidity probe fault. This results when DE sediments aggregate and cake upon the probe. These discharges could be minimized if a self cleaning mechanism were installed.

48 The RDVF should also be kept in optimal repair according to Section 13 of the Komline RDVF

Manual to reduce. Labatt London maintenance practices should reflect the specifications provided by the manufacturer.

3.3.4 Bright Beer

Bright Beer tanks may be a potential point source for BOD loading of the brewery effluent due residual beer volumes after product transfer to packaging. The sewer selected for analyze of

Bright Beer wastes had inputs from several other contributories. As such, the values were not particularly useful. Quantifying bright beer tank residuals should be considered an objective for future sampling programs as this point source could not be adequately described over the duration of this project.

3.3.5 Packaging

In 2008, packaging consumed approximately 745 000 m3 of water, and accounted for 43.7% of

Labatt London's total water usage. Water demand is incurred by bottle washers, line lubricant, vacuum fillers, jetters, and pasteurizers. There are three bottling lines (Line 1, 4, & 5), one canning line (Line 3), and a keg Line (Line 2). Assuming no evaporation or leakage, all water consumed by packaging becomes wastewater.

The packaging wastewater can be characterized as having a significant BOD (1000 to 2000 mg/L) with TSS and TP contributions below the maximums established by the municipality. BOD contributions are primarily attributed to the various processes that result in beer discharged to drain, most notably brand changes, fobbing, and breakages. From the week of April 20-25, 2009, it was projected that the bottle, canning, and keg lines lose approximately 2.1%, 1.1%, and 2.3% of finished beer by volume respectively. Brand changes account for a significant portion of this loss, however much of it is recovered from the Butts Tank and sent back to carbonation to be

49 refurbished. Non-blendable specialty beers that contain potential allergens (i.e. potassium metabisulphate), flavoured adjuncts, or high colour must be discharged directly to the floor drains. To ensure there is no mixing of the chase brand, a DCW rinse and bowl flush must follow.

The specialty brands include:

Bud Light Lime (Lime flavour additive)

Dealcholized Blue (KMS preservative)

Selection Dealcholized (KMS)

PC Red Dealcholized (KMS)

PC Blonde Dealcholized (KMS)

Compliments Dealcholized (KMS)

Lakeport Ale (KMS)

Lakeport Honey Lager (Honey Additive)

Lakeport Red (High colour)

Budweiser Select (Low AE)

Bass Pale Ale (High colour/AE)

Approximately 21 hL are discharged to drain per brand change. Labatt personnel estimate these brand changes amount to 315 m3 beer per year sent to drain. Diversion of this waste stream from drain to rebate beer would reduce BOD.

The bottle soakers also incur a significant BOD loading. This may be derived from residual beer, label pulp, glues, and resins maintained on the return bottles. BOD from labels may be reduced by:

• Employment of low BOD pigments, labels, and adhesives;

• Label size reduction (i.e. Sleeman);

50 • Painted labels (i.e. Steam Whistle); and

• Avoiding pulp formation by employing alternate label removal strategies.

Reducing the screen size on the Butt Sprayer effluent which discharges to the soaker pit is also worth investigating. The bottle washers consumed approximately 236 622 m3 of water in 2008.

Pasteurizers consumed approximately 158 699 m3 in 2008. This waste stream is very clean, so there is potential for reuse.

Brand Changes on the Bottle Lines may also be an area for reducing wastewater nutrient loading.The extent of lateral mixing between different brands of beer during a Brand Change has not been investigated. It is possible that the magnitude of mixing may vary between different beers (i.e. dissimilar beers versus similar beer). This may be a consideration for future optimization. Due to a lack of information regarding the extent of mixing in the lines during a

Brand Change, it is common practice to waste more beer that may be necessary to preserve product integrity. This may present an opportunity for Rebate Beer reduction in favour for increased finished product at no additional cost.

One Labatt packaging operator believes that greatest potential for optimization of the packaging process would come from allowing the fillers to chase out existing volume in the beer lines with de-aerated carbon filtered water (DCW). This should be done after brands that require a line flush as all volume in the lines (16 to 24 hL) then goes to drain. Any brands with flavouring (i.e. lime, honey) cannot be reclaimed, and are sent directly to drain. This would result in less wasted mixed beer.

Minor increases in liquid usage may also be available the actual volume between each tank and each filler were determined, and increase the number of dozen for tank changes that are further away from the fillers and minimize the volume of beer line pack after the last brand has been pushed back.

51 The Keg Line exhibits significant BODs loading from returns and quality control practices. While in operation, the keg line is two to ten times more concentrated than the bottle and canning lines(2492mg/L to 13884 mg/L), however the quantity of water used is significantly smaller than the other packaging lines (15 377 m3 in 2008). The majority of the keg line BOD contributions are attributed to the volume of residual beer in return kegs, quality control samples, brand changes, and low fills. Labatt personnel estimate 33 903 hL of beer per year are sent to drain from these sources. A recommendation to conduct a survey on the volume of beer in return kegs was made. Not enough data are available to confidently determine whether running domestic, US, or Bass Ale production makes a significant difference on BOD loading. It appears that domestic production results in the highest effluent concentrations, followed closely by US and Bass Ale.

Line lube is continuously sent to floor drains even while to line is idle. The keg sump pit does not exhibit any significant wastewater loading while it is offline.

3.3.6 Powerhouse

The effluent C02 scrubber water discovered to be a significant and previously unrecognized

BOD5 contributor. The source of the BOD is speculated to result from the volatilization of beer stone in the Fermenters. The scrubber water has a sweet aroma, and contains no solids and very

little phosphorous. This is to be expected as the influent C02 scrubber water is de-aerated carbon filtered water. There could be potential to reuse this water source. Further investigation should be conducted to determine the suitability for reuse in certain applications. For example, the scrubber water may serve as sparging water for use in either the lauter tun or whirlpool. It may also prove a useful as a clean in place (CIP) rinse water. Assuming 2008 water usage values and city water costs of $1.00 per m3, reuse of this relatively clean waste stream could save

$13,600/yr in raw water usage.

52 3.3.7 Clean In Place

The duration for the various CIP washes were slightly modified on RS-View between April and

September '09. For example, the duration of the sanitizer rinse for the Vertical Fermenters was decreased from 41 minutes to 18 minutes in total. These changes are actually fairly common throughout the course of the year as operators aim to increase water efficiency while maintaining optimum performance of the CIP cycle. The effect on the wastewater effluent should be accounted for when future changes are made.

3.3.8 Main Effluent

The brewing process results in batch discharges that result in a highly variable main effluent composition. All sampled locations, with the exemption of the pasteurizers and boiler blow

down, were in exceedance of the maximum WDB BODs concentrations of 300 mg/L (see Figure

7). The average BOD5/COD ratio for the main effluent 0.69 ± 0.16 which is on the high end of literary values of 0.65 ± 0.05. (Brito, et al. 2007). The ratio indicates a high concentration of organics, as BOD/COD > 0.3. The wastewater is thus readily decomposable, and suitable for biological treatment.

53 Labatt London Main Effluent Oxygen Demand *09

5000 •4500 S 4000 | 5500 e 5000 | 2500 ~ 2000 | 1500 3 IOOO • 6005 500 0

Month

Figure 7: COD and BOD5 Loading of Brewery Effluent May 2009 to February 2010

TSS in the main effluent is primarily associated with the brewing side. TSS concentrations are consistently over the WDB maximum of 350 mg/L (see Figure 8). Yeast and spent DE are the largest contributors. TSS concentrations were notably higher when the RDVF was not performing well resulting from spent DE discharged directly to drain. Yeast slugs from

Fermentation, Krausening and Aging enter the waste stream whenever vessels are manually cleaned or CIP'd. The tank bottoms are discharged directly to the floor drains. This waste volume is roughly approximated at 5 hL per vessel; it may vary dramatically between vessels type. By design vertical tanks have a smaller ratio in the volume of waste to tank capacity than horizontal tanks.

54 Labatt London Main Effluent Solids Loading '09 ?snn

• TSS

• VSS

y * ./

Month

Figure 8: TSS and VSS Loading of Brewery Effluent from May 2009 to February 2010

S»« Count Cumulative (H»)

200-224 462 193 joe .tux 193 224-252 420 us 294 4-Ml t ; 366

252-283 322 134 SSI* - Hi t • i so: 263-31? 298 12 4 tn.t - sir 62 7 317-356 218 8 1 »i7-»M.r •H 71 e 956-399 220 9 2 358.7 • JM.I m m 80S 3S9-449 131 55 3M.1 • 447.7 • m 864 448-502 65 3.5 447.7 Mi.4 m I 89 S 5C2-364 78 3 3 502.4 - 563.7 m i 93 2 564-632 48 2.0 568.7 • 4)2.5• 95 3 632-710 SO 2.1 *32.8 • T9S.6 • 97 4 710-796 22 09 70S.4 - TMJ i 983 756-893 12 0.5 796.2 • 193.4 i 96.8 t>*j - 1902 5 0.2 883.4 -1002.4 99 0 11)02-1125 7 OJ 1002.4-1124.T 99 3 1125- 126«2 1 U 1 1124.? • 1261.9 994 1262- 14lt> 9 04 1261.9 • 1416.9 88? M16- 15t» 3 0 1 1415.S • 1598.7 H9fe

1S89-1783 3 0 1 1SI8.7.1712.5 100 0 1/83-2000 0 oa 17M.S-209# iUUl) > 200C 0 0 3 > 2080 100 0

Total Count: 2397 Mean: 328 microns Standard Deviation: 159 microns

Figure 9: Labatt Brewery Main Effluent Particle Size Distribution

55 Total phosphorous loading is primarily attributed to the brewing side. There were only three noted exceedances from the 10 mg/L maximum during the sample period (see Figure 10). The vast majority of the phosphorous originates from the malt in the beer. Phosphorous accumulates within the tank bottoms. Krausen tank bottoms owned a TP concentration ranging from 112 to 568 mg/L, identifying it as the most concentrated point discharge to sewer tested.

There is also significant phosphorous loading from post-acid washes during CIP operations.

The effluent pH was generally within the 6.0 to 10.5 range specified in the WDB with one exception in September (see Figure 10). The pH adjustment system may require some fine

tuning. It is presently set to dose with C02 or NaOH when the pH exceeds exactly 6.0 or 10.5 to save on chemical. It is recommended that the range be at least somewhat narrowed to allow for some redundancy to prevent the effluent pH from going off spec. It is also suspected that the rapid mixer off the effluent pit may not be performing optimally.

Labatt London Main Effluent Total Phosphorous Loading and pH *09

•as K 0 c 1-i.O B 10.0 IP

• T= • pi-

J? /

Month

Figure 10: TP Loading and pH of Brewery Effluent May 2009 to February 2010

End-Of-Pipe treatment is recommended to secure immediate BOD5 reductions to below acceptable concentrations. As process improvements continue into the future, end-of-pipe

56 treatment would still be useful as a polishing step. A Biological Packed Tower Pilot Test has been proposed to accurately assess the aerobic microbial treatment capacity. The suggested design would modify the existing pH adjustment reactors to accommodate a robust, low footprint biological alternative for brewery effluent treatment. The BOD/TKN ratio is greater 100 (much greater than 5), suggesting that aerobic decomposition would dominate over nitrification

(Metcalf and Eddy, Wastewater Engineering: Treatment and Reuse: Fourth Edition 2003). As long as the wastewater is kept aerobic, very few aromatics would be generated by this system.

57 Chapter 4: Added Value from Rebate Beer in Anaerobic Digestion

This phase of the research was designed to investigate the feasibility of using rebate beer (RB) as a co-substrate for the anaerobic digestion (AD) of dairy manure (DM). Rebate beer is a non­ food grade product that consists of expired market returns and/or mixed brands of beer reclaimed from beer packaging lines in the brewery. Rebate beer must be disposed of as a liquid waste by the brewers. The disposal of rebate beer to sanitary sewers without pre-treatment is not often an economically viable option because of high-strength sewer surcharges (HSS)

incurred from BOD5 exceedences associated with the high sugar content of beer. Anaerobic digestion is a renewable energy alternative that uses microbial action to convert organic matter from waste into biogas, and stabilized fertilizer. Dairy manure is a common primary agricultural material input for anaerobic digesters, however their performance is somewhat limited by a carbon deficiency. Carbon is often considered the growth-limiting substrate for the family of microorganisms in the anaerobic digestion of dairy manure; it is required for microbial synthesis of volatile fatty acids (VFA) by acetogenic bacteria, which in turn is food for the biogas producing methanogens (Lyberatos and Skiadas 1999). Rebate beer has high concentrations of readily accessible carbon for microorganisms manifested by soluble carbonaceous biochemical oxygen

demand (cBOD5) concentrations that are in excess of 12 000 mgo2/L (see Chapter 3). Rebate beer is classified as a category 2 non-agricultural source material (NASM), and may currently be utilized in Ontario anaerobic digesters in quantities up to a maximum 25% by volume under the

Nutrient Management Act (NMA). Positive results from testing RB as a co-substrate for the anaerobic digestion of dairy manure could establish grounds to create a mutually beneficial partnership between brewers and farmers:

• Brewers would have an affordable alternative disposal route for RB; and

58 • Dairy farmers operating anaerobic digesters would gain a faster return on investment

from increased biogas generation.

An experimental procedure was devised to test the suitability of RB as a co-substrate. A preliminary biochemical methane potential (BMP) test was conducted to validate progression toward bench-scale digester operation. The bench-scale digester was a single phase continuously stirred tank reactor (CSTR) composed of cylindrical PVC digester and maintained at mesophilic temperatures (35°C) by a temperature controller. The digester was fed daily with a mixture of rebate beer and dairy manure. A 50% DM/50% RB and 75% DM/25% RB blend (by volume) were tested against a 100% DM control feed stock. The digester was fed 500mL of

3 diluted feedstock to achieve an organic loading rate of 1 kgvs/m /day. A sample was taken daily from the digester, and various parameters, including temperature, pH, alkalinity expressed as

bicarbonate (HC03), volatile fatty acids (VFA), total solids (TS) and volatile solids (VS), were measured, either daily or twice weekly. Gas volume produced during anaerobic digestion was recorded by a wet-tipped gas meter. The quality of the biogas was determined with a Gas

Chromatograph.

All blends were found to have a volatile solids (VS) reduction that exceeded 29% for the majority of the digester's operation. Increases in methane yields were observed with the addition of rebate beer. Stable digester operation was achieved for both the 75% DM/25% RB and 100%

DM control blends as the ratio of volatile fatty acid to bicarbonate (VFA/HC03) ratio decreased to below 0.3; effluent VFA concentrations were reduced to near zero. The 50% DM/50% RB produced the highest quantity of biogas however the digester performance was less stable than the other blends. Digester instability was shown through longer acclimatization periods to

achieve steady state operations, and in VFA/HC03 ratios that would occasionally climb too high, which would inhibit methangenesis and ultimately cause digester failure. This study indicates

59 that successful mesophilic anaerobic digestion of DM with RB as a co-substrate is both achievable and sustainable in maximum volumes allotted by the NMA (25% RB). The optimum blend likely lies between 25% and 50% RB in DM. Further study to narrow this range may be desired if the legislation changes to reflect an increase in the allowed volume.

4.1 Introduction to Anaerobic Digestion

Anaerobic digestion is the multi-variant biological decomposition of organic waste. The following section provides basic background data on the current theory, innovations, and

Ontario legislation involved anaerobic digestion, and presents the overall objectives of the study.

4.1.1 Background on Anaerobic Digestion

Anaerobic digestion uses microbial processes in the absence of oxygen to convert organic wastes into a valuable biogas and stable, liquid fertilizer (Demirer & Chen 2005). The biogas

principle components are 50-75% methane (CH4) and 25-50% carbon dioxide (C02), and may be burned for onsite heating or energy generation (Archer, et al. 2005). The digester effluent is ideal liquid fertilizer for agricultural land application. Compared to the undigested feedstock, the digestate is a higher quality fertilizer with more stable, bioavailable forms of nitrogen in ammonia, and phosphorous, as orthophosphates. The digester effluent also exhibits improved hygienic conditions in terms of ground pathogen and odour reduction (Lusk 1995).

The three major steps of anaerobic digestion are illustrated in Figure 11; Hydrolysis,

Acetogenesis, and Methanogenesis. During the first stage, complex organic compounds, e.g. fat, cellulose, and proteins, are hydrolyzed into small soluble compounds by various groups of decomposing bacteria. Followed by this is the second stage, during which the soluble organic compounds are fermented into acetate, organic acid, and hydrogen, which will be converted

60 into methane and carbon dioxide eventually in the third stage. During the anaerobic process,

the electrons in the biodegradable organic matter (i.e. BOD5) are transferred to methane, in which carbon is in its most reduced oxidation state, and the organic matter is stabilized.

Theoretically, one gram of BOD5 can generate 0.B5 L of CH4 under standard conditions (i.e. STP at T = 20°C and P = 1atm) (Nebot, et al. 1995).

Complex Particulate Organic Matter

Carbohydrates Proteins Fats

Amino Sugars Acids Long Chain Fatty Acids

^ Propionate Butyrate Valerate

Acetate Hydrogen

Methane

Figure 11: The carbon flow in the methane production process

Anaerobic digestion can be employed to treat a large group of organic compounds. However, compared to highly structured materials, for example, the woody and agricultural residues, anaerobic digestion is more suitable to process poorly structured, easily degradable wastes, such as food waste and sewage sludge.

Co-digestion, also called co-fermentation, is the simultaneous anaerobic digestion of different substrates (co-substrates) with a primary substrate at much higher relative proportions within an existing digester. Co-digestion can be employed in agriculture, to digest the co-substrates

61 composed of manure and energy crops to improve biogas energy output. It also demonstrates great potential for digesting some organic wastes with sewage sludge.

The improved biogas production through co-digestion results from the balanced non-lignin carbon to nitrogen (C/N) ratio and micronutrient. Unbalanced C/N ratio inhibits the anaerobic digestion efficiency due to the formation of ammonia nitrogen (TAN) and volatile fatty acids

(VFAs), which, if they accumulate too much in the digester, would inhibit the activity of methanogens, the bacteria responsible for methanogenesis. For example, the C/N ratio of sewage sludge is typically between 6/1 and 16/1,while organic waste has a much higher content of organic carbon content, with the C/N ratios of about 30/1 or higher. The optimal C/N ratio for anaerobic digestion should be in the range of 20-32 where the maximum methane production per loading rate is around 25 (Hills and Roberts 1981). Therefore, the combination of the beer and dairy manure could result in a better-balanced C/N, as well as nutrient recipe, and consequently leads to enhanced biogas production.

In addition, the production of biogas through anaerobic co-digestion offers significant advantages over other forms of waste treatment, including:

• Less biomass sludge is produced in comparison to aerobic treatment technologies;

• Successful in treating wet wastes of less than 40% dry matter;

• Pathogen inactivation. There is often less viable e-coli and fecal coliform in digestate

than in raw manure, rendering it safer for land application. This is especially true for

multi-stage digesters and/or if a pasteurization step is included in the process;

• Minimal odour emissions as 99% of volatile compounds are oxidatively decomposed

upon combustion, e.g. H2S forms S02; and

• The effluent slurry produced (digestate) is an improved fertilizer in terms of both its availability to plants and its rheology.

62 4.1.2 Biogas Facilities

Farms and vineyards are well suited to develop biogas facilities. Biogas systems better stabilize traditional fertilizers such as dairy manure for land application while potentially turning a profit from fuel energy generation.

The feasibility of independently operated digestion facilities are constrained by capital costs compared to the financial returns on biogas from available feedstock. Historically, only large establishments would generate sufficient manure or food wastes to be able to afford such a facility. However the push for green energy and institution of the Feed-in Tariff (FIT) model in

Ontario has allowed the establishment of smaller facilities. The Ontario Power Authority (OPA) proposed FIT rates for 20-year contracts range from $0.16 CAD/kWh (Canadian dollars) for biogas projects less than 500 kW to $0,104 CAD/kWh for projects over 10 MW (Greer 2009).

The Ontario Biogas Systems Financial Assistance Program ran from September 2008 to March

2010. It was an $11.2 million investment that offered grants to farmers and agri-food businesses to develop operational biogas systems for the production of clean energy. Up to 27 new,

Ontario based biogas systems were initiated by this program alone. These systems reduce electricity costs and green house gases, improve waste diversion rates, and contribute to local economies (OMAFRA 2010).

The Ontario government has allowed biogas facilities to accept up to 50% Non-Agricultural

Source Materials (NASM) as feedstock for anaerobic digesters under the General Nutrient

Management Regulation (O. Reg. 267/03). NASM includes yard waste, fruit and vegetable peels, food processing waste, pulp and paper biosolids and sewage biosolids. Proper spreading of these materials on farmland returns essential nutrients to the soil to help foster new plant growth. The Ontario Ministry of Agriculture, Food, and Rural Affairs (OMAFRA) issues certificates

63 of approval (CoA) for NASM application to agricultural land. A NASM Plan would have to be developed.

NASM and Soil testing results are used to complete the NASM Plan for calculation of the maximum land application rate. Soil samples collected from the proposed application area on behalf of the farmer are required to determine background soil pH, metal, and nutrient concentration. Requirements for NASM material testing are dependent upon the prescribed category of NASM. Category 1 materials do not require sampling and analysis, unless the material is intended to be spread at a rate that exceeds 20 tonnes per hectare/year. In such cases, NASM sampling for nitrogen and phosphorus is required; the soil must be tested for soil pH, phosphorus and potassium. Category 2 and 3 materials require testing for metals, nutrients, and possibly other parameters, (e.g., fats, oils & grease, sodium) (0. Reg. 267/03). All material sampling and laboratory analysis must be performed by an accredited laboratory for the particular analysis to ensure the material meets all quality standards as outlined in 0. Reg.

267/03. NASM generators must provide the Farmers or other Certified Plan Preparer with NASM testing results prior to the receipt of NASM.

4.1.3 Rebate Beer as a Co-Substrate

The affordable disposal of rebate beer is an issue for breweries across Canada. Rebate beer is unmarketable product resulting from unblendable beer reclaimed from within brewery operations, and from expired product returned to the brewer by vendors. Rebate beer does not meet the product's food quality standards for consumption. Approximately 3% of all beer produced by a brewery ends up as rebate beer. For a large brewery, this may translate into upwards of 10,000 m3 of rebate beer per year. Rebate beer has a COD:TN ratio of about 120:1

and is high in BOD5 (>12 000 mg/L); this ratio may vary between different brands. Its disposal to sanitary can overload wastewater pre-treatment systems and may result in violations to

64 environmental legislations. Depending on the jurisdiction, costly high strength sewage surcharge fees may be applied by the municipality. Source separation and alternate disposal is desirable in most industrial brewing facilities. Rebate beer shipment to pharmaceutical manufacturers for further distillation into medical grade alcohols is a common disposal method; however, due to transportation costs, feasibility and scales of economy are limited to distance between facilities.

The residual fermentable sugars and alcohol content make rebate beer a desirable feedstock for the production of pharmaceutical-grade alcohol. Brewers may be able to turn a marginal profit off rebate beer if transportation costs can be minimized. Labatt London reports a Hamilton based company accepted rebate beer at a net cost of $20.00/m3 to the brewer.

Rebate beer may be potentially utilized as a co-substrate for anaerobic digestion facilities using dairy manure in the local farming communities. The proposed disposal method of rebate beer would be mutually beneficial to both the brewer and the dairy farmer. Rebate beer qualifies as a

Category 2 NASM under 0 .Reg. 267/03. The carbon-nitrogen ratio of dairy manure in terms of

COD to TKN is between 20:1 and 25:l(Burke 2001).The bioavailable non-lignin carbon to khedjal nitrogen ratio of screened dairy manure is 8:1 (Hills and Roberts 1981). The C:N:P ratio for anaerobic biological treatment is about 250:5:1 (Metcalf and Eddy 2003).

4.1.4 Digester Objectives

The objective of this project is to evaluate the effectiveness of rebate beer as a co-substrate with dairy manure in bench scale anaerobic digester. A 50% DM/50% RB and 75% DM/25% RB feed stock will be tested and compared to a 100% DM baseline. Effectiveness will be measured in terms of biodegradability, process continuity, and impact of varying organic loading rate on biogas production and the quality of biogas.

65 4.2 Bench Scale Digester Project Methodology

The following section details the experimental methodology for bench scale digestion of dairy manure with rebate beer as a co-substrate. This investigation required the construction and operation of small bench scale anaerobic digester. A secure source of dairy manure and rebate beer was established:

• Dairy manure was obtained from the Elora Dairy Cattle Research Station; and

• Rebate beer was provided by Labatt Brewery in London Ontario.

Several analytical procedures were performed throughout the investigation on both feedstock and digestate to monitor overall digester performance. Additional information is located in the Appendix B.

4.2.1 Source of Waste

Fresh dairy manure was collected from the Elora Dairy Research center every two months throughout the duration of the project. Elora Dairy used a free stall arrangement, straw bedding, and dry scrape manure removal. The healthy milking or 'wet' cows were fed a regiment detailed in Table 6.

Manure was collected directly from the sump pit in 18L buckets, transferred to the University of Guelph, and stored in a laboratory refrigerator at 4°C. The dairy manure was put through a Waring Laboratory

Blender from General Electric on low in 1L increments for 30 seconds intervals to cut up large strands of straw. Blending eased digester feeding by reducing clogs at the feed port.

66 Table 6: Dairy Cow Feeding Regiment

Feed Ingredient TMR (35 kg) TMR (35 kg) As Fed Dry Matter Straw 1.3 1.3

Haylage (31.17% dry matter) 20 6.25

Corn Silage (36.48% dry matter) 17.1 6.25

High Moisture Corn 7.3 4.97

Dairy Supplement (Pellet) 5.0 5.00

TMR Totals 50.7 23.8

Bottled rebate beer was provided by Labatt London. The 351mL bottles of assorted brands were stored at room temperature.

4.2.2 Preliminary Biochemical Methane Potential Test

A Biochemical Methane Potential (BMP) test was completed as a precursor to the bench-scale anaerobic digester project. The BMP test simulates a closed small-scale anaerobic batch reactor in a sealed 500 mL

Wheaton bottle. It provides insight into toxicity and the amount of biogas that may be produced from a given substrate. In effect, it would determine the overall viability of rebate beer as a co-substrate with dairy manure as the BMP test. The BMP procedure outline is Owen et al, 1978. This method was simple, inexpensive, and provided a good indicator of suitability of constituent for anaerobic digestion (Owen, et al. 1978).

The organic loading rate (OLR) and hydraulic rentention time (HRT) were control variables for the bench

3 scale digester. A volatile solids concentration of 2 kgvs/m was used for the BMP test. The VS for beer

and dairy manure were 3.00% (approximately 31 300mgvs/L) and 10.8% (approximately 108 000 mgvs/L) respectively. The proportions of stock solution, innoculum, and rebate beer to dairy manure added in the BMP test is recorded in Table 7.

67 Table 7: Amount of substrates required for BMP test of rebate beer and dairy manure

Assay Sample # Stock Sol. Inoculum Distilled Rebate Dairy Identification Duplicates Vol Volume Water Beer Manure (mL) (mL) (mL) (g) (g) 1 Control (Blank) 2 90 20 90 0.00 0

2 DM 100%/RB 0% 2 90 20 90 0.00 4

3 DM 75%/RB 25% 2 90 20 90 3.25 3

4 DM 50%/RB 50% 2 90 20 90 6.5 2

5 DM 25%/RB 75% 2 90 20 90 9.75 1

6 DM 0%/RB 100% 2 90 20 90 13 0

Total Assay Required: 12

Biogas volumes were measured with a water displacement device (Demirer, et al. 2000). The measurements were initially completed on a daily basis when gas generation rate was high. The frequency of measurement was scaled back to every 3 to 5 days to achieve at least 10 mL between measurements.

Biochemical Methane Potential: Dairy Manure and Rebate Beer OOK Dairy Manure s 100% Rebate Beer '€ «>25X Dairy Manure -8 75* Rebate Beer 0hm & —* *w50% Dairy Manure 3 f 50% Rebate Beer I5 is. -*-75% Dairy Manure » 25% Rebate Beer 1 3 - *100% Dairy Manure E 3 0% Rebate Beer V

10 IS 20 25 30 35 40 Time (day)

Figure 12: BMP of dairy manure and rebate beer

68 The RB and DM BMP test validated the progression toward Bench-scale anaerobic digester operations.

There were no observed toxic effects to the microorganisms as each blend produced biogas. From the results of the BMP assay presented in Figure 12, it was evident that rebate beer assisted in biogas generation from dairy manure early on. The 50% DM/50% RB and 75% DM/25% RB blends appeared to be the most appropriate proportions as they produced a higher cumulative volume of biogas than the

100% DM baseline. The 25%DM/75%DM blend and 100% RB biogas cumulative volumes were overtaken by the 100% DM baseline toward the end of the experiment.

4.2.3 Digester Design

A cylindrical 8"0 PVC digester was used to conduct the experiments. The experimental apparatus is illustrated in Figure 13. Particulars of several components are listed in Table 8. The unit had an active capacity maintained at 12 L with 3L of head space. The digester contents were continuously mixed by an externally mounted variable-speed DC motor. The custom stainless steel shafts were rotated at a speed of approximately 45-60 rpm. The mixing system was sealed with oil seals to maintain anaerobic conditions. The digester was heated to a constant 35°C with 4.6m electrical heating cable controlled by a local thermocouple and temperature controller. The heating cable was wrapped around the exterior of the vessel and fixed to the side with aluminum tape; a 10cm gap was maintained between each successive wrap. A reflective 5mm thick insulation was fitted over the sides of the digester over the heat tracer wire. The volume of the gas produced during digestion was recorded by a patented wet-tip gas meter. Biogas quality samples were collected in a 500 mL tedlar bag during steady state operation by temporarily exchanging the connection to the wet-tip gas meter for a 1 hr. The digester was outfitted with two 1"0 ball valves for sampling; a feed port on the topside, and an extraction port at the bottom for withdrawing effluent. A sample of digested mixed waste was withdrawn daily from the bottom of the digester after recording the volume of gas produced. Feedstock was added by gravity through the feed port in the top of the digester.

69 Tip Counter

Oil Seals & Bearings

Wet-Tip Gas Meter Gas Transfer Line Digestate Level Heating Cable

Mixing Shaft Temperature Controller Insulation

Thermometer - Thermocouple

Sample Port

Figure 13: Schematic drawing of lab digester

As a safety precaution, the digester was placed in a flat 25L bin for secondary containment to capture all

digestate if the vessel were compromised. An H2S monitor was also placed near the digester as it operated in the open directly on the lab bench. Ideally, the digester unit would have been placed under a designated fume hood to control the aromatics released during feeding.

70 Table 8: Primary Digester Components List

Unit Supplier Cat. No.

Love Controller Model RA 505335 Cole Parmer 02110-82

IKA Variable Speed Mixer 115 VAC Cole Parmer 50705-00

Thermometers 25 to 125°F Cole Parmer 08127-15

Heat Tracer 120VAC 40A, 15W/ft @ 50F Drexan 010811

The feed stocks for the digesters were blended in relevant proportions according Table 9 prior to use.

The finished feedstock was transferred into labelled 18L feed buckets, and stored at 4°C.

Table 9: Digester Feedstock Blends

Dairy Manure Rebate Beer Average Number of (% by volume) (% by volume) Storage Time Blends Made 25% 75% Not Tested 0

50% 50% 16 days 7

75% 25% 62 days 4

100% 0% 77 days 1

It should be noted that the proportions of rebate beer to dairy manure for the BMP test were mass based as opposed to volume based for the bench scale anaerobic digester operations. The switch was made as the Nutrient Management Act restricts the permitted amount of NASM in agricultural anaerobic digesters based on volume, not mass. This means that the BMP results for the 75% DM/25%

RB blend would be more representative to the results expected for the 50% DM/50% RB blend under for the bench scale as the density of both manure and beer is approximately equal to 1g/mL.

4.2.4 Start up Procedure and Operation

The initial start-up of the digesters involved using 8 L of actively mesophilic digested sludge from the

City of Waterloo wastewater treatment plant as an inoculum, and adding 5 L of mixed waste to the

71 digester. After the feed and digested sludge were placed in the laboratory digester, temperature was rapidly raised to 35°C. No feed stock was added to the digester for a period of 2-4 days while the performance of the digesters were monitored for both process stability (pH, volatile fatty acid, alkalinity, and gas production) and solid reduction parameters. The design daily feed sludge (see Table 9 for recipe) was added to the digester as gas production increased and organic acid concentration level dropped to below 3000mg/L.

500 mL of well mixed effluent was drawn from the sampling port at the bottom of the digester once a day and set aside for analysis. The digester was then fed 500 ml of diluted feedstock at an organic

3 loading rate of 1.0±0.1kgVs/m /day. The specified volume of mixed feedstock was determined using equation 8, and diluted to 500 mL with warm tap water.

OLRXVD X10 Vfeed = -7T [81

Where Vfeed = Volume of feedstock in mL

VD = Working volume of digester (12L)

Cvs = Volatile solid concentration of feedstock (mg/L)

3 OLR = Organic Loading Rate (1kg vs/m /day)

Feedstock volumes for the 50% DM/50% RB, 75% DM/25% RB, and 100% DM trials were 220mL, 180mL, and 130mL respectively. Digestate height within the digester was monitored monthly using a dipstick, and adjusted accordingly with tap water to maintain the design hydraulic retention time (HRT) of 24 days.

The water level in the wet tip gas meter had to be maintained at a constant depth to maintain unit's accuracy. The wet tip gas meter was calibrated monthly by injecting air into the gas transfer line with a

72 100 mL syringe. The water within the wet-tip flow meter is adjusted to a pH of 4 with H2S04 to inhibit microbial growth; intrusion of tipper water into the digester would thus disrupt the anaerobes and biogas generation. To prevent siphoning of water from the wet tipper, the feed port at the top of the digester had to be opened at the time of withdrawal.

The bench scale digester had a several manageable operational challenges to overcome, regarding biogas leakage, the wet tipper, and temperature control. Biogas leakage through the mixing shaft and seals was an issue. An air tight seal in the digestion vessel is essential for the wet tipping bucket to record accurate volumes of the generated biogas. During a leak, biogas will bypass the wet tipper as it no longer has to overcome the small amount of static pressure head (<0.25 psi) in the unit. The result is zero additional volume recorded after the leak formation. Leaks were tested for using soapy water during all low volume reads. Bubbles indicate a leak. Repairs were completed accordingly. Through experience, it was deemed necessary to change oil and grease seals on a monthly basis. The digester usually requires a day or two to recover once the vessel is opened. The vessel may be lightly pressurized with a 1:1 CO2/N2 gas (<0.25 psi) to assist in stubborn leak identification. This practice is only recommended as a last resort, as it impedes digester recovery. The trapping of gases seems to inhibit the anaerobes metabolism.

The original Vaseline seal around at flanged connection the top of the digester was not sufficient to provide an air tight seal with any additional pressure applied to the digestion vessel (i.e. leak testing).

Channelling of biogas would occur through the Vaseline and leaks would develop as a result. This issue was effectively resolved in August 2010 when the Vaseline seal was replaced with a custom 1/8" thick pure gum rubber gasket.

Reduced registered gas volume may also result from water in the line between the wet tipper and the top of the digester as a result of siphoning. The gas line was inspected daily to ensure they were

73 relatively free of water and condensation. If water were present, the gas line was disconnected, drained, and blown out with compressed air until dry before being reattached.

