Attachment_B_SOW.DOCX

Attachment B

PRODUCT MANAGER – SOLDIER CLOTHING & INDIVIDUAL EQUIPMENT

SOLDIER ENHANCEMENT PROGRAM

MO

UNTAIN SAFETY RESEARCH (MSR) GUARDIAN HAND PUMP

PURIFIER SPECIFIC TEST PLAN,

NSF PROTOCOL P248, APPENDIX B

DECEMBER 2018

1. PURPOSE. The purpose of this testing is to evaluate the microbiological reduction performance of the MSR Guardian hand pump. This Purifier Specific Test Plan (PSTP) details testing of the device, including the technology and design, the challenge water chemical and microbial composition, the exact laboratory procedures planned, and follows NSF Protocol P248, Military Operations Microbiological Water Purifiers, Appendix B (2012).

2. REFERENCES. Appendix A contains a list of references used in developing this plan.

3. BACKGROUND. This project is directed by Product Manager – Soldier Clothing and Individual Equipment (PM-SCIE) under the Soldier Enhancement Program (SEP). The U.S. Army Public Health Center (APHC) will support this testing as the Government Review Agency (GRA). The GRA will evaluate the final laboratory reports for device performance and comparison to protocol requirements.

4. DEVICE INFORMATION.

4.1 Device Identification. The MSR Guardian hand pump (Figure 1) is a compact water purifier using a hollow-fiber ultrafiltration membrane for waterborne pathogen reduction. Is was designed to supply microbiologically-safe water to a 4-person military team. Manufacturer instructions can be found in Appendix B.

4.2 Treatment. The device uses a hand pump to draw water from the source, through the membrane and into a product water container of the user’s choice. Waterborne pathogen reduction is achieved through mechanical size exclusion only. The device does not contain adsorptive technologies.

Figure 1. MSR Guardian Hand Pump Table 1. Device Information.

5. TESTING SPECIFICATIONS.

5.1 Test Event. This testing will take place over a 10-day long test event, simultaneously challenging 3 replicate devices.

5.2 Test Waters. Test waters shall follow NSF Protocol P248 and are shown below in Table 2. Test water shall be made fresh daily using laboratory-grade reverse osmosis/de-ionized (RO/DI) water as the base then adjusted as necessary to meet Protocol specifications. Adjustments will be made using the materials stated in Section 3.4 of NSF Protocol P248 and shown in Table 2 below. All test waters shall be sufficiently mixed during the test to prevent particulate settling.

5.3 Flowrate and Capacity. The testing flow rate will approximate 2.5 L/min by pumping the device 60-80 strokes/min by hand. A mechanical arm will not be used to pump the device. This stroke rate will be maintained throughout the test to the best ability of the technician. As the device clogs the pumping effort will increase until 60 psi is reached due to membrane clogging and water squirts from the pressure relief valve. At that point, the membrane is fouled and requires replacement the membrane cartridge will be replaced and testing will continue.. The membrane shall not be replaced during a sampling event.

Table 2. Test Water Characteristics.

General Filter Challenge Adjustment

5.4 Analytical Procedures. Following daily test water preparation the water quality will be analyzed per the methods shown in Table 3.

Table 3. Analytical Procedures.

5.5 Microbiological Challenges. In accordance with the requirements of Table 2-1 of NSF Protocol P248, the pathogens used to challenge the device and the required reductions are shown in Table 4. During testing, the concentration of each organism introduced to the water will vary according to the following sequence:

5.5.1 Test waters will meet the specification shown in Table 2. It is anticipated that the device capacity will be about 600 L, resulting in a daily production rate of 75 L. The first 150 L (2 days) tested will meet General Test Water (GTW) specifications then the next 150 L tested will meet Filter Challenge Test Water (CTW) specifications. Once 300 L is produced GTW and CTW will alternate each day, processing 75 L daily.

5.5.2 Test waters shall have microbiological addition according to the concentrations shown in Table 4. Since MS2 and fr must be kept separate to allow for individual quantification, the organisms shall be serially challenged at each sample point using the following pairings:

Challenge A – MS2 and Cryptosporidium parvum combined Challenge B – Escherichia coli and fr combined

5.5.3 At each sample point (except first draw and post stagnation) 2 L of Challenge A test water shall be processed to waste followed by 3 L of Challenge A to be collected then aliquots drawn for analysis. One influent sample shall be collected as representative of Challenge A covering all replicate devices. These procedures will be repeated for Challenge B test water. Additionally, the turbidity of each device effluent shall be measured during the 2 L waste of Challenge B. The first draw sample will follow the above procedures except that 3 L will be collected for analysis without discarding the first 2 liters. Post stagnation sampling is described below in Section 6.10.

