SOUR Test Standard Operating Procedure

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SOUR Test SOP template

Instructions

Ohio EPA has created this Specific Oxygen Uptake Rate (SOUR) Test Standard Operating
Procedure (SOP) template to help you develop a SOUR test SOP. Use of this template will help
ensure that your SOP addresses all the elements required in your NPDES permit, the Ohio
Administrative Code, and the Code of Federal Regulations.

Tips for completing this template:

 This template is designed for use by all wastewater treatment plants meeting vector
attraction reduction for biosolids using the SOUR test option, VAR-4.
 This template was developed in Microsoft Word so that you can easily add tables and
additional text.
 To make it easier to complete, this template generally uses BLUE TEXT where the
operator is expected to enter information.

While Ohio EPA has made every effort to ensure the accuracy of all instructions and guidance
contained in this template, the actual obligations of regulated facilities are determined by the
relevant provisions of the permit, not by this template. Ohio EPA welcomes comments on this
template at any time and will consider those comments in any future revision of this document.
This template was updated 11/2023.
Revision Date Revision Notes Document Number
12/2023 Created. 1.00

Comment:
 This instructions page can be deleted when you save your version of the SOUR test SOP.
 This Standard Operating Procedure is intended to be useful for YOUR specific facility. Please update
BLUE TEXT to information specific to your facility and add or rearrange text as needed.

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 SOUR Test Standard Operating Procedure
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SOUR Test

Standard Operating Procedure

Vector Attraction Reduction Option #4

For Biosolids Treatment

INSERT FACILITY NAME

NDPES #INSERT NPDES NUMBER

INSERT FACILITY STREET ADDRESS

INSERT FACILITY STREET ADDRESS LINE 2

Version Date Significant Changes Responsible Person

SOUR001 4/17/2023 Original version INSERT AUTHOR NAME

Contents
1. Background...............................................................................................................................................................4

1.1 Applicability.........................................................................................................................................................4

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2. Conducting a SOUR Test.........................................................................................................................................5

2.1 SOUR Test Summary............................................................................................................................................5

2.2 Equipment............................................................................................................................................................5
2.3 Sample Collection................................................................................................................................................5
2.4 Total Solids...........................................................................................................................................................6
2.5 Dissolved Oxygen (DO) Procedure.....................................................................................................................7
2.6 Calculations.........................................................................................................................................................7

2.6.1 oxygen consumption rate..............................................................................................................................7

2.6.2 specific oxygen consumption rate.................................................................................................................8
2.6.3 temperature correction.................................................................................................................................8

2.7 Documentation.....................................................................................................................................................9
2.8 Calibration and other QA/QC..............................................................................................................................9

3. Frequently Asked Questions..................................................................................................................................10

4. Additional Resources..............................................................................................................................................12

Ohio EPA Contacts..................................................................................................................................................13

Appendix 1. Sample Frequency Table......................................................................................................................14
Appendix 2. SOUR Calculator Worksheet...............................................................................................................15
Appendix 3. Definitions............................................................................................................................................16

Comment:
 After editing SOP content, please update the Table of Contents above by clicking anywhere on the table and
selecting “Update Table.”

1. Background
Before sewage sludge can be beneficially used via land application, it must be treated to meet
Class B Biosolids standards for:
 screening
 metals concentrations
 pathogen reduction
 vector attraction reduction

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The Specific Oxygen Uptake Rate (SOUR) test is one of 10 options that can be selected to meet
vector attraction reduction for Class B biosolids in Ohio.

The SOUR test is a measure of oxygen consumption rate, measured in milligrams of oxygen per
hour per gram of total solids (dry weight basis). In other words, the SOUR test shows how fast
the bugs are consuming food. A low SOUR indicates that the sludge is stabilized.

To achieve this option, the SOUR result must be equal to or less than 1.5 (mg/g)/h.

1.1 Applicability

This test is valid for aerobically digested sludges only.