The temperature controller would occasionally malfunction. The manufacturer suggested it was likely due to an inductive load at the output. Periodically current feedback from the output would cause the controller relays to short circuit. The unit would then have to be sent back to the manufacturer for repair or replacement. This anomaly may have been averted by retrofitting the temperature controller with a snubber. The digester required 2 to 3 weeks to move from stable psycrophillic to stable mesophilic operating conditions. Both the mixer and the temperature are turned off during any digester maintenance. Turning off the temperature controller and mixer before making any adjustments to the unit seemed to reduce the occurrence of temperature controller failure.

The mixing shaft would occasionally work itself loose from the automatic mixer. After time, grease from the bearing works its way up the shaft into the chuck of the mixer. This would slowly cause the shaft to slide downward. The paddles would eventually hit the either the thermometer or the thermocouple sensor and stall the mixer. No mixing resulted in thermal stratification of the digestate within the digester, but had little notable impact on biogas generation. The top of the mixing shaft and the inside of the chuck of the mixer was thoroughly cleaned with a kim-wipes and a cotton swab before retightening. Proper shaft alignment was also important. Bearings were re-greased periodically.

74 4.2.5 Digester Sampling Plan

Table 10 shows the sampling frequency of each parameter during start-up and once steady state conditions were established for each operating condition.

Table 10: Frequency of analytical parameters

Sampling Parameters Analytical Method Frequency

Temperature Thermometer Daily

PH pH Probe Daily Gas production Wet-Tip Gas Meter Daily

Biogas quality Gas Chromatographer 2/steady state

Alkalinity (Anderson & Yang, 1992) 2/week

Total Volatile Fatty Acids (Anderson & Yang, 1992) 2/week

Solids (TS, VS, TSS, and VSS) Standard Methods 2540 D 2/week

tCOD & sCOD Standard Methods 5220 D 4/steady state

BOD, cBOD Standard Methods 5210 B 4/steady state

Total Phosphorous Standard Methods 4500-P E 4/steady state

Total Potassium ALS Labs (Ref: EPA 200.8) 4/steady state

Nitrite, Nitrate Dionex Ion Chromatographer 4/steady state

NH3-N Ammonia Probe 4/steady state

C/N Ratio 4/steady state

During start-up periods regular monitoring was performed to characterize process stability and volatile solid reduction. The pH, Volatile Fatty Acid (VFAs) and alkalinity of the digester effluent were measured to monitor the digester stability.

Total solids (TS) and volatile solids (VS) removals, and gas production were tested as the primary indicators of solid reduction. Once each system had achieved steady-state, samples were taken from the influent and effluent of each digester on four different days and analyzed for the additional parameters listed in Table 10. All measurements were determined in duplicate at minimum. Biogas quality

75 measurements were to determine the methane and carbon dioxide content. Hydrogen sulfide (H2S) tests were conducted randomly for lab safety purposes.

4.2.6 Physical and Chemical Analyses of Digestate

A variety of physical and chemical analyses were performed in this study to assist in the quantification of digester performance. Unique handling of samples pertaining to a particular analytical procedure is detailed in this section. Referenced analytical procedures from Standard Methods (APHA, 1995) are detailed in Appendix C. Digested effluent was thoroughly mixed prior to analysis. Analysis was completed on undiluted feedstock; the reported results were adjusted to reflect actual concentrations going into the digester with dilution factor the applied volatile solids loading rate.

Temperature

The temperature of the anaerobic digester effluent was determined immediately after collection during each experiment using standard alcohol based thermometer. The operational temperature off the temperature controller and an additional inline thermometer were also recorded during sampling. A 3°C discrepancy was typically observed between the three values. This is likely due to the calibration of the thermometers. It may have been an indicator of limited thermal stratification in the digester. The effluent temperature was considered the most representative for unit operation. Faster mixing or improved paddle design may be able to homogenize the internal temperature.

PH

The pH of the anaerobic digester effluent was determined immediately after sample collection. A pH meter with a glass electrode was used for the analysis. Samples were stirred continuously until the probe registered a stable pH. Three point calibrations were completed monthly. The pH of the feedstock

was recorded exclusively during VFA/HC03 analysis.

76 Solids

The total solids and volatile solids concentrations of the feed and digested sludge were determined by placing a known volume of each sludge type in an aluminum evaporating dish and following the procedures established in the Standard Methods (APHA, 1995). The mass of the empty evaporating dishes was recorded in a table. Digester effluent was measured out in 50mL aliquots with a standard

50mL glass graduate cylinder. Transfer of the material was done as quickly as possible to inhibit settling of solids in the graduated cylinder. A 5mL plastic cast baking spoon was used to measure out the feed as the material was too thick to determine with a more accurate volume transfers technique. Feedstock was heaped over the measuring spoon and gently patted down with a scoopula to remove air bubbles.

The flat end of a clean scoopula was used to cut excess feedstock from the top and sides of the measuring spoon. The curved end of the scoopula was used to dislodge the sample into the evaporating dish. Evaporating dishes with sample were then measured and placed into to an oven at 103°C for drying overnight. Total solids were those remaining after the water was evaporated. The volatile solids were determined by massing the dried residue before it was placed in a furnace at 550°C for sixty minutes. The ash was cooled in a desiccator before weighing with an analytical balance.

Total suspended and volatile suspend solids was determined using Standard Methods 2540 D. A 5-6 mL volume of sample was placed upon a Whatman 1.2 pirn glass fiber filter paper on a vacuum pump using the same volume transfer techniques as stated above.

Total and Volatile solids were reported in both solids percentage by mass and mg/L. The percentage value is more accurate as is negates the error induced by the volume measurements. However, the mg/L value is more familiar from a wastewater background. The

Total Solids Calculation:

77 Oven Dried Mass - Mass of Crucible Initial Sample Mass - Mass of Crucible X Initial Sample Mass - Mass of Crucible Sample Volume ^ Solids Percentage(%) Sample Density (mg/L)

Volatile Solids Calculation:

Oven Dried Mass - Furnace Mass Initial Sample Mass - Mass of Crucible Initial Sample Mass—Mass of Crucible Sample Volume Solids Percentage(%) Sample Density (mg/L)

The suspended solids calculations were completed using equations 9 and 10 with the modification that the filter paper was included in the crucible mass (i.e. mass of crucible plus filter paper).

Alkalinity and Volatile Fatty Acids

Effluent volatile fatty acids (VFA) and alkalinity concentrations are a performance indicator for digester

operation. Alkalinity is expressed as bicarbonate (HC03). The VFA to bicarbonate ratio must generally fall below 0.3 to limit acetogen growth and prevent acidification of the digestate.

The alkalinity and total fatty acids concentrations were estimated according to the technique of

Anderson and Yang (1992). In the method, a 50 mL effluent sample is subjected to a two-point titration with 0.1N sulfuric acid in a lOOmL glass burette to a pH of 5.1 and 3.5. For the titration, the sample was placed atop a stir plate on its highest setting in a 250mL glass beaker along with a magnetic stir bar to promote uniform mixing of the acid. The initial pH of the sample was recorded along with the volumes of acid required to meet the two end points. The pH was only recorded if it remained stable for at least

10 seconds. As a result, each trial took approximately 20 minutes to perform. These values were input into a program described by Anderson and Yang the relevant concentrations. The alkalinity and organic

acids were reported as the concentration in milligrams per liter (mg/L) as bicarbonate (HC03 ) and acetic acid (ch3cooh), respectively. The method has a 96% recovery of VFA and bicarbonate, and is generally less than ImM/L in most titrations. Organic acids other than VFA such as citric and lactic acids and serious interfere with VFA/Bicarbonate results. Orthophosphates may also interfere with bicarbonate values to a lesser extent (Anderson & Yang, 1992).

78 Gas Production and Biogas Composition (CH4, C02)

The volume of biogas was measured every day using a wet-tip gas meter. The wet-tip gas meter was calibrated monthly. Biogas samples for methane analysis were initially obtained by swapping the gas transfer line from the wet-tip gas meter to a 500 mL tedlar bag for a 1hr period at least twice during the end of steady state operation before a new feeding regiment. The use of tedlar bags on these units disrupted digester operation for a period of a few days even if only connected for the 1hr duration. To rectify this, biogas samples were withdrawn directly from the gas transfer line in 30 mL aliquots with a

30mL gas-tight syringe (Hills and Roberts 1981). Biogas quality samples must henceforth be drawn by a syringe to reduce the impact on the digester.

The biogas composition was measured using an Aligent Technologies (6890N) gas chromatographer (GC) equipped with a TCD detector. Separation of the analytes was performed with a J & W Scientific CS-

Carbon Plot column (30m length B 0.32mm I.D., 1.5 micron film thickness). The GC oven temperature program was: 30°C for 7 min. Argon was used as the carrier gas, with a constant column flow of 1.5

mL/min. The detector temperature was 150°C. A standard curve of various mixtures of pure CH4 and C02 was created to project the concentrations of these biogas constituents from the peak areas produced by the chromatogram. No correction was made for water vapour.

Oxygen Demand

Biochemical oxygen demand (BOD5) analysis utilized Standard Methods 5210B. Three separate dilutions from each sample were used. Samples required dilution to ensure oxygen consumption in the 300mL bottles over 5 days was between 30% - 90%. The effluent samples dilution factors were typically 500,

100 and 1500. Influent dilution factors were typically 5000, 10000 and 15000. Glucose glutamic acid

(GGA) tests were completed to verify method integrity.

Total and Soluble Chemical oxygen demand (COD) analysis utilized Standard Methods 5220D. lOmL acid washed glass vials. Soluble COD was determined from 50 mL of sample centrifuged at 3000 RPM and 4°C

79 for 20 min. The supernatant was passed through a washed 1.2 p.m glass fiber filter paper. Analysis was performed on the filtrate. This methodology had a strong linear correlation of absorbance to COD

concentrations within the range of 100 and 1000 mg02/L. Samples outside this range returned false low

values. Samples with concentrations above 1000 mg02/L were diluted to fall within this range to ensure accurate results.

The effluent and feedstock required dilutions with de-ionized water by factors of 10 and 100 respectively at a minimum. The absorbance was measured at 600nm using an Aquamate

spectrophotometer (Thermo: Electron Corportation). 500mgO2/L KHP standards were run with regularly to validate sample results.

Ionic Species

Total Phosphorous analysis utilized Standard Methods 4500PE. The digester effluent and feedstock samples both required dilution with de-ionized water by a factor of 100. The samples were also subjected to heat and acid digestion.

Total Potassium was sent out to ALS Laboratory in Waterloo Ontario for analysis. The sample size was limited 50 to lOOmL based on availability, and stored for a maximum of 10 days at 4"C after submission to allow multiple days to be submitted at once. Results were received within one to two weeks of submission.

Ammonium nitrogen (NH4-N) was measured using an ammonium probe option from the pH meter. The pH of a 50 mL sample was increased to above 11 with 10N NaOH reagent added dropwise. A magnetic stir bar was employed to mix the sample. The pH probe was then switched to the ammonium probe, and the function on the pH meter was changed from pH to

mV. The most negative value in mV was recorded and compared against a standard curve to determine a concentration.

80 Nitrates (N03 ) and nitrites (N02 ) were analyzed using a Dionex Ion Chromatographer (ICS-2000). Total

Nitrogen was measured using a Shimadzu Total Organic Carbon Analyzer (TOC-V CSH) with the Total

Nitrogen Measuring Unit (TNM-1) option. Sample pretreatment was required as both devices are may be clogged by particles larger than 1 nm. A 100 mL sample was centrifuged at 3000 RPM and 4°C for 20 min. The supernatant was passed through a washed 1.2 urn filter and the filtrate collected. The filtrate was again passed through a 0.45 nm filter before analysis.

4.3 Digester Results and Discussion

This section describes the feedstock characteristics, followed by performance results for the bench scale anaerobic digestion trial for the London project. Further information can be viewed in Appendix D.

4.3.1 Feed Characteristics

The general composition of the digester feed stocks are presented in Table 11. The variation in solid contents for both TS and VS values was relatively small with a standard deviation of 10 % about the mean.

81 Table 11: Average Characterization of Undiluted Feedstock

50% DM* 75% DM 100% DM Parameter Units 50% RB 25% RB 0% RB Theoretical Dilution Factor Unitless 2.27 2.78 3.85

Amount of Manure Added (mL/day) 110 135 130

Amount of Beer Added (mL/day) 110 45 0

pH Unitless 5.88 6.64 6.91

Total Solids (mg/L) 67,800 82,900 104,000

Total Suspended Solids (mg/L) 53,300 74,400 99,000

Volatile Solids (mg/L) 55,400 69,500 88,200

Volatile Suspended Solids (mg/L) 49,800 62,800 86,800

Total COD (mg/L) 96,900 70,900 60,800

Soluble COD (mg/L) 68,900 31,300 21,500

BODs (mg/L) 33,100 31,900 19,300

Bicarbonate (mgN/L) 25.3 80.7 93

VFA (mgN/L) 168 131 75

Ammonia-N (mg/L) 4,450 5,150 6,027

NO3 (mg/L) 2.94 133 62.7

NO2 (mg/L) 1.98 3.8 113.9

Total Phosphorous (mg/L) 526 270 310

Total Potassium (mg/L) 1800 1600 1600

*Data is from the second trial of the 50%RB/50%DM feedstock

All parameters of the feed sludge were measured during steady state digester operation. Some

parameters such as TS/VS, pH, and VFA/HC03" were measured more frequently.

82 The dairy manure used in this project was representative of other anaerobic digestion studies with a

12.1% TS content and 10.3% VS content. Raw dairy manure typically has a total solids content of about

15%, 83% of which is volatile solids (Krich, et al. 2005). Most of the solids from DM existed in the suspended solid form. Nearly all solids in rebate beer were dissolved. Rebate beer has 3.2%TS (~33,300

mgrs/L), and 3.0%VS (~31,300 mgvs/L). In other words, 93.9% of the solids in beer are volatile. The solids in the combined feed stocks are primarily from the dairy manure. The average TS/VS of the dairy manure used in all blends is representative of the 100% DM baseline (see Table 11).

3 Warm tap water was used for the dilution of the feed stocks to maintain the lkgVs/m /day organic loading rate. Approximately 130 ± 5mL of dairy manure diluted with the appropriate volume percentage of rebate beer and tap water was present in each 500mL volume of digester influent. It was determined that the theoretical dilution factor (Equation 11) was larger than the actual dilution factor (Equation 12).

Ideally these dilution factors would be equal. Accurate volume transfers of small amounts of dairy manure are difficult; the material is heterozygous, viscous, and clings to the side of transfer vessels. The actual dilution factor, which is based on tested feedstock volatile solids concentrations, is therefore more applicable to adjust the data set.

Theoretical Dilution Factor = [11]

Where: Vw = Volume of water (mL);

VDB = Volume of dairy manure (mL);

VM = Volume of rebate beer (mL).

C,VS Undiluted Feedstock Actual Dilution Factor = [12] C,VS Diluted Feedstock

Where: Cks undiluted Feedstock = Volatile Solids Concentration of undiluted feedstock (mg/L);

Cvs undiluted Feedstock = Volatile Solids Concentration of diluted feedstock (mg/L).

83 Each tested parameter has a slightly different dilution factor.

The average TS concentrations in the rebate beer blends weighted with those of the dairy manure are estimated to be 3.4% greater than the actual TS concentrations in the mixed 75%DM/25%RB feed.

Similarly for VS, the estimated was 6.1% higher than the actual mixed blend. The discrepancy may be accounted for in the heterogeneous nature of dairy manure and in the error introduced during the volume transfers in the mixing process. However, this difference could also be an indicative of microbial metabolism during storage.

The prepared feedstock containing rebate beer would occasionally acidify during storage. This was likely exacerbated by the traffic around the refrigerator with the frequent opening and closing of the doors.

The combined feedstocks were remade every 1 HRT (24 days) to reduce acidification. The

50%RB/50%DM blend was made slightly more frequently at 15 days to avoid the pH slipping below 5.1, as this would render the VFA/hco3 titration method inapplicable. This phenomenon could be avoided in a reproduction of this study if fresh feed stock was prepared on a daily basis instead of in bulk.

4.3.2 Steady-State Digester Performance

The digester was operated on each feedstock at an organic loading rate of lkgVs/m3/day for a minimum of two months (2 HRTs) until steady state operation was achieved. A summary overall digester performance is presented in Table 12.

The temperature and mixer RPM were fairly consistent across all three trials. The mixer RPM had to be increased near the end of the second 50%DM/50%RB trial to prevent it from stalling. This is responsible for the higher standard deviation noted in 50%DM/50%RB mixer speed.

A higher methane concentration has been observed in scenarios where the feedstock is heavily diluted

with process water (Krich, et al. 2005). The solubility of C02 in water is much higher than that of CH4.

This is not the case with the rebate beer, as higher methane percentages were noted where lower

84 process water volumes were applied. This is a strong indication that rebate beer increases biogas quality as well as quantity.

Table 12: Summary of Digester Operations

50% DM 50% RB 75% DM, 25% RB 100%DM, 0% RB Parameter Average Std Dev. Average Std Dev. Average Std Dev. Start Date 14-0ct-10 21-Jul-09 12-Jun-10

End Date 28-Jan-ll ll-Jun-10 13-0ct-10

Mixer Speed (RPM) 53 25 50 3 57 2

Temperature fC) 34.7 0.6 36.4 1.2 35.3 0.6

Digestate pH 7.07 0.03 7.27 0.11 7.28 0.14

Biogas Vol. (L/day) 13.5 8.5 8.3 5.2 7.0 4.6

Methane Cone. (%) 55.0% 54.7% 2.2% 47.5% 4.4%

Net Energy (kJ/day) 679.4 415.4 113

The observed methane concentrations were lower than expected. Dairy manure alone typically has a

CH4% of 55% to 65% (Hills and Roberts 1981). The maximum recorded methane percentage was only

55% for the 50% DM/50% RB blend. The lower methane concentrations for the 100% DM/0% RB may be due to the longer holding times as fresh dairy manure exhibits higher methane potential. Free ammonia is assumed to inhibit methanogenesis in many anaerobic digestion models (Lyberatos and Skiadas 1999).

The higher concentration of NH3 in the 100% DM blend may also have attributed to the lower than expected methane value. Gas collection and analysis techniques may have also introduced a degree of experimental error.

The enthalpy of reaction (AH) for the combustion of methane gas in pure oxygen using Equation [13] yields -807kJ/mol.

85 -74.&kJ I mol 0 kJImol -393.5kJImol -2A2kJ!mol CHa + 202 —A-* C02 +2H20(g) [13]

The average net energy in kJ/day was calculated in Table 12: Summary of Digester OperationsTable 12 using Equation [14] as a basis of comparison for the activity of each blend.

E pQYAH [14] M where: E = Daily energy production in biogas per litre of feedstock (kJ/day)

3 p = Density of methane (1819gCH4/m at STP)

AH = Enthaply of reaction for combustion of methane (-807kJ/molCH4)

M = molar mass of methane (16.043gCH4/mol)

Q = Daily biogas flow rate (m3/day)

Y = Percentage of methane in biogas (molCH4/molbi0gas)

Co-digestion of other biomass wastes is typically not expected to augment the dairy waste methane potential by more than 10% to 20% by volume (Krich, et al. 2005). However, it is evident from Table 12 that the anaerobic digester feed stocks with increased beer to manure ratios yields significantly higher methane potential. Biogas volumes were 19% and 93% for 75% DM/25% RB and 50% DM/ 50% RB respectively compared to dairy manure alone, and the net energy values were 4 to 6 times higher.

Figure 14 shows the overall all methane production of each blend when normalized by the volatile solids destruction.

86 Mean Steady-State Methane Production

• 50%DM/50%RB

m 75%DM/25%RB

• 100%DM/0%RB

Figure 14: Methane production normalized by volatile solids destruction

The digester pH was consistent and stable around 7.0 throughout the entire duration of each trial, even though the feed stock with rebate beer was acidic, with a pH as low as 5.1. Digesters fed with manure exhibit a self-regulation of the pH attributed to the interplay of generated ammonia with VFA (Lyberatos

and Skiadas 1999). NH3 is liberated with the digestion of organic acids (acetate, VFA), and also associated with an increase in pH (Hjorth, et al. 2009).

Net NH3 was created in the 75%DM/25%RB and 100%DM trials, and a net loss was noted in the 50%

DM/50% RB blend. The overarching trend is a decrease in NH3 in digester effluents from rebate beer containing-blends; however the It was expected that the 50%DM/50%RB blend would also have generated ammonia.

87 Table 13: Summary of Chemical Parameters in Digester Effluent

50% DM 50% RB 75% DM, 25% RB 100%DM, 0% RB Parameter Average Std. Dev. Average Std. Dev. Average Std. Dev. TS (mg/L) 22,000 1,890 22,600 3,760 26,600 1,150

TSS (mg/L) 21,400 4,810 17,200 2,230 23,300 2,150

VS (mg/L) 18,500 1,740 16,900 3,900 19,400 1,250

VSS (mg/L) 17,600 3,820 14,100 1,900 20,300 2,380

Total COD (mg/L) 26,000 11,500 10,500 10,200 - --

Soluble COD (mg/L) 5,280 787 3,290 746 6,030 193

BOD5 (mg/L) 2,020 367 4,480 1,830 4,240 397

VFA (mgN/L) 3.5 2.5 9.2 12.8 3.3 1.5

Bicarbonate (mgN/L) 74.2 4.4 107 21 86.5 4.7

VFA/HC03 Ratio 0.05 0.034 0.076 0.089 0.041 0.020

Ammonia-N (mg/L) 1,800 2,200 32,400 15,100 51,400 63,300

Nitrate (mg/L) 0.545 - 32.6 6.0 0.4 0.2

Nitrite (mg/L) 0.125 - 201 28.2 9.4 1.0

Total P (mg/L) 184.1 46.7 32.7 5.5 90.0 19.3

Total K (mg/L) 720 ~ 607 37.9 <1000 0

The VFA was nearly entirely consumed for 100% DM, 75% DM/25% RB, and the second 50%RB/50%RB trial. As such, the VFA/BA ratio was much lower than 0.3 for the 75% and 100% blends; values indicative

of a well-stabilized product. The 50%RB/50%DM VFA/HC03 ratios were high for the first trial, and resulted in periodic acidification of the system. This was attributed to an unstable source of dairy manure and was remedied with help of Elora Dairy. The second trial was much more successful and had a stable VFA/hco3 ratio for 4HRTs. The VFA/HCOj data at these temperatures are in agreement with the

88 substantial reduction in VS. As VFA was virtually zero in the effluent of the 75% and 100% DM blends, they may have been the growth limiting substrate for these trails. The ideal amount of rebate beer co- substrate likely lies between 25%-50%.

The BOD5 and sCOD removal rates were similar for each trial ranging from 71.9% to 89.5% (see Figure

15). The trend in digester oxygen demand reduction closely mirrors the trends in VS reduction and methane percentage. The tCOD removal rate for the 50%RB/50%DM blend was 39%, which was significantly less than the removal rates for the 75%DM/25%RB and 100%DM trials of 85% and 90% respectively. This may be attributed to either experimental error, or that carbon is no longer the limiting substrate for the 50%DM/50%RB trial; the microbes would therefore not work as aggressively to reduce the insoluble fraction of COD.

Comparison of Oxygen Demand Reduction Between Different Feedstocks

100%

• 50%DM/50%RB

* 75%DM/25%RB

• 100%DM/0%RB

Soluble COD BOO

Figure 15: Comparison of the oxygen demand reduction from digested feed stocks with varying dairy manure to beer ratios during steady state operation of bench scale anaerobic digester

The average reduction of solids and key chemical parameters through the digester is summarized in

Table 14.

89 Table 14: Summary of Digester Performance through Reduction of Physical and Chemical Parameters

Parameter 50% DM/50% RB 75% DM/25% RB 100% DM

TS Reduction (%) 32% 51% 28%

TSS Reduction (%) 44.1% 11.6% 35.8%

VS Reduction (%) 40% 56% 29%

VSS Reduction (%) 46.7% 14.0% 36.1%

Total COD Reduction (%) 39.0% 85.1% -

sCOD Reduction {%) 82.6% 89.5% 71.9%

BOD5 Reduction (%) 86.1% 86.0% 78.0%

Ammonia Reduction (%) 8.1% - -

Total P Reduction (%) 20.4% 87.9% 71.0%

The solids reduction from the feedstock to the digester effluent is provided as an indicator of digester performance. Previous studies established that 41.6% of dairy waste is biodegradable (Morris 1976).

This should be considered the upper limit for total solids reduction through the digester for the control blend of 100% DM/0% RB. The recorded 28 ± 6% decrease in TS for the 100% DM trial is a further indication of some mechanism of anaerobe inhibition at work in the control blend that was not present for the trials with rebate beer. The 75%DM /25% RB blend exhibited the best solids removal of the three feeds. The volatile solids reduction was 56 ± 7% for the duration of the trail; total solids were reduced by

51 ± 7%. The 100% DM blend VS were reduced by 29% ± 5%. The 50% DM/50% RB blend had very inconsistent solids removal with 40 ± 14% for VS removal and 32 ± 15% for TS removal (see Figure 16).

90 Comparison of Solids Destruction Between Different Feedstocks

60%

• 50% DM/50% RB 2 30% * 75% DM/25% RB • 100% DM/0% RB

Total Solids Volatile Solids

Figure 16: A comparison of the solids destruction from digested feed stocks with varying dairy manure to beer ratios during steady state operation of bench scale anaerobic digester

The shape of Figure 16 with a local maximum at 75% DM/25% RB is similar to rise and fall of %VS removal with increasing to non-lignin carbon to khedjal nitrogen ratio as recorded by Hills and Roberts

1981. This trend is observed in %sCOD and %BOD5 removal (see Table 13).

4.4 Recommendations for Anaerobic Digestion of Brewery Wastes

This study demonstrated the benefits of RB as carbon co-substrate for dairy manure in a bench scale anaerobic digester. The next logical progression in the advancement of this research would be pilot scale operations. Steadily increasing the percentage of RB in an anaerobic digester that initially operated on

100% DM would likely help the digester achieve steady state more quickly.

Rebate beer is not the only brewing byproduct that is suitable as co-substrate as a co-substrate for the anaerobic digestion of dairy manure. Brewer's grains and spent yeast are also suitable candidates.

However these materials are generally designated to more lucrative disposal options, such as animal feed supplements. Each brewer's grains, spent yeast, and rebate beer may also be suitable materials for conversion into ethanol and other biofuels (see Chapters 2 and 3). The optimum byproduct disposal

91 method is always be constrained by the current economy and legislation. Disposal methods should be re-evaluated whenever one of these factors change. Presently, if the AD facilities are in close proximity to the point of generation, AD should be considered as a feasible option for most food wastes.

The pressurization of the digester above the 0.25 psi exerted by the water level in the wet tip gas meter impacted digester operation. This was observed while during pressure tests for leaks, and extraction of biogas samples with tedlar bags. Biogas samples removed by negative pressure with a syringe did not result in zero biogas generation the next day. Opening and closing the lid of the digester and exposing it to atmospheric pressure for short periods of time did not affect digester operations. It is possible that increased positive pressure in the digestion vessel promotes the solubility of various biogas constituents back into the digestate to inhibit anaerobic metabolism. It would be interesting to examine the effect of variations in operating pressure on a digester performance related to the solubility of gases. Methane

and C02 have different water-gas partitioning coefficients; C02 is more soluble than CH4. As such, some

C02 may remain in the liquid portion.

Intensive studies into the operational effects of fluctuations of pressure within a digestion vessel have not yet been investigated. Recognized issues of pressure on biogas are isolated to gas turbine operational requirements, and safety (i.e. ensuring that the tops of digestion vessel don't blow off). Such an investigation could provide insight to optimize methane percentages in biogas while maintaining the activity of the anaerobes. This could increase the efficacy of industrialized anaerobic digestion operations.

If the digestate were to be dewatered to create a solid fertilizer, the high ammonia concentrations may

an environmental issue. NH3 may be removed from anaerobically digested effluents through the forced

precipitation of magnesium ammonium phosphate hexahydrate (MgNH4P04-6H20), commonly called struvite (Uludag-Demirera, et al. 2008). Struvite is a valuable plant nutrient source for nitrogen and

92 phosphorus since it releases them slowly and has non-burning features because of its low solubility in water.

This study demonstrates that rebate beer is ideal for promoting biogas production in anaerobic digesters; it may also be suitable as a feedstock for other fuel conversion techniques. The Krausen and

Aging tank bottoms along with rebate beer maybe excellent candidates for distillation into fuel-grade ethanol. The Molson Coors Brewery in Golden Colorado successfully instituted this practice in 1996 in conjunction with the engineering firm Merrick and Company. The brewery produces 16 million barrels of beer and 3 million gallons of fuel grade ethanol per year for a $6 million US investment in the ethanol facility (Miller 2008).

Waste beer is also a suitable stock for Virent's Aqueous Phase Reforming (APR). This process generates hydrogen and/or hydrocarbons from biomass without volatilizing water, which represents major energy savings compare to other biofuel technologies. Waste beer already contains the aqueous phase carbohydrates necessary of APR. Reformation occurs when liquid passes over a metal alloy catalyst - some of which generally contains nickel (Ni) and ruthenium (Ru) with trace elements of platinum. The type of catalyst determines whether methane, propane, or hydrogen is produced. The technology was extended in 2006 to produce gasoline, biofuel, and jet fuel. The operating temperature is 200°C, which is quite low for a reforming technology. An average of 25% of the hydrogen produced by the process is used to maintain the operating temperature, leaving 75% available for other uses. The low operating temperature also minimizes undesirable decomposition reactions. APR is conducted at pressures

(usually 15-50 bar) where the hydrogen rich effluent can be effectively purified using pressure-swing adsorption or membrane technologies. APR uses one simple phase separator to separate the hydrogen gas from the liquid. It accomplishes this at a higher purity than conventional digestion, gasification, or steam reforming methods that may require up to five to six vessels. The hydrogen produced is suitable for use in fuel cell technology because of the low CO concentrations (30 ppm) that would otherwise

93 quickly poison fuel cell catalysts. Constant flow operation allows conversion to occur in a matter of minutes. Overall, this energy conversion method is highly efficient, emission neutral, low temperature, constant flow, compact, and has very low carbon monoxide generation. Capital and operating costs of this system are unknown (RENEW 2004). Virent in collaboration with Royal Shell Oil is currently operating a biogasoline demonstration plant in Madision Wisconson that produces 10 000 gallons of gasoline a year.

4.5 Anaerobic Digestion Project Conclusions

Successful steady state mesophilic anaerobic digestion was achieved at an organic loading rate of

3 lkgvs/m d for each blend. This study suggested that this ratio of rebate beer to manure resulted in increased biogas generation and higher methane yields. As a result, 4 to 6 times the energy was produced for the 75% DM/25% RB and 50% DM/50% RB blends compared to the 100% DM blend alone at the prescribed OLR. This would translate into a faster ROI for dairy farmers operating AD facilities.

The optimal RB content in DM by volume likely lies between 25% - 50% according to the collected biogas data. The first trial of the 50% rebate beer blend suggested that the beer-to-manure ratio is perhaps too high, as it resulted in periodic acidification. VFA concentrations in the digester effluent steadily increased proportionally with the percentage of beer. However, both the second 50%DM/50%RB, the

75% DM/25%RB and the 100%DM digester effluents showed negligible, near zero VFA concentrations.

Higher ratios of DM/RB exhibited better sCOD and BOD removal. The effluent sCOD and BOD was nearly

the same across all blends despite the fact that the influent sCOD and BOD5 concentrations were considerably higher with the addition of rebate beer. tCOD concentrations increased with of the addition of rebate beer as shown with the switch from the 75% DM/25 RB blend to the 50% DM/ 50%

RB blend. This is also followed by a decrease in the TS destruction, and may be an indicator of reduced hydrolysis.

94 Each scenario was able to reduce the influent VS concentrations by at least 29% for the effluent.

However, the magnitude of steady state solids destruction was much more variable with increased proportions of beer to manure. Feed stocks with beer co-substrate took longer to achieve steady state operations; they are more organically active with a higher percentage of bio-available nutrients.

Degradation of this material occurs considerably faster in rebate beer blends than with dairy manure alone which is apparent in the preliminary BMP study. As such, the digester effluent from DM/RB blends were more vulnerable to swings in chemical composition. More frequent digestate monitoring may be required to compensate. However, this issue may not be as prevalent if the study were conducted on a larger scale as a small digestion vessel volume would be more sensitive to change.

95 Chapter 5: Biological Packed Tower Retrofit Pilot

5.1 Introduction to BioTower Pilot

Labatt Brewery in London, ON has been investigating low area footprint wastewater treatment

technologies for BOD5 reduction. Many brewers acknowledge that biological wastewater treatment

(BWT) technologies are the most cost-effective and efficient methods known for BOD5 removal (T.

Goldammer 2008). However, the character of brewery wastewater presents several barriers for BWT that must be addressed.

5.1.1 Problem Definition

Brewery wastewater effluent is batch discharge in nature. Labatt London's average effluent flow rate is

3 5000m /day. Untreated, it exhibits high BOD5 concentrations in excess of 2000 mg/L, Total Suspended

Solids (TSS) averaging 600 mg/L, and a highly variable pH (from 2 to 12). The BODs is primarily derived from waste beer and brewery byproducts such as post-runoff and yeast. Clean-ln-Place (CIP) chemicals for vessel washing, bottle cleaning residuals, and beer residuals have significant influence over the final effluent pH. The local municipal WWT plant applies a flow-weighted High-strength Sewage Surcharge

(HSS) fee to treat industrial effluents with BOD, TSS, and Total Phosphorous (TP) concentrations higher than 300 mg/L, 350 mg/L and 10 mg/L respectively. In addition, the municipality established an acceptable effluent discharge pH range between 6.0 and 10.5 to protect sanitary infrastructure. Labatt

London adjusts effluent pH to within the accepted range; however there is no BOD or TSS removal. As such, Labatt London is charged $1 million+ in HSS fees annually. 80% to 90% of this cost is attributed to

BOD; its reduction would result in significant savings.

A robust pH control system is crucial to maintain the large volume of effluent within city limits. Given the water intensive nature of brewing, the pH adjustment of wastewater may be a considerable expense

96 to brewers. Labatt London's existing pH adjustment system is somewhat overdesigned, and perhaps not utilized to its fullest potential. The pH adjustment system consists of four large equalization reactors in series. Reactors 1 and 2 both have a 90m3 volume, while reactors 3 and 4 have a 60m3 volume. It is

designed to maintain effluent pH within the legislated discharge range with minimal acid (C02) and base

(NaOH) dosing. Labatt exceeds pH discharge guidelines a few times each quarter as the system "set points" are at the extremes of the permitted pH range. This maximizes chemical savings, but does not provide an adequate preventative factor of safety. Besides narrowing the system's pH set point range, a more effective inline mixer with throttled variable speed dosing pumps could reduce the instances of pH exceedance. This could significantly increase system responsiveness, and reduce the number of equalization tanks required.

5.1.2 Retrofit Opportunity

An equalization tank rendered redundant presents an opportunity for potential BOD5 removal. The tank could be converted into an aerobic biological-packed tower (BioTower) with a minor retrofit to incorporate a biological treatment component. The BioTower is an Attached Growth Process; microorganisms fix themselves to the surface of a packing media to form a biofilm. Biofilms are considerably more resistant to environmental fluctuations compared to conventional activated sludge processes, and making them a superior candidate for brewery wastewater treatment.

To test the feasibility of the retrofit, a pilot-scale BioTower project was built and operated at Labatt

Brewery from November 2010 to February 2011. For biological wastewater treatment at Labatt, the pH set point range of the existing system would have to narrow from '6.0 to 10.5' to at least '6.0 to 8.0'.