Table 4. Microbiological Concentration and Required Reductions.

1 If target influent concentrations are not achieved, the influent concentration must be sufficient to demonstrate required reduction to be considered as a data point.

5.6 Acceptable Reduction Deviation. In accordance with Section 3.8.3.2 of NSF P248 (reference 1), the water purifier must continuously meet or exceed the log reduction requirements shown in Table 4, except for the following acceptable allowance. Up to 10% of influent/effluent sample pairs may vary from the reductions required in Table 4 by:

This allowance applies to 6 sampling points (stagnation is excluded), resulting in 18 sample pairs per organisms across the 3 replicate devices. It is acceptable that 1 sample pair for each organism fail to meet the required reduction, but be within the allowance stated above. In all cases, the geometric mean of all microbiological reductions must meet or exceed the requirements of Table 4. Allowances beyond those stated due to identified issues are evaluated on a case-by-case basis.

5.7 Microbiological Sample Processing and Enumeration.

5.7.1 All microbiological samples shall be analyzed at the appropriate number of dilutions to cover the range of organism counts possible. Each dilution shall be enumerated in triplicate. The report will show the three enumerations selected from the appropriate dilution for influent and effluent samples. The geometric mean of the three enumerations will be used to calculate log10 reductions. For E.coli analysis, the IDEXX Colilert Quanti-tray 2000 method shall be used. For this method, analyzing an un-diluted and a 1000x diluted effluent sample (both in triplicate) is considered sufficient when the influent bacteria count is between 1 x 107 - 1 x 108 CFU/100 mL. This will provide a reportable count for reductions greater than 1.6 log10.

5.7.2 Production and assay procedures for the microbial challenges shall be conducted according to the following methods:

1. Escherichia coli (ATCC 11229) shall be prepared using the method specified in Asburg, E.D., Methods of Testing Sanitizers and Bacteriostatic Substances; in Disinfection, Sterilization, and Preservation (Seymour S. Block, ed., 1983), or equivalent. The samples will be assayed using the IDEXX Colilert Quanti-Tray 2000 Method for detection of total coliforms and E. coli. This method utilizes the technology of the Enzyme Substrate Test and is based on Standard Methods for the Examination of Water and Wastewater: Section 9221 - Multiple- Tube Fermentation Technique for Members of the Coliform Group (20th Edition). MPN will be determined from the IDEXX-provided table and reported without confidence intervals.

1. Coliphage MS-2 shall be prepared and assayed using the method in Annex A, Section A.8.2.2 of NSF/ANSI 55: Ultraviolet Microbiological Water Treatment Systems; or equivalent method.

1. Coliphage fr shall be prepared and assayed using the method in Annex A, Section A.8.2.2 of NSF/ANSI 55: Ultraviolet Microbiological Water Treatment Systems with the modification of E. coli ATCC 19853 as host in place of E. coli 15597; or equivalent method.

1. Cryptosporidium parvum oocysts will be ordered prepared from Bunch Grass Farms or an alternate approved vendor. Samples will be enumerated using the test method in Annex A of NSF/ANSI 53: Drinking Water Treatment Units – Health Effects (2005).

5.8 Reporting. A final report shall be prepared detailing the testing procedures, data collection, data analysis, quality assurance and quality control (QA/QC), deviations from this test plan or NSF Protocol P248 (if any), and if the laboratory has reason to question the validity of the reported data.

5.9 Good Laboratory Practices. Testing should adhere to ISO 17025 and follow generally-recognized good laboratory practices.

5.10 Quality Assurance and Quality Control. Laboratory QA/QC procedures will be followed. Specific QA/QC samples shall include (not all inclusive):

5.10.1 A daily positive and negative control for each microorganism.

5.10.2 A Pre-test Contamination Check as described below in Section 6.5.

6. TESTING PROCEDURES. The following describes discrete testing procedures that will be followed during testing and may not be shown in the order in which they will be performed.