Other required conditions:

 sludge in the digesters has a percent solids between 0.5% and 2%

 sludge has not been deprived of oxygen for more than 2 hours before the test
 The temperature of the sludge in the digester is between 10°C and 30°C

This test is not appropriate for:

 sludge that has been dewatered

 composted sludge
 anaerobic sludge (including sludge from an anaerobic lagoon)

 heat or air dried sludge

Comment:
If the sludge at your facility does not meet the required conditions for the SOUR test, there are other ways that
vector attraction reduction can be achieved. Please see the Additional Resources section to contact your local
2. Conducting a SOUR Test
Ohio EPA biosolids district representative or review the Pathogens and Vector Attraction in Sewage Sludge
guidance document.

2.1 SOUR Test Summary

The sludge sample is oxygenated. Temperature and dissolved oxygen (DO) are measured every
minute for 15 minutes. Total solids of an aliquot are determined. The SOUR is then calculated
from test results.

This SOP follows Standard Methods 2710 Part B, Oxygen-Consumption Rate as required by
OAC 3745-40-09(A) and 40 CFR Part 503.8.

2.2 Equipment

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 500 mL container
 300 mL BOD bottle
 DO probe (calibrated) or respirometer (calibrated)
o If a respirometer is used, follow manufacturer instructions instead of the DO
measurement procedures in this SOP
 Magnetic stirrer and stir bar
 Stopwatch
 Thermometer (if not incorporated into the DO meter)
 Equipment to measure total solids, if conducted on-site

2.3 Sample Collection

Location: Describe sample location. For example – east side of aerobic digester 2, close to the
mixer.

Collection Methods:

 The digester should be well mixed and aerated prior to sample collection.
 Collect several grab samples over about 5 minutes and composite the samples.
 Composite sample size should be at least 500 mL.
 Once collected, the sample should be kept under aeration and at the same temperature as
in the digester. Larger sample volumes or insulated containers can help maintain sample
temperature.

Timing and Frequency:

 The sample shall be collected no more than 15 minutes before starting the SOUR test.
 The test shall be conducted several days before biosolids are removed from the facility
for land application.
 Due to naturally high variability in SOUR conditions, the arithmetic mean of seven
successive SOUR tests over a 2-3 week period shall be used to indicate average
performance.
 The test should be completed FILL IN MINIMUM NUMBER OF TESTS PER YEAR
times per year. If using this test to meet regulatory requirements for vector attraction
reduction in biosolids, this test shall be conducted at the same frequency listed for metals
and nutrients in the 581 monitoring table of the NPDES Permit.
o For example, if only one set of SOUR tests per year is required, start performing
the tests 2-3 weeks before land application is planned to begin.

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2.4 Total Solids

Total solids should be determined by following Standard Method 2540 Part G. Aliquot size is
typically 25mL.

Total solids should be reported in both g/L (dry weight) and % total solids.

 g/L will be used in the SOUR calculations.

 If % total solids is between 0.5% and 2.0%, the test is valid.

Comment:
 Close readers may notice that Standard Method 2710B indicates that volatile solids be determined, instead
of total solids as in this SOP. US EPA has provided guidance that total solids should be used when
demonstrating vector attraction reduction via the SOUR test.

2.5 Dissolved Oxygen (DO) Procedure

Before beginning this DO procedure, the DO sensor should be calibrated. This DO procedure
should be started within 15 minutes of sample collection.

1. Fill a 500mL bottle ¾ of the way full of sludge.

2. Increase the DO concentration by either shaking the bottle vigorously for 5 minutes or
bubbling air or oxygen through it.
3. Place a magnetic stir bar in a clean 300 mL BOD bottle.
4. Pour sludge from the oxygenated 500mL bottle into the 300 mL BOD bottle. Fill the
bottle completely to the top.
5. Immediately insert the DO probe into the BOD bottle, making sure that the probe tightly
seals the sludge from the atmosphere.
6. Activate the magnetic stirrer. Note that adequate mixing is essential.
7. Wait about 30 seconds for the DO readings to stabilize.
8. Record an initial temperature and DO.
a. If the DO is less than 4 mg/L at the start of the test, start the test over. Shake the
bottle more vigorously for a longer period of time and repeat the test beginning
with a higher DO level.
9. Start the timing device. Every minute for the next 15 minutes, record the DO and
temperature.
a. If the DO probe does not include an integrated thermometer, record the
temperature only at the beginning and end of the test.

