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Microbiology Animal Food Project Testing Track 3 Accelerated Sample Preparation for Microbial Pathogen Detection in Pet Food Samples

Award Number: U19FD007110
ORGANIZATION: FOOD AND DRUG ADMINISTRATION
OPDIV: FDA
AWARD CLASS: COOPERATIVE AGREEMENT
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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Accelerated Sample Preparation for Microbial Pathogen Detection in Large Number of Pet Food Samples Abstract We are submitting a proposal for Discipline A: Microbiology, track 3 for Animal Food Product testing with Dog treat food/Salmonella as a hazard to Commodity pairs. Here we combine different expertise from 2 laboratories, Laboratory of Renewable Resources Engineering (LORRE) and the Office of Indiana State Chemist (OICS), to assemble a proposal consisting in animal food sample collection, accelerated food sample preparation to bring microorganisms of interest to detection levels, FDA approved loop mediated isothermal amplification (LAMP) for Salmonella screening in animal food, and whole genome sequencing as a method for confirmation of pathogen isolates. We have further optimized our sample preparation approach, as a follow up to 2015 FDA challenge award, and we now can simultaneously concentrate and recovery microorganisms of interest in 4 samples, bringing their numbers from initially not detectable, due to be present at initial levels of 1CFU/g or lower, to detection levels. This accelerated processing of samples is complete in a time frame of about 4 hours. It consists of a microbial short enrichment in selective medium combined with an enzyme treatment, followed by a pre-filtration step to remove large particles and microfiltration to concentrate and recovery selected microorganisms to detection level. The enzyme treatment helps to break down possible biofilms formation and particles that may clog pores in the hollow fiber membrane during microfiltration. Enzymes selected are based on food matrices. In the case of the samples proposed here, proteases possible combined with lipases will be used. We have shown the enzymes do not affect microbial viability, when in the presence of their respective substrates. While we have focused on Salmonella as a microorganism of interest, our developed approach can also be applied to other microorganisms. The Office of Indiana State Chemist (OISC) microbiology laboratory has major expertise in sample collection for animal food testing. It has also many years of experience in microbiological method development, testing for antibiotic concentration, and bacteria enumeration and identification, including several years of pathogen screening for Salmonella, Listeria monocytogenes, Campylobacter and Escherichia coli (STEC) in animal feed. Last year, the OISC microbiology laboratory contributed for testing in a national wide Salmonella outbreak in pet treat pig ear in Indiana. The proposed number of samples to be tested here is 250 samples pairing dog treat/Salmonella. Expected outcomes are accelerated sample preparation, increased speed and sensitivity of sample testing, and initial insights of potential recovering and detection of difficult to culture microorganisms.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2024 ( Subtotal = $104,140 )
2024	2024	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Food and Drug Administration Research	000	6	7/24/2024	NON-COMPETING CONTINUATION	$104,140
														Subtotal = $104,140

Issue Date FY: 2023 ( Subtotal = $110,000 )
2023	2023	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Food and Drug Administration Research	004	5	9/25/2023	NON-COMPETING CONTINUATION	$0
2023	2023	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Food and Drug Administration Research	003	5	6/30/2023	NON-COMPETING CONTINUATION	$110,000
2023	2022	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Food and Drug Administration Research	002	4	2/28/2023	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$0
2023	2022	PURDUE UNIVERSITY	2550 NORTHWESTERN AVE STE 1900	WEST LAFAYETTE	IN	47906	TIPPECANOE	USA	Food and Drug Administration Research	000	3	11/1/2022	NON-COMPETING CONTINUATION	-$109,188
2023	2022	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Food and Drug Administration Research	001	4	11/9/2022	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$109,188
														Subtotal = $110,000

Issue Date FY: 2022 ( Subtotal = $109,993 )
2022	2022	PURDUE UNIVERSITY	2550 NORTHWESTERN AVE STE 1900	WEST LAFAYETTE	IN	47906	TIPPECANOE	USA	Food and Drug Administration Research	001	3	8/12/2022	NON-COMPETING CONTINUATION	$0
2022	2022	PURDUE UNIVERSITY	2550 NORTHWESTERN AVE STE 1900	WEST LAFAYETTE	IN	47906	TIPPECANOE	USA	Food and Drug Administration Research	000	3	7/22/2022	NON-COMPETING CONTINUATION	$109,993
														Subtotal = $109,993

Issue Date FY: 2021 ( Subtotal = $110,000 )
2021	2021	PURDUE UNIVERSITY	155 S GRANT ST	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Food and Drug Administration Research	001	2	7/7/2021	NON-COMPETING CONTINUATION	$110,000
2021	2020	PURDUE UNIVERSITY	155 S GRANT ST	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Food and Drug Administration Research	000	1	10/30/2020	NEW	$0
														Subtotal = $110,000

Issue Date FY: 2020 ( Subtotal = $110,000 )
2020	2020	PURDUE UNIVERSITY	155 S GRANT ST	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Food and Drug Administration Research	000	1	9/8/2020	NEW	$110,000
														Subtotal = $110,000

Grand Total All Awards = $544,133

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Microbiology Animal Food Project Testing Track 3 Accelerated Sample Preparation for Microbial Pathogen Detection in Pet Food Samples

Award Number: U19FD007110

ORGANIZATION: FOOD AND DRUG ADMINISTRATION

OPDIV: FDA

AWARD CLASS: COOPERATIVE AGREEMENT

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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