6.1 Device Conditioning and Pre-Test Contamination Check. Perform device conditioning and a pre-test contamination check to ensure the device is free of environmental contamination. Results from this test must be received prior to Test Day 1. Using RO/DI lab water, pump 2 L through the device to waste then collect the next 3 L, aliquot, and analyze for MS2, Cryptosporidium, E.coli, and fr. Analyze influent for the same organisms. Analyze all samples in triplicate without dilution. This water does not count towards the testing capacity.

**If results for any organism are positive contact the test sponsor before proceeding to Test Day 1.

6.2 Device Integrity Test. Immediately after device conditioning and at the end of testing on the final day check each filter for membrane integrity. Follow manufacturer instructions (summarized below).

1. Fill a clear wide-mouth bottle 1/3 with RO/DI water and attach to the bottom of the device.

2. Disconnect the dual hose from the device.

3. Turn the device upside down and orient the inlet/outlet barbs away from you and over a sink.

4. Pump the device 5-10 strokes until only air exists the outlet barb.

5. Place and securely hold a finger over the outlet barb (smaller hole) while pumping the device 5-10 complete strokes (1 complete stroke is pump handle out and in).

6. Continue to hold finger over outlet barb and observe for 30 seconds to see if air bubbles form in the bottle. If any bubbles are noticeable pump the device 5 more strokes and continue to observe the bottle for another 30 seconds. If bubbles continue to form the device is compromised.

**If any devices fail the integrity check place them aside for return to the manufacturer. Select a spare device and repeat Sections 6.1 Device Conditioning and Pre-Test contamination check and 6.2 Integrity Test procedures. Only devices that pass the integrity check are to proceed on to testing.

6.3 Test Water Preparation and Analysis. Testing requires various test waters, including RO/DI, unseeded GTW or CTW, and GTW or CTW seeded with microbiological Challenge A, Challenge B, or background bacteria.

6.3.1 Prepare test water in accordance with Table 2, using GTW during days 1, 2, 5, and 9, and CTW during days 3, 4, 8, and 10. Test water will be created and analyzed each morning of testing to ensure it meets Table 2 requirements. At the end of each day the test water will be analyzed for temperature, turbidity, and pH.

6.3.2 At sample points add microbiological Challenge A or Challenge B to the test water and analyze device influent and effluent samples to determine log reduction achieved by the device. Analyze the device effluent turbidity from the 2 L waste of Challenge B.

6.3.3 Between sample points seed test water with background bacteria in accordance with Table 2 and verify influent bacteria count. Do not analyze device effluent for background bacteria.

6.3.4 Where appropriate, collect a single influent sample as representative of the influent water quality to all replicate devices on test.

6.4 Sample Point Procedures.

1. Seed test water with Challenge A (MS2 and Cryptosporidium) to levels shown in Table 4.

2. Pump Challenge A test water through the device, discarding the first 2 L of product to waste. Collect the next 3 L of product water, aliquot, and analyze for Challenge A organisms in triplicate at appropriate dilutions.

3. Sample Challenge A-seeded influent water and analyze for Challenge A organisms in triplicate at appropriate dilutions.

4. Seed test water with Challenge B (E.coli and fr) to levels shown in Table 4.

5. Pump Challenge B test water through the device, discarding the first 2 L of product to waste. Collect the next 3 L of product water, aliquot, and analyze for Challenge B organisms in triplicate at appropriate dilutions and turbidity.

6. Sample Challenge B-seeded influent water and analyze for Challenge B organisms in triplicate at appropriate dilutions.

6.5 Non-Sample Point Procedures. Pump appropriate test water as specified in Table 2 through the device to waste. This water contains background bacteria but is not seeded with any other organisms.

6.6 End of Day Procedures. At the end of each test day perform the following:

1. Analyze and record influent test water for temperature, turbidity, and pH.

2. Remove the influent hose from the test water and pump the device until no water is produced. Cover the end of the effluent tubing with tin foil to minimize the potential for contamination.

6.7 Post Stagnation Sampling Point. Pump 3 L of unseeded CTW water through the device. Collect the entire 3 L, aliquot and analyze for MS2, Cryptosporidium, E.coli, and fr in triplicate without dilution.

6.8 Cleaning Procedures. The device continuously cleans the membrane by flushing about 10 percent of the influent water to waste. No additional cleaning is required.