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2.6 Calculations

Ohio EPA recommends using Ohio EPA’s SOUR Calculator to automatically calculate the
SOUR (see Appendix 2). SOUR can be calculated manually in three steps:

2.6.1 oxygen consumption rate

Plot observed DO readings (mg/L) versus time (minutes) on arithmetic graph paper or in an
Excel worksheet and determine the slope of the line of best fit. The slope is the oxygen
consumption rate in milligrams per liter per minute (mg/L)/min. In the following example, the
oxygen consumption rate is 0.0856 (mg/L)/min.

8.5

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f(x) = − 0.0855882352941177 x + 8.11691176470588
DO (mg/L)

7.5

6.5

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0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Time (min)

2.6.2 specific oxygen consumption rate

Use the total solids (g/L) measured from Section 2.4 and the oxygen consumption rate
((mg/L)/min) from Section 2.6.1 in the following equation to determine SOUR:

SOUR T°C (mg/g)/h =

2.6.3 temperature correction

Oxygen uptake rates are affected by temperature. SOUR results must be adjusted by the average
temperature that the test was conducted at using the following formula:

Where T is the average temperature of the sewage

sludge during the SOUR test; and 7
Where θ = 1.05 if T > 20°C or
Where θ = 1.07 if T < 20°C
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Results

If the SOUR is less than or equal to 1.5 (mg/g)/h, the SOUR test has been passed.

If the SOUR is greater than 1.5 (mg/g)/h, the SOUR test has been failed. This sludge CANNOT
be beneficially used until a vector attraction reduction option is achieved.

2.7 Documentation

Remember - if there are no records of something, then it did not occur!

This SOP, when filled in for the permitted facility, meets the requirements of 3745-40-04(C)(4)
(b) and 3745-40-09 (C)(3)(c) for a standard operating procedure for SOUR testing.

Records, including calibration records, should be kept for at least 5 years or in accordance with
the permittee’s record policy, whichever is greater.

The following are required to be included on Annual Sludge Reports if using VAR-4 (SOUR
test):

 Dissolved oxygen readings taken every minute over a fifteen-minute period

 Calculations and temperature records showing that each test was corrected to 20°C
 Total solids for the sewage sludge sample
 The SOUR calculations for each test. For example, if the monitoring frequency required
is once per year, a minimum of seven SOUR tests and the arithmetic mean of the seven
tests shall be included in the ASR.
 Note that if the Ohio EPA SOUR Calculator is used, a pdf of the completed worksheet
fulfills all these Annual Sludge Report requirements.

2.8 Calibration and other QA/QC

Calibrate the DO probe according to Standard Methods 4500-O Part G. This will likely instruct
the user to follow the manufacturer’s procedures exactly. If a zero DO calibration is
recommended in the manufacturer’s calibration guide, do that as well.

Duplicates are required for both total solids and DO measurements.

 Total solids: 10% of samples should be checked via a duplicate.

 DO: due to high variability, the SOUR test should be conducted 7 times over 2-3 weeks.

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Interference with the DO probe may be caused by:

 Lower cell sensitivity due to prolonged use of membrane electrodes in waters containing
such gases as hydrogen sulfide (H 2S). Response: frequently change the electrolyte and
membrane and calibrate the membrane electrode.
 Membrane fouling due to bacterial or algal growth, soap, foam polymers, grease, and
non-aqueous-phase liquids. Response: replace the membrane.