BWT systems predominantly require a stable, neutral pH that may range from 6 to 8 for aquatic microorganisms to function optimally (Wang, Pereira and Hung 2009). The increase pH adjustment chemical consumption costs would be offset by HSS savings from BOD reduction. The pilot would help determine the feasibility of the system BioTower retrofit in terms of:

97 • Breadth of pH range for optimal biological treatment, and

• Ideal Hydraulic Retention Time (HRT).

The pH ranges to be tested were 7.0 ±0.3, ±0.5, and ±1.0. This would provide an insight into the biofilm tolerance to pH fluctuations. The HRT would be varied from 14 to 35 minutes. This would give an idea into the number of tanks that should be converted into BioTowers (max. 3). The least sensitive condition was tested first (pH range of 7.0 ±0.3 and HRT of 35 min). Over the operation of the pilot, the HRT would then be decreased and pH range increased providing successful operation at the previous condition. A

minimum BOD5 removal of at least 50% was desired before moving onto a new operating condition. The

resulting information may be applied to determine the junction where savings from BOD5 reductions would exceed the expense of additional chemical dosing, and whether a retrofit is economically feasible of a 2 year return on investment (ROI).

BioTower pilot operations at Labatt London were terminated in February 2011 as only a maximum of

20% COD removal was achieved at the least sensitive operating conditions. Data indicated that the HRT for the tested BioTower arrangement was not sufficient to provide adequate contact time for significant, reproducible COD reduction. Although the pilot trial was unsuccessful overall, there are several conclusions that may be drawn from the pilot and associated lab testing to advance the progression of wastewater treatment initiatives at Labatt. This chapter will present the details of the BioTower pilot and summarize several major useful findings.

5.1.3 Background on Attached Growth Processes

An attached growth process (AGP) utilizes biofilm for the biological degradation of organics in wastewater. A 'biofilm' describes a community of diverse of aquatic micro-organisms (aerobic and facultative bacteria, fungi, algae, protozoa, etc.) that adhere to a surface media. Biofilm growth is desirable in wastewater treatment applications to stimulate nutrient removal through microbial action.

98 The operating conditions of the trickling filter define the overall dynamics of the microbial community, and thus the removal efficiency of the system.

Facultative bacteria are the predominating organisms in trickling filters

• Achromobacter, • Flavobacterium • Pseudomonas • Alcaligenes

Within the slime layer, where adverse conditions prevail with respect to growth, the filamentous forms

Sphaerotilus natans and Beggiatoa will be found. The fungi present are also responsible for waste stabilization, but their role is usually important only under low-pH conditions or with certain industrial wastes. Rapid fungal growth can clog filters and restrict ventilation.

Identified Fungi:

• Fusazium • Mucor • Penicillium • Geotrichum • Sporatichum • Various Yeasts

Algae grow in the presence of sunlight in the exposed regions on an AGP vessel. Algae do not take a direct role in waste degradation, and may be considered a nuisance due to their potential for fouling the media surface - which may generate odours.

Protozoa are predominately of the ciliate group, including Vorticella, Opercularia, and Epistylis. Their function is to feed on the biological films. The result is a decrease in effluent turbidity while the biofilm is maintained at a higher growth state.

Microorganisms travel to the media surface by both hydrodynamic and mass transfer mechanisms.

Surface attachment occurs through the production of a polysaccharide binding material. The rate and location of attachment have a significant effect on microbial ecology within the biofilm (Patwardhan

99 2003). The porosity of the biofilm itself dramatically decreases with increasing thickness. Oxygen diffusivity through the biofilm is inversely related it's density. Diffusivity was found to decrease by a factor of five toward the bottom of the biofilm (Fu 1994).

The slime layer thickness can reach depths up to 10 mm. Organic materials from the liquid absorb onto the biological film or slime layer. The outer layer of the biofilm (0.1 to 0.2 mm) is fully aerobic. Oxygen is fully consumed after further increases in biofilm depth, and an anaerobic environment is established at the surface of the packing. As the biofilm thickness increases, the substrate is utilized before it can penetrate the inner biofilm depth. Bacteria in the slime layer witch to an endogenous respiration state and lose their ability to cling to the packing surface. The liquid washes the slime off the packing, and a new slime layer starts to grow in phenomena known as sloughing. Sloughing is a function of organic and hydraulic loading on the filter. Hydraulic loading accounts for shear velocities, and the organic loading rate the metabolism of the slime layer.

Performance of the biofilm is often diffusion-limited. Substrate removal and electron donor utilization occurs within the depth of the attached growth biofilm and subsequently the overall removal rates are a function of diffusion rates and the electron donor and electron acceptor concentrations at various locations in the biofilm. Limiting DO concentrations are generally higher in attached growth processes than suspended growth (Metcalf and Eddy, Wastewater Engineering: Treatment and Reuse: Fourth

Edition 2003).

The optimal temperature range for growth is between 10 and 36°C. Biofilm layer decay arises from several mechanisms including detachment, auto-digestion, and protozoan grazing. The rate of substrate removal increases exponentially with increasing biofilm thickness to a local maximum (Characklis 1982).

This critical value is termed the active thickness, and is dependent on the substrate concentration in the reactor. At low substrate concentrations, filamentous organisms prevail.

Rate of substrate removal (Rs):

100 Mmt* S C [15]

where: Umax = maximum specific growth (T1)

S = Concentration of growth limiting substrate (ML 3)

3 Ks = Saturation constant (ML )

C = Concentration of dissolved oxygen (ML 3)

3 Kc = Oxygen inhibition coefficient (ML' )

X = Initial biomass concentration (ML3)

Yx/S= Maximum biomass yield coefficient (M cell formed per M substrate consumed)

Rate of oxygen uptake (Rc):

[16]

1 where: ke = Biomass oxygen utilization rate (T )

Yc/s= (M cell formed per M oxygen consumed)

The wastewater influents to AGP treatment systems generally require primary clarification. Biological treatment results in biomass production and sludge. AGP effluents are generally conveyed to sedimentation tank for solids removal.

Quantifying the biomass in the system is not possible. The attached growth is not uniformly distributed in the random pack AGPs, the biofilm thickness can vary, the biofilm solids concentration may range from 40 to 100 g/L, and the liquid does not uniformly flow over the entire packing surface area, which is referred to as the wetting efficiency (Metcalf and Eddy, 2003). Therefore only broad parameters can be used to quantify loading criteria for design and operating specifications, including:

3 -1 • Volumetric organic loading - the mass of BODs applied per volume of media (ML ! );

2 1 • Unit area loadings - the mass of BOD5 per unit surface area of packing material (ML" !"" ); and

101 • Hydraulic application rates - the wastewater flow rate per cross sectional area of AGP vessel

perpendicular to flow (LT1).

5.2 BioTower Pilot Methodology

Assembly of the Biological Packed Tower began on Monday, November 8, 2010. The pilot went through several permutations as operational obstacles presented themselves. The engineering drawing for the final arrangement is presented in Appendix E. The following section will describe the methodology that governed the management strategy for the BioTower pilot.

5.2.1 Experimental Apparatus

Biological Packed Tower Design

The BioTower column was a vertically mounted length of clear PVC pipe with a diameter of 20 cm and a height of 150cm (47L). The packed volume was 33L with an active volume of 30L (wastewater volume absent of media). In the bottom of the BioTower, 2.5L of washed quartz gravel sat atop a fine mesh screen to serve as a coarse air diffuser. The gravel had 50% void spaces.

The spherical packed media was placed in a random configuration on top of the gravel. The media was polypropylene Jaeger Tri-Packs®. Each 5.1 cm diameter ball had a geometric surface area of 157 m2/m3 and displaced a 6.25 ml volume of water (Jaeger Products Inc. 2004). There were 300 balls in total. The media was held down by a 10 kg steel weight wrapped in plastic to prevent them from floating.

The air supply for aeration was obtained by placing a T into the compressed air line that works the

pneumatic C02 valve for Reactor 3. The air flow rate was periodically adjusted to maintain the dissolved oxygen (DO) concentration in the BioTower effluent between 2.0 and 4.0 mg/L. This effluent DO range is typical for wastewater treatment applications as it ensures fully aerobic conditions while minimizing operating costs due to aeration. A hand-held DO probe was used to measure the DO concentration at the outlet to ensure that adequate air was supplied to the biomass in the column.

102 Neutralized wastewater was pumped from the buffer tank to a port that sat above the gravel at the bottom of the BioTower. The wastewater would travel up through the column media, and exit out the top through a Vi" line, and directed through a hose into a floor drain. A static wastewater level was maintained in the column by installing a system of check valves for backflow prevention. The wastewater flow rate out of the BioTower was controlled with 3 separate valves. Samples of the treated wastewater will be collected at the outlet of the column and sent for analysis of key process parameters.

Out water flow back to flume dowiwtresLni Airflow

Chemical feed flow

In water flaw from flume upstream Figure 17: Biological Packed Tower Schematic

Equalization Tank

The brewery wastewater pH required further neutralization for biological treatment to be viable. A 1200

L tote originally used to store lime flavouring was supplied by Labatt. This served as an equalization tank.

Brewery wastewater effluent was obtained by attaching to a 2" valve at the top of Reactor 4. Brewery effluent flowed via gravity through a 2" hose through into the top of the equalization tank (see Figure

18).A coarse screen with %" mesh intercepted the wastewater before it entered the tote. Coarse filtration was necessary to prevent clogging within the system. Without the filter, cigarette butts and paper pulp would occasionally plug intakes to the pumps, resulting in unit shutdown. Fine screens have been successful in plastic packing media to prevent fouling of the packing within the BioTower column

103 (3 mm mesh would have been ideal). A 2.5 cm diameter overflow line was installed 12.5 cm from the top of the buffer tank. The effective working tank volume was about 1000L.

A PHCN-37 pH controller was installed to actuate two peristaltic pumps for pH adjustment in the equalization tank. A pH probe mounted in the center of the buffer tank, 5" from the bottom provided the input to the pH controller. The pH controller would actuate one of two peristaltic pumps outfitted with 6mm 10 C-Flex tubing that dosed chemical from 20L HDPE reservoirs. The pH adjustment chemicals

were 5% H2S04 for acid, and 5% NaOH for base. The peristaltic pumps were set at 30 RPM to prevent overdosing; this rate was determined through experimentation. Chemical was pumped into the influent of the buffer tank whenever the wastewater went outside the set points programmed into the pH controller. The pH adjustment chemical reserves were monitored and topped up daily.

A 34" recirculation line was installed that withdrew wastewater from the bottom of the tank, and pumped it to the top. This would ensure that the contents of the buffer tank were thoroughly mixed. A valve was place at the end of the recirculation loop for to obtain sample of the equalization tank contents for wastewater analysis.

A float switch was installed to cut the power to the pH controller, recirculation, and BioTower column pump once the wastewater level in the buffer tank ever dipped below 20cm. This would prevent damage to the pumps, and eliminate sporadic chemical dosing as the probe cannot register the correct pH if not completely submerged. Pilot operations would resume as normal once the equalization tank depth surpassed the 50 cm mark.

104 Figure 18: Equalization Tank Arrangement

Biofilm Acclimatization

A healthy biofilm is elementary to an attached growth wastewater treatment process. Municipal wastewater treatment is an excellent seed to help establish biofilm development. 40L of secondary clarifier water from the Guelph WWTP was first obtained on Monday, November 15. Mixed liquor was poured into the packed column until the media was completely submerged. The column was then aerated for a period of five days to help establish the development of a biofilm. The dissolved oxygen concentration during acclimatization was maintained at 8.0 mg/L to ensure aerobic conditions and inhibit the development of nitrifiers (Metcalf and Eddy 2003). The wastewater was constantly re­ circulated through the column from the bottom to the top. The equalization tank was completely filled during this period to work out any bugs in its operation. The BioTower was reacclimatized a second time

January 31, 2011 after a week of inactivity due to operational problems.

Control Variables

The set points on the pH controller were adjusted to 6.7 and 7.3 respectively with a hysteresis set to the minimum of 0.1. The RPM for the base/acid peristaltic pumps was adjusted to 0.30 to prevent overdosing.

105 The retention times to be tested during the pilot test were to range from 14 to 35 minutes. The HRTs were selected to represent the number of reactors that would require conversion in full scale operation

(maximum of three reactors out of four).

5.2.2 Daily Maintenance

The pH, temperature, and DO of the influent to the column and the effluent from the column were measured and recorded by Labatt personnel on a daily basis. The daily routine and all recorded data from the daily maintenance on the BioTower Daily Maintenance sheets provided by UoG (see Appendix

F).

5.2.3 Sampling Analysis

Laboratory analysis on the BioTower samples was completed at UoG for COD, TSS/VSS, TP, TN and pH.

Standard laboratory analysis procedures for UoG are available for review in Appendix C. COD was

chosen over BOD5 as it is a faster test (4 hrs opposed to 5 days) with a strong correlation to BOD5. The bod5/COD ratio used by Labatt is 0.53, as supported by literature. The reactor 4 wastewater effluent was regularly analyzed for COD and TSS by Quality Control at Labatt Brewery's as part of their Best

Management Practices These values were obtained to supplement the data set developed at UoG. It was noted that the COD values reported by Labatt were consistently lower than the values reported by

UoG as the test methods differ. Labatt used high range HACH vials (0-15,000 mg02/L) whereas Guelph

uses Standard Methods 5220D (0-1000 mg02/L) with a five time sample dilution. UoG sample data was considered for this entry as duplicates and standards were run as part of UoG lab procedure. TSS analysis was similar between both labs. It is also noteworthy that the COD of the Reactor 4 effluent is considerably higher than the values reported from the Labatt London 2009 Wastewater Characterization

(see Chapter 2), nor did the sample for that study occur during this portion of the year.

106 5.3 Results and Discussion

After 3 months of operations and numerous lost time delays, it was determined that the HRT of the pilot was not sufficient to allow for a significant COD reduction. Delays were attributed to:

• Technical and operational challenges (see Section 5.3.1);

• Frequent changes in the daily maintenance technician roster; and

• Staff availability.

Many of these challenges were resolved toward the end of the pilot trial. The COD removal between the equalization tank and BioTower was limited from 4.3 to 19.8% at the best case scenario. This limited reduction is not sufficient to justify a retrofit of the existing pH reactors. Labatt London would likely have to expand their wastewater treatment facilities to incorporate a biological component. A summary of the BioTower daily maintenance and performance summary is available in Appendix F.

The average pH of the reactor 4 influent into the equalization tank was 7.98 (as = 1.23) for the duration

of the pilot, and ranged from 5.86 to 10.14. 5% H2S04 and 5% NaOH consumption over the life of the pilot averaged 4.4 l/day and 6.1 L/day respectively; more caustic was consumed on average than acid, despite the average caustic pH of the Reactor 4 wastewater. This is indicative of microbial action within

the equalization tank as metabolism releases C02 into the wastewater.

The equalization tank was able to maintain system pH within the set points 90% of the time. The

average operational pH was 6.83 (o5 = 0.34). The system was out of the pH range 5 times. Of those five failures, three were early where the pH probe was not submerged in the wastewater. This was remedied with a float switch. Extensive sludge accumulation in the bottom of the equalization tank was responsible for other two instances. When the pH probe made contact with the solids, it produced a false acidic reading of 5.30, when in fact the wastewater may have been extremely caustic.

107 The average effluent pH of the BioTower column was 7.54 (os = 0.53), and was consistently higher than the pH of the equalization tank. It may be a symptom of inadequate mixing of caustic within the equalization tank, or an indicator of the biofilm characteristics. An increase in pH in biological treatment systems occurs either where denitrifying bacteria cause the release of hydroxyl (-OH) ions, or with the consumption of volatile fatty acids by methanogens. Both nitrification and methanogenesis occur in oxygen depleted environments. Anoxic and anaerobic zone may exist within the slime layer wedged between the media surface and an aerobic zone exposed to the wastewater.

5.3.1 Operational Issues

Flow Rate

The flow control system proved inadequate to provide a constant effluent flow rate through the

BioTower column. A constant HRT was supposed to be one of the control variables for the BioTower pilot. A static HRT is only possible were the active volume of the BioTower and effluent flow rates are constant (see [17).

HRT = ^ 1171

where: HRT = Hydraulic retention time (min)

V = Static volume of wastewater in the BioTower column (L)

Q = Wastewater flow rate from equalization tank (L/min)

Both of these parameters were not fully static throughout the course of the pilot, although they were adjusted back to the set points on a daily basis. The flow rate would decrease an average of 50% over a

24 hr period from the 0.85 L/min required for the 35 min HRT through the column. The average flow rate ended up being 0.61 L/min, which translates to an HRT of 49 min - considerably longer than what the maximum scaled up retrofit would allow. This means that the reported COD reduction for the pilot is somewhat overstated for the 35 min HRT trial. A single speed reciprocating pump was used to deliver

108 wastewater from the equalization tank to the BioTower. The volume of water in the water in the equalization tank exerted hydraulic pressure at the inlet of the pump. Whenever the volume in the equalization tank dropped because of a clog at the influent, the flow rate out the BioTower would decrease proportional to the drop in pressure head minus the head supplied by the pump. This would frequently occur whenever there was a clog in the equalization tank inlet.

The pilot would have benefitted from a variable speed pump actuated by feedback from a flow meter to ensure constant effluent flow rate and HRT. The system could also have been gravity fed by elevating the equalization tank. With an uninterrupted wastewater supply, the overflow line would have been sufficient to ensure constant tank depth were always consistent via.

Sludge Production and Solids Entrainment

Clogging at the influent of the buffer tank was a chronic problem. The gate 2" valve was opened just enough so that wastewater would just start to exit through the overflow. Debris would occasionally collect at the lip of the valve and result in a clog. The buffer tank had enough capacity to hold water for

8 hours of normal pilot operation at the longest HRT after the event of a clog. After the buffer tank reached the minimal water level, the pilot would turn off. From that point onward, the contents of the

BioTower column would slowly begin to drain back into the buffer tank. There were two check valves placed in series to prevent backflow; however it still occurred to a degree. The impact of clogging was mitigated by having personal check up on the pilot twice a day - once at the beginning and once at the end of a working day. The clog could easily be removed by actuating the valve.

Sludge build up in the buffer tank was also an issue. This problem wasn't identified until late into the project. After 10 weeks of operation, over 5" of thick sludge had accumulated in the bottom of the equalization tank. Sludge began to contact the pH probe. This caused false low reads in the wastewater pH and caustic over dosing. The long term complication was in sludge entrainment into the recirculation line and column feed water line. This was a problem as the equalization tank output was at the very

109 bottom of the tank. The sludge was very fine and viscous in consistency which allowed it to become easily entrained. The sludge composition was likely spent diatomaceous earth bleed through from the rotary drum filter, spent yeast from fermentation and aging tank bottoms wash out, and packaging wastes (paper pulps), all of which exhibit a very high organics content. As a result, in the wastewater sampling, the equalization tank wastewater loading was generally higher the Reactor 4 composites. The discrepancy in concentrations was initially attributed to placement of the Reactor 4 auto-sampler and the batch discharge nature of wastewater which could have account for large fluctuations in solids over time. The tank was manually cleaned out once the problem was identified, and sludge depth was monitored on a weekly basis. A simple way to avoid sludge entrainment in future pilots that require equalization tanks would be to draw wastewater from a higher level within the tank.

Nitrogen

A noted nitrogen deficiency could have inhibited the extent of COD removal. The approximate Carbon to

Nitrogen to Phosphorous (C:N:P) ratio in wastewater for aerobic metabolism should be 100:5:1. C, N and

P may be represented by BOD, TN, and TP respectively. The actual C:N:P ratio determined from testing in 2009 was 1770:14.4:11.3; this may be reduced to 100:0.81:0.61. However, the highest recorded TN reduction through the BioTower column was a mere 15%. To combat this potential problem, 50 g of

ammonium nitrate (NH4N03) were added to the equalization tank as a daily nitrogen supplement.

Calculations suggested that upward of 200 gNH4N03/day would have been required to satisfy the ideal

C:N:P ratio, but UoG chose to act conservatively. It should be noted that the best COD reductions during

the pilot were on the days with NH4N03 addition, but it was not a substantial improvement. The

maximum observed COD removal before NH4N03 addition was 10.6% and 19.8% after NH4N03 addition.

TN was always present in the effluent, with or without the nitrogen supplement in the range 12 to 34

mgTN/L.

110 A further investigation into the possibility of nitrogen deficiency, a 24 hour time series analysis to test

COD reduction on 1L grab samples of brewery wastewater with varying concentrations of NH4N03 was conducted. The brewery wastewater was aerated over a 24 hr. period. There was no addition of microorganisms, such as polyseed, to the wastewater as it was determined through observation that viable microorganisms were already present within the brewery wastewater; the results therefore would also provide insight into the benefits of simple aeration of the wastewater. DO concentrations

were maintained above 2.0 mg02/L. Samples were withdrawn, acidified to a pH below 2 with H2S04, and analyzed for COD at the 0,1, 2,4, 8 and 24 hr interval. The results are present in Figure 19.

COD Reduction Time Series Analysis No Polyseed 22-Mar-ll 4500

-•-0 mg NH4N03/L —•""20 mg NH4N03/L

•"*r 40 mg NH4N03/L -*-60 mg NH4N03/L

0 4 8 12 16 20 24

Time (hr)

Figure 19: Results of COD reduction 24 hr. time series in brewery wastewater with ammonia nitrate supplements for investigation into the possibility if a nitrogen deficiency affecting biological wastewater treatment

The time series analysis suggests that nitrogen addition can substantially improve COD reduction up to

50% over a 24 hr HRT; however there is no distinct improvement in the shorter retention times (<2hrs).

The nitrogen deficiency is apparent to some degree, and should be kept in consideration when pursuing future biological wastewater treatment.

Ill There are several methods in which nitrogen concentrations in brewery wastewater can be improved.

Recirculation of the BioTower effluent back to the pilot head works may improve COD removal.

Recirculation dilutes inlet substrate concentrations while reducing available carbon, thus improving the

C:N:P ratio. Performance deteriorates when recirculation occurs above a given rate (Metcalf and Eddy,

2003). Anaerobic biological treatment may be more effective than aerobic treatment as it requires less nitrogen (C:N:P of 250:5:1 opposed to 100:5:1). More nitrogen could be included in the wastewater through slight adjustment to brewing processes, such as switching from phosphoric acid to nitric acid for

CIP processes.

Oxygen

The dissolved oxygen (DO) concentrations in the BioTower effluent were subject to daily changes. The average DO was 3.02 mg/L however concentrations dropped below the minimum recommended 2 rogDo/L 40% of the time. This limit was established to prevent the system performance from becoming oxygen limited. The airflow rate was adjusted weekly as necessary (i.e. low DO concentration over successive days). The COD reduction may have been more significant if the DO could have been consistently maintained above this minimum. The final DO concentration is a function of flow rate, temperature, BOD loading, and microbial activity. Better control over these variables would lend to a more consistent effluent DO concentration.

Temperature Fluctuations

Temperature is directly tied to the solubility of gases, and rate of metabolic processes. Biofilm processes such as trickling filters are less effective at lower temperatures. The final BioTower effluent temperatures fluctuated from 9.5 to 23.6°C over the duration of the pilot, and could change as much as

10°C within a 24 hr period. This is still within the optimal growth range for biological wastewater treatment. However in a few instances the wastewater may have been even colder; there were lost time days for pilot operations in January when pipes froze. Rapid temperature changes may have stressed the

112 biofilm. Thermal control and operation during summer months are recommended for any future biological treatment pilot projects.

5.4 Recommendations for End-of Pipe Treatment

A high rate solids removal technology is highly recommended for this waste stream. Many units are available that can treat industrial effluents with flow rates up to 14 000 m3/day. The removal of 40 to

70% TSS often results in a corresponding 10 to 30% reduction in BOD5 for many waste streams. Solids removal would:

• Reduce the High Strength Sewage Surcharge;

• Cut maintenance costs associated with sludge removal in pH adjustment reactors;

• Potentially reduce occurrence of pH exceedance by removing colloidal particles that may

deposit on probes; and

• Provide a form of primary clarification that by convention should make biological treatment

more viable.

The project budget did not allow for the inclusion of a pilot-scale high-rate solids removal technology,

(i.e. a centrifuge belt press) in parallel operation to the BioTower pilot as desired upon project conception. An attempt was made by UoG to have US Centrifuge process 25L of brewery wastewater in one of their pilot centrifuges in Indianapolis to develop some figures for TSS and BOD reduction with a centrifuge belt press (see Appendix G). The sample was collected from the Labatt London Brewery, transported and received at US Centrifuges pilot facility within a 24 hour period. The treated effluent was to be tested at Heritage Environmental in Indianapolis for COD, TP, and TSS. Unfortunately due to poor sample handling and mismanagement, the results from the US Centrifuge test were inconclusive, and funding was not available for a second attempt. Continued pursuit into solids removal technologies is highly encouraged.

113 5.5 Conclusions

The BioTower Pilot was not able to satisfy the project objectives. It appears that the conversion of three of the four pH adjustment reactors to biological packed towers would not provide enough of a hydraulic retention time for significant COD removal. Significantly more space than the 210m3 provided by

reactors 2, 3, and 4 would be required to begin to meet BOD5 reduction targets with an aerobic AGP.

Aerobic AGP for biological treatment is still one of the most compact and effective methods for BOD reduction. However the results of this pilot suggest that an additional facility with a larger wastewater holding capacity would be necessary at Labatt London. More land would be required to house these tanks, as there is no additional room for expansion.

There is room for improvement in the experimental design of the BioTower pilot. Further study into the

BioTower retrofit may be warranted if the following components can be address:

• Fully automated system to reduce required labourer hours. Tight controls are required over all

aspects of biological treatment as varying flow, pH, temperature, solids, and nutrient loading

can complicate the performance of the system;

• Installation of PLC systems and data loggers for tighter management of control variables;

• Operation over the spring/summer months to avoid freezing issues, or better insulation of the

wastewater effluent room;

• Primary clarifier addition for solids removal;

• Sludge management strategy in place for equalization tanks; and

• Nitrogen deficiency remedied through either supplementation, a switch from divosheen

(phosphoric) to nitric acid for acid wash CIPs, or by considering anaerobic over aerobic.

A repeat trial with the above addressed may yield a stronger performance from this wastewater treatment technique.

114 Exploration into high rate solids removal technology for primary treatment is strongly recommended at

Labatt London. There are several units available on the market that should be able to drop the brewery

effluent TSS to within municipal specifications with some BOD5 removal, such as belt filter presses- centrifuge combinations. The mean particle diameter size is 328. Sonication valves maybe reduce particle size below the 1.2 |im diameter mark; the cut-off that defines a total suspended solid. Solid removal would also cut down on the frequency for cleaning out sludge from the pH adjustment reactors.

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119

i Glossary of Terms

Acids: an acid cleaning agent removes mineral deposits (e.g. beer stone) that do not dissolve in alkaline cleaning agents. 1.0% saltpeter acid is the most frequently used.

Adjuncts: substances such as syrup or molasses added during the brewing process that provides fermentable sugars in addition to those from malt.

Aerobic: catabolic processes that require oxygen as the electron acceptor for the synthesis of ATP.

Anaerobic: catabolic process that use substances other than oxygen as the electron acceptor for the synthesis of ATP.

Biochemical Oxygen Demand: A wastewater parameter indicative of the magnitude of dissolved oxygen uptake required by microorganisms to decompose the readily available organic matter.

Brand Change: The process of switching a packaging line from one brand of beer to another

Beer Pack: Mixed beer in the transmission pipe from Bright Beer to Packaging, called so as one brand pushes or chases the other.

Beer Stone: A mixture of calcium oxalate and organic material that deposits on container surfaces during brewing operations, also known as beer scale.

Bright Beer: Finished beer awaiting packaging

Butts Tanks: A vessel that stores the mixed volume of beer (beer pack) that occurs in the conveyance lines from the Bright Beer Tank to the Filler Bowl as a result of a Brand Change.

Caustics: An alkali cleaning agent such as caustic soda lye, polyphosphates and silicates used to remove organic dirt. They also emulsify, dissolve proteins and kill some bacteria.

Chemical Oxygen Demand: A wastewater parameter indicative of the amount of oxygen necessary to oxidized all organic compounds present.

120 Chill-haze: Large proteinaceous colloids formed at low temperatures through the aggregation of hydrophilic proteins with a phenolic substance (tannins) as the coagulating agent (Gan, et. al., 1998).

Chill haze increases turbidity and is often undesirable in finished beverages. The selection of grains with low-water soluble nitrogen content reduces its formation.

Cold break: Trub separation after wort cooling

Crown: A beer bottle cap.

Diatomaceous Earth (DE): a light, friable sedimentary soil consisting of siliceous diatom remains, also known as kieselguhr. The typical chemical composition is 86% silica, 5% sodium, 3% magnesium, and 2% iron. With high thermal resistance and a characteristic high porosity, DE is an attractive, naturally occurring filter media and absorbent.

Facultative: a qualifying adjective indicating that an organism is able to grow in either the presence or absence of an environmental factor (for example, "facultative aerobe").

Fermentation: catabolic reactions producing ATP (adenosine triphosphate) through substrate level phosphorylation in which organic compounds serve as both primary electron donor and ultimate electron acceptor. Yeast (saccharomyces cerevisiae) fermentation in anaerobic conditions produces

alcohol (or ethanol) and carbon dioxide (C02) as a byproduct. Aerobic conditions yield mostly C02.

Filler Bowl: An automated carousel styled bottle filling unit used for the industrial bottling of beer.

Fly Ash: Darker beer recipes occasionally require slightly burnt malt that elicits this fine particulate in the flume of kilns.

Fobbing: Overfilling/foaming of a bottle of beer during the filling operation to minimize the oxygen transfer into the bottled beer before capping.

Green Beer: Recently fermented, un-aged beer that hasn't reached maturity.

Granules: a self aggregating, robust pellet of self-immobilized bacteria (~2-7 mm) that has a regular, dense and strong microbial structure with high biomass retention, otherwise known as granular sludge.

121 Bacteria in digesters form these clusters in response to external stressors (hydrodynamic shear forces, a high ration of height to diameter of the reactor, frequent repetition of feast and famine conditions). The interior is dominated by large cavities and rod-shaped bacteria, while the outer surface is laced with fungi and filamentous microorganisms. They have a good settling ability, are shock and toxic loading resistant.

Grist: milled malt.

Gruit: an old fashioned herb mixture for bittering and flavouring beer popular before the extensive use of hops.

Hectoliter: The common volumetric unit of measurement used in brewing operations. One hectolitre

(hL) is equivalent to 0.1 m3.

Hops: derived from the female cone (also known as the strobiles) of the hop pant. Hops impart a characteristic bitter flavor in beer that counter-balances the sweetness of malt. It behaves both as a biological stabilization agent that help coagulates colloidal proteins, and as an antibiotic which favours the activity of yeast.

Jetter: A nozzle that sprays a fine stream of de-aerated carbon filtered water over the mouth of a filled, uncapped bottle leaving the filler bowl to increase the fobbing effect.

Kieselguhr: (see Diatomaceous Earth)

Krausen: A method to carbonate beer in which wort is added to the fermented/finished beer to achieve natural carbonation

Lauter Tun: An insulated vessel with a screened bottom (a 0.7 to 1.1 mm wire mesh) where mash

(generally at 78°C) is separated into clear liquid wort and residual spent grains. The spent grains form a filter cake responsible for clarification of the wort drawn from the bottom of the lauter tun.

Lucilite: amorphous silica used for filtration of rough beer (see diatomaceous earth).

122 Malt: The resulting product of cracking a cereal grain to release sugars by natural enzymatic action through a process of germination by soaking in water, followed by rapid drying (kiln drying) with hot air to reduce moisture content to around 4 per cent. The hotter the grain is dried, the less diastatic/enzymatic power of the finished malt. Barely is the most popular malt grain employed in brewing. The malting process is may be completed offsite from the brewery.

Mash: The product of combining mixed milled grains (commonly malt with supplementary grains such as corn, sorghum, rice, rye, or wheat) and water in an insulated vessel (the mash tun) maintained at a constant temperature (typically 45°C, 65°C, or 72°C). The enzymes in the malt reduce starches in the grains into simple sugars (primarily maltose) that may be utilized by yeast.

Pasteurization: the killing of micro-organisms in aqueous solutions by heat. Traditionally in the US, only beer in cans and bottles is pasteurized after it has been packaged. The process involves running the vessel through a hot water spray (approximately 60°C) for two to three minutes. This allows the beer to be stored at room temperature for a period of 120 days, or at 4°C for 6 to 9 months as opposed to 45-60 days unpasteurized.

Pitching: the addition of yeast to unfermented wort.

Plato Scale: An empirically derived scale to measure the density of beer wort in terms of percentage of extract by weight at a reference temperature of 17.5°C. This correlates to the amount of fermentable material present in wort. The scale expresses density as the percentage of sucrose by weight; wort measured at 12° Plato has the same density as water - sucrose solution containing 12% sucrose by weight denoted as 12% Brix.

Pre-coating: a water intensive process of preparing a filter vessel with a uniform deposition of diatomaceous earth (DE) by circulating a DE and clean water slurry at a high flow rate over the filter leaves.

PRO: Post Run-Off

123 Racking: a brewing term that encompasses chilling, filtration, centrifugal separation, and carbonation processes. It results in the refinement of green beer from fermentation into a finished product.

Rough Beer: Beer that has not been through clarification/filtration.

Saltpeter acid: otherwise known as potassium nitrate (KN03), a naturally occurring stone salt commonly used in acid washes for CIP processes in brewing. It is also the chief constituent of gun powder.

Sparge: water sprayed over spent grains in the lauter tun to reclaim residual liquor.

Spent grains: are the discarded husks of cereals that result from starch removal during the mashing process. They have the consistency of oatmeal mixed with sawdust. Occasionally spent grains are utilized as a coarse filter for wort during lautering (in a strain master). Around 140 kg of spent grains are generated per m3 of wort produced in a well-designed and efficiently-working brewhouse. A water content of 80 percent is typical. Spent grains may be sold as animal feed.

Staging: A large warehouse for storage of packaged beer ready for delivery into the market place.

Stillage: See spent grains.

Thermophilic Anaerobes: otherwise known as thermophiles, a type of bacterial organism that thrives at relatively high temperatures (45°C - 80°C). Some are anaerobes that use the sulfur instead of oxygen as an electron acceptor during respiration. Some are lithotrophs that oxidize sulfur to sulfuric acid as an energy source, thus requiring the microorganism to be adapted to very low pH (i.e., it is an acidophile as well as thermophile). The thermophiles in anaerobic digesters would be classified as facultative or moderate thermophiles that typically operate at a neutral pH of 7.

Trub: the material along with the hops debris left in the boil kettle or hop back after the wort has been transferred and cooled. Brewers generally prefer that the bulk of the trub be left in the kettle rather than stay in contact with the fermenting wort. Although it contains yeast nutrients, its presence can impart off flavors in the finished beer. Trub contains a large proportion of polyphenols from the malt.

124 Typical trub has a dry matter content of 15-20 per cent. Trub has a high BOD5, around 110,000 mg/kg wet trub. Trub is separated from the wort in whirlpool vessels.

Underback: also known as the PRO tank, it is a vessel used to store recovered mash liquor from sparging the lauter tun before being sent to the wort kettle.

Wort: a liquid consisting of unfermented malt sugars extracted from the mashing process that is utilized by brewing yeast to produce alcohol. Bitter hops may be added to the wort and boiled to impart additional flavor and aroma.

Yeast: a facultative microorganism used to ferment beer. Excess yeast is produced during the fermentation process. Around 20-40 kg of yeast slurry is produced for each cubic metre of beer. This has a very high BOD value, in the range of 120,000-140,000 mg/l.

