SOP on Media growth Promotion, Inhibition and Sterility Check

March 26, 2016September 8, 2024 Pharmaceutical Guidanace

SOP on Media growth Promotion, Inhibition, and Sterility Check

Objective

To lay down the procedure for Media Growth Promotion, Inhibition, and Sterility Check.

Scope

This SOP is applicable for Media Growth Promotion, Inhibition, and Sterility Checks in the Microbiology Laboratory of (Pharmaceutical Company Name).

Responsibility

Chemist or above of QC laboratory.

Head – Microbiology Section.

Accountability

Head – Quality Control.

Abbreviations and Definitions

QC: Quality Control

GPT: Growth Promotion Test

SCDM: Soyabean Casein digest Medium

FTM: Fluid Thioglycollate medium.

Procedure

Check all the sterile lots of the media (Prepared as per the directions given ‘Preparation of Sterile media SOP No.: for its sterility, ability to promote the growth of specific organisms, and Growth Inhibitory property.

Prepare Culture suspension as per SOP No.:

Inoculate portions of the test media in plates/tubes/bottles with 0.1 ml of specified microbial culture suspension containing 10-100 CFUs.

For the Selection of Organisms and method of Inoculation, follow Table-1.

Use any one of the organisms for the Growth promotion Test for Media.

Use one or more environmental isolates if available, preferably in the media used for environmental monitoring, sterility test, media used for enumeration tests of water, products, etc.

Incubate the inoculated media at recommended temperatures specified for the organism as per Table 2 and Table 3.

Similarly, incubate the uninoculated media as Negative control for sterility check at a specified temperature for a time specified as per Table 2 and Table-3.

Retain the Negative controls meant for sterility test under incubation for 14 Days at 22.5+2.5oC for SCDM and at 32.5+2.5oC for Fluid Thioglycollate medium.

Use any one of the organisms for inoculating SCDM and one aerobic and one anaerobic organism for inoculating FTM used as a positive control in Sterility Test respectively from Table-1.

There should not be any microbial contamination present in the Sterility check portions of media by the completion of the incubation period.

Observe and note the observation in the record as per Annexure No.:

Write the interpretation by comparing the acceptance criteria mentioned in table 1.

Record the details of Media GPT, Sterility, and inhibition tested for each autoclaved media Lot as per Annexure No.:

Perform GPT on each received lot of dehydrated media and note down the results as per Annexure. No.

Growth should be clear and visible in the form of turbidity in the case of Liquid media and distinct colony formation in the case of Agar media and the results shall be comparable to the previously approved batch of the media.

Perform all inoculations aseptically underclass ‘A’ LAF cabinet in the subculturing area.

Perform Microbial recovery for solidified media to demonstrate the suitability of the test method.

Discard all the used media/tubes/bottles after completion of the test as per SOP No.: QC- (SOP on Decontamination and Disposal of Used Media).

Discard the used media plates/tubes/bottles even before the complete incubation period, if the acceptance criteria are met.

Table 1: Growth-promoting and inhibitory properties of media

1. Media: Soyabean-Casein

Digest Agar
Medium
Test Microbial culture:
S.aureus (ATCC 6538)
Property: Growth promoting
Method of inoculation: Pour Plate
Acceptance criteria: More than 70% of initial count of culture suspension used.
Test Microbial culture: B. subtilis (ATCC 6633)
Test Microbial culture: S. abony (ATCC 14028)
Test Microbial culture: E. coli ( ATCC 8739)
Test Microbial culture: C.albicans(ATCC 10231)
Test Microbial culture: A. niger (ATCC 16404)

2 Media :Potato Dextrose Agar

Test Microbial culture: C.albicans( ATCC 10231)
Test Microbial culture: A.niger(ATCC 16404)
Property: Growth promoting
Method of inoculation: Pour Plate
Acceptance criteria: More than 70% of the initial count of culture suspension used.

3 Media: Sabouraud

Dextrose Agar
Media (or)
Potato Dextrose
Agar Medium
Test Microbial culture: C.albicans (ATCC 10231)
Test Microbial culture: A.niger( ATCC 16404)
Property: Growth promoting
Method of inoculation: Pour Plate
Acceptance criteria: More than 70% of the initial count of culture suspension used.