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3. Frequently Asked Questions

1. What should I do if DO drops below 1 mg/L during the test?

The sludge has become oxygen-limiting and the SOUR test is no longer valid. Start the test
over at Step 2. Shake the bottle or bubble in air longer to start at a higher DO level, ideally
4.0 mg/L or above.

2. How long can I wait before starting the test?

The SOUR test should be started within 15 minutes of collecting the sample. If the test
cannot be started within 15 minutes, the sample may be tested within 2 hours of collection if
the sample is maintained at digester temperature and oxygenated. If these conditions are not
maintained, a new sample should be collected.

3. How often do I need to do the SOUR test?

Your facility should conduct a set of SOUR tests at least as often as metals are monitored in
sludge per the 581 monitoring table in the NPDES permit. Because SOUR can have high
variability, the SOUR test should be conducted seven times over the course of 2-3 weeks to
obtain an average SOUR value.

4. Can I land-apply sewage sludge that did not pass the SOUR test?

Generally, no. Sewage sludge must meet a vector attraction reduction option from the list at
OAC 3745-40-04 before it can be land-applied as Class B biosolids. If the SOUR test is not
passed, a different vector attraction reduction option must be used, such as immediate
incorporation or injection. Note that if the sewage sludge has been tested for SOUR seven
times in succession and the average value is below 1.5 (mg/g)/h, the sewage sludge has
passed the SOUR test.

5. My DO probe can autocorrect DO readings to 20°C. Do I still need to do a temperature

correction?

Yes. The SOUR is designed to capture performance of the digester and adjust that to
uniform temperature standards. Because the uptake rate changes based on temperature, you
could never know the “true” performance of your digester at its operating temperature if an
auto-adjusted DO value is used.

6. What if my sludge is not between 0.5% and 2% total solids?

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If the solids content is greater than 2% but less than 4%, dilute the sample to 2% solids in
the BOD bottle using decant from the digester or deionized water. If the solids content is
less than 0.5%, sludge can be thickened to less than 2%, provided that thickening takes
place over fewer than 2 hours and digester temperature is maintained. If the solids content is
greater than 4%, the SOUR test is not appropriate.

7. What if sludge in my aerobic digester is not between 10°C and 30°C?

Sludges outside of this temperature range do not meet test requirements. Because
temperature changes in digested sludge cause short-term instabilities in oxygen uptake rate,
sludge samples may not be heated or cooled to meet the temperature requirement.

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4. Additional Resources
Ohio EPA biosolids SOUR SOP requirements, Ohio Administrative Code (OAC) 3745-40-09(C)
(3)(c) https://codes.ohio.gov/ohio-administrative-code/rule-3745-40-09

Ohio EPA biosolids webpage

https://epa.ohio.gov/divisions-and-offices/surface-water/permitting/biosolids-program

Pathogens and Vector Attraction in Sewage Sludge. EPA/600/R-22/194, January 2023.

https://cfpub.epa.gov/si/si_public_record_Report.cfm?dirEntryId=356976&Lab=CESER

SM 2710B. Oxygen-Consumption Rate in Standard Methods for the Examination of Water and
Wastewater, 23nd Edition

SM 2540G. Total, Fixed, and Volatile Solids in Solid and Semisolid Samples in Standard
Methods for the Examination of Water and Wastewater, 23nd Edition

SM 4500-O Oxygen (Dissolved) in Standard Methods for the Examination of Water and
Wastewater, 23nd Edition

Specific Oxygen Uptake Rate Calculator and Instructions https://epa.ohio.gov/divisions-and-

offices/surface-water/permitting/biosolids-program

US EPA biosolids page https://www.epa.gov/biosolids

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Ohio EPA Contacts

Please contact your district biosolids coordinator with questions or comments. Up-to-date contact
information is available at the Ohio EPA Biosolids webpage at https://epa.ohio.gov/divisions-
and-offices/surface-water/permitting/biosolids-program