125 126 | Appendix A | Brewery Wastewater Characterization Sample Detail

127 LOCATION: 1- Brewhouse and Packaging Lines 1,4, & 5

SAMPLE TYPE: GRAB S COMPOSITE X

DATES: April to July '09

QUANTITY: 4

BOD- 2370 mgA VSS- 132 mg/L ° 5- (±1252) (±108) 2.4 mg/L 9.4 (±0.6) P (±1.8)

DRAWING AutoCAD Drawing: Level 1 West Brewing Side.

Manhole at which the brewhouse and packaging

lines 1, 4, and 5 empty. There is continuous Figure 1A: Manhole outside packaging line 4 wastewater flow. Can line beer changeovers cause a significant spike in BOD, TSS, and TP. There are three shifts per day. Depending on production, there are 3 to for brand changes for the bottle line per shift, and 1to 6 brand changes DESCRIPTION: for the can line per shift.

Samples were drawn from a weighted 1 L plastic sample jar lowered on a nylon rope.

Figure IB: Packaging Sewer detail LOCATION: 2 - Brewhouse and Packaging Line 1

SAMPLE TYPE: GRAB V COMPOSITE X

DATES: April to July '09

QUANTITY: 5

1894 mg/L 69.1 mg/L (±627) (±17.2)

1177 mg/L PARAMETERS: BOD- VSS- 34.4 mg/L B0Ds' (±360) VSS- (±8.9) 2.5 mg/L 10.3 TP' (±0.8) PH- (±0.3)

DRAWING AutoCAD Drawing: Level 1 West Brewing Side

Manhole very close to the Brewhouse. The Figure 2A: Manhole in sidewalk, NW corner of Brewhouse wastewater appears considerably higher than expected, suggesting there may be an additional contributing source.

There are significant spikes in BOD during line 1 brand line changes and PRO and Trub discharge DESCRIPTION: to drain.

Samples were drawn from a weighted 1 L plastic sample jar lowered on a nylon rope.

Figure 2B: Brewhouse sewer depth detail LOCATION: 3 - Packaging Line 1

SAMPLE TYPE: GRAB >/ COMPOSITE V (x2)

DATES: 22-APR-09

QUANTITY: 5

1059 mg/L 62.2 mg/L (±505) (±31) BOD- 931 mg/L 33.1 mg/L PARAMETERS: BO°S (±637) VSS' (±7.6)

TP: 1',6",?fL pH: (±0.6) K (±0.7) AutoCAD Drawing: Ground Floor West Packaging DRAWING Figure 3A: Manhole outside Reception, NE corner of Brewhouse Side

Manhole directly outside Administration. Exclusively bottle line 1 wastewater. Bottle Washers, Pasteurizer, Vacuum Pumps (see locations 15, 16, &17), and floor drains are primary contributing sources.

Two composite samples were obtained by an DESCRIPTION: ISCO automatic water sampler. Four samples per hour over a four hour period were collected in each case.

Grab samples were drawn from a weighted 1 L Figure 3B: Packaging Line 1 sewer depth detail plastic sample jar lowered on a nylon rope. LOCATION: 4 - Powerhouse C02 Scrubber Water

SAMPLE TYPE: GRAB • COMPOSITE X

DATES: April to July '09

QUANTITY: 3

2709 mg/L 0.0 mg/L (±232) (±0.0) AVERAGE 1756 mg/L 0 0 m6/L CHEMICAL BODs: VSS: 5 (±260) (±0.0) PARAMETERS: 0.06 mg/L ,, 5.28 TP: (±0.05) PH: (±0.25)

DRAWING AutoCAD Drawing: Level 1 West Brewing Side

Sample Spigot in the bottom of the C02 scrubber

Tank. The C02 Scrubber removes C02 and odours generated by the Fermenters. De-aerated Carbon Filtered Water is used as the scrubbing water. Wastewater is discharged to drain continuously. The flow rate is a relatively consistent 1.9 m3/hr. During low production periods, the flow rate may DESCRIPTION: be reduced to 0.9 m3/hr. Operations are

controlled by RS View. C02 is collected off the Fermenters for reuse in other processes (i.e. Aging Tank purging after CIP). Dimethyl sulphides (DMS) gases are expunged. BOD is from Beer Figure 4A: Open sample spigot on C0 Scrubber Tank in Powerhouse Stone. Samples were drawn from the spigot at 2

the bottom of the C02 Scrubber vessel. LOCATION: $ 5 - Powerhouse Boiler Blow Down Water § CO CD 3 SAMPLE TYPE: GRAB COMPOSITE X TEL 5' oo on DATES: April to July '09 C 3 3 o> QUANTITY: 3 < o rnn- 93 mg/L 2.0 mg/L (±84) (±2)

2 L m g /L PARAMETERS: BOD,: f^5( VSS: °° „ l 5 (±6.4) (±0.0) 1.76 mg/L 11.9 (±0.5) P (±0.28)

DRAWING AutoCAD Drawing: Level 1 West Brewing Side Ul i There are two Blow down water streams. The CO 2. composition of the water is monitored and controlled by Powerhouse Operators through RS E. o $ View. Boiler blow down water is held at 350°F Q_ O while in operation, and cooled to 140°F before € 3 sent to drain. The boiler discharges for 20-30 $O) H- seconds per shift. Bottom blow down is fD DESCRIPTION: completed twice a day to remove un-dissolved solids in the boiler mud drum. The valve is opened for about 10-15 seconds. Boiler water is usually slightly basic, lately is has been high due to condensate contamination.

The boiler blow down water was drawn from the Figure 5A: Boiler blow down sample line (orange label) in o> sample line in the Powerhouse Lab. o~ Powerhouse Lab sink a> LOCATION: 6 - Aging and Fermentation

SAMPLE TYPE: GRAB COMPOSITE X

DATES: 22-APR-09

QUANTITY: 4

COD: TSS: (±334) (±27.6) . 653 mg/L 32.1 mg/L PARAMETERS: B0Ds- (±396) VSS" (±21.1) 9.4 mg/L 6.3 (±0.5) P (±1.0)

DRAWING CAD Drawing: Level 1West Brewing Side

Figure 6A: Manhole outside door leading to Chip washers Filtration

Manhole immediately outside Chip Washer door, close to sample location 12. Contributing sources from Chip Washer, Powerhouse, Carbonating DESCRIPTION: Room, and Central CIP return.

Grab samples were drawn from a weighted 1 L plastic sample jar lowered on a nylon rope.

Figure 6B: Fermentation sewer depth detail $ LOCATION: 7 - Vertical Fermenter Clean In Place Wastewater SE 0)CO 3 "EL SAMPLE TYPE: GRAB X COMPOSITE •/ 5' 00 00 c DATES: April 3 3 QJ QUANTITY: *3 (1 if no standard deviation provided) *< o

INITIAL BURST

COD: 115000 mg/L TSS: 3500 mg/L

BODs: N/A VSS: N/A •sj I < TP: 25 mg/L PH: 4.21 Figure 7A: Bottom of Vertical Fermenter. April samples obtained from floor drains. oo> PRE RINSE 0) —s 3 0) 1517 mg/L 197 mg/L 3 COD: TSS: r-+ (±879) (±134) rt> PARAMETERS: 904 mg/L 223 mg/L o B0Ds: VSS: -o (±609) (±140) 5.0 mg/L 10 TP: PH: (±1.3) (±2.0)

POST ACID CYCLE RINSE

790 mg/L 346 mg/L COD: TSS: (±864) (±) 39 mg/L 19 mg/L B0D : VSS: 5 (±68) (±) cu Figure 7B: CIP Effluent Pit behind curtain in front of Chip Washer 32.0 mg/L 3.5 crO) TP: pH: (±13.3) (±2.1) o3 Q_ O 3 PARAMETERS pit in the basement near Carbonating. SANITIZER RINSE DESCRIPTION CONT'D rnn- 37.0 mg/L 65 mg/L (±38.4) (±83)

B0D : 3 5 /L VSS: 6 7 m L s ( ±5.07) g/ 3.97 mg/L 5.90 (±0.30) P ' (±0.82)

POST RINSE

rnn- 39 mg/L 200 mg/L (±18.5) (±280)

/L VSS: BODs: °(+™o) 2.80 mg/L

2.49 mg/L 6.27 (±1.74) P (±0.34)

DRAWING CAD Drawing: Level 1 West Brewing Side

Vertical Fermenter Clean In Place is a two hour procedure with several steps. Cone yeast is recaptured. The initial burst is the bottom of the cone. Pre rinse is recycled CIP water sent to drain. The vessels are then acid washed (1.13% Divosheen). The Post Acid Rinse is sent to drain. DESCRIPTION: Sanitizer rinse is chlorinated water. The final rinse is de-aerated carbon filtered water. Labatt cleans an average of 3 Fermenters per day.

Grab samples were coordinated with RS View

during drawn from a weighted 1 L plastic sample Figure 7C: CIP Effluent Pit jar lowered on a nylon rope from the CIP effluent o 3 Q. O 3 $ LOCATION: AutoCAD Drawing: $ 8 - Aging Tank Manual Clean DRAWING <•> Ck> 3 "D_ SAMPLE TYPE: GRAB COMPOSITE X Aging tanks are manually cleaned before a CIP is 5' initiated, very similar to Krausen Tanks (11). oo Averages of 5 tanks are cleaned per day. Residual c DATES: JUNE to AUGUST 3 3 beer (~5 hL per tank size) is drained to the floor. CD QUANTITY: 1 The vessels are then periodically rinsed out with «< 0 city water (1 minute on, 2 minutes off) until the «X) INITIAL BURST 1 effluent runs clear. The process takes DESCRIPTION: approximately 17 minutes to complete. City COD: 100415 mg/L TSS: 3768 mg/L water is supplied by 2" lines at 60 to 80 psi.

00 B0Ds: 102034 mg/L VSS: 3768 mg/L Aging tank CIP applies warm acid (Divosheen I > 00, 1.13% @ 31°C) rinse. CIP wastewater was from 5* TP: 41 mg/L pH: 4.03 oo the acid wash post rinse, and a sanitation rinse. QJ•H The process takes 1hour to complete. 3 CLEAR RINSE cu 3 C COD: 3225 mg/L TSS: 765 mg/L QJ_ PARAMETERS: Q rD O) B0D5: 2089 mg/L VSS: 765 mg/L 3

TP: 3.86 mg/L pH: 5.00

CIP BACK FLUSH

COD: N/A TSS: 0 mg/L

B0D5: 48 mg/L VSS: 0 mg/L

O) O" TP: 2.5 mg/L pH: 5.12 Figure 8A: Aging Tank QJ o 3 Q_ O 3 LOCATION: 9 - Rotary Drum Filter Pit

SAMPLE TYPE: GRAB COMPOSITE X

DATES: 22-APR-09

QUANTITY: 2, 2, and 5 respectively

BEER FILTER

8934 mg/L 23942 mg/L COD: TSS: (±3699) (±26868) 6454 mg/L 3176 mg/L BOD : VSS: s (±4181) (±3273) 18.01 mg/L 5.64 TP: pH: (±6.64) (±0.08) Figure 9A: Beer Filter

DE SLURRY

8794 mg/L 19047 mg/L COD: TSS: (±981) (±3606) PARAMETERS: 6601 mg/L 2648 mg/L BOD : VSS: 5 (±1420) (±628) 14.3 mg/L 5.13 TP: pH: (±8.9) (±0.77)

RDF FILTRATE TO EFFLUENT PIT

3927 mg/L 7414 mg/L COD: TSS: (±2555) (±7420) 2960 mg/L 3780 mg/L BOD : VSS: 5 (±1947) (±6408) Figure 9B: Rotary Drum Filter 3.4 mg/L 6.1 TP: pH: (±3.4) (±0.7) DRAWING: AutoCAD Drawing: Level 2 West Brewing Side CO o> Solids are reclaimed from diatomaceous earth 3 -o_ (DE) Slurry generated from dumping the beer 5' cm filter by a rotary drum vacuum filter (RDVF). Beer on beer filter uses 80 kg of DE c is filtered at -2°C. The 3 as a pre-coat. An injection of 0.065 kg of DE is per 3 hL of filtered beer prevents channelling. A max. 4500 hL of beer is run before the filter cake is O<£> dropped. City water is sprayed for 30 minutes in I bursts to dislodge the old cake. Each filtration washout is 50 hL, and occurs 3 times per day during peak production. There is a turbidity meter on the beer filter effluent that governs flow diversion to either the DE Slurry tank for lO 1 treatment by the RDVF, or directly to drain. a

The RDVF filter media is a 150 urn nylon fabric DESCRIPTION: c a> with 100 mesh/ inch. The filtrate empties to the 3 DE Effluent Pit, along with water for priming the pump. This wastewater results is a significant point source for solids loading. The RDVF filter mesh size was too large for the application at the time of sampling negatively affecting removal efficiency. This manifested an excess of TSS in the Figure 9C: DE Effluent Pit with 300 hL DE Slurry Storage Tank in filtrate. Background. Beer filter washout was sampled from the bottom of the beer filter with a weighted 1 L There are four effluent pipes in the effluent pit. From left to right, plastic sample jar lowered on a nylon rope. DE they are 1) Overflow, 2) RDVF Filtrate, 3) Vacuum Pump Seal Water, Slurry was obtained from the filtrate effluent pipe and 4) DE Slurry from RDVF vat.

at the bottom of the tank. The RDVF filtrate was CD O" collected from the effluent pit while the unit was a> in operation. o 3 Q_ O 3 LOCATION: 10 - Surplus Yeast Tanks

SAMPLE TYPE: GRAB V COMPOSITE X

DATES: 22-APR-09 to July '09

QUANTITY: 3

5178 mg/L 589 mg/L LO (±3841) " (±593) _ __ 2332 mg/L 168.5 mg/L PARAMETERS: BODs: (±1161) VSS" (±167)

Tp. 4.45 mg/L 6.49 (±0.71) P " (±0.67)

DRAWING AutoCAD Drawing: Level 2 West Brewing Side Figure 10A: Surplus Yeast Tanks

Manhole between Surplus Yeast Collection Tanks. It is at least 30 ft deep. This manhole to receives wastewater from various different source, including the Keg line, floor drains from Krausen, Vertical Fermenter floor drains, Cellar 1 Fermenters and Aging Cellar 14, Yeast Room, DESCRIPTION: Cellars 2 & 4, and the tower above cellar 3. Wastewater was highly variable in appearance from one sampling event to the next.

Grab samples were drawn from a weighted 1 L plastic sample jar lowered on a nylon rope. Figure lOB: Manhole Location under Surplus Yeast Tanks LOCATION: AutoCAD Drawing: Level 3 West Brewing Side CO 11- Krausen Tank Manual DRAWING OJ 3 Manual Clean. C02 Evacuation takes *g^ SAMPLE TYPE: GRAB X COMPOSITE S 5* approximately one hour. The Krausen tank OQ m bottoms (~5 to 10 hL) are then sent to floor c DATES: August to September 09 3 drains. The bottoms consist of beer, yeast, and 3 QJ wood chip sediments. City water is applied in a QUANTITY: 2 series of 10 to 15 minute bursts to facilitate o to removal, followed by 1 to 5 min with the water INITIAL BURST off to allow draining. Large tanks require 5 to 7 iterations. At the halfway mark, an operator 153 lOOmg/L 103774 mg/L COD: enters the tank to manually split the chips with a (±76600) (±6422) squeegee to further dislodge beer, and then burst 94700 mg/L 97299 mg/L BODs: rinsing is resumed. Once the effluent runs clear, I (±23400) (±6330) the beech chips are removed and transferred to t; a> 200 mg/L 5.24 TP: the Chip Washers. The Krausen Tanks are then c:cn (±124| pH (±0.23) CIP'd. Labatt London averages 19 tanks cleaned 3 —H st per week. Tank sizes vary from 560 to 1450 hL. O) 1 RINSE DESCRIPTION: 3

17625 mg/L 10639 mg/L The CIP cycle consisted for Krausen and Aging cu COD: 3 (±141) (±52) Tanks consist of a pre-rinse, a caustic wash C PARAMETERS: 9L 10804 mg/L 10008 mg/L followed by a post rinse, a sanitizing wash Q BODs: ro (±193) (±13) followed by a post rinse, and a final post rinse. Q> 3 39.4 mg/L 5.54 The final rinse is recycled as the pre-rinse for the TP: (±12.8) P (0.10) next CIP cycle.

2nd RINSE For the Manual Clean, grab sample were obtained from Tank 206 and 1402 every time the 10175 mg/L 5251 mg/L COD: hose was reapplied. (±4349) (±1241) CD crQJ 4978 mg/L 5058 mg/L Krausen CIP samples were drawn from the CIP BODs: (±1332) (±1242) o effluent pit (See Vertical Fermenters). The CIP 3 CL 24.5 mg/L 5.82 effluent is assumed to be representative of aging O TP: 3 PH tanks as well. |4.1±) ' 0.08 o 2 GO 3rd RINSE 0)

11388 mg/L 6442 mg/L COD: era3 (±3977) (±3457) co c 6504 mg/L 6173 mg/L BOD : 3 s 3 (±3811) (±3116) CD 22.0 mg/L 5.78 TP: O (±2.6) (0.22) ID

FIRST SLUG AFTER CHIPS SPLIT

COD: 56938 mg/L TSS: 56986 mg/L Hi i BODs: 40035 mg/L VSS: 53 990 mg/L

PARAMETERS C:in TP: 21.1 mg/L pH: 5.28 FD CONT'D 3 -HQJ Figure 11A: Krausen Tank 3 4™ RINSE ^r

QJ 2740 mg/L 1267 mg/L 3 COD: C (±1769) (±836) Q 1386 mg/L 1234 mg/L n> BOD5: o> (±685) (±806) 3 7.0 mg/L 6.65 TP: (±4.2) "" (±0.33)

5™ RINSE

382 mg/L COD: TSS: QJ (±138) (±74.9) O" CD 914 mg/L 365 mg/L BOD5: (±136) (±63.1) O3 Figure 11B: Krausen tank interior with hose applied Q. 3.08 mg/L 7.19 O TP: 3 (±2.98) PH" (±0.1) LOCATION: 11 - Krausen Tanks CIP

SAMPLE TYPE: GRAB X COMPOSITE V

DATES: August to September 2009

QUANTITY: 2

PRE RINSE

8571 mg/L 4420 mg/L COD: TSS: (±1480) (±5283) 3303 mg/L 890 mg/L BOD5: VSS: •Wi'iiii — (±4592) (±291) wmm Figure 11C: Krausen Tank Manual Clean sample Profile 17.7mg/L 7.52 TP: PH: (±0.90) (±2.84)

POST CAUSTIC RINSE

1277 mg/L 363 mg/L COD: TSS: (±171) (±105) PARAMETERS: 254 mg/L BOD : 789 mg/L VSS: s (±8) 11.6 mg/L 13.29 TP: PH: (±7.22) (±0.02)

CI02 RINSE

COD: 103 mg/L TSS: 17 mg/L Figure 11D: Separated sample (Chips split)

BODs: N/A VSS: 16 mg/L

TP: 1.5 mg/L pH: 11.52 LOCATION: 12 - Main Effluent before Reactors (SAN 34)

SAMPLE TYPE: GRAB V" COMPOSITE X

DATES: 22-APR-09

QUANTITY: 4

2863 mg/L 1019 mg/L (±1070) (±1048) 1699 mg/L 1272 mg/L PARAMETERS: B0Ds- (±348) VSS" (±1172) 4.78 mg/L 7.95 (±3.62) P (±1.74) Figure 12A: Manhole outside of doors to Reactors DRAWING AutoCAD Drawing: Level 1West Brewing Side

Manhole SAN 34 is the primary junction before collective brewery wastewaters enter the reactors to be discharged to sanitary. DESCRIPTION:

Grab samples were drawn from a weighted 1 L plastic sample jar lowered on a nylon rope.

Figure 12B: Main effluent sewer depth detail (SAN 34) LOCATION: 13 - Keg Line Sump Pit

SAMPLE TYPE: GRAB •/ COMPOSITE X

DATES: 22-APR-09

QUANTITY: 6

COD" 7373 mg/L TSS- 117mg/L (±3180) (±44)

4536 mg/L PARAMETERS: BOD- VSS- 81 mg/L B0Ds- (±2115) VSS" (±56) 4.5 mg/L 10.17 (±0.5) P (±0.91) Figure 13A: Keg Line running Domestic Brands DRAWING AutoCAD Drawing: Ground Floor Keg Line

Sump pit is approximately 2.5ft x 3.5 ft, located in SE corner of Keg Line. Wastewater is very warm. Line lubricant, CIP wastewater, waste beer and spills are contributing sources. Beer from return kegs, spillage, and brand changes are responsible for significant BOD loading. BOD loading changes depending on the brand being processed. DESCRIPTION: Kegs range in size (15, 20, 30, and 58 L). Domestic kegs are full silver, US brands have a blue stripe, and Bass Ale is orange. Foreign kegs have a white stripe. The nutrient loading may vary depending Figure 13B: Keg Line Sump Pit on the brand and volume being run. When the line is idle there were negligible nutrients detected in the keg sump (no exceedances). LOCATION: 14 - Reactor 4

SAMPLE TYPE: GRAB */ COMPOSITE V"

DATES: July to October '09

QUANTITY: 6

2333 mg/L 900 mg/L (±1355) (±411)

PARAMETERS: Rnn • 1797 mg/L \/c<- 568 mg/L B°°5- (±602) VSS" (±235) 11.4 mg/L 6.13 (±3.6) P (±0.75)

DRAWING AutoCAD Drawing: Level 1 West Brewing Side Figure 14A: Wastewater Reactors

The final area before wastewater is discharged to the municipal sanitary system. A pH adjustment occurs before the wastewater enters the reactor. Composite samples obtained from Labatt automatic sampler on top of reactor 4. 6 aliquots DESCRIPTION: are drawn over a 24 hr period. Grab samples were obtained from a sample spigot at the bottom of Reactor 4. Composite samples generally had greater nutrient loading than grab samples.

Figure 14B: Reactor 4 sample spigot (yellow handle) LOCATION: 15 - Packaging Line 1: Bottle Soaker Pit & Butt Sprayer

SAMPLE TYPE: GRAB • COMPOSITE X

DATES: September to October '09

QUANTITY: 2

rnrv 4231 MG/L TCC- 179 MG/L (±4358) (±31) . 1573 mg/L 144 mg/L PARAMETERS: Figure 15A: Line 1 Bottle Washer B0D5" (±827) VSS" (±18) 3.18 mg/L 12.34 (±1.06) P (±0.08)

DRAWING AutoCAD Drawing: N/A

The soaker pit receives wastewater from the return bottle washing process. Butt sprayer water, Sodrox BW caustic cleanser, and sanitizing Figure 15B: Line 1Soaker Pit grate by Butt Sprayer

water (CI02) are the main constituents. The blue colour of the wastewater is derived from the pigments from old bottle labels dissolved by the DESCRIPTION: caustic cleaner. The wastewater flow rate is approximately 0.9m3/hr (Labatt 2007 WW Study)

Samples were obtained from the Bottle Soaker Pit using a collection cup device from the Chemical Storage in Bottle Line 1.

Figure 15C: Soaker Pit Wastewater. LOCATION: 16 - Packaging Line 1Pasteurizer

SAMPLE TYPE: GRAB

DATES: September to October '09

QUANTITY: 2

81 mg/L 0.0 mg/L (±115) (±0.0)

PARAMETERS: BOD- 39m8/L VSS- 00mg/L B0Ds- (±56) VSS" (±0.0)

TP- 017m6/L pH: 770 (±0.08) p (±1.66)

DRAWING AutoCAD Drawing: N/A

The pasteurizers are a series of thermally Figure 16A: Top of Line 1 Pasteurizer regulated water baths that gradually increase the temperature of bottled beer from 21°C to approximately 64.1°C at the midpoint (chamber 6) for residual enzyme inactivation, and then gradually cools the bottle back to room temperature (~25°C). Water is supplied to the DESCRIPTION: entire pasteurizer at a rate of 0.9m3/hr (Labatt 2007 WW Study). Approximately 159,000 m3 of water were used by the Line 1, 3, 4, and 5 pasteurizers in 2008.

Samples were obtained at the top of the pasteurizer using a collection cup device from the Chemical Storage in Bottle Line 1. Figure 16B: Pasteurizer depth detail LOCATION: 17 - Packaging Line 1: Vacuum Pumps

SAMPLE TYPE: GRAB COMPOSITE X

DATES: September to October '09

QUANTITY: 2

1326 mg/L 4.6 mg/L (±476) (±1.7)

592 mg/L 4 4 m L PARAMETERS: rnr> • \/cc. 8/ B0Ds- (±270) VSS" (±1.9) 0.61 mg/L 6.41 (±0.06) M ' (±0.88)

DRAWING AutoCAD Drawing: N/A Figure 17A: Bottle Line 1 Vacuum Pump outside Filler Bowl

The vacuum pump is situated immediately outside the filler bowl. The effluent empties directly to a floor drain. The pictured vacuum filler (left) replaced the sampled vacuum pump. This replacement has likely effected water use DESCRIPTION: and effluent quality.

Samples were obtained at the floor drain of the vacuum pump using a collection cup device from Chemical Storage in Bottle Line 1.

Figure 17B: Vacuum Pump Sample Location at floor drain LOCATION: 18 - PRO Tank 0) 3 "D_ SAMPLE TYPE: GRAB V COMPOSITE X 5' OQ CO October c DATES: 3 3 OJ QUANTITY: 1 O V£> COD: 17450 mg/L TSS: 420 mg/L

PARAMETERS: B0Ds: 12400 mg/L VSS: 411 mg/L

TP: 19 mg/L pH: 6.06

DRAWING AutoCAD Drawing: N/A 00 I Post Runoff (PRO) is the liquid obtained off the -u20 mash tun and tauter tun after sparging spent O CD grains until the resulting filtrate registers a 3 minimum 1° Plato. Sparge water below 1° Plato is discharged directly to drain. PRO is also generated from the sparging the whirlpool after wort clarification and hops addition. Some of this sparge water is sent directly to the PRO Tank, DESCRIPTION: while the bottoms are sent to the trub tank. Here they further separate. The supernatant in the Trub tank is sent to PRO. The solid trub is mixed Figure 18A: PRO Tank in Basement of Brewhouse with sample spigot with spent grains on lower left a> Q>a~ Samples were obtained directly from the spigot on the PRO tank. Tank level is recorded by RS o 3 Q_ view in the Brewhouse. The PRO Tank has a 25 hL O 3 capacity, and held 9 hL at the time of sampling. o z | Appendix B | Labatt Wastewater Process Flow Diagram

150 LEGEND PROCESS INPUTS

WASTEWATER

PRODUCT STREAM ROT. DRUM - BYPRODUCT C02 —~(4) I— -an SCRUBBER FILTER ® SAMPLE LOCATION ADJUNCT

BR:GHT BEER

KRAUSEN/ BRIGHT BREWHOUSE FERMENTING FILTRATION AGING BEER BUFFER TANK Mil CAUSTIC

t

) ) BOTTLE PKG. ^6- POWER FILLER REJECT REBATE BEER LINE 1 HOUSE

_/v I t t T i t CAN WASTEWATER PKG. LINE 3 TREATMENT «O POST RUNOFF

BORHER

KEG RETURNS

BOTTLE PKG. KEG LINE LINE 4 & 5

SPENT GRAINS

YEAST

ENGINEERING APPROVALS DATE PLANT LONDON 04'10 DEPARTMENT DATE APP'O DRAWN JOB TITLE PROPOSAL FOR WASTEWATER CHARACTERIZATION I ••'BLr.OsS W-JCR ocs't? SJM PROCESS BREWERY WW. PFD v.'1 i.nTtlA. ON r-t K SHaStdb DWG. TITLE : 13 V.AI..V5.-: S'ROW =" Good things brewing CHECK •,;\v BCVA WC,«!MWS--.91^lLSitMC,.::: • UTU.ITIES DWG. NO. 3 2 4 0 0 0 0 0 4 JV.V.J! I. • M .-.i.-n. ! School of Engineering PROJ MGER SCALE: NTS PLANT CLASS AREA SHEET REV , | Appendix C | Standard Methods For Laboratory Analysis

152 Standard Methods Chemical Oxygen Demand S. Massen

Chemical Oxygen Demand (COD)

5220 D Closed Reflux, Colorimetric Method

From Standard Methods for the Examination of Water and Wastewater

Updated with Annotations

Tuesday, May 26, 2009

General Discussion:

a. Principle: Most types of organic matter are oxidized by a boiling mixture of chromic and sulphuric acids. A sample is refluxed in strongly acid solution with a known excess of potassium

dichromate (K2Cr207). After digestion, the remaining unreduced K2Cr207 is titrated with ferrous

ammonium sulphate to determine the amount of K2Cr207 consumed and the oxidizable organic matter is calculated in terms of oxygen equivalent. Keep ratios of reagent weights, volumes, and strengths constant when sample volumes other than 50 mL are used. The standard 2-h reflux time may be reduced if it has been shown that a shorter period yields the same results.

Colorimetric reaction vessels are sealed glass ampoules or capped culture tubes, Oxygen consumed is measured against standards at 600 nm with a spectrophotometer.

b. Interferences and limitations: Volatile straight chain aliphatic compounds are not oxidized to any appreciable extent. This failure occurs partly because the volatile organics are present in the vapour space and do not come in contact with the oxidizing liquid. Straight chain aliphatic

compounds are oxidized more effectively with silver sulphate (Ag2S04) is added as a catalyst.

However Ag2S04 reacts with chloride, bromide, and iodide to produce precipitates that are oxidized only partially. The difficulties caused by the presence of the halides can be overcome

largely, though not completely, by complexing with mercuric sulphate (HgS04) before the

refluxing procedure. Although 1 g HgS04 is specified for 50 mL sample, a lesser amount may be used where sample chloride concentration is known to be less than 2000 mg/L, as long as a 10:1

ratio of HgS04:CI' is maintained. Do not use the test for samples containing more than 2000 mg Cl'/L. Techniques designed to measure COD in saline water are available.

Nitrite (N02 ) exerts a COD of 1.1 mg O^mg N02-N. Because concentrations of N02" in waters

rarely exceed 1 or 2 mg N02—N/L, the interference is considered insignificant and usually is

ignored. To eliminate a significant interference due to N02", add 10 mg sulfamic acid to the reflux vessel containing the distilled water blank.

Reduced inorganic species such as ferrous iron, sulphide, manganous manganese, etc. are oxidized quantitatively under the test conditions. For samples containing significant levels of these species, stoichiometric oxidation can be assumed from known initial concentration of the interfering species and corrections can be obtained from the COD value obtained.

153 Standard Methods Chemical Oxygen Demand S. Massen

Volatile organic compounds are more completely oxidized in the closed system because of longer contact with the oxidant. Before each use inspect culture tube caps for breaks in the TFE liner. Select culture tube size for the degree of sensitivity desired. Use the 25 x 150 mm tube for sample with low COD content because a larger volume sample can be treated.

Materials & Equipment: Apparatus:

• Digestion vessels: Preferably use borosilicate culture tubes, 16x100 mm, 20x150 mm, or 25 x 150 mm, with TFE-lined screw caps. Alternatively, use borosilicate ampoules, 10 mL capacity, 19 to 20 mm diameter.

• Heating Block: cast aluminum, 45 to 50 mm deep, with holes sized for close fit of culture tubes or ampoules.

Reagents:

• Digestion Solution: Add to about 500 mL distilled water 10.216 g K2Cr207, primary standard

grade, previously dried at 103°C for 2 hr, 167 mL conc. H2S04, and 33.3 g HgS04. Dissolve, cool to room temperature, and dilute to 1000 mL.

• Sulfuric Acid Reagent: Add Ag2S04, reagent or technical grade, crystals or powder, to conc.

H2S04 at a rate of 5.5 g Ag2S04/kg H2S04. Let stand 1to 2 days to dissolve Ag2S04.

• Sulfamic Acid: Required only if the interferences of nitrites is to be eliminated (see above).

• Potassium Hydrogen Phthalate (KHP) Standard: Lightly crush and then dry potassium hydrogen

phthalate (HOOCC6H4COOK) to constant weight at 120°C. Place in desiccator for 15 min before use. Dissolve 425 mg in distilled water and dilute to 1000 mL. KHP has a theoretical COD1 of

1.176 mg 02/mg and this solution has a theoretical COD of 500 mg O^L. This solution is stable when refrigerated for up to 3 months in the absence of visible biological growth.

Stock Chemicals for COD 5220-D

1 Milli-Q Water MW=18.01 g'mol" , H20, 18.2

H20 Mega'fi Resistance Mercury Sulfate Powder 7783-35-9 M63-500 HgSCU MW = 296.65 g'mol"1 Potassium Dichromate Orange Crystalline Powder 7778-50-9 1 P188-500 K2Cr207 MW = 294.18 g'mol Potassium Hydrogen Phthalate (KHP) Crystalline Powder (99.99%) 877-24-7 1 AC17712-1000 HOOCC6H4COOK MW = 204.2236 g'mol" Silver Sulfate Powder 10294-26-5 1 S190-100 Ag2S04 MW = 311.799 g-mol" Sodium Hydroxide, 10N Liquid SS255-4 1310-73-2 NaOH MW = 40.00 g'mol"1 (4L) Sulfuric Acid 7664-93-9 Liquid, 95-98% Conc. 3512964

154 Standard Methods Chemical Oxygen Demand S. Massen

Stock Chemicals for COD 5220-D

1 | H2SQ4 1 I MW = 98.075 g'mol | (4L, F-grade)

Procedure:

c. Treatment of samples: Wash culture tubes and caps with 20% H2S04 before first use to prevent contamination. Refer to Table 15 for proper sample and reagent volumes. Place sample in culture tube or ampoule and add digestion solution. Carefully run sulphuric acid reagent down inside of vessel so an acid layer is formed under the sample-digestion solution layer. Tightly cap tubes or seal ampoules, and invert several times to mix completely. Caution: Wear face shield and protect hands from heat produced when contents of vessel are mixed. Mix thoroughly before applying heat to prevent local heating of vessel bottom and possible explosive reaction.

Place tubes or ampoules in the block digester or oven preheated to 150°C and reflux for 2 hours. Cool to room temperature and place vessels in test tube rack.

TABLE 15: SAMPLE AND REAGENT QUANTITIES FOR VARIOUS DIGESTION VESSELS

Sulfuric Digestion Total Final Sample Acid Digestion Vessel Solution Volume (mL) Reagent (mL} (mL) (mL)

Culture 16x100 mm 2.5 1.5 3.5 7.5 tubes: 20x150 mm 5.0 3.0 7 15.0

25x150 mm 10.0 6.0 14 30.0

Standard 10 mL ampoules 2.5 1.5 3.5 7.5

Note: The University of Guelph - School of Engineering uses the HACH 16x100 mm round bottom glass tubes with screw tops. The glass tubes must be thoroughly washed after each use. The preferred cleaning method is:

i. Dump the spent COD sample into a glass waste jar labelled appropriately to identify the chemicals present.

ii. Fill each glass tube with distilled water. Shake it to suspend any precipitates. Quickly invert the test tube 180° so the meniscus/surface tension of the fluid prevents the fluid from pouring out. This will only work for 16x100 mm glass tubes with the screw tops. Allow the precipitates to settle to the bottom. Tilt the glass tube slightly to break the surface tension and allow the distilled water and precipitate to drain into the waste jar. Repeat until there are no visible sediments in the test tube.