4 Media: Plate Count Agar Medium

Test Microbial culture: P. aeruginosa( ATCC 9027)
Property: Growth promoting
Method of inoculation: Pour Plate
Acceptance criteria: More than 70% of initial count of culture suspension used.
Test Microbial culture: S. aureus (ATCC 6538)
Test Microbial culture: Salmonella.abony (ATCC 14028)
Test Microbial culture: E. coli( ATCC 8739)

5. Media: Soyabean –Casein Digest Medium (Other than sterility test)

Test Microbial culture: P. aeruginosa( ATCC 9027)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: Visible growth should be observed and compared to previously tested and approved Media.
Test Microbial culture: S. aureus (ATCC 6538)
Test Microbial culture: C. albicans (ATCC 10231)
Test Microbial culture: A. niger( ATCC 16404)
Test Microbial culture: B. subtilis( ATCC 6633)

6. Media:Soyabean –Casein Digest Medium (for sterility test)

Test Microbial culture: C. albicans (ATCC 10231)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: Visible growth should be observed and compare to previously tested and approved Media.
Test Microbial culture: A. niger ( ATCC 16404)
Test Microbial culture: B. subtilis ( ATCC 6633)

7. Media :Fluid thio-glycollate Medium

Test Microbial culture: Cl.Sporogenes(ATCC 19404)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: Visible growth should be observed and compare to previously tested and approved Media.
Test Microbial culture: S. aureus (ATCC 6538)
Test Microbial culture: P. aeruginosa( ATCC 9027)
Test Microbial culture: M. luteus (ATCC 9341)

8. Media :Fluid Lactose Medium

Test Microbial culture: Salmonella. abony (ATCC 14028)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: Visible growth should be observed and compare to previously tested and approved Media.
Test Microbial culture: E. coli( ATCC 8739)

9 Media :Levine Eosin-Methylene Blue Agar Medium

Test Microbial culture: E. coli( ATCC 8739)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Metallic sheen under reflected light and a blue-black appearance under transmitted light.

10 Media : MacConkey Agar

Test Microbial culture: E. coli ( ATCC 8739)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Brick-red may have a surrounding zone of precipitated bile.

11 Media :Fluid Selenite-Cystine Medium

Test Microbial culture: Salmonella. abony (ATCC 14028)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: A characteristic growth on Selective agar Media*
Test Microbial culture: S. aureus (ATCC 6538)
Property: Inhibitory
Method of inoculation: Addition
Acceptance criteria: No growth or No Characteristic growth*

12 Media :Fluid Tetrathionate Medium

Test Microbial culture: Salmonella.abony (ATCC 14028)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: A characteristic growth on Selective agar medium*
Test Microbial culture: S. aureus (ATCC 6538)
Property: Inhibitory
Method of inoculation: Addition
Acceptance criteria: No growth or No Characteristic growth*

13. Media :Bismuth Sulfite Agar Medium

Test Microbial culture: Salmonella. abony (ATCC 14028)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Black or green colonies
Test Microbial culture: S. aureus (ATCC 6538)
Property: Inhibitory
Method of inoculation: Pour plate/
Method of inoculation: Spreading/
Method of inoculation: Streaking
Acceptance criteria: No growth or No Characteristic growth

14. Media :Brilliant Green Agar Medium

Test Microbial culture: Salmonella abony (ATCC 14028)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Small, transparent, colorless or pink to white opaque colonies (frequently surrounded by pink to red zone
Test Microbial culture: Salmonella.aureus (ATCC 6538)
Property: Inhibitory
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: No growth or No Characteristic growth

15.Media :Xylose-Lysine-Desoxycholate Agar Medium

Test Microbial culture: Salmonella. abony(ATCC 14028)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Red-colored colonies, with or without black centers
Test Microbial culture: E.coli ( ATCC 8739)
Property: Inhibitory
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: No growth or No Characteristic growth

16. Media :Rappaport Vassiliadis

Test Microbial culture: Salmonella. abony (ATCC 14028)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: Visible growth should be observed and compare to previously tested and approved Media.
Test Microbial culture: S. aureus (ATCC 6538)
Property: Inhibitory
Method of inoculation: Addition
Acceptance criteria: No growth or No Characteristic growth

17. Media: Cetrimide Agar Medium

Test Microbial culture: P. aeruginosa( ATCC 9027)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Generally greenish colonies
Test Microbial culture: E. coli ( ATCC 8739)
Property: Inhibitory
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: No growth or No Characteristic growth

18.Media :Pseudomonas Agar Medium for the Detection of Fluorescin

Test Microbial culture: P. aeruginosa( ATCC 9027)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Generally colorless to yellowish colonies.
19 Media :Pseudomonas Agar Medium for detection of Pyocyanin
Test Microbial culture: P. aeruginosa( ATCC 9027)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Generally greenish colonies