6.9 End of Test / Device Clogging. It is desired to perform a sampling event as the device clogs (defined as the flowrate decreasing to 25% of initial flowrate). Follow one of the following two procedures based on device flowrate:

1. Device flowrate does not drop to 25% prior to the 75% sampling point: Continue with test procedures and scheduled sampling points. Follow Day 10-1 procedures to test until flowrate drops to 25%, initiate 100% sampling point, then end test.

2. Device flowrate drops to 25% prior to the 75% sample point: At this time, seed the current test water and perform a sampling point, as described in Section 6.4. This sampling point becomes (replaces) the next scheduled sampling point (skip the next sampling point but still process all scheduled water volume). Replace the device filter cartridge and continue with test procedures, including any future sampling points. Follow Day 10-2 procedures to collect the final 100% sampling point at 590 – 600 L, then end test. If flowrate drops to 25% on Day 10 prior to reaching 590 L, initiate the 100% sample point then end test.

7. DAILY TESTING SUMMARY.

DAY 0

1. Perform DEVICE CONDITIONING AND PRE-TEST CONTAMINATION CHECK (6.1).

2. Perform device INTEGRITY TEST (6.2)

DAY 1

1. PREPARE and ANALYZE General Test Water (GTW) without background bacteria (6.3).

2. Perform SAMPLING POINT PROCEDURES for Challenge A and B to collect 0% sample (6.4). This is a First Draw Sample. For Challenge A, do not filter any to waste; collect entire 3 L batch for analysis. For Challenge B, discard the first 2 L to waste then collect the next 3 L for analysis.

3. SEED an appropriate volume of GTW with background bacteria and collect sample to analyze bacteria concentration (6.3).

4. Perform NON-SAMPLE POINT PROCEDURES to process 67 L to waste (6.5).

5. Perform END OF DAY PROCEDURES (6.6).

Daily volume filtered = 75 L, cumulative 75 L.

DAY 2

1. PREPARE and ANALYZE GTW with background bacteria (6.3).

2. Perform NON-SAMPLE POINT PROCEDURES to process 75 L to waste (6.5).

3. Perform END OF DAY PROCEDURES (6.6).

Daily volume filtered = 75 L, cumulative 150 L.

DAY 3

1. PREPARE and ANALYZE Challenge Test Water (CTW) without background bacteria (6.3.

2. Perform SAMPLING POINT PROCEDURES for Challenge A and B to collect 25% sample (6.4).

3. SEED an appropriate volume of CTW with background bacteria and collect sample to analyze bacteria concentration (6.3).

4. Perform NON-SAMPLE POINT PROCEDURES to process 65 L to waste (6.5).

5. Perform END OF DAY PROCEDURES (6.6).

Daily volume filtered = 75 L, cumulative 225 L

DAY 4

1. PREPARE and ANALYZE CTW without background bacteria (6.3).

2. SEED an appropriate volume of CTW with background bacteria and collect sample to analyze bacteria concentration (6.3).

3. Perform NON-SAMPLE POINT PROCEDURES to process 65 L to waste (6.5).

4. Perform SAMPLING POINT PROCEDURES for Challenge A and B to collect 50% sample (6.4).

DAY 5

1. PREPARE and ANALYZE GTW without background bacteria (6.3).

2. SEED an appropriate volume of GTW with background bacteria and collect sample to analyze bacteria concentration (6.3).

3. Perform NON-SAMPLE POINT PROCEDURES to process 65 L to waste (6.5).

4. Perform SAMPLING POINT PROCEDURES for Challenge A and B to collect 60% sample (6.4).

DAY 6 – Stagnation DAY 7 – Stagnation

DAY 8

1. Perform POST STAGNATION SAMPLING (6.7)

2. PREPARE and ANALYZE CTW with background bacteria (6.3).

3. Perform NON-SAMPLE POINT PROCEDURES to process 72 L to waste (6.5).

4. Perform END OF DAY PROCEDURES (6.6).

Daily volume filtered = 75 L, cumulative 450 L

DAY 9

1. PREPARE and ANALYZE GTW with background bacteria (6.3).

2. Perform SAMPLING POINT PROCEDURES for Challenge A and B to collect 75% sample (6.4).

3. Perform NON-SAMPLE POINT PROCEDURES to process 65 L to waste (6.5).

4. Perform END OF DAY PROCEDURES (6.6).

Daily volume filtered = 75 L, cumulative 525 L

FOLLOW ONE OF THE TWO DAY 10 PROCEDURES BELOW.