155 Standard Methods Chemical Oxygen Demand S. Massen

iii. Submerge the glass tubes and caps in an acid bath consisting of 20% sulphuric acid and distilled water for at least 2 hours. Remove the materials from the acid bath with plastic tongs, and thoroughly rinse with distilled water. Invert the glass tubes in a test tube rack and allow them to air dry. Store the cleaned glass tubes in a dry, clean area away from chemicals or active lab equipment.

d. Measurement of dichromate reduction: Invert cooled samples, blank, and standards several times and allow solids to settle before measuring absorbance. Dislodge solids that adhere to container wall by gentle tapping and settling. Insert unopened tube or ampoule through access door into light path of spectrophotometer set at 600 nm. Read absorbance and compare to calibration curve. Use optically matched culture tubes or ampoules for greater sensitivity; discard scratched or blemished glassware.

e. Preparation of calibration curve: Prepare at least five standards from potassium hydrogen

phthalate solution with COD equivalents from 20 to 900 ng 02/L. Make up to volume with distilled water; use same reagent volumes, tube, or ampoule size, and digestion procedure as for samples. Prepare calibration curve for each new lot of tubes or ampoules or when standard prepared in 4a differ by > 5% from calibration curve.

Calculation:

COD as mg O2/L = mg 02 in final volume x 1000/ mL sample

Precision and Bias: Forty-eight synthetic samples containing potassium hydrogen phthalate and NaCI were tested by five laboratories. At an average COD of 193 mg Ojl in the absence of chloride, the standard

deviation was + 17 mg O2/L (coefficient of variation 8.7%). At an average COD of 212 mg 02/L and 100 mg Cl'/L, the standard deviation was ± 20 mg O2/L (coefficient of variation, 9.6%).

156 Standard Methods Chemical Oxygen Demand S. Massen

Laboratory Form for COD Tests Client: Sample Date: Analysis Date: Lab Tech:

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157 Standard Methods Biochemical Oxygen Demand Appendix C

5-Day BOD Test

Standard Methods 5210 B

From Standard Methods for the Examination of Water and Wastewater

Updated with Annotations:

Friday, June 18, 2010

General Discussion: The biochemical oxygen demand (BOD) determination is an empirical test in which standardized laboratory procedures are used to determine the relative oxygen requirements of wastewaters, effluents, and polluted waters. The test has its widest application in measuring waste loadings to treatment plants and in evaluating the BOD-removal efficiency of such treatment systems. The method consists of filling with diluted and seeded sample, to overflowing, an airtight bottle of specified size and incubating it at the specified temperature for 5 days. Dissolved oxygen (DO) is measured initially and after incubation, and the BOD is computed from the difference between the initial and final DO. Because the initial DO is determined shortly after the dilution is made, all oxygen uptake occurring after this measurement is included in the BOD measurement.

BOD = no inhibition of nitrogenous oxygen demand CBOD = inhibition of nitrogenous oxygen demand -Warm chilled samples to 20°C before analysis. -Ideally begin analysis within 6h of collection.

Materials & Equipment: Apparatus:

• Incubation Bottles: 250-300mL capacity, glass-stopper bottles with a flared mouth are preferred. Clean with detergent, rinse, drain. Use water seal & place foil cap over mouth of bottle to reduce evaporation of water seal during incubation;

• Air Incubator: Thermostatically controlled, maintained at 20 ± 1°C, excludes all light to prevent photosynthesis;

• Diffuser stone;

• Oxygen-sensitive membrane electrode: polarographic or galvanic, with appropriate meter; • Stir Plate;

• 'M' Parafilm. Reagents:

• Phosphate Buffer Solution: Dissolve the following in 500mL distilled water, and dilute to 1L:

1. 8.5g KH2P04

158 Standard Methods Biochemical Oxygen Demand Appendix C

2. 21.75g K2HP04

3. 33.4g Na2HP04-7H20

4.1.1% NH4CI

The pH should be 7.2 without further adjustment;

• Magnesium sulphate solution: Dissolve 22.5g MgS04-7H20 in distilled water and dilute to 1L;

• Calcium Chloride Solution: Dissolve 27.5g CaCI2 in distilled water and dilute to 1L;

• Ferric Chloride Solution: Dissolve 0.25g FeCI3-6H20 in distilled water and dilute to 1L;

• IN Acid Solution: For neutralization of caustic waste samples. Slowly, and while stirring, add 28mL conc. Sulphuric acid to distilled water. Dilute to 1L;

• IN alkali solution: For neutralization of acidic waste samples. Dissolve 40g sodium hydroxide in distilled water, and dilute to 1L;

• Sodium sulfite solution: Dissolve 1.575g Na2S03 in lOOOmL distilled water. This solution is not stable: prepare daily;

• Nitrification inhibitor, 2-chloro-6-(trichloromethyl) pyridine. Nitrification Inhibitor 2579-24 (2.2% TCMP), Hach Co. or equivalent;

• Glucose-glutamic acid solution: Dry reagent-grade glucose and reagent-grade glutamic acid at 103°C for lh. Add 150mg glucose and 150mg glutamic acid to distilled water and dilute to 1L. Prepare fresh immediately before use;

• Ammonium chloride solution: Dissolve 1.15g NH4CI in about 500mL distilled water, adjust pH to 7.2 with NaOH solution, and dilute to 1L. Solution contains 0.3mg N/mL;

• Source water for preparing BOD dilution water: Use demineralised, distilled tap, or natural water for making sample dilutions. Must be free of heavy metals, specifically copper, and toxic substances, such as chlorine, that can interfere with BOD measurements.

Stock Chemicals for BOD5 5210-B

Ammonium Chloride Powder 12125-02-9 1 A661-3 NH4CI MW = 53.49 g.mol" Calcium Chloride Anhydrous Powder 10043-52-4 1 C614-500 CaCI2 MW = 110.98 g.mol" Ferric Chloride Powder 10025-77-1 1 188-100 FeCI3*6H20 MW = 270.32 g.mol' Dextrose (D-Glucose) Anhydrous Powder 50-99-7 D16-1 C6H12O6 MW = 180.18 g.mol"1 Glutamic Acid Crystalline Powder 56-86-0 1 A125-100 H00CCH2CH(NH2)C00H MW = 147.13 g.mol" Potassium Phosphate Dibasic Anhydrous Powder 7758-11-4 P290-500 K2HPO4 MW = 174.18 g.mol*1 Potassium Phosphate Monobasic Powder, 99% Pure P285-3 7778-77-0 1 KH2P04 MW= 136.09 g.mol (3kg)

159 Standard Methods Biochemical Oxygen Demand Appendix C

Stock Chemicals for BOD5 5210-B

Magnesium Sulfate 10034-99-8 M63-500 MgS04«7H20 1 MW=18.01 gTTiol' , H20, Milli-Q Water 18.2 Mega'fl Resistance Powder, Nitrification Inhibitor 2533-34 HACH Formula 2533 Sodium Hydroxide, ION Liquid SS255-4 1310-73-2 NaOH MW = 40.00 g'mol"1 (4L) Sodium Phosphate Dibasic Heptahydrate Powder 7782-85-6 1 S-373 Na2HP04«7H20 MW = 268.07 g.mol" Sodium Sulfite Powder, for CI" neut. 7757-83-7 1 S430-500 Na2S03 MW = 126.04 g.mol" Sulfuric Acid Liquid, 95-98% Cone. 3512964 7664-93-9 1 H2SO4 MW = 98.075 g.mol" (4L, F-grade)

Procedure: 1. Sampling and Storage:

Samples for BOD analysis may degrade significantly during storage between collection and analysis, resulting in low BOD values.

(i) Grab samples: If analysis is begun within 2h of collection, cold storage is unnecessary. If analysis is not started within 2 hr of collection, keep the sample at or below 4°C from the time of collection. Begin analysis within 6 h of collection; when this is not possible because the sampling site is so distant from the laboratory, store at or below 4C and report the length and temperature of storage with the results. In no casestart analysis more than 24 h after grab sample collection. When samples are to be used for regulatory purposes make every effort to deliver sample for analysis within 6 h of collection.

(ii) Composite samples: Keep samples at or below 4C during compositing. Limit compositing period to 24 h. Use the same criteria for storage of grab samples, starting the measurement of holding time from end of compositing period. State storage time and conditions as part of the result.

2. Sample Pretreatment:

(i) pH adjustment: For highly acidic or alkaline samples: neutralize to pH 6.5-7.5 with solution of sulphuric acid or sodium hydroxide of such strength that the quantity of reagent does not dilute the sample by more than 0.5%. For the 300 mL jars, add no more than 1.5 mg/L of neutralizing agent.

(ii) Temperature: Bring samples to 20±3°C before making dilutions.

160 Standard Methods Biochemical Oxygen Demand Appendix C

(iii) Nitrification inhibition: Add 3 mg 2-chloro-6-(trichloro methyl) pyridine (TCMP) to each 300mL bottle before capping, or add sufficient amounts to the dilution water to make a final concentration of lOmg/L.

(iv) Chlorine residual: For samples that contain residual chlorine, allow exposure to light for 1-2

h (often occurs naturally during sample transport and handling), or add Na2S03 solution.

Determine required volume of Na2S03 solution on a 100 to 1000 mL protion of neutralized

sample by adding 10 mL 1 + acetic acidic or 1 + 50 H2S04, 10 mL potassium iodide (Kl)

solution (lOg/lOOmL) per 1000 mL sample and titrating with Na2S03 solution to the starch

iodine end point for residual. Add to neutralize sample the proportional volume of Na2S03 solution determined by the above test, mix, and after 10 to 20 min check sample for residual chlorine.

Note: Excess Na2S03 exerts an oxygen demand and reacts slowly with certain organic chloramine compounds that may be present in chlorinated samples.

(v) Toxic substances: Samples containing certain toxic metals often require special study and treatment.

3. Preparation of dilution water:

(i) Place desired volume of water in a suitable bottle and add lmL each of phosphate buffer,

MgS04, CaCI2 and FeCI3 solutions/L of water.

(ii) Seed dilution water (Guelph, ON, WWTP aeration basin 2), if desired.

(iii) Test and store dilution water.

(iv) Before use, bring dilution water temperature to 20°C.

(v) Saturate with DO by shaking in a partially filled bottle or by aerating with organic-free filtered air. Dissolved concentration should be above 7.5 mg O^L before using dilution water for BOD tests.

4. Dilution water check: (if nitrification inhibition is used)

(i) Store seeded dilution water in a darkened room at room temperature until oxygen uptake is at or below 0.2 mg/L.

(ii) Check stored dilution water to determine whether sufficient ammonia remains after storage. If not, add ammonium chloride solution to provide a total of 0.45mg ammonia/L as nitrogen.

Note: The UoG ammonium probe should register less than 14.5 mV.

(iii) If dilution water has not been stored for quality improvement, add sufficient seeding material to produce a DO uptake of 0.05-0.1 mg/L in 5 days at 20°C. The DO uptake in 5d at 204C should not be more than 0.2mg/L and preferably not more than 0.1 mg/L.

161 Standard Methods Biochemical Oxygen Demand Appendix C

5. Glucose-Glutamic Acid Check:

(i) Periodically check dilution water quality, seed effectiveness, and analytical technique by making BOD measurements on a mixture of 150mg glucose/L and 150mg glutamic acid/L as a standard check solution. Tests should be performed in triplicate.

(ii) Determine the 5d 20°C BOD of a 2% dilution {3 mg/L) of the glucose-glutamic acid (GGA) standard check solution (20 mL GGA solution/L seeded dilution water, or 6.0 mL/300-mL bottle). The resulting average BOD for the three bottles, after correction for dilution and seeding, must fall into the range of 198 ± 30.5 mg/L. If the average falls outside this range, evaluate the cause and make appropriate corrections. Consistently high values can indicate the use of too much seed suspension, contaminated dilution water, or the occurrence of nitrification; consistently low values can indicate poor seed quality or quantity or the presence of a toxic material. If low values persist, prepare a new mixture of GGA and check the sources of dilution water and source seed.

6. Seeding:

(i) Seed source: domestic wastewater, undisinfected effluents from biological waste treatment plants, activated sludge, etc. Generally, 1 to 3 mg/L of settled raw wastewater or primary effluent or 1:10 dilution of mixed liquor/300mL bottle will provide a suitable amount of microorganisms. Do not filter seed suspension before use. Agitate the seed suspension during transfer. Do not add seed directly to wastewater samples if they contain materials (or a pH) that are toxic before dilution. Always record the exact volume of seed suspension added to each bottle.

Note: The University of Guelph - School of Engineering obtains their seed from the Gueiph Wastewater Treatment Plant Aeration Basin 2. A sample line from this basin in located in Prof. Zhou's membrane sample shed. The sample line must be flushed for at least 5 minutes before obtaining a sufficient volume of the seed for BOD analysis.

Allow the seed to settle in the collection container for 30 minutes or until there are visible strata. The supernatant (which is relatively clear) is used as the seed water. 3L of stock seed volume generally yields 1 L of seed water from this location. Carefully decant the supernatant into a separate vessel. Do not draw in materials from the settle portion of the seed volume to avoid excess nutrient introduction. Pipette the desired portion directly from the stock seed volume to the BOD sample jars. University of Guelph generally uses 2 mL of seed per 300 mL BOD bottle.

(ii) Seed control: Determine seed DO uptake. Make dilutions of seed such that the largest quantity results in at least 50% DO depletion. A plot of DO depletion, in mg/L, vs. mL seed should present a straight line for which the slope indicates DO depletion per mL of seed. The DO-axis intercept is oxygen depletion caused by the dilution water, and should be less than 0.1 mg/L. To determine a sample DO uptake subtract seed DO uptake from total DO uptake. The DO uptake of seeded dilution water should be between 0.6 and 1.0 mg/L, but the amount of seed added should be adjusted from this range to that required to provide the GGA check results of 198 ± 30.5mg/L. For example, if 1 mL of seed suspension is required to achieve 198 ± 30.5mg/L in the GGA check, then use 1 mL in each BOD bottle receiving the test wastewater.

162 Standard Methods Biochemical Oxygen Demand Appendix C

7. Dilution technique:

(i) Make several dilutions of prepared sample to obtain residual DO of at least 1mg/L, and DO uptake of at least 2 mg/L after 5d incubation.

(ii) An analysis such as COD may be correlated approx. with BOD & serve as a guide in selecting dilutions.

(iii) In the absence of prior knowledge, use the following dilutions: 0.01 to 1.0% for strong industrial wastes; 1to 5% for raw and settled wastewater; 5 to 25% for biologically treated effluent; 25-100% for polluted river waters.

(iv) Using a wide-tip volumetric pipette, add the desired sample volume to individual BOD bottles of known capacity.

(v) Fill BOD bottles at least two-thirds full of dilution water without entraining air.

(vi) Add appropriate amounts of seed suspension and nitrification inhibitor (if not added directly to the stock dilution water) to the individual BOD bottles or to the dilution water. When a bottle contains more than 67% of the sample after dilution, nutrients may be limited in the diluted sample and subsequently reduce biological activity. In such samples, add the nutrient, mineral, and buffer solutions directly to the diluted sample at a rate of 1 mL/L (0.30 mL/300-mL bottle) or use commercially prepared solutions designed to dose the appropriate bottle size.

(vii) Dilute to final level with dilution water so that insertion of stopper will displace all air, leaving no bubbles.

(viii) For dilutions greater than 1:100, make a primary dilution in a graduated cylinder before making final dilution in the bottle.

(ix) If the membrane electrode method is used for DO measurement, prepare only one BOD bottle for each dilution.

(x) Determine initial DO on this bottle and replace any displaced contents with dilution water to fill the bottle within 30 min of preparing the dilution.

(xi) Stopper tightly, and water-seal with Mill-Q or dilution water. Cover stopper and flared mouth of bottle with one square of 'M' Paraflim to reduce evaporation of water seal, and incubate for 5 days ± 6 h at 20°C.

(xii) Rinse DO electrode will Milli-Q between determinations to prevent cross-contamination of samples.

8. Determination of initial DO:

Use the membrane electrode method (Section 4500-0.G) to determine initial DO on all sample dilutions, dilution water blanks, and where appropriate, seed controls:

163 Standard Methods Biochemical Oxygen Demand Appendix C

(i) Effect of temperature on electrode sensitivity, (jiA/mg/L):

Log = 0.43 mt + b

Where:

t temp, °C

m constant that depends on membrane material

b constant that depends on membrane thickness

Determine values of <|> and m at one temperature (0 and t0), then calculate the sensitivity at any desired temperature as follows:

Log = log 4»o + 0.43 m (t-t0)

Construct calibration curve: temperature on x-axis versus electrode sensitivity on y-axis, should be a straight line passing through y-axis above zero.

(ii) Effect of ionic strength on electrode sensitivity:

Log 0

4>s = sensitivity in salt solution

4>o = sensitivity in distilled water

Cs = salt concentration (preferably ionic strength)

ms = salting-out coefficient

Calculate sensitivity for any value of Cs once (j>0 and ms are determined. Construct calibration curves at different temperatures by plotting concentration on x-axis (in M or mg/L) versus electrode sensitivity.

(iii) Calibration: Calibrate membrane electrodes by reading against air or a sample of known DO concentration (determined by iodometric method), as well as in a sample with zero DO (e.g.

Add excess sodium sulfite, Na2S03, and a trace of cobalt chloride (CoCI2) to distilled water to bring DO to zero).

Note: Whenever a DO probe is used, it may be necessary to take the readings several times in succession until a final consistent value is obtained. This is especially important if the preliminary DO reading is below 2 mg/L Regardless, it is advisable to take measurements at least twice, more if there is a discrepancy between the initial readings.

9. Dilution water blank:

(i) Use a dilution water blank as a rough check on quality of unseeded dilution water and cleanliness of incubation bottles.

164 Standard Methods Biochemical Oxygen Demand Appendix C

(ii) Together with each batch of samples incubate a bottle of unseeded dilution water.

(iii) Determine initial and final DO. The DO uptake should not be more than 0.2 mg/L and preferably not more than 0.1 mg/L.

10. Incubation:

(i) Incubate at 20 ± 1°C BOD bottles containing desired dilutions, seed controls, dilution water blanks, and glucose-glutamic acid checks. Water-seal bottles.

11. Determination of final DO:

(i) After 5 d incubation, determine DO in sample dilutions, blanks and checks. Only bottles, including seed controls, giving a minimum DO depletion of 2.0 mg/L and a residual DO of at least 1.0 mg/L after 5 d of incubation are considered to produce valid data, because at least 2.0 mg 02/uptake L is required to give a meaningful measurement of oxygen uptake, and at least 1.0 mg/L must remain throughout the test to ensure that insufficient DO does not affect the rate of oxidation of waste constituents. Exceptions occur for reporting purposes only when the depletions for tests using undiluted samples in all bottles fall below 2.0 mg/L and when the residual DO in all dilutions is less than 1.0 mg/L.

Calculations:

(i) When dilution water is not seeded:

/ D\-D2 BODs, mg/L =

(ii) When dilution water is seeded:

where: Dj = DO of diluted sample immediately after preparation, mg/L

D2 = DO of diluted sample after 5d incubation at 20°C, mg/L P = decimal volumetric fraction of sample used 8j = DO of seed control before incubation, mg/L

B2 = DO of seed control after incubation, mg/L / = ratio of seed in diluted sample to seed in seed control = (% seed in diluted sample)/(% seed in seed control) If seed material is added directly to sample or to seed control bottles:

/ = (volume of seed in diluted sample)/(volume of seed in seed control) Report results as CBOD if nitrification is inhibited.

When all dilutions result in a residual DO < 1.0, selct the bottle having the lowest DO concentration (greatest dilution) and report:

{Dl B BOD5, mg/L> ~ 2)/

165 Standard Methods Biochemical Oxygen Demand Appendix C

Precision and Bias:

There is no measurement for establishing bias of the BOD procedure. The glucose-glutamic acid check prescribed is intended to be a reference point for evaluation of dilution water quality, seed effectiveness, and analytical technique. Single laboratory tests using a 300-mg/L mixed glucose-glutamic acid solution provided the following results:

Number of months: 14 Number of triplicates: 421 Average monthly recovery: 204 mg/L Average monthly standard deviation: 10.4 mg/L

In a series of interlaboratory studies conducted by the US EPA in 1986, each involving 2 to 112 laboratories (and as many analysts and seed sources), 5-d BOD measurements were made on synthetic water samples containing a 1:1 mixture of glucose and glutamic acid in the total concentration range of 3.3 to 231 mg/L. The regression equations for mean value X, and standard deviation S from these studies were:

X = 0.658 (added concentration, mg/L) + 0.280 mg/L S = 0.100 (added concentration, mg/L) + 0.547 mg/L

For the 300 mg/L mixed primary standard, the average 5d BOD would be 198 mg/L with a standard deviation of 30.5 mg/L. When nitrification inhibitors are used, GGA test results falling outside the 198 ±30.5 control limit quite often indicate use of incorrect amounts of seed. Adjust amount of seed added to the GGA test to achieve results falling within this range.

166 0) 3 | BOD Bottle Volume: 300 mL Y/Z 300/fY/Z)j Start: / / End: 02 Consumption 1 BOD Q_ o> CL fD rt 3" B O a.on

Blank A 0 n/a n/a n/a B 0 n/a n/a n/a Seed A n/a n/a n/a B n/a n/a n/a Average seed

ro 1 1A 4 oo* Sample ID: 1B 4 3* 3 4/22/2009 @ 7:45AM 3C 4 3 a. 4 4A 4 Sample ID: 4B 4 Sample Time: _/__/_ @ : 4C 4 s 5A 4 Sample ID: 5B 4 Sample Time: __/_/_ @ 5C 4

6 6A 4 Sample ID: 6B 4 > Sample Time: _/__/_ @ 6C 4 T5 "D 3 Q. X* O Standard Methods Total Suspended and Volatile Solids Appendix C

Total Suspended and Volatile Solids

Standard Methods 2540 D

From Standard Methods for the Examination of Water and Wastewater

Updated with Annotation

Tuesday, May 26, 2009

General Discussion:

a. Principle: A well mixed sample is filtered through a weighted standard glass-fiber filter and the residue retained on the flter is dried to a constant weight at 103 to 105C. The increase in the weight on the filter represents the total suspended solids. If the suspended material clogs the filter and prolongs filtration, it may be necessary to increase the diameter of the filter or decrease the sample volume. To obtain an estimate of the total suspended solid, calculate the difference between total dissolved solids and total solids.

b. Interferences: Exclude large floating particles or submerged agglomerates of nonhomogeneous materials from the sample if it is determined that their inclusion is not representative. Because excessive residue on the filter may form a water-entrapping crust, limit the sample size to that yielding no more than 200 mg residue. For samples high in dissolved solids, thoroughly wash the filter to ensure removal of dissolved material. Prolonged filtration times often resulting from filter clogging may produce high results owing to increased colloidal materials captured on the clogged filter. Highly mineralized water with a significant concentration of calcium, magnesium, chloride, and/or sulphate may be hygroscopic and require prolonged drying, proper desiccation, and rapid weighing. Materials & Equipment:

• Drying Oven: for operation at 103 to 105°C;

• Muffle Furnace: for operation at 550°C70 mm 0 glass microfibre filter disk (Whatman® 934- AH™, Cat. No. 1827 070);

• Vacuum pump;

• Analytical Balance, capable of weighing 0.1 mg;

• Evaporating dishes: Aluminum or stainless steel planchet or Gooch crucible;

• Erlenmeyer Flask;

• Porcelain funnel, 90 mm diameter;

• Tweezers. Procedure:

1. Obtain the combined mass of a glass fiber filter disk placed on an aluminum or stainless steel planchet with a scale accurate to at least 0.1 mg.

168 Standard Methods Total Suspended and Volatile Solids Appendix C

2. Insert the filter disk with wrinkled side up in filtration apparatus. Apply vacuum and wash disk with three successive 10 mL portions of distilled water. Continue suction to remove all traces of water.

3. Pipette a recorded volume of sample onto the cleaned filter disk. Continue suction to remove all traces of water, and discard washings. Remove filter from filtration apparatus and transfer to an aluminum or stainless steel planchet as a support. Alternatively, remove crucible and filter combination if a Gooch crucible is used.

4. Dry in an oven at 103 to 105°C for at least 1hr.

5. Cool in desiccators to balance temperature and weigh. Repeat cycle of drying or igniting, cooling, desiccating, and weighing until a constant weight is obtained, or until weight loss is less than 0.5 mg between successive weighing. Store in desiccators until needed. Weigh immediately before use.

6. If volatile solids are to be measured, ignite at 550 ± 50°C for 15 min in a muffle furnace, and repeat step 5.

Calculation: TSS m^ - B) x 1000 L sample volume, mL where: A = weight of filter + dried residue, mg and B = weight of filter, mg.

Precision and Bias:

The standard deviation was 5.2 mg/L (coefficient of variation 33%) at 15 mg/L, 24 mg/ (10%)at 242 mg/L, and 13 mg/L (0.76%) at 1707 mg/L in studies by two analysts of four sets of 10 determinations each.

Single-laboratory duplicates analyses of 50 samples of water and wastewater were made with a standard deviation of differences of 2.8 mg/L.

169 Standard Methods Total Suspended and Volatile Solids Appendix C

Laboratory Form for Total Suspended and Volatile Solids Client: Sample Date: Analysis Date: Lab Tech:

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170 Standard Methods Total Phosphorous Appendix C

Total Phosphorus

Standard Methods 4500-P E

From Standard Methods for the Examination of Water and Wastewater

Updated with Annotations

Tuesday, June 17, 2010

General Discussion:

Total phosphorus is commonly performed for wastewater, and can range from 4-12 mg/l in municipal sources (Metcalfe & Eddy, 2003). This method is based on Standard Methods procedure 4500-P, (APHA, 2007), and makes use of persulfate digestion and ascorbic colorimetric methods. Precision and bias: using this method, an error of 5-10% is typical for solutions of 0.1 - 7.0 mg/l P. The minimum detectable concentration is 10 ng/l P.

Standard curves using the University of Guelph equipment produced most reproducible values when the standard concentrations were maintained below 2.0 mg/L. When previous data on a sample were available, dilutions with Mill-Q water were made accordingly.

The first portion of this standard operating procedure (SOP) describes the method to be used to analyze for each analyte in SMP, and is followed by lab data sheets to be pasted into the researchers lab book for both calibration curves and regular analyses.

Materials & Equipment:

Apparatus:

• Autoclave;

• Spectrophotometer: With infrared phototube for use at 880nm;

• 3mL Quartz Cuvette;

• Eye Dropper;

• 100 mL Burette;

• Acid washed glassware: Use acid-washed glassware for determining low concentrations of phosphorus. Phosphate contamination is common because of its absorption on glass surfaces. Avoid using commercial detergents containing phosphate. Clean glassware with hot dilute HCI and rinse well with distilled water. Preferably, reserve the glassware only for phosphate determination, and after use, wash and keep filled with water until needed. If this is done, acid treatment is required only occasionally.

171 Standard Methods Total Phosphorous Appendix C

Reagents:

• Phenolphthalein Indicator;

• 5N H2S04: 70mL concentrated sulfuric acid dissolved to 500mL with distilled water;

• H2S04 solution (10.8N): Add 300mL concentrated H2S04to ~600mL distilled water and dilute to 1L with distilled water;

• Solid Potassium Persulfate (K2S208);

• IN NaOH: Dilute stock ION NaOH;

• Potassium Antimonyl Tartrate Solution: Dissolve 1.3715g K(Sb0)C4H406-l/2H20 in 400mL distilled water in a 500mL volumetric flask & dilute to volume. Store in a glass-stoppered bottle at 4°C. Alternatively, stock solutions can be ordered through Fisher Scientific.

• Ammonium Molybdate Solution: Dissolve 20g (NH4)6Mo7024-4H20 in SOOmL distilled water. Store in a glass-stoppered bottle.

• Ascorbic Acid: O.IM: Dissolve 1.76 g ascorbic acid in lOOmL distilled water. The solution is stable for about 1week at 4°C.

• Combined Reagent: Mix the above reagents in the order provided using the following proportions for lOOmL of the combined reagent:

1. 50mL5NH2S04

2. 5ml potassium antimonyl tartrate solution

3. 15mL ammonium molybdate solution

4. 30mL ascorbic acid solution

Let all reagents reach room temperature before they are mixed. Mix after addition of each reagent. If turbidity forms in the combined reagent, shake and let stand for a few minutes until turbidity disappears before proceeding. The reagent should be light yellow in appearance, and is stable for 4h;

• Blank Reagent: Start with 50 mL of 5N H2S04, add 15 mL of ammonium molybdate solution. Ensure solutions are at room temperature before addition. This solution will be added to the extract and used as a blank to compensate for any turbidity in the fitrate;

• Stock Phosphate Solution: Dissolve in distilled water 219.5mg anhydrous KH2P04 and dilute to 3_ lOOOmL; lmL = 50 |ig P04 P;

• Standard Phosphate Solution: Dilute 50mL stock phosphate solution to lOOOmL with distilled water; lmL = 2.5^g P;

172 Standard Methods Total Phosphorous Appendix C

Stock Chemicals for Total Phosphorous 4500-PE

Ammonium Molybdate (VI) Tetrahydrate Crystalline powder 12054-85-2 1 A674-500 (NH)4Mo7024*4H20 MW = 1235.86 g.mol" Antimony Potassium Tartrate Liquid 11071-15-1 1 5872-32 K2[Sb2(C4H406)2]-3H20 MW = 667.86 g.mol Ascorbic Acid White Crystalline Powder 50-81-7 1 A62-500 c6h8o6 MW = 176.13 g.mol" Phenolphthalein Indicator Liquid 5600-16 7732-18-5 1 C2oHI404 MW = 318.33 g.mol" (500 ml) Potassium Persulfate Powder 7727-21-1 1 P282-500 k2s2o8 MW = 270.32 g.mol" Potassium Phosphate Monobasic Powder, 99% Pure P285-3 7778-77-0 1 kh2po4 MW = 136.09 g.mol" (3kg) 1 MW=18.01 g.mol" , H20, Milli-Q Water 18.2 Mega 'fi Resistance Sulfuric Acid Liquid, 95-98% Cone. 3512964 7664-93-9 1 h2so4 MW = 98.075 g.mol" (4L, F-grade) Sodium Hydroxide, ION Liquid SS255-4 1310-73-2 NaOH MW = 40.00 g.mol"1 (4L)

Persulfate Digestion Method

Note: This procedure is required for all wastewater samples. Standards and blanks must be carried through the digestion procedure as well.

1. Add 1 drop phenolphthalein indicator to ~50mL sample.

2. If a red colour develops, add 10.8N H2S04 dropwise to just discharge colour.

3. Add lmL of 10.8N H2S04 solution.

4. Add 0.5g solid K2S208

5. Heat for 30min. in autoclave at 121°C (98-137 kPa).

6. Cool, add 1 drop indicator. Using a burette, titrate to a faint pink colour with IN NaOH. Approximately 15 mL of IN NaOH are required depending on the pH.

7. Make up to lOOmL with Milli-Q water by adding the volume calculated from the following:

Volume Milli-Q = (100ml total) - (50 ml initial) - (1ml H2S04) - (burette volume)

= 49 ml - burette volume

173 Standard Methods Total Phosphorous Appendix C

Note: This dilution step has been elaborated upon from the original Standard Method to increase overall precision.

8. Split the sample into two 50 mL volumes. One will be used as a turbidity correction with blank reagent, the second will be used for with the combined reagent for total phosphorous determination.

9. Determine absorbance using ascorbic acid method.

Ascorbic Acid Method 1. Pipet 50mL sample into a 125mL Erlenmeyer flask.

2. Add 0.05mL (1 drop) phenolphthalein indicator.

3. If a pink colour develops add 5N H2S04 drop-wise to just discharge the colour.

4. Add 8mL combined reagent and mix thoroughly.

5. After at least 10 min but no more than 30 min, measure absorbance of each sample at 880 nm, using reagent blanks as the reference solution.

6. Correction for turbidity or interfering colour: Natural colour of water generally does not interfere at the high wavelength used. For highly coloured or turbid waters, prepare a blank by adding all reagents except ascorbic acid and potassium antimonyl tartrate to the sample. Subtract blank absorbance from absorbance of each sample.

7. Preparation of calibration curve: Prepare individual calibration curves from a series of 6 standards within the phosphate ranges indicated below:

Approximate P range Light Path, (mg/L) (cm) 0.30 - 2.0 0.5 0.15 -1.30 1.0 0.01 - 0.25 5.0

Use a distilled water blank with the combined reagent to make photometric readings for the calibration curve. Plot the absorbance versus phosphate concentration to give a straight line passing through the origin. Test at least one phosphate standard with each set of samples.

Calculation:

mg P/L = me P (in approx. 58mL final volume) x 1000

mL sample

174 Standard Methods Total Phosphorous Appendix C

Laboratory Forms for Performing Standard Curves

Client: Sample Date: Analysis Date: Lab Tech:

1 Blank 1 0 50 2 2 10 so 3 3 20 50 4 4 30 50 5 5 7 B 9 10 Volume of Milli-Q to add = 49 mL - Volume of NaOH added

(0.2195)

1 Blank IP 2 3 t 5 6 7

B - 9 10 Volume of Milli-Q to add = 49 mL - Volume of NaOH added

175 Standard Methods Total Phosphorous Appendix C

References:

American Public Health Association. Standard Methods for the Examination of Water and Wastewater. 2007. 21st Edition. Edited by Andrew D. Eaton, Lenore S. Clesceri, Eugene W. Rice and Arnold E. Greenberg. Washington, D. C:, 2007.

Metcalf, Eddy. Wastewater Engineering: Treatment and Reuse: Fourth Edition. New York: McGraw Hill, 2003.

176 Anaerobic Digestion Study Acclimatization Period Appendix D

| Appendix D

Supplementary Bench-Scale Anaerobic Digester Performance Data

177 Anaerobic Digestion Study Acclimatization Period Appendix D

Appendix D Introduction

This Appendix is supplementary data for Chapter 4: Added Value from Rebate Beer in Anaerobic Digestion. It details period of acclimatization period before steady state analysis, and provides insight into how long a digester would take to stabilize once the feeding regiment is changed to incorporate rebate beer.

Anaerobic Digestion Supplementary Results and Discussion

The 50% DM/50% RB blend was run between March and July 2009, and again between October and January 2011 (Figure 20). The reason the 50%RB/50%DM trial was run twice is that a consistent and reliable supply of dairy manure was not secured for the first attempt. The varying manure sources were speculated to have contributed to the highly variable solids destruction noted in the results. This variable was removed for the second attempt at this trial. Digester Operation: 50% Dairy Manure, 50% Rebate Beer

• pH —•—Biogas Production (L/day) —it—Temperature (C)

Figure 20: Digester operation on trial 1 of the 50% DM/50% RB feedstock

The first trial of the 50% DM/50% RB used 250mL of feedstock diluted with 250mL of tap water, and produced an average of 12.2 L/day biogas. Steady state was never achieved. The amount of biogas generated varied greatly from day to day. The digestate pH dropped sharply over the course of a week early in May to from 7.2 to 5.5 following a large increase in biogas generation. This indicates system instability.

178 Anaerobic Digestion Study Acclimatization Period Appendix D

Digester Operation: 50% Dairy Manure, 50% Rebate Beer

Biogas Production (L/day) Temperature (C)

Date

Figure 21: Digester operation for trial 2 of the 50%RB/50%DM feedstock

The second attempt at the 50%RB/50%DM trial used 220mL of feedstock and 280ml of tap water. It produced an average of 13.5 l-Biogas/day. Steady state was achieved fairly quickly, however the solids destruction never leveled off. This was attributed to the high biological activity of the feedstock, despite it being stored in the fridge. The two zero points result from when the feed port was incidentally left open over night, allowing the generated biogas to escape to atmosphere.