20. Media :Vogel-Johnson Agar Medium

Test Microbial culture: S. aureus (ATCC 6538)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Black colonies Surrounded by yellow zone
21 Media :Baird-Parker Agar Medium
Test Microbial culture: S. aureus (ATCC 6538)
Property: Growth promoting
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: Green purple colonies, surrounded by clear zones 2 or 5 mm

22. Media : Mannitol-Salt

Agar Medium
Test Microbial culture: S. aureus (ATCC 6538)
Property: Growth promoting
Method of inoculation: Pour plate/ Spreading/Streaking
Acceptance criteria: Yellow colonies with yellow zones
Test Microbial culture: E. coli( ATCC 8739)
Property: Inhibitory
Method of inoculation: Pour plate/Spreading/Streaking
Acceptance criteria: No growth or No Characteristic growth

23. Media :Sabouraud Dextrose Broth

Test Microbial culture: C. albicans (ATCC 10231)
A. niger (ATCC 16404)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: Visible growth should be observed and compared to previously tested and approved Media.

24.Media : Reinforced Agar medium

Test Microbial culture: Cl. Sporogenes (ATCC 19404)
Property: Growth promoting
Method of inoculation: Pour plate
Acceptance criteria: More than 70% of the initial count of culture suspension used

25. Media : Reinforced broth

Test Microbial culture: Cl.sporogenes(ATCC 19404)
Property: Growth promoting
Method of inoculation: Addition
Acceptance criteria: Visible growth should be observed and compared to previously tested and approved Media

26. Media : ** R 2A Medium

Test Microbial culture :
P. aeruginosa( ATCC 9027)
S. aureus(ATCC 6538)
Salmonella.abony(ATCC 14028)
E. coli ( ATCC 8739)
Property: Growth promoting
Method of inoculation: Pour plate
Acceptance criteria: More than 70% of the initial count of culture suspension use.

*- To be confirmed by streaking on one of Salmonella selective media (i.e. Bismuth Sulfite Agar

Medium / Brilliant Green Agar Medium / Xylose-Lysine-Desoxycholate Agar Medium)

**-For R2A incubate the inoculated media at 22.5 + 2.5^OC for maximum up to 5 days

Table 2: Culture conditions for Microbial recovery

Name of the Organism: A. niger (ATCC 16404)
Incubation Time: 5 days
Incubation Temperature: 22.5 + 2.5^OC

Name of the Organism: C. albicans(ATCC 10231)
Incubation Time: 3 days
Incubation Temperature: 22.5 + 2.5^OC

Name of the Organism: S. Typhimurium (ATCC 14028)
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: P. aeruginosa ( ATCC 9027)
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: E. coli ( ATCC 8739)
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: S. aureus( ATCC 6538)
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: B. subtilis (ATCC 6633)
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: Cl. sporogenes (ATCC 19404)
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: B. vulgatus (ATCC 19404)
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: M. luteus(ATCC (ATCC 8482)
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: Environmental isolate-Bacteria
Incubation Time: 48 hours
Incubation Temperature: 32.5 + 2.5^OC

Name of the Organism: Environmental isolate-Yeast
Incubation Time: 3 days
Incubation Temperature: 22.5 + 2.5^OC

Name of the Organism: Environmental isolate-Mold
Incubation Time: 5 days
Incubation Temperature: 22.5 + 2.5^OC

Table 3: Incubation conditions for Broth Medium

Medium: FTM

Name of Organism: Cl. sporogenes(ATCC 19404)
Incubation Time: 3 days
Incubation Temperature: 32.5 + 2.5^OC
Name of Organism: P. aeruginosa ( ATCC 9027)
Incubation Time: 3 days
Incubation Temperature: 32.5 + 2.5^OC
Name of Organism: S. aureus ( ATCC 6538)
Name of Organism: B. subtilis (ATCC 6633)
Incubation Time: 3 days
Incubation Temperature: 32.5+ 2.5^OC

Medium: SCDM

Name of Organism: B. subtilis (ATCC 6633)
Incubation Time: 3 days
Incubation Temperature: 22.5 + 2.5^OC
Name of Organism: A. niger (ATCC 16404)
Incubation Time: 5 days
Incubation Temperature: 22.5 +2.5^OC
Name of Organism: C. albicans(ATCC 10231)
Incubation Time: 5 days
Incubation Temperature: 22.5 + 2.5^OC

Medium: RAPPAPORT VASSILIADIS

Name of Organism: Salmonella abony(ATCC 14028)
Incubation Time: 1 days
Incubation Temperature: 32.5 + 2.5^OC
Name of Organism: Salmonella aureus(ATCC 6538)
Incubation Time: 1 days
Incubation Temperature: 32.5 + 2.5^OC