Program Supervisor Northeast District (NEDO)

Betsy Sheerin John Micklewright
Betsy.Sheerin@epa.ohio.gov John.Micklewright@epa.ohio.gov
614-644-2150 330-963-1285
Northwest District (NWDO) Central and Southeast Districts
Catherine Robertson (CDO and SEDO)
Catherine.Robertson@epa.ohio.gov Hannah Hurdley
419-352-8461 Hannah.Hurdley@epa.ohio.gov
614-644-2033
Southwest District (SWDO)
Kristen Diesburg Whitney White
Kristen.Diesburg@epa.ohio.gov Whitney.White@epa.ohio.gov
937-285-6442 614-705-1141

Appendix 1. Sample Frequency Table

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Amount of Sewage Sludge Generated

Minimum frequency of required monitoring
(dry U.S. tons per calendar year)

Greater than zero (0) but less than three hundred

Once per year
twenty (320)

Greater than or equal to three hundred twenty (320)

Once per quarter
but less than one thousand six hundred fifty (1,650)

Greater than or equal to one thousand six hundred

fifty (1,650) but less than sixteen thousand five Once every 2 months
hundred (16,500)

Greater than or equal to sixteen thousand five

Once per month
hundred (16,500)

Minimum frequencies to demonstrate vector attraction reduction. This table can be found at
OAC Rule 3745-40-09 https://codes.ohio.gov/ohio-administrative-code/rule-3745-40-09

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Appendix 2. SOUR Calculator Worksheet

If completed, this excel-based worksheet for each test is the only supporting information that needs to be
attached to Annual Sludge Reports to document VAR-4. This sheet can be printed out and filled in while
conducting the test. The calculator with instructions can be found on the Ohio EPA Biosolids webpage at
https://epa.ohio.gov/divisions-and-offices/surface-water/permitting/biosolids-program

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Appendix 3. Definitions

Aerobic digestion is the biochemical decomposition of organic matter in sewage sludge into carbon
dioxide and water by microorganisms in the presence of oxygen.

Beneficial Use means the placement of class B or bulk exceptional quality biosolids onto a beneficial use
site through the spraying or spreading of biosolids onto the surface of the beneficial use site, the injection
of biosolids below the surface of the beneficial use site, the incorporation of biosolids into the soil, for the
purpose of providing an agronomic benefit, or the distribution of exceptional quality biosolids that do not
satisfy the definition of bulk exceptional quality biosolids.

Biosolids means sewage sludge or mixtures containing sewage sludge that have been treated for
beneficial use.

Composite sample means a sample that is comprised of a minimum of six grab samples, collected at
such times and locations and in such a fashion, as to be representative of the facility's sewage sludge or
biosolids.

Oxygen consumption rate (oxygen uptake rate, OUR) is the rate of oxygen usage by microorganisms in
a biological system.

Oxygen-limiting means the level of dissolved oxygen is rate-limiting when it is the factor that controls
the rate of oxygen uptake by the microorganisms in a sample.

Sewage sludge is solid, semi-solid, or liquid residue generated during the treatment of domestic sewage
in a treatment works. Sewage sludge includes, but is not limited to, domestic septage; scum or solids
removed in primary, secondary, or advanced wastewater treatment processes; and a material derived from
sewage sludge. Sewage sludge does not include ash generated during the firing of sewage sludge in a
sewage sludge incinerator or grit and screenings generated during preliminary treatment of domestic
sewage in a treatment works.

Specific oxygen uptake rate (SOUR) is the mass of oxygen consumed per unit time per unit mass of
total solids (dry weight basis) in the sewage sludge.

Total solids are the materials in sewage sludge that remain as residue when the sewage sludge is dried at
103°C to 105°C accordance with part 2540G of the "Standard Methods for the Examination of Water and
Wastewater.”

Vector attraction is the characteristic of sewage sludge that attracts rodents, flies, mosquitos, or other
organisms capable of transporting infectious agents.

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