The 75% DM/25% RB blend was run from July 2009 to June 2010 (Figure 28). The duration of this trial was extended because of outstanding issues with the temperature controller.

179 Anaerobic Digestion Study Acclimatization Period Appendix D

Solids Destruction: Solids Destruction: 50% Da iry Manure, 50% Rebate Beer 50% Dairy Manure, 50% Rebate Beer (8««d on Theoretical Dilution factor) • • J *000 -V- *• w • #*4 •fl» • 20000 11 »\ • V ... If vn. • «#• * * <> % \ \ "% % % % % \ \ \ % \ \ % X DM* VA y E Figure 22: Solids analysis for the 50% DM/50% RB Figure 23: Solids destruction for the 50% DM/50% RB

Solids Destruction: Digester Solids Analysis: 75% Dairy Manure, 25% Rebate Beer 75% Dairy Manure, 25% Rebate Beer _ 70% 60000 3 = ^ 50000 • • •• « | % 60% E. 40000 ••••• ••• •» • i ? * » • | 30000 • „ ?»- , ; „ ¥ —Influent TS I# 50", * •• S 20000 • »* ,* -J*». • » c | 40% . S 10000 * * -Influent VS ft) si • « V 30% -VS S " - Effluent TS a.

Date Figure 24: Solids analysis for the 75% DM/25% RB Figure 25: Solids destruction for the 75% DM/25% RB

Digester Solids Analysis: Solids Destruction: 100% Dairy Manure 100% Dairy Manure

45000 50% 5 40000 » * } .35000 " • » •** . "O ?e 45% £ 30000 * J • * • •- • • * - I * 40% S 25000 • * • * • II " 20000 ^ • ••• «* • Influent \ 2* 35% A # cV 3X 1 15000 • Effluent N £ « 30% • T5 | IOOOO : 4 2 25% • VS 3 5000 * Influent 1 0 • Effluent" 20% v.. '<-/• V "V

Date

Figure 26: Solids analysis for the 100% DM Figure 27: Solids destruction for the 100% DM

180 Reference to Figure 24 and Figure 26 illustrates little variation difference in influent TS and VS concentrations over time. The difference in TS and VS destruction during anaerobic digestion between the 75%DM/25%RB and 100%DM blends is 23±7% and 17±7% respectively. This difference is significantly greater than the suspected solids destruction during storage. It may then be said with confidence that dairy manure mixed with 25% rebate beer exhibits better solids destruction during mesophilic anaerobic digestion than dairy manure alone.

Digester Operation: 75% Dairy Manure, 25% Rebate Beer

• pH -•—Biogas Production (L/day) —^--Temperature (C)

40

35 ... 30 a> •o 25 3 C 20 OBIS 5 15 10 5 0 % \ \ \ Date

Figure 28: Digester Performance on the 75% DM/25% RB blend

The 75% DM/25% RB (Figure 28) produced an average of 8.3 L/day biogas. The bearings for the mixing shaft were replaced September 18th, which solved the leaking problem that began in early September. Steady state was achieved with this blend. The system pH never dropped below 7.0 throughout the entire trial.

The 100% DM baseline was run from June to September 2010 (Figure 29). Steady state operation was identified when, in order of importance, the effluent pH, VFA/Bicarbonate, solids destruction, temperature, and gas generation were consistent for 2 weeks or greater. Gas generation should ideally be consistent, although it is sometimes difficult to delineate between biogas leakage and digester failure. The gas volumes dropped to zero when leaks formed, however biogas is still being generated. The pH must be maintained between 7.0 and 8.0, and should only fluctuate by ±0.1 during steady state. The VFA/Bicarbonate ratio must be below 0.3. The temperature must be maintained in the mesophilic range. The volatile solids are substrate on which the anaerobes feed, and

181 Digester Operation: 100% Dairy Manure

Biogas (L/day) —A—Temperature (C)

Figure 29: Digester Performance on the 100% DM blend

The 100% DM produced an average of 7.0 L/day biogas. Steady state at mesophilic temperatures was established at the end of July. The pH never dropped below 6.9.

From Figures 1 to 3, it is evident that the higher the ratio of beer to dairy manure yeilded the greatest average amount of biogas per day. However, biogas generation for the 100% dairy manure was the most consistent of the three blends trialed, indicating the greatest system stability.

Figure 30 to Figure 32 represents the total volatile fatty acid concentrations (TVFA) and bicarbonate alkalinity that were measured in this study.

182 Effluent VFA and Bicarbonate: 50% Dairy Manure, 50% Rebate Beer 100 f 80 So £. 60 J 40 (0 £ 20 • VFA I 0 N Bicarbonate O u <9. % %

Date

Figure 30: Effluent VFA and Bicarbonate Concentrations of the 2nd 50% DM/50% RB trial

Effluent VFA and Bicarbonate: 75% Dairy Manure, 25% Rebate Beer 200 mJ Z 150 So E c 100 '

Date

Figure 31: Effluent VFA and Bicarbonate Concentrations of 75% DM/25% RB trial

183 Effluent VFA and Bicarbonate: 100% Dairy Manure

-•—Bicarbonate

Date

Figure 32: Effluent VFA and Bicarbonate Concentrations of 100% DM trial

184 50%DTV1 / 50%RB Effluent Technician's Notes: MlXft! Mixer Mixer Mi te Mixer New Feed Slopped Stopped Stopped Stopped Stopped Parameter 14-Oct 15-Oct Ib-Uct 1 < -Oct 18-Oct 19-Oct 20-0ct 21 -Oct 22-Oct 23-Oct 24-Oct 25-Oct 26-Oct 27-Oct 28-Oct 29-Oct 30-0ct 31-Oct 01-Nov 02-Nov 03-Mov General Sample Time hour 14:00 17:45 16:45 19:20 19:10 15:00 10:40 10:22 12:50 14:18 18:40 17:25 15:45 16:45 14:05 18:46 15:47 21:00 13:20 16:10 12:07 Elapsed Time days 1.17 1.16 0.96 1.11 0.99 0.83 0.82 0.99 1.10 1.06 1.18 0.95 0.93 1.04 0.89 1.20 0.88 1.22 0.68 1.12 0.83 HMixer Speed RPM 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 ITemperature *F 95 95.36 95 95 95 95 96.8 95.9 95 95 95 94.1 95.9 95 94.1 95 95 93.2 95 95 95 : *C 35.0 35.2 35.0 35.0 35.0 35.0 36.0 35.5 35.0 35.0 35.0 34.5 35.5 35.0 34.5 35.0 35.0 34.0 35.0 35.0 35.0 Tinntrwi1 tjlfltfly DUU16IRi RA 64 14 54 I 39 73 63 65 DO 59 59 57 60 56 61 63 60 53 63 Gas Production mL 7800 14600 12600 13000 13200 12200 12200 12800 12800 11800 2800 11800 11400 12000 11200 12200 12600 12000 10800 10600 12600 I 7.8 14.6 12.6 13 13.2 12.2 12.2 12.8 12.8 11.8 2.8 11.8 11.4 12 11.2 12.2 12.6 12 10.8 10.6 12.6 Normalized gas production L/dav 6.646154 12.62703 13.14783: 11.73668 13.29231 14.76303 14.88814 12.96203 11.60705 11.12042 2.366978 12.44635 12.25075 11.52 12.6 10.20602 14.38658 9.857387 15.86939 9.480745 15.15789 Cumulative Gas production L 7 19 44 57 72 87 100 112 123 125 138 150 161 174 184 199 208 224 234 249 Digestate Height cm ml aMjaagggillz 35.6 mumi am : * *S' * #»»& mmm. mw"" 8iogas Yield ^9vs 0.283137 0.537933 0.56012 0.500002 0.566275 0.62893 0.63426 0.552204 0.49448 0.473749 qioSilj 0.530321 0.521902 0.490771 0,536781 0.434879 0.612976 0.645752

rog«N/L 81.72637 81.2771 mg-N/L 2.098296 3.012285 75 0.037062

mg/t 17074 12412 IOrganic Loading Oxygen Demand m rog/L .-ta-.i ISCOD (SSx25-50 dilution) mai ggrr 5rev; n' -T k mg/L 1 'I T I m9fl- mg/L m9/L mg/L mg/L |Total Phosphorus mg/L

Influent This data is for undiluted F««d

Water Volume mL 320 i 3201 320| 3201 3201 320 I 3201 320 320 i 3201 320 I 320; 320 320 320 I 320 320: 320 I 3201 320 320 Feed Volume ml 180 180 180 180 180 180 180 180 180 160] 180 180 160| 180 180 r~ 1601 1801 180 L 180| 180 180 Ideal Feed Volume mL g|||||g|g gjjggpsfjd mi jgSftgaMlm mm%wm 1™W*W8m 2» Dairy Manure 75% 75% I 75% I 75% I 75% 75% 75% 75%| 75% I 75% 75% s 75% 75% 75% 75%; 75% 75% 75% 75% I 75%l L Beer 25% I 25% I 25%| 25% I 25% I 25% I 25% 25% 25%]I 25% 25% I 25%| 25%| 25% 25% II 25% 25% 1 25% 25% 25% 25% Hydraulic Retention Time days WSISMmM Mm 23.08965 jHHg] •Md |§^?g&gftI i •mEH ii psisii mam isiiiii §nv mmwm mmm sn*i nasi ®§ssjsMm mmwM iiiisig mmm •«* » * - > s •"r-rv®** .r - -«w.fxubn ^'uasgmt

KSZ2 ~BE2Zg ~ 'If | ' .VKSII ' i:?;i wmmmi «S"5X 7 ' ?I1 •y>'-y3rj " : sn |4§^|ll|

** _ - -- — — — ~ ^ " ffflBli SI3I11 iPlllii!PiSII a. J- I - '-*£ •« ' V - ; Stlllit iiSSlii wini iiikipS iniH slllSpt ii@i£n litlSilH siiik llilljii® Siiii 13SS ilSiS SSS S31I1IllSfi .'z*4J»» v - *;* «*>* *** if 4 -tJZlSfr*" J" * *

185

50%DM / 50%RB

Technician's Notes:

Sample Time hour 20:25 17:47 18:10 15:00 21:10 16:37 18:00 0:00 18:40 17:20 16:50 11:49 17:00 16:20 22:05 9:00 20:05 20:00 18:30 20:00 21:30 Elapsed Time days 1.14 0.89 1.02 0.87 1.26 0.81 1.06 1.25 0.78 0.94 0.98 0.79 1.22 0.97 1.24 0.45 1.46 1.00 0.94 1.06 1.06 Mixer Speed RPM 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 56 Temperature •F 93.2 93.2 95 93.2 95 95 95 94.1 94.1 94.1 94.1 93.2 93.2 92.3 95 95 95 93.2 94.1 95 95 •e 1 34.0 34.0 35.0 34.0 35.0 35.0 35.0 34.5 34.5 34.5 34.5 34.0 34.0 33.5 35.0 35.0 35.0 34.0 34.5 35.0 35.0 Tipping Bucket j 78 72 73 68 75 69 73 75 66 73 69 64 70 73 72 59 77 68 70 71 0 |Gas Production mL ] 15600 14400 14600 13600 15000 13800 14600 15000 13200 14600 13800 12800 14000 14600 14400 11800 15400 13600 14000 11949.3 0 1 L 1 15.6 14.4 14.6 13.6 15 13.8 14.6 15 13.2 14.6 13.8 12.8 14 14.6 14.4 11.8 15.4 13.6 14 11.9493 0 Normalized oas production L/day 13.73945 16.17473 14.37047 15.6672 11.9337 17.02828 13.80433 12 16.97143 15.45882 14.09362 16.18262 11.51342 15.01714 11.61681 25.94198 10.53492 13.64739 14.93333 11.2464 0 Cumulative Gas production L 560 576 590 606 618 635 649 661 678 693 707 723 735 750 762 787 798 812 827 838 838 Oigestate Height cm iafeBiteifa US— 1MEM *'jf|8p888? I <>.•&.. \J 'mm WMfWt Biogas Yield L/tfvs 0.585324 0.689071 0.612206 0.667449 0.508396 0.725433 0.588088 0.51122 0.723011 0.658572 0.600412 0.689407 0.490491 0.639756 0.494896 1.105172 0.448805 0.581402 0.636185 0.479116 0 Acidity/Alkalinity I? "06 Bicarbonate mo*N/L 72.64095 73.10232 71.36753 ~ — VFA mq*N/L QE553 5.55649 7 W>?4, VFA/Bicarbonate fop. <0. t) 07MI1 0 11)71?® ••••• EES! TS mg/L 17909.4 18568 «B9M VS ma/L 13623 14363 15530 14320 1472* organic Loading Svs'day

5 day BOD rtxvl '-mm - * , *' BI.'.'.'.M Bl'1 V

vnmor

I HiHHS IN- ii i.jrs£ \ « ; i*«^®s»s§s triS&,3*iK;

Influent This data is for undiluted F««d Unit 25-^Jov 26-f\.ov 27-Nov 28-Mov 29-Nov 30-Nov 01-Ooc 02-Dec 03-(?ec 04-Dec OS-Doc 06-Dcc 0?-Dec 08-Dcc 09-Dcc 10-Dcc 11-Dec 12-Doc 13-Dec 14-Dec 15-Dec

••&!]•••&] ••££!] ••E53 *»k. •ililllh. •ES3 HIES ••£££] , : • ' v::;sf'"r v". -fSBS! ••' "T* :."v~ ' illifi f > , "" ~l •' " " ' •.HVZHI " ' x?~- ^ ... _, „ mmmsmi /rvy3^"5 - -. • •"•S3F< J - Z,*i-*-•.•?*•<• " *n ™- -Jid'^SLC : »" * ' •« vsp^S-f- - • ' ^ • • ~ ' «.'yD/.'v-' - • • « y- -' f-. •»>» . •••.t - i«» •).<^ *

IKBEST " KIH KHj •E&3F:;. -5|llp- 5§2ji " •• . • "Bgdgai-' r •• '•EEE •»•'"VV1 - •»>»IIM ' BEHHJ ' K».V»:!|- BH.'Al' kl t ' • '?nnsa *• 'BEES 'KEE3

_-•> - • *s - <• "* ; ", •"• % & mftmm spasi ISPSIP lapses *«««mmms p«s«i mmmm mmmx mmms wwsbi »pssh IIHIS» sssiii ibshpsi MMM wetKM ~ ' » v,-^, r'" -" ..... < . N -- " • ,'T ' - " " • y'r •• < '- • •"•?& - "r • '/- \*m • W.r»'VJ - ' x" - • T ~ -v• . rr- "v » 1 »--.»• v. - * ,• .*vis* p^ « S >• ' r ^•TWWIIIIMIIIIW ffifT w >*&*""' Tm • -• -• -* x,si<- •*-<•xz-vzmmtmmm *. U :., JW"'.. v<^ , 50%DM / 50%RB Effluent Ud Left Technician's Notes: Op«n

Sample Time hoty 17:20 18:10 11:15 17:40 16:15 10:30 10:55 16:55 10:00 22:00 23:59 19:00 0:00 22:27 20:35 16:50 21:45 21:30 8:10 8:40 8:20 Elapsed Time days 0.83 1.03 0.71 1.27 0.94 0.76 1.02 1.25 0.71 1.50 1.08 0.79 1.21 0.94 0.92 0.84 1.20 0.99 0.44 1.02 0.99 Mixer Speed RPM 56 56 56 56 56 56 56 0 56 51 0 0 0 0 0 0 0 0 0 0 0 Temperature •F 94.1 95.9 95 95 95 95 95.9 95 94.1 93.2 90.5 91.4 91.4 91.4 91.4 91.4 92.66 93.2 93.2 93.2 93.2 34.5 35.5 35.0 35.0 35.0 35.0 35.5 35.0 34.5 34.0 32.5 33.0 33.0 33.0 33.0 33.0 33.7 34.0 34.0 34.0 34.0 I *c I [Tipping Bucket I 69 75 62 86 69 66 74 78 67 89 0 59 65 61 62 60 59 59 58 70 72 iGas Productk>n ml | 11612.7 12622.5 10434.6 14473.8 11612.7 11107.8 12454.2 13127.4 11276.1 14978.7 0 9929.7 10939.5 10266.3 10434.6 10098 9929.7 9929.7 9761.4 11781 12117.6 L 11.6127 12.6225 10.4346 14.4738 11.6127 11.1078 12.4542 13.1274 11.2761 14.9787 0 9.9297 10.9395 10.2663 10.4346 10.098 9.9297 9.9297 9.7614 11.781 12.1176 Normattzad gas production Uday 14.05234 12.19893 14.65934 11.42042 12.34117 14.60752 12.24167 10.50192 15.84155 9.9858 0 12.53179 9.053379 10.97511 11.31463 11.968 8.241365 10.03422 21.96315 11.54057 12.28827 Cumulative Gas production L 852 864 879 890 902 917 929 940 956 966 966 978 987 998 1010 1022 1030 1040 1062 1073 1086 Dkjestate Height cm "HfWTWITIflf iMHBinnrai ifssmspg WMSSSM w MifP? •IjjSiPlfij li!i!B!IIE Biogas Yield L/9vs 0.598653 0.519695 0.624513 0.466529 0.525755 0.622305 0.521516 0.447399 0.674876 0.425412 0 0.533875 0.385689 0.467558 0.482022 0.509857 0.351096 0.427475 0.935667 0.491648 0.523501 Acidity/Alkalinity

ma*N/L 77.54759 75 17668 *P 92R32 mg«N/L 2.2810051}^ iv- fl * | 11 !' I r vFA/Bicarbonate fo 0 03fl414il^* * I >' ^Tw! »' EBEE33 18164 20331 15441 13451 11*41 14824 11S14 s»4*1 Organic Loading 9vs/day h 77

5 day 8QD SCOD t'SS *25-50 diiurbn; mg/L 1 ITCOO

TSS fng/^^^ VSS mg/L Ammonia mg/L Nitrate mg/l | 1 Nitrite mg/l I TK mg/l. 1 Total Phosphorus mg/L I

Influent This data is for undiluted F«ed 1?-Dcc 18-Dcc 19-DPC 20-DCC 21-0ec 22-Doc 23-Dec 24-pec 25-DPC 2&-DPC 27-Dcc 28-Dec 29-DPC 30-Dec 31-Dec 01-Jan 02-Jan 03-Jan 04-Jan 05-Jan

I ••E33 WRtEEh ••E33 ••E33 ••EH] HHii ^

»< * ' ~ >! — '.' >!* * ^

- , - ' v ; 1 f: • -- • v* > 1- Z* ?• *. - -^41H - . - -t ' i- ev '• -v- -iprr- ' - >)* •|KEIB m 11 •EH3 •EMU IKE3S •EH3 •an - "rfHEEJ ~as«ic.v ;HE!ED •EES

Oxygpn Demand

v . „F » - . ;S"zcs•*" • ** " ™_" ~r •"rv B-'W *„ 1 •-™- • ,i»- r -- • ii- ~ *">• "* J.

~S ' ~ ~ S ^ ° -X ' «• * ' ..*P*tr "* - ",i; ' * - T ~ • ifl -•> - " . " 7 ••' •*, ?" - •-- "* t •"*'» - - y -•!»«*.'" »•$?•*,, vltiMMmiMHWI Total Phosphofus mg/L , iUf * „ uai.v* i ,".«m;•.sszutritsu^s.i^r,. j:i

188 50%DM / 50%RB Effluent 4th HRT Technician's Notes: New

Sample Time hour 8:15 18:39 22:30 16:30 17:35 20:45 18:00 16:30 13:00 21:15 19:45 19:45 9:35 19:33 18:00 18:20 21:00 20:00 17:00 21:45 lapsed Time days 1.00 1.43 1.16 0.75 1.05 1.13 0.89 0.94 0.85 1.34 0.94 1.00 0.58 1.42 0.94 1.01 1.11 0.96 0.88 1.20 Mixer Speed RPM 0 0 0 87 90 87 90 87 90 85 86 86 86 85 86 86 86 80 86 81 [Temperature •F 93.2 95 95.9 95 95 95 95 95 96.8 95 95 95 95 95 95 95 95 95.9 95 95 34.0 35.0 35.5 35.0 35.0 35.0 35.0 35.0 36.0 35.0 35.0 35.0 35.0 35.0 35.0 35.0 35.0 35.5 35.0 35.0 I *C |Tipping Bucket I 57 85 83 67 75 84 89 80 78 85 86 77 97 128 99 93 97 84 81 105 I Gas Production mL 1 9593.1 14305.5 13968.9 11276.1 12622.5 14137.2 14978.7 13464 13127.4 14305.5 14473.8 12959.1 16325.1 21542.4 16661.7 15651.9 16325.1 14137.2 13632.3 17671.5 i i i 9.5931 14.3055 13.9689 11.2761 126225 14.1372 14.9787 13.464 13.1274 14.3055 14.4738 12.9591 16.3251 21.5424 16.6617 15.6519 16.3251 14.1372 13.6323 17.6715 Normalized gas production L/day 9.626525 9.980581 12.03783 15.0348 12.07734 12.48931 16.91712 14.3616 15.36866 10.64595 15.43872 12.9591 28.32307 15.22132 17.81206 15.43749 14.69259 14.75186 15.57977 14.75186 Cumulative Gas production L 1095 1105 1117 1132 1144 1157 1174 1188 1203 1214 1230 1243 1271 1286 1304 1319 1334 1349 1364 1379 Dtqestate Heighl cm iKismiKMiaini WfBMP*1 niMiia &wm. HHHKh* IMi \ Biotas Yield L/QVS 0.410106 0.42519 0.512832 0.640508 0.514515 I 0.532065 I 0.720698] 0.611328 0.654731 0.453536 0.657715 0.552079 1.20661 0.648454 0.758824 0.657663 0.625929 0.628454 0.663724 0.628454

73.37597 2.46464? 0.033589

164i>4 IB '31 21346 134 b9 15"W 61On 15520 Wd»y

1733.75 1878 "5 1912 *> 2•>*><» 15f |SCOP t'SS x25-50 dilution'; WMMmU ITCOO (SS <100-200d-luisor 242*4 3*273 24853 2W9 19457

12-B5 26 1375? 51 14695 1*24? 5 15417.79 10639 41 114113 12*17 S 13 58 48 12519.4

[Total Phosphorus" 171,13 247.76 165.3C 125.52 210.54

Influent This data is for undiluted F«d mi I Water Volume mL 280 2801 280j 280 280! 280 280 280 280 280 2801 280 280 280 280 280 280 280 280 280 Feed Volume mL 220 220 220 220 220 220 220 220 220 220 220 220 220 220 220 220 220 220 220 220 j Ideal Feed Volume mL MIMi HI ISHMI " ; J Dairy Manure 75%I 75%| 75% 75% 75% 75% I 75% 75% 75% 75% 75% [ 75% 75% 75% 75% 75% 75% 75% 75% 75% Beer ! 25% | 25%l 25% 25%| 25% 25% I 25% 25% 25% 25% 25% 25%| 25% 25% 25% 25% 25% 25% 25% 25% |HydraiAcRetentionTim^^_

[Bicarbonate VFA ma*N/L •pflVFA/Bicarbonate sop. <0 5i 1 EIOH 1

•frs ma/L 1 nvs ma/L | El Actual Oraonic Loadkto Rate ko^/mW j

•S5davBOD ma/L j USCOD (SS x3Q ma/L | tBlCOD^SSx^O^OOdifu^ rng/^^l mSL| VSS ma/L fffm 51931 V ** t >. Ammonia ma/L IMM r-V-- • . * .! §11111111111 I r__ Nitrate mo/L pffPP rt%?r — IT" Nitrite mn/l IflnMt MjMB li't. | r| *4* „ J . *!r. • _ * TK mo/L HUB Iff * * ' j Total Phosphorus mo/L HH IBMBtLli *[• ' J i •« 50%DM / 50%RB Effluent Technician's Notes:

^•samote Time how 22:45 15:00 15:15 ^HbJaosed Time day* 1.04 0.66 1.01 Mixer Speed RPM 65 66 85 ^•Temoerature •F 95 95 95 35.0 35.0 35.0 • "C I BMTippino Bucket 1 145 167 4 Production mL | 24403.5 28106.1 673.2 24.4035 28.1061 0.6732 ^•Normalized oas production l/day 23.42736 41.51055 0.66626 ^•Cumulative Gas production L 1403 1444 1445 ^•Dioestate Heioht cm HBiogas Yield ^9vs 0.996045 1.768419 0.028384

19 pH 7.09II 7.131I 7.121

|[Organic Loading 1IS day BOD mg/L H SCOD tSS x25-50 dilution) mg/L I|TCO^S^<1G^20t^ijjuii0tt> 1 TSS mg/L 1 vss mo/t a Ammonia mg/L 1 Nitrate mg/L i Nitrite mg/L B TK mg/L 1 Total Phosphorus mq/L Influent This data is for undiluted F«gd

Feed Volume (dew Feed Volume Pairy Manure

unc Retention rime

mq»N/L mg-N/L vFA/Bicart>onate ;op

Actual Oraanic Loadina Rate Oxygen Demand |5 day BOD mg/L IsCOD i'SS x30 dilution. mo/L ITCODiS^D^O^jiutions mg/L

rss mg/L Ivss mg/L I Ammonia mg/L r j [Nitrate mg/L t. iNHrite mg/L "i— j TK mg/L l|Total Phosphorus mg/L

190 75% DM/25% RB

Effluent

Technician's Notes:

Parameter 21-Jul 22-Jul 23-Jul 24-Jul 25-Jul 26-Jul 27-Jul 28-Jul 29-Jul 30-Ju! 31 -Jul 01-Auq 02-Aug 03-Aug 04-Aug 05-Aug 06-Aug 07-Auq 08-Aug 09-Aug 10-Aug General sample rm 13:00 U:20 10 30 10:30 12:12 10:30 18:10 11:42 17:20 21:00 17:30 17:00 13:25 13:05 13:30 10:30 10:00

Mixer Speed Temperature

ripping Bucket Gas Production 24120 19170 19170 21240 18180 7470 11160 10530 5480 13230 11610 10440 11610 24.12 18.18 11.16 10.53 13.23 11.61 10.44 11.61 Normalized gas production 16.16105 22.81388 21.24 16.97743 0.755245 5.783226 10.7136 7.9806316 6.159696 4.638316 4.340093 1.896565 1.286809 5.607692 13.71766 13.41634 1.41188 11.93143 11.85702 Cumulative Gas production Pigettate Height 37 Bkxias Yield (not accurate) B.7555045 mm mm 1.725201 AcidityfAlkalinity 7.356 7.514

03753 vFA/Bicarbonate

2 3 1 19705 Organic Loading Svs'day

5 day CBOD SCOD ;SS *25-50 dUutiors) TCOD (5S xl00-200 diiutioi 3300

Ammonia

Total Phosphorus

Influent

21-Jul 22-Jul 23-Jul 24-Jul 25-Jul 26-Jul 27-Jul 23-Ju! 29-Jul 30-Jul 31 -Jul 01-Aug 02-Aug 03-Aug 04-Aug 05-Aug 06-Aug 07-Aug 08-Aug 09-Aug 10-Aug

I—B33 : ^^^^3 ^HE33 IHEEE3 BBES9 IHES3 HHES3 HHES3 HES3 HHESuS IHE53 wrnmm mmmm WMB iH tUl^ Acidity'Alkalmity IliSS SiSilt pj^ii dnis liim wMM Mmmi IllSijiiraii iSlilii

•'""I • 1.U . 3 > •> "as UIEEE3 -HH , -hi i ] mwHma USB* Oxygon Demand • - - "EE^O™ ~ " "~j .. --- — l»TW*:.l

191 75% DM/25% RB

Effluent

Technician's Notes: in

«r Broke 5n«> Df.vne'.i <2L'

Sample Time hour 10:15 11:00 11:08 10:45 19:30 13:20 14:30 15:3C 124J 13:15 14:35 17:40 12:30 14:49 9:00 18:00 14:30 10:00 9:3( 14:30 11:15 BapMdTime day* 1.01 1.03 1.01 0.98 1.36 0.74 1.05 1.04 0.89 1.02 1.06 1.13 0.78 1.10 0.76 1.38 0.85 0.81 0.98 1.21 0.86 Mixer Speed RPM 50 51 50 51 51 50 50 51 51 51 46 47 52 47 48 48 48 48 51 48 50 Temperature *F 98 96 98 98 97.75 98 98 98 98 98 98 98.25 98.5 98.25 98 98 98 95 96.8 96.8 95 •c 36.7 36.7 36.7 36.7 36.5 36.7 36.7 36.7 36.7 36.7 36.7 36.8 36.9 36.8 36.7 36.7 36.7 35.0 36.0 36.0 35.0 Tipping Bucket 105 113 108 115 146 159 146 30 81 100 107 120 101 116 106 148 190 70 128 102 110 Gas Production mL 9450 10170 9720 10350 13140 14310 13140 2700 7290 9000 9630 10800 9090 10440 9540 13320 17100 6300 11520 9180 9900 L 9.45 10.17 9.72 10.35 13.14 14.31 13.14 2.7 7.29 9 9.63 10.8 9.09 10.44 9.54 13.32 17.1 6.3 11.52 9.18 9.9 Normalized gas production L/day 9.352577 9.861818 9.666298 10.517996 9.629313 19.25832 12.53086 2.592 8.233412 8.816327 9.123158 9.570462 11.58372 9.520963 12.59175 9.687273 20.01951 7.753846 11.76511 7.597241 11.4506 Cumulative Gas production L 227 ^^237 247 257 267 286 299 301 310 319 328 337 349 358 371 381 401 408 420 428 439 K! Bjooa^jej^no^cturate^^^^^l'"* 2.61946 2.696968 0.725965 5 386085 2.680485 rams flslirfl 7T71RS3 mm MHH MHI pH 7*66 7 1*7 7*81 7*27 7 *11 7 **fi 7*0* 7 *73 738 "Pi •'H!! •73!! •7S •7HJ 7 **9 •fS

VFA IgHEEa SITCTrg] VFA/B itszna »131223 USD CG2HQ i TS ^EjZiZ] VS •BSS Ofgani IMJ BiiUJ ELKIBUS: Oxygen Demand 5 day ' BE3B • v:ini SCOl TCOC EE9ET3 nraai

TSS VSS | g»|.; jsggijgjij ipSlili Slpjijil^ilil sillliif fefi|3 lliiiiis fiplill MSii ifilllipi BHBB Ammo Nitrate Nitrite TK TotalF

Influent

Parameter 11-Aug 12-Aug 13-Aug 14-Auq 15-Aug 16-Aug 17-Aug 18-Aug 19-Aug 20-Aug 21-Aug 22-Aug 23-Aug 24-Aug 25-Aug 26-Aug 27-Aug 28-Aug 29-Aug 30-Aug 31-Aug

!BIK!i]^^B^HpESHKQ3H|KI3HHB^BpB^HBKEIHKBHBK03 j ^ WMMMiJ •ME233BBE33HKS3HHE£x3HK33BK£E]^K23I^E33 ^K32^E23^H33^BE23^KIZ]HKI33BBE33HK23 ••£<£]••££• I • -s j» : . - Acfdity-'Alkalmity SsSE^ssE^ssiiSSS |^EE3 ... >• M3B *"*13J KS3 SSH3 |JJE|E| tg^^B1 IHCHII HUES ITIII •E33! «.*--i.W*U - - s9HlTIE3 Ul'/'ilJ Hr^l Oxygen Demanq lEEESE dnzs ggirun • '••m w» wm »•>'-''>1 •riiin

® WMiWMM i mmm%m wmmmm. mmmmm mmmmm mmmmm mmmmm m&mmm $mmmm mmm&m mmmmm mmmmm mmmmm

192 75% DM/25% RB

Effluent CaShrated VVMt« Tipptng mould SUCK* CK$>et»*r Technician's Notes: rt2 fnL5 • 'I«W Fttri Op*n«rf.

ArKii!» S! ma*N/L 117J2854 109.9433 44 6.534251 6.248007 4 4 VFA/Bicarbonate *c 0.055712 0.056829

23033 13370

Oxygen Demand 5 day CBOu SCOD iSS X25-S0 dilution) TCOO {5Sx*00-2 dilution)

Ammonia Nitrite m

TrtaH^Phosghonjs

Parameter Unit 01-Sop 02-Sop 03-Sop 04-Scp 05-Sep 05-Sep 0~-Sep 0S-5CD C?-Sop 10-Sop ^ 1-SOD 12-Sop 13-Sep 14-Sep 15-Scp 16-Sep 17.Sep 18-Scp 19-Sep 20-Sep 21-Sep Feed

^ *. * K* ' H wSMm ifSlfil P

Oxyqen Demand t E2sT»IE2E

Steady State

193 75% DM/25% RB

Effluent

Technician's Notes:

Parameter 22-Sep 23-Sep 24-Sep 25-Scp 26-Scp 27-Sep 28-Sep 29-Sep 30-Sep C1-Oct 02-0et 03-0ct 04-0ct 05-0c1 06-0ct 07-0ct 08-0ct 09-0ct 10-0ct 11-Oct 12-Oct Genera! sample lima 10:20 12:10 15:55 i4:oo 13:40 15:34 13:20 14:20 18:17 17:02 16:11 i/:50 13:20 19:33 16:4a n:oo 13:20

Temperature

ripping BucXot Gas Production 12052 1500 12972 11960 10120 11040 11224 10764 13892 10120 7084 8216 12.052 12.972 10.304 11.96 11.408 10.12 11.04 9.476 11.224 10.764 13.892 10.12 7.084 18.216 9.476 14.904 Normalized gas production 11.34306 10.68387 11.21903 11.19831 12.12845 10.57112 11.15835 10.5984 8.136816 10.19077 11.63611 10.55618 13.248 11.03391 1.4566 0.00094 .31422 13.06254 Cumulative Gas production Dtp*state Height Yield mot accurate!