Medium: FTM

Name of Organism: Environmental Isolate- Bacteria
Incubation Time: 3 days
Incubation Temperature: 32.5 + 2.5^OC

Medium: SCDM

Name of Organism: Environmental Isolate- Bacteria
Incubation Time: 3 days
Incubation Temperature: 22.5 + 2.5^OC

Medium: SCDM

Name of Organism: Environmental Isolate-Yeast
Incubation Time: 5 days
Incubation Temperature: 22.5 + 2.5^OC

Medium: SCDM

Name of Organism: Environmental Isolate Mold
Incubation Time: 5 days
Incubation Temperature: 22.5 + 2.5^OC

Forms and Records (Annexures)

Media growth Promotion, Inhibition and Sterility Check
Growth Promotion Test Report

Distribution

Master copy – Quality Assurance
Controlled copies – Quality Assurance, Production, Quality Control, Stores, Engineering, and Human Resource

History:

Date	Revision Number	Reason for Revision
	00	NEW SOP

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NewportAuburn, Newquay, Newry, NGERULMUD, NIAMEY, NICARAGUA, NICOSIA, NIGER, Nigeria, Nizhniy Novgorod, Nizhny, Noida, Norfolk, Norfolk Island, North, North Ayrshire, North East Derbyshire, North East Lincolnshire, NORTH KOREA, North Lanarkshire, North Lincolnshire, NORTH MACEDONIA, North Somerset, North West, North Yorkshire, Northamptonshire, Northern, Northern Ireland, Northern Territory, Northumberland, Northwest Territories, Northwich, Norway, Norwich, Nottingham, Nottinghamshire, NOUAKCHOTT, Nova Scotia, Novara, Novgorod, Novosibirsk, Nu`ayjah, Nunavut, Nuoro, NUR-SULTAN, Nuremberg, Oakland, Old Al Ghanim, Old Al Hitmi, Old Al Rayyan, oman, Omsk, Onaiza, ontario canada, OntarioToronto, Oran, Orenburg, Oristano, Orkney Islands, Oryol, OSLO, Otago, Ottawa, OUAGADOUGOU, Ouargla, Oxford, Oxfordshire, Padua, Paisley, Pakistan, PALAU, Palermo, PALESTINE, PANAMA, PANAMA CITY, PAPUA NEW GUINEA, Para, PARAGUAY, Paraiba, PARAMARIBO, Parana, Paris, Parma, Patna, Pavia, Pembrokeshire, Penrith, Penryn, Penza, Penzance, Pernambuco, Perth, Perth and Kinross, peru, Perugia, Pesaro and Urbino, Pescara, Peterborough, Philippines, PHNOM PENH, Piacenza, Piaui, Pimpri, Pisa, Pistoia, Plymouth, Poland, Poole, Pordenone, PORT LOUIS, PORT MORESBY, PORT OF SPAIN, PORT VILA, PORT-AU-PRINCE, Porto Portugal, PORTO-NOVO, Portsmouth, portugal, Potenza, Powys, Prague, PRAIA, Prato, Preston, PRETORIA, Prince Edward Island, PRISTINA, Pskov, Pune, PYONGYANG, Qatar, Quebec City, Queensland, QUITO, Ragusa, Raipur, Rajkot, Ramsey, Ranchi, Ras Abu Aboud, Ras Al Khaimah, Ras Lafan, Ravenna, Rawdat Al Hamama, Rawdat Al Khail, Rawdat Egdaim, Rawdat Rashed, Reading, Redcar and Cleveland, Reggio Calabria, Reggio Emilia, Regina, Relizane, Renfrew, Renfrewshire, Repton, REYKJAVIK, Rhondda Cynon Taf, Rieti, RIGA, Rimini, Rio de Janeiro, Rio Grande, Ripon, Riverside, RIYADH, Romania, Rome, ROSEAU, Rostov, Rotterdam, Rovigo, Rumeilah, Russia, Rutland, RWANDA, Ryazan, Saharanpur, Saïda, SAINT GEORGE'S, SAINT JOHN'S, SAINT KITTS & NEVIS, SAINT LUCIA, Saint Petersburg, SAINT VINCENT & THE GRENADINES, Sakhalin, Salem, Salerno, Salford, Salisbury, Saltash, Samara, SAMOA, SAN JOSE, SAN MARINO, SAN SALVADOR, Sandhurst, SANTIAGO, SANTO DOMINGO, SÃO TOMÉ & PRÍNCIPE, SÃO TOMÉ SAUDI