DAY 10-1 (FILTER NOT YET REPLACED)

1. PREPARE and ANALYZE CTW without background bacteria (6.6).

2. SEED an appropriate volume of CTW with background bacteria and collect sample to analyze bacteria concentration (6.6).

3. Perform NON-SAMPLE POINT PROCEDURES until the flowrate drops to 25% of Day 1 flowrate (about 0.625 L/min) (6.5).

4. Perform SAMPLING POINT PROCEDURES for Challenge A and B to collect 100% sample (6.4). Since device flowrate is significantly reduced, collect the minimum required volume for analysis, while ensuring the required 2 L 10x void volume to waste is followed. Slow the pumping rate, as necessary, to produce water as the device clogs.

5. Perform device INTEGRITY TEST (6.2)

6. End of Test Daily volume filtered = 75 L (estimated), cumulative 600 L (estimated)

OR

DAY 10-2 (FILTER PREVIOUSLY REPLACED)

1. PREPARE and ANALYZE CTW without background bacteria (6.6).

2. SEED an appropriate volume of CTW with background bacteria and collect sample to analyze bacteria concentration (6.6).

3. Perform NON-SAMPLE POINT PROCEDURES to process 65 L to waste (6.5).

4. Perform SAMPLING POINT PROCEDURES for Challenge A and B to collect 100% sample (6.4).

5. Perform device INTEGRITY TEST (6.2)

6. End of Test Daily volume filtered = 75 L, cumulative 600 L

Table 5. Sampling Schedule.

Day/ Volume (L) Sample Point Volume (L)

Day 1 1 – 75 0% First Draw 1-8 L

Day 2 76 – 150

Day 3 151 – 225 25% 151 – 161 L

Day 4 226 – 300 50% 290 – 300 L

Day 5 301 – 375 60% 365 – 375 L

Day 8 376 – 450

Day 9 451 – 525 75% 451 – 461 L

Day 10 526 – 600 (est.)

100% 590 – 600 L

OR

Clogging Point 590 – 600 L (est.)

8. CONCLUSIONS. This test plan provides detailed procedures for testing against NSF Protocol P248, Military Operations Microbiological Water Purifiers, Appendix B (2012). Adherence to recommended procedures will allow for objective determination of the microbiological reduction capabilities of the tested devices and will provide an indication of the capabilities provided to the Warfighter during military operations.

9. POINT OF CONTACT. The point of contact for this test plan is Mr. Art Lundquist, U.S. Army Public Health Center, arthur.h.lundquist.civ@mail.mil, 410 436-3919.

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APPENDIX A

REFERENCES

1. NSF International, 2012. NSF Protocol P248 Military Operations Microbiological Water Purifiers, NSF International, Ann Arbor, Michigan.

2. American Public Health Association (APHA), American Water Works Association (AWWA), and Water Environment Federation (WEF). (2006) Standard Methods for the Examination of Water and Wastewater, 21st Edition.

· 4500-H+ pH Value, B. Electrometric Method

· 4500- Cl Chlorine (Residual), G. DPD Colorimetric Method

· 2550-Temperature

· 2540-Solids, C. Total Dissolved Solids Dried at 180oC

· 2320-Alkalinity, B. Titration Method

· 5310-Total Organic Carbon (TOC), C. Persulfate-Ultraviolet or Heated-Persulfate Oxidation Method

· 5310-Total Organic Carbon (TOC), B. High-Temperature Combustion Method

· 9222-Membrane Filter Technique for Members of the Coliform Group, B. Standard Total Coliform Membrane Filter Procedure

3. Disinfection, Sterilization, and Preservation. Ed. Block, SS. Philadelphia: Lea & Febiger, 1983.

4. International Organization for Standardization. (1997) ISO 12103 Part 1.A2: Road vehicles - Test dust for filter evaluation, Arizona test dust, Fine Test Dust.

5. U.S. Environmental Protection Agency. (1993) EPA Method 180.1: Determination of Turbidity by Nephelometry.

6. NSF International/ANSI, 2013. Standard 53: Drinking Water Treatment Units – Health Effects. NSF International, Ann Arbor, Michigan.

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