108 6431 98.81424 mq-N/l 7.855847 VFA/Bicarbonate «o (J.U044S 0.079501

26764 24?3 &&0S >44 ° pmy ij *****% _ 1

Oxygen Demand » ? ~ . ,r

SisiiStlflSlSa mm i«pigjj ifflilis - > . . • 4 sr Sis liisiBnil *".,"'3' ~ ..is:r-jESir -• - i^jnCMSB ' "4S-; .... j a• ,4K.-tfaSIBUHMtZXrZkXKUMHiBBS»5

Influent

Feed Volume Ideal Feed Volume Dairy Manure

raulic Retention Time

mg-N/L mg-N/L VFA/Bicwbonate :cp

Actual nic Loadin •K'KM'rl

SdayCBOD SCOO (SS >30 dilution TCOO l3S *400-500 dilution 1 vss rrxj/L Ammonia mo/L Nitrate mo/L Nitrite mq/U TK mq/L Total Phosphorus JStL

194 75% DM/25% RB

Effluent

Technician's Notes:

Sample rime

Mixer Speed

Tipping Bucket 5as Production

Normalized gas product ton Cumulative Gas production Dige state Height % Yield mot accurate) m

ma*N/L 93 71714 mo»N/L 6 4hGfl1S xl»li' V*Z\

•EZaD i WEESi•Mi- h Oxygen Demand SCOt5 day Sg S*Si* |e^*||i» Si*8 S*i|||^*1 ll| 1TCOt

;SSL IALLLilllli liilil I3SI1KiSB iillil 8111B SiSii SMSS wmm SSRSiSls* siilfiSi msmiliiii H HBH wmm

Influent

Parameter Unit 13-Oct U-Oct '5-Oct 16-0ct 17-Oct 13-Oct 19-Cc? 2C-0ct 21-Oct 22-Oct 23-Oct 24-Oct 25-Oct 26-Oct 27-Od 2 3-OCT 29-Oct 30-0ct 31-Oct 01-Nov 02-Nov Feed

Acidity'Alt-alinity

Oxyqen Demand

Steady State

195 75% DM/25% RB

Effluent

Technician's Notes:

Unit 03-Nov 04-Nov 05-Nov 05-Nov 07.Nov 08-Nov 09-Nov 10-Nov 11-Nov '2-Nov 13-Nov 74-Ncw 15-Nov 16-Nov 17-Nov 18-Nov 19-Nov 20-Nov 21-Nov 22-Nov 23-Nov

Sample Time hour 18:30 17:45 19:08 19:13 19:37 17:27 20:39 19:02 19:33 21:00 20:00 20:00 22:20 19:10 21:37 18:55 15:02 21:20 17:10 15:50 18:55 Bapsed Time days 1.03 0.97 1.06 1.00 1.02 0.91 1.13 0.93 1.02 1.06 0.96 1.00 1.10 0.87 1.10 0.89 0.84 1.26 0.83 0.94 1.13 Mixer Speed RPM 51 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 51 51 51 Temperature f 97.7 97.7 97.7 97.7 97.7 96.8 98.6 98.6 98.6 99.5 98.6 98.6 98.6 97.7 97.7 97.7 97.7 97.7 96.8 96.8 97 *C 36.5 36.5 36.5 36.5 36.5 36.0 37.0 37.0 37.0 37.5 37.0 37.0 37.0 36.5 36.5 36.5 36.5 36.5 36.0 36.0 36.1 Tipping Bucket 24 71 78 38 52 68 58 60 54 0 4 14 8 2 73 0 20 0 3 12 0 6a* Production mL 2208 6532 7176 3496 4784 6256 5336 5520 4968 0 368 1288 736 164 6716 0 1840 0 276 1104 0 L 2.208 6.532 7.176 3.496 4.784 6.256 5.336 5.52 4.968 0 0.368 1.288 0.736 0.184 6.716 0 1.84 0 0.276 1.104 0 Normalized gas production L/day 2.141091 6.74271 6.784924 3.483903 4.705574 6.876824 4.708235 5.91869 4.863304 0.384 1.288 0.670785 0.211968 6.093913 2.195195 0.333983 1.168941 Cumulative Gas production L 968 985 989 996 1001 1007 1012 1012 1012 1013 1014 1014 1020 1020 1023 1023 1023 1024 1024 BBMJZI — 9£1 Dioestate Height cm ami IBSS 2II5S liHiEiy MSM Biooas Yield mot accurate) L-'9*Voay ^fBBP 5 o.059358 f.706777 0.09354Z 0.327396

7 4 loH I 7.191 7171 ? 0| ?

sf tsssKfiSif tissjssgp fmtvfaa siiai mhsbr msmm mmmm BPisiiti bbsibi wsb msmm wmmm sspwss wksbsbi

tilf:?i ISF'tOiISBSJSis es:«;il«Rc;-^} sassjp SBSSPW SPSPiR53iS|i S3S ssfss mrnm mmmm ismxm mmms imsm mmm wmm mmm wsmm msssm sasaaii E*»sfi fssm mmssm mssm Oxygen Demand 71*4 r - - - • " J-5 - %*..•. - •« " ,V;> * „ » « < * *| ,V >» „ ' ^ l> r2 1 i N ^ J 11_1„ I

: ; ' w "F •;"• • pii ts® feissSij sffiW i l ®f :i? sSisft ®Sf SS (SiBfl SSIsli siSUs sspsisistsfiSf ifilsfiiBiisiii ssifciiis^BSs T, - ^ _5. , - . -V.-^--j , 5". * . -r :; r r— ---,4: .x; ~ •' " • •' . - • f - . Fr-^'xrc -j-^e nam mmmmm mmmmmmummmmmmmsm wmmm wmmmmMmmmmmm :

Influent

Water Volume mL 320 320 320 320 320 3201 320 320 320 320 320 320 320 320 320 320 320 320 320 320 320 Feed Volume mL 180 180 180 180 180 180^ 180 180 id 180 180 180 180] 180 180 180 180 180 180 180 180 Ideal Feed Volume mL •1 mmm mm BBS MM MM a—H WMmm read 11Mmmwt iDairy Manure I 75% 75% 75% 75% 75% 75% 1 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% lewr 25% 25% 25% 25% 25% 25%! 25% 25% 25% 1 25% 25% 25% 25% | 25% 25% 25% 25% 25% 25% 25% 25% ulic Retention Time Acidity-Alkalinity mg*N/L mg»N/L

mg/L mg/L MActual Oragnic Loading Rale I mg/L mg/L

mg/L mg/L mg/L mg/L ITK mg/L [Total Phosphorus mg/L

196 75% DM/25% RB Effluent

Technician's Notes:

m [Sample "Time hour 23:48 22:35 20:50 19:55 18:55 13:55 17:38 17:55 18:00 18:00 21:00 21:15 21:00 20:07 20:50 21:30 19:44 21:45 13:00 14:00 21:00 Oepsed Time days 1.20 0.95 0.93 0.96 0.96 0.79 1.15 1.01 1.00 1.00 1.13 1.01 0.99 0.96 1.03 1.03 0.93 1.08 0.64 1.04 1.29 Mixer speed RPM 51 51 51 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 51 51 50 Temperature •F 97.7 96.8 100.4 97.7 97.7 98.6 97.7 97.7 97.7 97.7 97.7 98.6 98.6 97.7 97.7 97.7 97.7 97.7 95 95 98 I *C I 36.5 36.0 38.0 36.5 36.5 37.0 36.5 36.5 36.5 36.5 36.5 37.0 37.0 36.5 36.5 36.5 36.5 36.5 35.0 35.0 36.7 iTipping Bucket I 0 0 0 0 183 98 188 143 11 102 10 92 22 0 0 33 76 87 123 111 63 |Gas Production mL I 0 0 0 0 16836 9016 17296 13156 1012 9384 920 8464 2024 0 0 3036 6992 8004 11316 10212 5796 L 0 0 0 0 16.836 9.016 17.296 13.156 1.012 9.384 0.92 8.464 2.024 0 0 3.036 6.992 8.004 11.316 10.212 5.796 Normalized gas production L/day 17.568 11.38863 14.97669 13.0025 1.008498 9.384 0.817778 8.376742 2.045305 2.953946 7.547586 7.383575 17.80879 9.80352 4.487226 Cumulative Gas production L 1024 1024 1024 1024 1042 1053 1068 1081 1082 1091 1092 1101 1103 1103 1103 1106 1113 1121 1138 1148 1153 Digest ate Height cm •Sflk| ug^oiy 222 22 5£S1 22 r.ioi /.i/i r.f«« / n 7 11 713| 7 12 714 7 131 7 Q9 7 121 ^ 7 15 7 15 7 15| J 141 ;«ytM mg*N/L 1 VFA/Bicarbonats ;oo. <0.5 '*v. ' 'fc **»**•> • v »JgH|

Organic Loading Ovs'day

5dayCBOD SCOO ^SS x2 50 dilution TCOD CSS x 5 00-200 dilution) m B Ammonia mg/L H Nitrate mg/L 9 Nitrite mg/L B TK mgfl. Total Phosphorus !Wk Influent

C Parameter 25-Nov 2 >-*' 01 -Doc G?-Ooc 0 3-Coc 04-Dec 05-DPC 06-DPC C7-DOC 03-DOC 09-DOC 10-DCC 11-DCC 12-DOC 13-DCC 14-Dec

Acidity;AIKahnity

s innwssssm muss sssiis ismm i AM FSSSIIIS MM WPSS? mmam Oxygen Demand

" -VX_t-V-.* --- - - '*

: " s *, ~ * ' » ~ * > ' i~\ ••& v ^ r^-TW*"" -<> * " * rxT?v ^ ' '• J }, ••- ' a : v :' * ~ ^ atjK iz,m ji^pnosgnorus^ ? VVV5« -*• - ««•««(! I •••

197 75% DM/25% RB

Technician's Notes:

Sample Time hour 18:55 9:40 9:40 20:00 20:00 20:00 14:00 21:00 17:00 11:00 18:00 21:30 16:00 15:5G 21:10 14:20 17:35 20:35 18:05 18:45 19:20 BepaedTTme days 0.91 0.61 1.00 1.43 1.00 1.00 0.75 1.29 0.83 0.75 1.29 1.15 0.77 0.99 1.22 0.72 1.14 1.13 0.90 1.03 1.02 Mixer Speed RPM 50 50 50 51 51 51 55 55 55 54 55 54 55 55 55 55 55 55 55 51 55 Temperature *F 98 98 98 97.7 97.7 95 69.8 97.7 95 95 95 95 98.6 98.6 98.6 98.6 98.6 98.6 98.6 98.6 98.6 •c 36.7 36.7 36.7 36.5 36.5 35.0 21.0 36.5 35.0 35.0 35.0 35.0 37.0 37.0 37.0 37.0 37.0 37.0 37.0 37.0 37.0 Tipping Bucket 103 0 17 0 0 0 0 122 51 0 0 6 36 65 0 6 0 0 0 84 0 Gas Production mt 9476 0 1564 0 0 0 0 11224 4692 0 0 552 3312 5980 0 552 0 0 0 7728 0 L 9.476 0 1.564 0 0 0 0 11.224 4.692 0 0 0.552 3.312 5.98 0 0.552 0 0 0 7.728 0 formalized gas production L/day 10.37676 0 1.564 8.689546 5.6304 0.481745 4.296649 6.021818 0 0.771728 7.519135 Cumulative Gas production L 1163 1163 1165 1165 1165 1165 1173 1179 1179 1179 1179 1184 1*90 1190 1190 1190 1190 ^190 1198 1198 Diqestate Heioht cm IMP §£MIR! flM Wf Biooas Yield mot accurate) »-'9w'a*y 0 6.438043 0 0 0 0.134927 o^2i8i45 0 0 2.105952 "

1 HHI PH I 7.151 7.131 7.14| 7.16| ' <9 t <3 I 7 1AJ 7 111 7 1g| 7 12 7 17 7.171 7 1fi| 7 14| 7 1A| 7 14| 7.14 • x ,c. ; >*es§t wmawa

!' . »MOTI

Oxygen Demand

vss Ammo r ; - - j. Nitrate ,t , .« Nitrite , ~ 3" •' - - TK ' '; f. • *. i * m .r -iisisfg:! 'I K:MMR•(••• V*- •;.2VK&SHK3MMB 4IK i*C°** I

Influent

Feed Volume Ideal Feed Volume Dairy Manure

rauic Retention Tim® Acidity/Alkaliriify

mq'N/L

Actual k: Load

5 day CBOP SCOP (SS >30 dilution) *400-500 dilution}

Nrtnte

Total Phosphorus

198 75% DM/25% RB

Effluent

Technician's Notes:

^•Samole Time hour 13:50 16:15 19:30 8:45 20:15 20:15 12:25 9:35 9:45 12:45 12:30 12:00 12:00 13:35 13:00 9:30 8:10 9:55 15:30 12:20 8:50 ^HElaoaed Time days 0.77 1.10 1.14 0.56 1.48 1.00 0.67 0.88 1.01 1.13 0.99 0.98 1.00 1.07 0.98 0.85 0.94 1.07 1.23 0.87 0.85 Mixer Speed RPM 55 51 55 51 51 51 51 55 55 55 55 55 55 55 55 55 0 38 36 36 36 ^•Temperature *F 97.7 96.8 97.7 96.8 96.8 98.6 98.6 97.7 97.7 98.6 97.7 95 95 97.7 97.7 97.7 96.8 96.8 97.7 97.7 97.7 H 36.5 36.0 36.5 36.0 36.0 37.0 37.0 36.5 36.5 37.0 36.5 35.0 35.0 36.5 36.5 36.5 36.0 36.0 36.5 36.5 36.5 ^9 Hoping Bucket 0 6 138 98 121 101 78 60 53 38 1 66 37 33 1 7 18 113 121 85 18 EBGas Production mL I 0 552 12696 9016 11132 9292 7176 5520 4876 3496 92 6072 3404 3036 92 644 1656 10396 11132 7820 1656 0 0.552 12.696 9.016 11.132 9.292 7.176 5.52 4.876 3.496 0.092 6.072 3.404 3.036 0.092 0.644 1.656 10.396 11.132 7.82 1.656 ^•Normalized gas production L/day 0.501502 11.1818 16.33087 7.525859 9.292 10.65303 6.258898 4.842372 3.107556 0.092968 6.201191 3.404 2.848104 0.094292 0.753951 1.753412 9.689476 9.031031 9.00864 1.938732 ^HcumUatrve Gas production L 1198 1199 1210 1226 1234 1243 1253 1260 1265 1268 1268 1274 1277 1280 1280 1281 1283 1293 1302 1311 1313 ^Hoiaestate Heioht cm TfflWH 1 1 1 1 ^BBiogas Y">eK3 mot accurate) ug^aavr IBS 0J2Tfi86 2 |MBM| •SPH 7.18 1 7.151I 7.15| 7.151L 7.121 7.12 - i 7.16 7.15 7.11 7.24 I 7311r 7.iaiI 7.21: ^^6 1 7.221 7.2*1I 7?l 7.24 I 7.25I t \ B>fl Bicarbonate mg*N/L 87.97527 M7679smsmmm 89.07939 i 86.99577 87.56336 r^rr 84 C3204 84 82 U9 $ y llVFA mg-N/L 4.435473 mmmmm 3.811767 imimr 2.456131 Si - -1 5.715479 asUllil ; 0.570675 17 4- 79918' 3 548949 4 660094 % I •HVFA/Bicarbonate ;cp. <0.5} 0.050417 ••MM MMttl 0.042943 0.027572 ff k 0.065698 •HUB ! 0.006517 £1. i-. 0 042057 0 05659 . .1- - \ M mg/L 218031 17009 mg/L 167191 166671 154301 12313 9W<|«y ~0555i "T335551 7.71495B 9±rdi[*j •> . I s 9?o65rrr >.1565

"a*- | SCOP (SS x25-50 "•a"- |TCOD (SS x; 00-200 dilution"

mg/L Ivss mg/L j 1 Ammonia mg/l. 1 Nitrate mg/L I I Nitrite mg/L j [TK mg/L I 1Total Phosphorus mg/L !

Influent

•waterVolume mL I 320i 3201 3201 3201 3201 320 320 I 320| 320 320 320 320 320 320 3201 320 320 320 320 320 320 ^•Feed Volume mL 180 I180I 180 180 180 I 1801 180 180 180 180 180l 180 180l 180 180 180 180 180 180 E9 Ideal Feed Volume mL jfggfgg awrattaww jggggBHB LLJFGJLGG NMGG MMWI BMBI mm pipmm I^MGI HJI3I Mm H E9 Dairy Manure 75% I 75%| 75% 75% 75% 75% i 7?% 75% 75% 75% 75%j 75% 75W 75% 75% 75% 75% 75% 75% •Beer ( 25%! 25%| 25% I 25%| 25% 25% 25% 25% I 25% 25% 25% 25% 25%| 25% 25%: 25% 25% 25% 25% 25% 25%

ff 115.9501 .i? *?.?** rats; mm mmr ma saaa. &4H 1Z&SSSt SItir.;.. BVFA mg-N/L 94.95058 128 3818 aU;«;: «?! ;vjy!U 158 2911 ffiiE.riiffli'in to***1* ..'tt-SSj M+m FR* IL-T* • JVFA/Bicart>onate ;OP. *:0.5) 0.818892 .wijjijr^TariigSK^i 1.009876 1 7734,6 3 013157 -

mg/L 81505 mg/L *8448 1 02672

|S day CBOD mg/L [SCOO (SS >30 dilution) mg/L > *400-500 dilution'i

mg/L vurnm yySBB Ivss mg/L 1 Lf* 1 > j mg/L I If" ' • ' .<•"* 1 I*1' .. J»-_ I Ammonia l—u-iL- [Nitrate mg/L E l-i-ss!—:- Wh =-.^7i fed 1 fix v"'J

' 4 iNitrtte mg/L 1 ?. I -*"> 3 ;• mg/L [ 5 ^ Vi' 1 p j 1 is iTotal Phosphorus mg/L [ mmm baBa] v

199 75% DM/25% RB

Effluent

Technician's Notes:

Sample Time

Mixer Speed

[Tipping Bucket Gee Production 8.556 10.488 6.716 13.616 9.476 11.776 20.884 Normalized gas production L/day 4.294179 5.085268 6.203077 11.0499 8.963039 9.603728 9.014103 8.337902 6.646763 10.45709 11.48942 12.68426 6.316927 11.14036 5.478834 3.703742 12.73153 5.87929 8.141671 15.41585 13.83301 Cumulative Gas production Dige state Height uiw®«y 2.510359 2.689o02 2.524661 2.335271 Acidity.'Alkalimty

mg'N/L mg»N/L BgEg EE3H1

rog/L 18.18:' 187971. J.." r 19134 ) rog/L 13574 ,.V V. .. 13260 •' ' - " 14424 ' wSay *>?**>(&

mg/L "*56 6667 ' z ^52^' Afytih,. - 1 • |SCOP {SS *25-50 dHufton) mg/L 1 . . ? ; FT. *33- j- I : MTCOO (SS x"jQO-200 g

Influent HSfiS! KSS3 HSBSq •tifliJ.j •mn HR39H •HXSS HHaS

Feed Volume ueai Feed Volume Dairy Manure

ulic Retention Time

Bicarbonate 46.47635 mg-N/L 201.5409 VFA/Bicarbonate icp.

MESS ° Ksni •B K3D

•g-:. - ~~ - " - Utjg riSS'ii SSSS £SSi SiSiS fei5S SISSl Sfett SllSS SiiiS SSfi HS5t SSSii ISStei Phosphorus mg/L 1 I

200 75% DM/25% RB

Technician's Notes:

16-Fcb 17-Feb 18-Feb tS-Feb 20-Feb 21-Feb 22-Feb 23-Feb 24-FeD 25-Ffb 26-Feb 2?-Feb 28-Feb 01-Mcir 02-Var 03-Mar 04-Var OS-Mar 00-Var O^-Mar

Sample Time hour 12:49 10:00 9:00 8:10 21:30 14:00 10:30 16:00 16:50 15:50 14:00 13:15 11:36 16:00 14:39 17:00 17:00 19:30 16:50 14:30 13:50 Elapsed Time days o.ea 0.89 0.96 0.97 1.56 0.69 0.85 1.23 1.03 0.96 0.92 0.97 0.93 1.18 0.94 1.10 1.00 1.10 0.89 0.90 0.97 Mixer Speed RPM 36 36 36 36 36 36 35 36 36 36 36 0 30 30 30 31 30 31 31 26 26 Temperature f 97.7 98.6 98.6 97.7 97.7 97.7 97.7 97.7 97.7 97.7 95 95 95 96.8 97.7 97.7 97.7 98.6 97.7 97.7 97.7 36.5 37.0 37.0 36.5 36.5 36.5 36.5 36.S 36.5 36.5 35.0 35.0 35.0 36.0 36.5 36.5 36.5 37.0 36.5 36.5 36.5 *C {Tipping Bucket j 177 176 198 201 270 144 122 89 56 51 41 22 139 120 84 78 130 103 68 63 60 |Gas Production rnL 1 18284 16192 18218 18492 24840 13248 11224 8188 5152 4692 3772 2024 12788 11040 7728 7176 11960 9476 6256 5796 5520 1 L ] 16.284 16.192 18.216 18.492 24.84 13.248 11.224 8.188 5.152 4.692 3.772 2.024 12.788 11.04 7.728 7.176 11.96 9.476 6.256 5.796 5.52 Normalized gas production L/day 24.05022 18.28744 19.008 19.15718 15.96857 19.26982 13.14029 6.661424 4.979114 4.896 4.08397 2.08929 13.73208 9.329577 8.188609 6.536015 11.96 8.582038 7.038 6.420185 5.677714 Cumulative Gas production L 1523 1541 1560 1579 1595 1615 1628 1635 1640 1644 1648 1651 1664 1674 I 16821 16881 1700 1709 1716 1722 1728 Digestate Height cm MM*MIfWf I f£a8B§8: fpn ii Bioaas Yield mot accurate) uy^aay f^§ff WW "aw" 2 613018 mmmmmTwos HHH mKM HUH 1HI ••• I ^•1•• •••• PH 7.38 7.36 7.3 7.33 | 7.3| 7.36| 7.36| 7.281 7.33 | 7.351 7.35 ^7Sf 7.36| 7.33| 7.35| 7.341 7.35! tST 7.34 7.35 7.34 Wd. i» IMtmS!mm mm flpiiSliSmmm mmmWMWM WMWMmmm mmm fffpiili §mm iiiPlflmmmi«lS mmm mmm mmm wmwmftlPfil! ilfitill wmwm

pipKJSI •E3Q |S|S£!IK3i| gjjpsf^|jj ii.vi.-:-B(Si Is llSiii i > ** mmsmm Oxygen Demand

*si fgggg §S®1| iii mmm mmm mmm'iw '- •«"• •.,; -- - - * v - „ ,- zr'n-mmr^r-r- , : msmmmwmmmmsmim TOWrhOS£hOfU^ LJ _l_ J_ JL J_ JL _l_ _1_ l i -1_ _L J- 1_ _I_ I L Influent

1 Water Volume mL 320 3201 320 320 320 320 320 320 3201 3201 320 320 320 320 320 320 320 320 320 320 320 Feed Volume mL 180 180 180 180 180 180 180 180 1801 1801 180 180 180 180 180 180 180 180 180 180 180 lldeal Feed Volume mL [Dairy Manure 75% 75% 75% 75% 75% 75% 75% 75% 75% [ 75% | 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% Beer 25% 25%| 25% 25% 25% 25% 25% 25% ' .'i 25% 25% 25% 25% 25% ?5% ?5% 25% 25% 25% 25% W Hydraulic Retention Time days v£'„J . L. ««! . » ?., ... w .1 ... 3 .i *a~. S *dk* **k. .* ' HW AcidttyAlkahmty

mq«N/L iVf A/Bicarbonate -op. <0.£) . H

mg/L Ivs mart. I lActua^raoni^oadir^Rtf^^

15 day CBOD mo/l I ISCOD i'SS >30 difiiliort} mqA. 1 ITCOO tss X400-500 dilution

Irss mg/L 1 |«y *?rf. &sus aaaas jtausb. inpas «'»• r! Ivss mo/L 1 -£656S i-il 1Ammonia mg/L [ «» I Nitrate mg/l [ iNrtrtte mg/L 1 ITK mg/L | •Total Phosphorus mq/L E

201 75% DM/25% RB

Effluent

Technician's Notes: St«ady State New Feed, Collected Gas 09-Vj' 10-f.*ar 11-'.

15:00 21:00 21:00 15:00 20:45 19:10 21:45 21:45 18:00 12:30 19:30 19:15 17:14 10:12 16:10 16:10 20:00 10:30 14:30 12:35

RPM

Gas Production 11408 11592 11132 10212 10396 11776 13432 12420 10.488 11.408 9.936 9.476 11.592 11.132 10.212 10.396 11.776 10.304 13.432 9.292 4.968 2.208 1.932 Normalized gas production 12.63467 9.351529 10.3423 10.05021 10.212 11.47145 12.56107 13.36735 10.39897 14.87495 13.55936 13.14389 0.736819 8.222897 1.892571 2.099683 Cumulative Gas production 1770 Digestate Height s Yield mot accurate}

7*9 7 7 « a i 5 209675 vFA/Bicarbonate {pp. <0.5 0.05

CUyqen Demand

m iwhk ESIE3 FEgnna

178.6 580.00 Total Phosphorus 30.83 Mil

Influent

Parameter un 10-V.ir H-P.\ir 12-Mar 13-V,v MA'ar 15-Mnr 16-Mar 17-Mar 18-^ar 19-^ar 20-Mar 21-Mar 22-'.*ar 23-Mar 24-Vjr 25-Mar 2S-Mar 27-'/ar 26-^ar 29-Mar

i2mm:

Acidity'Alkalmity •EEKAH ~~ ' :f? !•!•»ml*\ -.'i ^

ME3D mrzm ? \ IP 1PPIPS|P 0*yqen Demand

fMi issSlfePflili

^KjED ISS iSS iiSii Sia **

* mmaa 81! f!!fPS":l 4PBBK|P9K«t WPBiS StSSi MMH « > * ' 1 •* * : is isssii fiftiig SiafifsiIh mi attn «JPttOSg>W\J^_ mfl/L - * , s - -5 \BEEED.?,. O V*" — '•

202 75% DM/25% RB

Effluent

Technician's Notes:

Sample Time hour 1620 14:30 16:00 18:00 17:00 21:00 18:00 19:30 9:00 10:15 12:00 14:30 14:40 19:00 20:30 23:45 16:00 14:00 20:58 18:00 20.00 BapsadTime days 1.16 0.45 1.06 1.08 0.961 1.17 0.88 1.06 0.56 1.05 1.07 1.10 1.01 1.18 1.06 1.14 0.68 0.92 1.29 0.88 1.08 Mixer Speed RPM 32 31 29 31 30 30 30 31 30 30 30 30 29 0 50 26 27 26 3 30 51 Temperature *F 97.7 97.7 96.8 96.8 95 95 96.8 96.8 96.8 96.6 96.8 96.8 97.7 96.8 96.8 96.8 96.8 96.8 98.6 95 97.7 •c 36.5 36.5 36.0 36.0 35.0 35.0 36.0 36.0 36.0 36.0 36.0 36.0 36.5 36.0 36.0 36.0 36.0 36.0 37.0 35.0 36.5 Tipping Bucket 18 19 13 103 104 103 103 112 99 113 114 110 132 102 28 29 29 31 34 26 59 Gas Production mL 1656 1710 1170 9270 9360 9270 9270 10080 8910 10170 10260 9900 11880 9180 2520 2610 2610 2790 3060 2340 5310 L 1.656 1.71 1.17 9.27 9.36 9.27 9.27 10.08 8.91 10.17 10.26 9.9 11.88 9.18 2.52 2.61 2.61 2.79 3.06 2.34 5.31 Normalized gas production L/day 1.432216 3.8176744 1.1011765 8.556923 9.766957 7.945714 10.59429 9.487059 15.84 9.666535 9.562718 8.966038 11.79807 7.776 2.371765 2.298716 3.854769 3.043636 2.371582 2.670048 4.901538 Cumulative Gas production L 1916 1920 1921 1930 1939 1947 1958 1967 1983 1993 2003 2012 2023 2031 2033 2036 2040 2043 2045 2048 2053 Dige state Height cm 37 ;'V «; mm IBBf Ens 11121 2S23E SEE - , . I, • IPSE! Biooas YieW mot accurate) u^aay 0.856/121 YS^ Byre™ T.3HW904 •i12732347 7325536 7.438/8/ TrS7555? 9.178864 $m§axi 1.M5226 T.7883S4 "7W5TO41 3.813383 mm PH | 734 7.35I 7.36I 7.281 7.3 7.3 7.36 7.331 7.35] 7.34 7.36 7.39 7.331 7.361 7.37 7.39 7.41 7.38 7.35I 7.36 7.4 •••••• • . • • • • • • : w - !->•*»*, v; * n- ^ T r * MM ii mgw*?§ wm wi if itSp iMtiii iMM wmzzw imiii 81311 fw&^m SI? .. . Oxygen Demand

ssise sswiiiil it8JB§(i ftBWBI SMMW tSSSlR!ws&m iSfliiSi

-*T -k^^-4-. • w - -i.: , . yi-.-jjsg .-' , " < "J* > 5?*," ,f«5 ,' , - , , _ ,, r !iissaS SiSSs iiiSSSs SssSs 6i

Influent

320 320 3201 3201 3201 320! 320 320 320 320 320 320 320 180 180 _1B0l 1801 1801 180 180 180 180 180 180 ^iwl npRsJn iIdeal Feed Volume I Dairy Manure 75% 75% 75%| 75% 75%[ 75%J 75% 75% 75% 75% 75% 75% 75% 2$^[ 25% I 25% 1 25%| 25% 25% 25% 25% 25% 25% 25% ««• i | Hydiwij^etenttor^im^^^

Bicarbonate mg-N/L Ml. jfc» • JVHfri IvFA mg-N/L I ,| VFA® cartonate ;cp.

mgrt. I Ivs mart. I

fgftfcT |SCOP (SS x3G dilution)

3s&p ssa& -ittfe. iyflwto wsjifea Jtetih Ivss mg/L 1 [Ammonia mg/L I [Nitrate mart. I I Nit rite mart. I ITK mart- I |Totai Phosphorus mart. I

203 75% DM/25% RB

Effluent

Technician's Notes:

••• Sample Time hour 10:33 9:50 14:30 12:06 14:00 15:00 15 00 17:30 14:00 18:00 11:30 20:50 8:45 16:40 17:40 18:00 18:00 20:00 14:40 12:00 20:30 EilpMd Time days 0.61 0.97 1.19 0.90 1.08 1.04 1.00 1.10 0.85 1.17 0.73 1.39 0.50 1.33 1.04 1.01 1.00 1.08! 0.78 0.89 1.35 Mixer Speed RPM 50 50 36 0 30 26 41 0 0 0 0 46 46 46 45 45 45 30' 30 30 31 Temperature *F 97.7 98.6 96.6 98.6 95 95 96.8 96.8 95 96.8 96.8 100.4 97.7 95 96.8 96.8 96.8 95 97.7 97.7 97.7 36.5 37.0 36.0 37.0 35.0 35.0 36.0 36.0 35.0 36.0 36.0 38.0 36.5 35.0 36.0 36.0 36.0 35.0 36.5 36.5 36.5 1 *C 1 1Tipping Bucket 1 31 27 52 4 6 7 7 0 0 0 0 0 0 0 0 0 0 122 98 108 148 |Ges Production mL | 2790 2430 4596.6 353.6 530.4 618.8 618.8 0 0 0 0 0 0 0 0 0 0 10784.8 8663.2 9547.2 13083.2 1 L I 2.79 2.43 4.5966 0.3536 0.5304 0.6188 0.6188 0 0 Q 0 0 0 0 0 0 0 10.7848 8.6632 9.5472 13.0832 Normalized gas production L/day 4.602062 2.504796 3.648464 0.392889 0.49149 0.594048 0.6188 0 0 0 0 0 0 0 0 0 0 9.9552 11.1384 10.7406 9.66144 Cumulative Gas production L 2057 2060 2064 2064 2064 2065 2066 2066 2066 2066 2066 2066 2066 2066 2066 2066 2066 2076 2087 2098 2107 Dioestate Height cm jlBi :gjB3ME.f 3 -• -- 4MI gfffPlM mm 2!§S11 HiiSiSiii BE Bioaas Yield mot accurate) ug^/oay 3.580391 {.948724 !• 0.382378 0462168 3.481425 o o 8 6 0 0 0 T'/isT!?' 8.S65S43

fpH 1 7.371 7.36 7.M | 7.381| 7.37 7.38 7.4 74 7* I 7 36| 7 39I I 7 49I 74 74 ^^^51 7 36 I 73fliI 7.341

SSI BSSfS BIRR!!Kgm F#SSfl3 fSSSlllPHlill SSI^SfS RBifili BBSISfS fSBISlilitigllf Hw?ils^%jj§f^fsfgfs

t >Jlggjjjg| Is Oxygen Demand ,r r ^ ,»._w ,, »t ^ '"

Mi IHiiill ilStiff WMmm fHIffffl tiMPS iiittli IfSIIill fWi^i # |1fl|i|Sip fiWfrllf IPVPIP*® §gg?p®;f e§ |?;fl;:^f| PllPl^

, , « ^ N? » ' , _ * ••;-Wr:1^*: IPfsiilpIP*"'^ llsSIP''i::l!IS:S|lillR.:i:'cr!:-r:'^1 !liSSf:?

* '"I' ft * : ,* • " ' s ?»• nil <' , .- «;*%W **,*<>, * iV If. ~ ii_pnos£horus_ rrtfl/L i iiiiiss®! lit "><

Influent

[Water Volume mL 320 320 320 320 320 320| 320 320 320 320 320 320 320 320 3201 320 320 320 320 320 320 Feed Volume mL 180 180 180 180 180 180 180 180 180 180 180 180 180 180 180| 180 180 180 180 180 180 Ideal Feed Volume mL Dairy Manure 75* 75% 75% 75% 75% 75% I 75% 75% 75% 75% 75% 75% 75% 75% 75%[ 75% 75% 75% 75% 75% 75% Beer 25% 25% 25% 25% 25% 25% I 25% 25% 25% 25% 25% 25% 25% 25% 25% I 25% 25% 25% 25% 25% 25% Hydraulic Retention Time days ...... ,.*la '.... Acidityr'Alkalimty

rng»N/L BVFA/Bicarbonate -;sp

••vs mq/L | KflAdui^raarM^^adin^at^^ K^N^J

mo/L 1 •SSCOD (SS x20 dilution) mol 1 52333355 ft|TCO^SS*40^500di{uti0r0

m * •HTSS wBSSiSA •Ivss ma/L I 1 ••Ammonia mft • \ • „ H Nitrate mo/L I v. . hi 4.^ l •9 Nitrite mg/L I Lv S 1 * j-ft .J HTK ma/L 1 """ Hi OUM Phosphorus mg/L I 33 rr-r-;

204 II; '

Ji 31 II H m

si 75% DM/25% RB

Effluent

Technician's Notes:

Parameter 01-Jun 02-Jun 03-Jun 04-Jun 05-Jun 06-Jun 07-Jun 08-Jurt 09-Jun 10-Jun 11-Jun General 1Sample Time hour I 16:15 15:20 16:1$ 16:25 8:30 8:00 9:20 18:45 15:40 19:25 17:30 iSapeed Time deys 0.99 0.96 1.04 1.01 0.67 0.96 1.06 1.39 0.87 1.16 0.92 iMixef Speed RPM 0 56 61 61 60 60 60 60 61 61 •Temperature *F 76.1 73.4 71.6 70.7 71.6 71.6 68 68 68.9 68 68.9 24.5 23.0 22.0 21.5 22.0 22.0 20.0 2Q.0 20.5 20.0 20.5 1 *C 1Tipping Bucket 1 79 78 72 67 46 60 34 75 47 62 45 1Gas Production mL 1 6983.6 6895.2 6364.8 5922.8 4066.4 5304 3005.6 6630 4154.8 5480.8 3978 1 L : 6.9836 6.8952 6.3648 5.9228 4.0664 5.304 3.0056 6.63 4.1548 5.4808 3.978 Normalized gas production L/day 7.032436 7.169017 6.130643 5.881953 6.067996 5.416851 2.847411 4.761696 4.767261 4.740151 4.32326 Cumulative Gas production L _2334 ^ 2341 ^ 2347 2353 ^ 2359 2364 2367 2372 2377 2382 2386 B— Dioestate Height cm ^BH •1mmrnrn BUS •ail IBS Biooas Yield mot accurate} L/g^aay 4.5/6143 4.720883 4jl4294 2*152/7 3.704586 37708915 3.363484

IPH 1 r7aa 7 35 7 32 7 3 \71a\ |Bicarbonate IVFA IvFA/B

Oxygen Demand 15 day CBOO I SCOP jSS *25-50 dilution) iTCOD (SS X10Q-2G0 dilution?