ARABIA, SARAJEVO, Saratov, Saskatchewan, Sassari, Savona, Sawda Natheel, Saxony, Scotland, Scottish Borders, SENEGAL, SEOUL, Serbia, Sétif, Seville, SEYCHELLES, Shagra, Sharjah, Sheffield, Shetland Islands, Shropshire, Sidi bel Abbès, Siena, SIERRA LEONE, Siliguri, Simaisma, singular, Skikda, SKOPJE, Slough, SLOVAKIA, SLOVENIA, Smolensk, Sofia, Solapur, SOLOMON ISLANDS, SOMALIA, Somerset, Sondrio, Souk Ahras, SOUTH AFRICA, South Australia, South Ayrshire, South Bucks, South Cambridgeshire, South Derbyshire, South East, South Gloucestershire, SOUTH KOREA, South Lakeland, South Lanarkshire, South Sardinia, SOUTH SUDAN, SOUTH TARAWA, South Tyrol, South West, South Yorkshire, Southampton, Southend-on-Sea, Southland, spain, Springfield, SRI JAYAWARDENEPURA KOTTE, Sri Lanka, Srinagar, St Albans, St Asaph, St Davids, St. Austell, St. Ives, St. John's, Staffordshire, Stirling, STOCKHOLM, Stockton-on-Tees, Stoke Poges, Stoke-on-Trent, Stuttgart, SUCRE, sudan, Suffolk, Sunderland, Surat, SURINAME, Surrey, SUVA, Sverdlovsk, Swansea, Sweden, Swindon, switzerland, Sydney, Syracuse, SYRIA, TAJIKISTAN, TALLINN, Tambov, TANZANIA, Taranto, TASHKENT, Tasmania, TBILISI, Tébessa, TEGUCIGALPA, TEHRAN, Telford, Telford and Wrekin, Teramo, Terni, Thailand, Thane, The Hague, The Pearl, THIMPHU, Thiruvananthapuram, Thuringia, Thurrock, Thurso, Tiaret, TIMOR-LESTE DILI, Tintagel, TIRANA, Tiruchirappalli, Tizi Ouzou, Tlemcen, TOGO, TOKYO, Tomsk, TONGA NUKUʻALOFA, Torbay, Trapani, Trento, Treviso, Trieste, TRINIDAD & TOBAGO, TRIPOLI, Truro, Tula, TUNIS, TUNISIA, Turin, turkey, TURKMENISTAN, TUVALU, Tver, Tyne and Wear, Tyumen, Udine, UGANDA, Ujjain, Ukraine, Ukraine. Dnipropetrovsk, Ulhasnagar, Ulyanovsk, Umm al-Qaiwain, Umm al-Quwain, Umm Bab, Umm Birka, Umm Ghuwailina, Umm Lekhba, Umm Qarn, Umm Salal Ali, Umm Salal Mohammed, United arab emirates, UNITED ARAB EMIRATES (UAE), united Kingdom, UNITED KINGDOM* (UK), united state, United States, UNITED STATES OF AMERICA (USA), URUGUAY, UZBEKISTAN, Vadodara, VADUZ, Vale of Glamorgan, Valencia, VALLETTA MARSHALL ISLANDS, VANUATU, Varanasi, Varese, Vasai Virar, VATICAN CITY, VATICAN CITY (Holy See), VENEZUELA CARACAS, Venice, Verbano-Cusio-Ossola, Vercelli, Verona, Vibo Valentia, Vicenza, Victoria, Vienna, VIENTIANE, VIETNAM HANOI, Vijayawada, VILNIUS, Visakhapatnam, Viterbo, Vladimir, Volgograd, Vologda, Voronezh, Wadi Al Banat, Wadi Al Sail, Wadi Al Wasaah, Wadi Lusail, Wakefield, Wales, Warrington, Warsaw, Warwickshire, Washington, Wellington, Wells, West Berkshire, West Dunbartonshire, West Lothian, West Midlands, West Sussex, West Yorkshire, West Yorkshire (Bradford–Leeds), Western Australia, Western Isles, Westland, Whitehaven, Whitehorse, Wick, Wiltshire, Winchester, Windsor and Maidenhead, Winnipeg, Wisbech, Wokingham, Wolverhampton, Worcester, Worcestershire, Workington, Wrexham, Wrexham Qatar, Wycombe, YAMOUSSOUKRO, YAOUNDE, YAREN DISTRICT, Yaroslavl, Yellowknife, YEMEN SANA'A ZAMBIA, YEREVAN, York, Yorkshire and the Humber, Yukon, ZAGREB, ZIMBABWE, Zürich

SOP on Media growth Promotion, Inhibition, and Sterility Check

Reason for Revision

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