Ivss" 1 Amrm>ni. |NKrat<»

mqfl- | Total Phosphorus

Influent

[Water Volume mL 320 320 320 320 320 320 320 3201 320 320) 320 Feed Volume mL 180 180 180 180 180 180 180 180 180 180 180 Meal Feed Volume mL Dairy Manure 75* 75% 75% 75% 75% 75% 75%l 75% i 75% 75%l 75% iBeer 25% 25% 25% 25% 25% % Hydijijj^etenHw^hn^^^ clay^^

{Bicarbonate mq*N/L - STI 1 VFA mg«N/L _. |VFA/Bicart)onate ;cp.

mg/L mg/L

mg/L ISCOO i'SS >30 dilution) mg/L

mgfl. Ivss mg/L 1 [Ammonia mg/L 1 AiU mgA. | [Nitrate -j&- ^ gm 1Nitrite mg/L 1 ITK mg/L 1 |Total Phosphorus mg/L 1

206 100% DM/0% RB

Effluent

Technician's Notes: ONLY No Feed DAIRY Mixer Off H-0 Only

[Sample Time [Elapsed Time days BMixer Speed BTempefature

iTippmg Bucket [Gas Production

[Normalized gas production [Cumulative Gat production iDtoestate Height •Biogas Yiekf (not accurate) IWday 3.79955871 2.307339 3.934565 3.940619 3.827612 4.206402 3.886271 3.802234 3.960516 3.928731 Acdity.'Alkahn ity

mg«N/L mo*N/L VF A/Bicarbonate

Organic Loading gvs/day

5 day CBOD SCOD (SS <25-50 dilution ICUU fbb <1002!j0 OilllllOni

Ammonia

Total Phosphorus

Influent This data is for undiluted Feed

Water Volume mL 370 370 370 370 370 370 370 370 370 370 3701 370 370 370 370 370 370 370 370 370 370 Feed Volume mL 130 130 130 130 130 130 130 130 130 130 130| 130 130 130 130 130 130 130 130 130 130 Ideal Feed Volume mL [Dairy Manure 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75%! 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% Beer 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 1 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% Hydraujj^etentjofrTjm^^^

'Bicarbonate mg*N/L I [vFA mo'N/L I i > Yi/n * Y MTI ii ny» r 11 • T jtit 11. !• ,^i«tin?Tnl I VFA/Bicarbonate . <0.5) I

4L WMVs mo/L I RSOragnic^edin^Rat^^^^^

Wjl'soil^CBOO mo/L | a •9SCOO i'SS x30 dilution) mgi I |MTCOD{S^4C^O^iju?!Gn^ ifefe'b Airi: J MPs*""" mart. I gassy &&& &s& mo/L 1 •pB Ammonia mg/L I SSft- •jpfTKN mo/L I •1Nitrate mo/L 1 ••Nitrite mo/L 1 mrK mq/L 1 ^Hfotai Phosphorus mm,, I

207 100% DM/0% RB

Technician's Notes:

[Sample Time hour 9:00 14:45 19:20 18:20 17:50 18:15 16:40 18:15 14:10 15:06 12:35 18:00 16:20 15:45 18:00 21:00 16:05 11:10 15:20 10:20 14:20 14:00 Elapsed Time days 0.81 1.24 1.19 0.96 0.98 1.02 0.93 1.07 0.83 1.04 0.90 1.23 0.93 0.98 1.09 1.13 0.80 0.80 1.17 0.79 1.17 0.99 Mixer Speed RPM 60 60 60 60 60 59 60 60 60 60 60 60 56 56 60 56 56 60 56 60 56 56 [Temperature *F 68.9 68.9 71.6 72.5 73.4 73.4 94.1 95 95 96.8 95.9 95 95 95 95 95.9 95 95.9 95 95 95 95 20.5 20.5 22.0 22.5 23.0 23.0 34.5 35.0 35.0 36.0 35.5 35.0 35.0 35.0 35.0 35.5 35.0 35.5 35.0 35.0 35.0 35.0 1 1 iTipping Bucket 1 30 56 91 81 90 69 271 224 183 237 154 150 131 23 3 0 52 53 83 61 87 83 |Gas Production mL I 2652 4950.4 8044.4 7160.4 7956 6099.6 23956.4 19801.6 16177.2 20950.8 13613.6 13260 11580.4 2033.2 265.2 0 4596.8 4685.2 7337.2 5392.4 7690.8 7337.2 L 2.652 4.9504 8.0444 7.1604 7.956 6.0996 23.9564 19.8016 16.1772 20.9508 13.6136 13.26 11.5804 2.0332 0.2652 0 4.5968 4.6852 7.3372 5.3924 7.6908 7.3372 Normalized gas production L/dav 3.264 3.9936 6.754482 7.471722 8.125277 5.995511 25.64849 18.57609 19.49386 20.16655 15.20837 10.81836 12.44461 2.083849 0.242469 0 5.781128 5.892304 6.251815 6.811453 6.592114 7.440541 Cumulative Gas production L 77 81 88 95 103 109 135 154 173 193 209 219 232 234 234 234 240 246 252 259 265 273 Dtoestste Height cm m&nbkm HMH IPIMMiP MfllffiBlllg WBMS i« liii «KIM Biogas Yield 'ftot accurate; uw«y 3.296198 4.032996 6.821113 7.545428 8.20543 6.054655 25.9015 18.75934 19.68617 20.36548 15.35839 11.00757 12.66226 2.120295 0.246709 0 5.919037 5.925665 6.287212 7.319722 7.084017 7.995753

PH 7.12 7.1 7.14 7.14 7.13 6.93 6.S1 6.86 7.43 7.45 7.47 7-5 748 7.51 7.65 7.42 7.43 7.43 7.41 7.38 7.34 Bicarbonate mo*N/L Elgin 86.67435 86.86827 VFA ma*N/L BEE3 3.67087 3.032768 0.042352 0.034912

TS mo/L vs mo/L Organic Loading flvs/elay

5 day CBOD mo/L SCOO ;SS <25-50 dilution) mo/L TCOO fSS <100-200 d-luhom

TSS mo/L vss mo/L Ammonia mg/L TKN ma/L Nitrate mo/L Nitrite mart. TK mo/L I Total Phosphorus mg/L HHt. • •:

Influent This data is for undiluted Feed

Wattr Volurrw mL 370 370 370 I 3701 370} 3701 370 3701 370 370 370 370 370 370 370 Feed Volume mL 130 130 130 13ol 1301 130 130 130 130 130 130 130 130 130 ^ 130 Ideal Feed Volume mL gggm •MMrngMaM __ ||§||§|gg §|§|||g$| Dairy Manure 75% 75% 75% 75% r 75%] 75% 75% 75% 75% 75% 75% 75% 75% 75% Beer 25% 25% 25% I 25% | 25% j 25% I 25% 25% I 25% 25% 25% 25% 25% 25% 25% I

roo-N/L IVFA mq»N/L 1 IVFA/Bicartoonate top. <05)

mo/L 10871 TS |vsl mo/L m I Oraonic Loading Rate 0.99022

5 day mo/L [SCOD .'SS r30 dilution) mo/L |TC0^^^4C^50^iUrtion^

TSS mo/L VSS mo/L Ammonia mo/L TKN mo/L Nitrate mo/L iNitrtte mo/L I ITK mon. ] |To(al Phosphorus

208 VK-t-My B'WveUy 100% DM/0% RB Effluent Withdrew Technician's Notes: Gas Sample 3 HRT mmm HQ [Sample Time [Elapsed Time day IMixer Speed [Temperature

iTipping Bucket 31 iGas Production 2740.4

[Normalised gas production L/day [Cumulative Gas production 495 [Digestate Height ~n [Biogas Yield (not accurate; L/^vj/day 0.107466 0.276961 0.206078 2.984964 4.437325 3.241287 3,668189 3.086546 3.077548 4.027288 Acidity/Alkalinity

rr»a*N/L 07106 EEHS1 mg'Na. 4.10669 Z. /34&&Z 4.510281 mn1 M VFA/Bicarbonate ( 0.046106 U.UJlZio 0.030401 0.051544 »»•«: ?i

27001 17603 IIIIil§l Bvs/day •H

5 day CBOD SCOD (So x25oQ diji/tfO mmsm TCOD fSS <100-200 dilution) 6430.47 6378.44 5785.4S

21348 24261 20077 Ammonia 60140.39 18289

<1000 <1000 Total Phosphorus

Influent This data is for undiluted Feed

[Water Volume mL 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 Feed Volume mL 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 I Ideal Feed Volume mL I Dairy Manure 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% iBeer 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% 25% •autic Retention Time Actditv'Alkahnity mg»N/L mg*NA. |VF A/Bicarbonate IOP. • wiwmm

mq/L

:KWiS S5Si:?®g BfiSli fefpsliS gfSWf ii igtipffl ISilSi!®! ilHlStS SiiJilil #§1S sBCES

•USB ESSBJ

mg/L 62.65438 1Nitrite TTWjfl- 1 113.9397 : TK mg/L iTotal Phosphorus mg/L 300.85 Ff r-i'? mn

210 100% DM/0% RB

Technician's Notes:

[Sample Time hour 16:30 17:05 16:50 16:35 16:00 12:30 16:00 20:00 17:00 17:10 17:00 15:15 18:00 20:00 9:15 9:20 19:35 15:01 15:55 15:10 16:45 19:0C [Elapsed Time dayt 1.30 1.02 0.99 0.99 0.96 0.85 1.15 1.17 0.88 1.01 0.99 0.93 1.11 1.08 0.55 1.00 1.43 0.81 1.04 0.97 1.07 1.09 Mixer Speed RPM 59 60 60 41 41 0 0 0 0 55 55 55 55 56 55 56 65 55 55 55 55 56 (Temperature *F 93.2 96.8 95 95 93.2 95 91.4 89.6 89.6 89.6 94.1 93.2 93.2 96.6 93.2 93.2 95 95 93.2 93.2 93.2 81.5 C 34.0 36.0 35.0 35.0 34.0 35.0 33.0 32.0 32.0 32.0 34.5 34.0 34.0 37.0 34.0 34.0 35.0 35.0 34.0 34.0 34.0 27.5 IiTippma Bucket ' 22 24 0 0 0 0 0 0 0 38 31 0 0 3 3 5 0 0 1 2 4 7 |Gas Production mL I 1944.6 2121.6 0 0 0 0 0 0 0 3359.2 2740.4 0 0 265.2 265.2 442 0 0 88.4 176.8 353.6 618.8 L 1.9448 2.1216 0 0 0 0 0 0 0 3.3592 2.7404 0 0 0.2652 0.2652 0.442 0 0 0.0884 0.1768 0.3536 0.6186 Normalized gas production Uday 1.4976 2.071257 0 0 0 0 0 0 0 3.336033 2.759564 0 0 0.2448 0.480362 0.440471 0 0 0.085205 0.182503 0.331716 0.56576 CumUalrve Gas production L 509 511 511 511 511 511 511 511 511 515 517 517 517 518 518 519 519 519 519 519 519 52C Dtoestate Hetaht cm M— HIBHI BBHBKBi IMiMlW jfMMUM wmmm wmmm i—inn mamma 18— 8MBP HMMIB SBHBi HK; LWday 1.618318 2.238217 0 0 0 0 0 0 0 3.604944 2.982006 0 0 0.264533 0.519083 0.475976 0 0 0.092073 0.197214 0.356455 0.611365 Biogas Yield (net accurate} BB B pH • ^ 7.08 7.09 7,09 7.09 7.02 7.1 6.99 7.04 7.05 7.07 7.06 7.06 7.02 7.05 7.05 7.06 7.05 7.04 7.01 •• • it. ~ i* v. s mmssm mmm mmssm. msmm msmm f m sm wmsm

Oxygen Demand 5 day' [SCOO ;SS *25-50 dHutian; - - „ - I I I I . V ' I '

TKN mg/L Nitrate mq/L Nitrite mat TK ma/L iTot^Phosghorus^

Influent This d«1* i* for undiluted Feed

(Water Volume mL 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 370 [Feed Volume mL 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 130 [Meal Feed Volume mL Dairy Manure 75* 75* 75* 75* 75* 75* 75* 75* 75* 75* 75* 75* 75* 75* 75% 75* 75* 75* 75* 75* 75* 75* Beer 25% 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25* 25*

EIVFA rr»a*N/L 1 KFLVFA/Bicarbonate

mg/L 1 Uvs ma/L | BJOgjmi^^adjnfi^at^^^^^

H|5>d»CB00 ma/L I ••SCOO (SS x30 dilution) ma/L 1 SJTCOOIS^O^OOdlJtion^

mg/L I Hvss ma/L 1 mq/L mq/L m9/L rnq/L mo/L [Total Phosphorus mg/L

211 100% DM/0% RB

Effluent

Technician's Notes:

Parameter 28-Sop 29-Sop 30-Soo O1-Oct 02-0ct C3-Oct G-l-Cct C5-Oct OS-Oct G7-Oct 08-0c1 OS-Oct tO-Cct 11-0ct 12-Oct 13-Oct General

iBapsedTime days •Mixer Speed iTemperature

|Tipping Bucket I Gag Production

INormalized gas production [Cumulative Gas production •Digestate Height |Bioga$ Yield (not accurate; L/flvs/day 0.361992 0.433251 0 1.901031 2.700775

mg»N/L mg-N/L

Loading Ovs/dsy Oxygen Demand

5COD -SS X2S-50 dilution rCOD (SS <100-200 d;fu!?on

rotal Phosphorus

Influent This data is For undiluted Feed

Water Volume mL 370 370 370 370 370j 370 3701 370 370 370 370 370 370 370 370 370 Feed Volume mL 130 130 130 130 130 13p| 130 130 130 130 130 130 130 130 130 Ideal Feed Volume mL HBBB 3BSI m| gggggm mum gmnggij mmgg j|g|j§|^g| nm^n 1Dairy Manure 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% 75% IBeer 25% 25% 25% 25% 25% ^25%J 25% 25% 25% 25% 25% I 25% 25% 25% 25% •auiic Retention Time

mg*N/L mg»N/L |VFA/Bicarbonate iop.

mg/t mg/L

|5 day CBOD I SCOP ?SS rZO dilution) mg/l. BTCOO (SS >400-500 dlkiSon)

•TSS mg/L Hvss mg/L RBHBjSt M Ammonia mg/L SH EBTKN mg/L ] ••Nitrate mg/L SNrtrite mg/L ^•TK mg/L I .(i ^•Total PhosDhorus mg/L

212 | Appendix E | BioTower Drawings

213 ~\

DATE UNIVERSITY OF GUELPH 1 ENGINEERING APPROVALS PLANT LONDON SCHOOL OF ENGINEERING 04/11 THORFFFLFIOUGH SUflDlNG THE ABOVE FLOOR PLAN SHOWS THE LAYOUT OF HE BIOTOWER PILOT IN LA8ATT LONDON'S DEPARTMENT DATE APPD DRAWN JOB TITLE PROPOSAL FOR WASTEWATER CHARACTERIZATION 0(1STING WASTEWATER TT^ATUEKT ROOM POP PH ADJUSTMFNT THE PLUMBING 90 STONE ROAO EAST ARRANGEMENTS FOR THIS PROJECT APE ENTAILED W AN ACCOMPANY SNG PROCESS FLOW GURPH • ONTARIO- N1G2W1 SJM 01AGRAM CANADA PROCESS SEaj&aib DWG. TITLE BIOTOWER PILOT FLOOR PLAN uooa nungs orewing CHECK

ga School of UTiLITiES IABATT BREWERIES OF CANADA DWG. NO. 3 2 4 0 o o o o 1 j LONDON - ONTARIO • CANADA J B Engineering PROJ. MGER SCALE: NTS PLANT CLASS AREA I SHEET REV^ LEGEND • COMPONENT LINE

• EUEC. +VE LEAD

- ELEC.-VE LEAD

PRODUCT STREAM

AC© PUMP 175mA 250V MAX SLOW BLOW BASE PUMP

COLUMN PUMP

RECiRC. PUMP 4.0A 250V MAX, NC RELAY 1 SLOW BLOW N BALL VALVE M CHECK VALVE 0 BUCKET 3,OA 250V MAX NC RELAY 2 SLOW BLOW ® pH PROSE

FLOOR ORAiN

RECEPTACLE

REACTOR 4 EFFLUENT

SAMPLE SAMPLE POINT 2 POINT 1

W CPVC HXhi^K1 • y{ • ya -ijKSr

AIR SUPPLY

DATE UNIVERSITY OF GU6LPH | ENGINEERING APPROVALS PLANT : LONDON SCHOOL Of SNGIN4ER94G i 04/11 THIS PROCESS FLOW DIA2RA THORNBSOUGH 9ULDMG DEPARTMENT DATE APPD BIOLOGICAL PACKED TOWER PI SO STONE fiOAC EAST ORAWN JOB TITLE I PROPOSAL FOR WASTEWATER CHARACTERIZATION 2010TOFSeRUARY»1! GyEL""H • ONTARIO- N1GZW1 SJM PROCESS SiaAcdb DWG. TITLE ! BIOTOWER PILOT PFD uooa (rungs premng UTILITIES WM School of LA8AH 8REWERKS OF CANADA DWG. NO. i 3 | 2 4 0 o o o o % LONDON - ONTARIO • CANADA B Engineering PROJ. MGER SCALE: NTS PLANT CLASS AREA I SHEET | REV^ BioTower Pilot Daily Maintenance Methodology Appendix F

| Appendix F | BioTower Maintenance and Performance Summary

216 BioTower Pilot Daily Maintenance Methodology Appendix F

The following methodology was applied for daily maintenance of the BioTower pilot:

1. Record the pH and Temperature of Reactor 4, and the equalization tank. Reactor 4 data is

displayed on the monitor labelled 'Reactor 4 pH Meter 1'. This meter is located next to the main

door for the reactor pit. The equalization tank pH displayed is displayed on the pH controller. If

the pH controller display indicated the equalization tank pH is below the set points and the pH

adjustment chemical reservoirs are full, move the pH probe up and down in the buffer tank to

dislodge stagnant pockets of water.

2. Record the pH, temperature, and dissolved oxygen level of the BioTower effluent. Collect a 500

mL effluent sample from the BioTower column (Sample Point 1) using the custom sample jar

with the two holes in the lid. Ensure the lid is secure, and there is no head space. If sample

container is not available, cover open container with parafilm. Insert a stir bar in the sample jar

and place it on a stir plate. Obtain the pH meter located on bench by reactor sample spigots.

First remove the protective probe covers, and rinse the ends with distilled water. Insert both the

pH probe and DO probe through the holes in the sample jar lid. Alternatively, pierce the top of

the parafilm with the probes. Ensure probe tips do not making contact with the stir bar. Switch

the stir plate on and set to medium speed. Turn the hand-held DO/pH meter on and press the

measurement button. The top row on the meter display indicates pH, top left hand corner show

temperature, and the bottom row indicates dissolved oxygen. Allow the reading to stabilize for 2

minutes, or until the AR symbol in the upper right corner stops flashing. Record the pH,

temperature, and DO on the sample data sheet. Remove the probes and rinse with distilled

water. Blot dry with a kim wipe. Insert DO probe back into white protective case. Place 5 drops

of pH electrode solution into black protective case for pH probe, then replace the pH probe

cover. Hang the probes vertically to store. Waste effluent sample and distilled water may be

disposed of in the floor drain by Reactor 4.

217 BioTower Pilot Daily Maintenance Methodology Appendix F

3. Record the volume of the chemical left in the 20L acid/base reservoirs using the gradations

marked on the side. Shine a flashlight through the side wall of the bucket to make the chemical

level more apparent. Replace with new, full bucket from storage if chemical is below 10L.

Personal protect equipment (PPE) including gloves, face shield, raincoat must be worn when

handling pH adjustment solutions. Replacement chemical supplies are replenished on a weekly

basis by UoG.

To make 5% H2S04 solution (1.94 N), slowly add 1 L of conc. H2S04 acid to 19 L of tap water as

the reaction is very exothermic. To make 20 L of 5% NaOH solution, add 2 L of 50% caustic

solution to 18 L of tap water. Always ensure chemical is added to water, not vice versa. This

order is important with the NaOH solution to prevent the formation of caustic aerosols, and acid

to ensure there is no boiling or splashing of chemical.

4. Record the effluent flow rate out of the red hose from the BioTower column using the stop

watch and 1L graduated cylinder. Start the stop watch the moment effluent collection begins for

a 30 second interval. Normalize to L/min (multiply by 2, divide by 1000).

5. Ensure that the two reciprocating pumps are on and operational. If the pumps are off, check

that the GFCI receptacle was not tripped. Reset if necessary. Quickly inspect all tubing for leaks

or clogs. Look for wastewater on the floor in the pilot area as an indicator of leaks. Ensure the

effluent port on the BioTower column and the supply line for Reactor 4 for the buffer tank clear.

Dislodge obstructions with a screwdriver or by backwashing with a quick burst from the hose.

Repair leaks if possible by tightening hose clamps, otherwise call emergency contacts. Pay

special attention to peristaltic pumps for acid and base dosing. If the tubing looks crimped,

move the tubing down so the pump head to makes contact with fresh tubing, and correctly

reload. Check to see that the air lines are secure as well.

218 BioTower Pilot Daily Maintenance Methodology Appendix F

6. Ensure the height of the wastewater in the buffer tank is above the minimum indicated depth.

This is the depth were the pH probe is completely submerged. If it is not, the pH controller and

column pump should automatically shut off to prevent the pH controller from incorrectly dosing

the buffer tank and wasting chemical. To remedy this situation, fully open the Reactor 4 gate

valve and let the tank fill up (takes approximately 5 minutes). Reduce the flow rate of Reactor 4

effluent by closing the gate valve until the flow rate into the buffer tank resembles that of the

flow rate out of the BioTower column. The flow rate would ideally be slightly higher. Back the

flow rate off further if the wastewater level in the buffer tank begins to trickle out the overflow

line.

7. Collect two 500mL sample every Tuesday and Friday, one sample from the BioTower Column

effluent (Sample point 1), and the other from the BioTower Column influent (Sample Point 2).

Store at 4°C in the laboratory wastewater fridge. Sample bottles are all pre-abelled. New bottles

are located in a bag under the staircase.

219 Appendix G | US Centrifuge Results

224 Biological Packed Tower Pilot Daily Maintenance Checklist | Emergenecy Contacts: Scott Massen (519 993 9024) Hamidreza Satsiai (519 588 6648) •C o s.• Emily Hahn-Tmka (519 319 8213) z a X (0.8 L/min) REACTOR 4 PARAMETERS BUFFER TANK PARAMETERS COLUMN PARAMETERS z c o s "5. Acid Vol.* Base Vol. * EfT. Flow Rata 4 o» 5 E Date Time o to» W IL) (Umin) Dissolved Oxygen 1 Temperature1 m (dd-minm-yy) (24-tiour) Reactor 4 pH1 Reactor 4 Temp.' (*C) Controllar pH 2 Column Effluent pH* •o 1 1 J (mg/L) rc> • 0 •o •c Pumps Running * BloTower Water Level Full Initial Final Mtal Final Initial Final < i 1 H- Air Line Secure Example 13:00 6.0- 10.5 -20 *6.9 - 7. f f6.5 16.5 2 20 *0.85 0.85 7.52 *2.0-4.0 15.4 V V V V s X

17-Nov-10 In Range 7.38 2.58 19.3 < - .. .. . 18-Nov-10 In Range 7.45 2.21 19.2 •i

19-Nov-10 In Range 0.9 7.36 0.38 19.4

25-Nov-10 In Range 8.35 •'v. 1.65 15.7 X V

26-Nov-10 In Range 7.72 1.35 13.8 V X Y >• %'i 27-Nov-10 In Range 7.88 3.26 14.0 V X .V .. 28-Nov-10 in Range 7.58 1.69 13.9 V X

29-Nov-10 - In Range S.1S 0.21 12.8 V X

30-Nov-10 16:18 8.18 ' 6.96 4 4 16 16 8.29 3.20 13.0 V V V V V X

01-Dec-10 10:20 7.49 7.00 3 3 13 13 0.78 7.45 1.58 19.8 V V V < V X

02-Dec-10 14:00 8.15 6.98 19.5 19.5 8 8 8.30 6.64 9.5 X V V V V X

03-Dec-10 11:40 10.14 7.05 13 13 0 0

15-Dec-10 15:30 7.01 23.0 6.91 9 9 17.5 17.5 0.82 8.38 8.93 13.2 V V v v \ X

17-Dec-10 11:56 8.42 23.0 7.11 0 20 8.5 20 N/A V V X V V X

18-Dec-10 14:30 9.95 19.0 7.53 6.5 20 0 20 0.78 7.21 5.66 10.4 V X X > X

20-DeC-10 11:25 7.94 23.1 7.00 0 20 8 8 0.61 8.55 3.78 15.7 jfft v X V V v X

. 30-Dec-10 12:45 8.54 22.4 7.11 4 20 0 20 0.94 ' V \ * \ \

31-Dec-10 12:36 7.43 23.7 6.84 20 20 10 10 0.00

220 Checklist | Biological Packed Tower Pilot Daily Maintenance Emergenecy Contacts: Scott Massen (519 993 9024) 3 Hamidreza Salslai (519 588 6648) c. u. o a. •> 1 Emily Hahn-Tmka (519 319 8213) z a• O so o» c REACTOR 4 PARAMETERS BUFFER TANK PARAMETERS COLUMN PARAMETERS z c 2 • £ • s 4 o> c s $ % E Date Tim* AcM Vol. * Base Vol. Eft. Flow Rate o 3 V) E o e c I Initial Final Inital Final Initial Final < £L z z £ m 5 s Example 13.00 6.0 - 10.5 -20 *6.9 - 7.1 16.5 16.5 2 20 *0.85 0.85 7.52 *2.0-4.0 15.4 V V •j V V V X

1• • .... 11-Jan-11 14:30 8.08 21.5 7.13 18 18 2 20 0.72 9.27 9,29 15.6 V S; V V V V

12-Jan-11 12:35 7 63 6.89 20 20 9 9 0.80 7.35 1.63 17.2

13-Jan-11 13:50 964 6.80 16 16 4 4 0.60 N/A 2.13 14.7

14-Jan-11 13:55 5.94 14 14 3 3 0.70 6.54 2.48 18.4 V V V V

'• 17-Jan-11 8:45 6.23 2 20 1 5

18-Jan-11 15:10 8.91 6.61 19 19 2 2 0.15 7.98 2.79 11.1 V V •J V V V

19-Jan-11 14;30 7.72 21.7 6.91 19 19 0 6 1.00 7.24 1.45 17.1 V V V \ \ X

20-Jari-11 15:20 6.39 20.9 7.11 18 18 1 1 0.71 6.90 2.08 14.5 V V V V V X

21-Jan-11 13:00 6.80 19.3 6.74 18 18 0.5 20 0.76 6.68 3.53 10.2 V V V V •1 V

MB III 1 • I• r I 24-Jan-11 13:00 7.48 14.8 OFF 0 - X X X X V X

25-Jan-11 13:30 6.75 20.8 6.75 0 20 0 20 1.00 7.11 1.65 12.3 X X X X V V

26-Jan-11 14:00 6.40 19.7 6.93 19 19 18 18 0.80 7.19 0.47 16.5 V V V V •J X

04-Feb-11 15:00 8.06 23.0 6.98 14 14 18 18 0.88 7.40 N/A 8.4 V \ V V V v X

05-Feb-11 13:55 9.15 16.7 6.73 14 14 17 18 0.08 0.86 7.43 N/A 13.6 X V V X V V V X

11:45 5.86 26.8 6.73 14 18 14 14 0.54 0.S6 7.40 2.61 17.6 X > X V > \ 06-Feb-11 *•

07-Feb-11 8:45 6.60 26.1 6.82 18 18 14 14 0.25 0.72 7.50 2.91 17.3 X V V X V X V X

08-Feb-11 8:30 7.58 22.2 7.22 9 20 3 20 0.77 0.77 8.16 1.59 16.2 X V V X V V X

09-Feb-11 8:45 8.95 22.2 6.86 16 16 11 20 0.40 0.85 7.41 2.08 ! 14.7 V V V V V V V X

10-Feb-11 8:45 7.16 17.3 6.82 10 18 18 18 0.50 0.81 7.42 3.08 13.8 X V V si V V V V

221 fO N> M G> NJ 0 i 1 5? -n T1 -n *n T| "T| T1 0) (D (D (D (T» CD (D (D (D (D cr cr cr cr cr cr il 7 7 7 I c?

(0 <0 s g

XI 5o H 0X 00•••• •o > o 0 1 m _

< 2 0) 5' r* -is s m o p H s 0> m (0

3 -.1

Addsd 50g of NH

Pumps Running 8

No leaks

No Clogs ft Tank Above Min. Depth ' £ 3' BioTower Water Level Pull

Air Line Secure <0.8 L/min)

500ml Sample1 Labatt WW Pilot Summary

pH HRT Setpoints (min)

7.0±0.3 35 Reactor 4 Effluent (QC Labatt) 2493 - - 492 - Recirulation 4318 ~ 18.04 634 634 N BioTower 4134 - 16.81 276 276 N 11-Jan-11 Reactor 4- Retire. Reduction -73.2% . - - . -28.8% - Retire. • BioTower Reduction 6.8% 56.4% 56.4% Total Reduction -65.8% i. : - 43.9% - 7.0±0.3 35 Reactor 4 Effluent (QC Labatt) 2536 - - 726 - Recirulation 4454 12.8 12.67 400 227 N BioTower 3980 15.1 10.77 240 186 N 14-Jan-11 Reactor 4- Retire. Reduction -75.6% •• _ ...... _ 44.9% Retire. - BioTower Reduction 10.6% -17.5% 14.9% 40.0% 17.9% Total Reduction -57.054 _ - 67.0% - 7.0±0.3 35 Reactor 4 Effluent (QC Labatt) 2291 - - 538 - Recirulation 3179 11.9 18.15 657 392 N BioTower 4619 29.9 14.67 1090 822 N 18-Jan-11 Reactor 4- Retire. Reduction -38.8% • :-. . . -22.2% ..... Retire. - BioTower Reduction -45.3% -150.8% 19.2% -85.8% -109.9% Total Reduction -101.6% - - -102.5% • - 7.0±0.3 35 Reactor 4 Effluent (QC Labatt) 3057 - - 476 ~ Recirulation 6487 - - ~ - N BioTower 5951 - - - - N 21-Jan-11 Reactor 4- Retire. Reduction -112.2% — #VALUEI - Retire. - BioTower Reduction 8.3% #VALUEI VVALUE! 0VALUE! Total Reduction -94.7% • - - MALUEi - 7.0±0.3 35 Reactor 4 Effluent (QC Labatt) 2947 - - 232 - Recirulation 2972 - 14.78 384 198 Y BioTower 2808 - 14.53 187 166 Y 06-Feb-11 Reactor 4- Retire. Reduction -0.8% - -65.7% _ Retire. - BioTower Reduction 5.5% .. 1.7% 51.5% 16.0% Total Reduction 4,7% . V.' «. . • - : 19M% . » 7.0±0.3 35 Reactor 4 Effluent (QC Labatt) 2621 - - 886 - Reactor 4 (Grab) 2699 26.5 16.88 321 223 N Recirulation 4402 39.0 23.72 294 241 Y 10-Feb-11 BioTower 4458 34.4 14.65 284 237 Y Reactor 4- Retire. Reduction •68.0% .. 66.9% Retire. - BioTower Reduction -1.3% 11.9% 38.2% • 3.3% 1.4% ; Total Reduction -70.1% - - 67.9% 7.0±0.3 35 Reactor 4 Effluent (QC Labatt) 2775 - - 631 - Reactor 4 (1hr. Composite) 4315 18.0 20.01 515 319 Y Recirulation 4026 14.6 19.36 374 314 N 13-Feb-11 BioTower 3625 12.3 13.19 186 186 Y Reactor 4- Retire. Reduction -45.1% 40.7% _ Retire. -BioTowerReduction 10.0% 15.5% 319% 50.3% 40.8% Total Reduction -30.6% • • 70.8% r : . Reactor 4 Effluent (QC Labatt) - - Reactor 4 3535 - - 399 309 Recirulation 5489 - - 284 217 BioTower (QC Labatt) 4091 - - - - 17-Feb-11 BioTower 4404 - _ . 236 203 Reactor 4- Retire. Reduction #DIV/0! •:- : —: : #D)V«)! Retire. - BioTower Reduction 19.8% tVALUE! nmjuB 6.5% Total Reduction mrnt "-r} UDIV/01 is-::'--' Reactor 4 Effluent (QC Labatt) - - - Reactor 4 3308 - - 132 129 Recirulation 4018 - - 269 234 21-Feb-11 BioTower 4158 - - 275 262 Reactor 4- Retire. Reduction #DIV/0l #CHV/0l - Retire. -BioTower Reduction -•u-asx. mmum MALUE! -12.2% Total Reduction #Divm UDIV/01 • -

223 | Appendix G | US Centrifuge Results

224 BOD5 • • 1 Blank 0 n/a n/a n/a 0 8.13 19.6 7.95 19.5 0.18 2.21 n/a Seed n/9; -:y: rj/a"-:.;;: ri/s 0.00 8.09 19.5 6.59 19.4 1.5 18.54141 n/a n/a n/a n/a 0.00 8.09 19.6 6.25 19.4 1.84 22.74413 n/a | Average Seed: | 1.67 20.64 n/a

A 5 2500 4 625 0.48 8.2 19.3 1.21 20 6.99 85.2439 3325 Bs-? 2000 4 SOO 0.60 8.04 19.5 0.44 20 7.6 94.52736 2965 C 5 1500 4 375 0.80 8.03 19.5 0.03 20 8 99.6264 2373.75 TOTAL SUSPENDED SOLIDS

I lO-Nov-lO 11A^^ 50 3.65005 3.708 ^3^663[ 1159 834 1 11B 50 3.42528 3.48838 3.45142| 1262 739.2 I

Comparison

Removal Sample Identification BOD5 TSS Removal mg/L mg/L Raw Brewery Effluent 3325 1210 University of Guelph (see above) Centiifuged 3000 9.77% 280 76.86% Heritage Environmental (see COA) Centrifuged + Polymer 2400 27.82% 400 66.94% Heritage Environmental (see COA)

225 Discussion

Grab samples were collected from the Reator 4 sample spigot at labatt, London on 12-0ct-10, and not sampled until one month aftwerward (between 4-Nov-10 and 12-Nov-10). US Centrifuge did not provide Heritage Environmental sufficent quantity to complete the solids tests (only 50 ml, requested that they give 1 L). Chris Boyle said that the small volume was because the samples were decanted, which was not what we specified. As a result, there was not enough sample for total phosphorous analysis.

The BOD5 results of 9.77% for 'centrifuged effluent' and 27.82% removal for 'centrifuged effluent with polymer1 are reasonable. Most of the BOD is soluble, thus solids removal would have little effect. This result is consistent with the Ocean Chemcials test in November 2009 where polyacrylomide was dosed with the Reactor 4 composite, and the coagulated solids were filtered out through a paper towel; this only resulted in 22.6% BOD5 removal. The descrepancy between the BOD results of the treated can be attributed to laboratory inexperience with this particular waste stream. Brewery effluent is stronger than typical wastewater streams, and laboratories commonly under-estimate dilution factors

US Centrifuge reported on 20-0ct-10 a 87.5% TSS removal. Independent tests indicate a maximum 77% removal rate. This may be attributed to the small volume of sample that was dropped off to the lab for analysis (only 50ml). Decanting is a solids removal technique, thus the documented solids removal efficiency an overestimate. Solids removal results are also inconsistent as one would expect more removal with polymer addition. The Ocean Chemicals experiment in November 2009 removed 94.6% - however those solids were filtered off. The average pore size of the paper towel was unknown.

Further discussion pending the reciept of the methodology applied to the treated effluent from US Centrifuge.

Heritage Environmental Note: Amount used for A898359 for TSS was 25 ml; A898360 was 13 ml. "Limited sample" was noted on the benchsheet. BOD result for both samples was taken from the 1:10 and 1:100 dilutions combined.

226