DEVELOPMENT OF A SANITIZING TREATMENT TO IMPROVE SAFETY AND QUALITY OF INDIANA

CANTALOUPE

Dr. Kevin M. Keener Indiana Horticultural Congress

01/19/2016

A 2014-15 Purdue University AgSEED* Funded Project

* This activity was funded, by Purdue, as part of AgSEED Crossroads funding to support Indiana's Agriculture and Rural Development”.

INTRODUCTION Ø Food Safety

High consumer demand for fresh fruit and vegetables. Fresh produce is or can become contaminated with pathogens:

Ø 2011 E. coli outbreak on fresh strawberries Ø 2011 Listeria outbreak on whole cantaloupes Ø 2012 Salmonella spp. outbreak Ø Salmonella outbreak on fresh tomatoes, February

2014 Ø 2014-15 caramel apples contaminated with Listeria

Ø Project Goal: Improve safety and quality of Indiana cantaloupe

INTRODUCTION – PLASMA Ø  Plasma Background

Ø  4th State of matter: Ionized gas consisting of ions, electrons, neutral species, and light (UV – visible)

Ø  Highly conductive, exists in equilibrium and non-equilibrium states

Ø  Constitutes 99% of the universe although limited occurrence on Earth

Effect of plasma treatment on PE surface. Testing ink balls up, forming drops on an untreated surface; while on the plasma treated surface, the ink forms a continuous film (adapted from Bardos and Barankova, 2010, with permission).

BACKGROUND - TYPES OF ATMOSPHERIC PLASMAS

4

n  Air plasma chemistry- Highly complex n  >75 species & ~500 reactions n  Occur on four different time scales: nano-, micro-, milli-, and seconds

VISION Develop an atmospheric pressure plasma technology

to decontaminate materials (food, pharmaceuticals, medical devices, packaging, etc.) without heat or chemicals.

PURDUE’S COLD ATMOSPHERIC PRESSURE PLASMA (PCAPP) TECHNOLOGY

< 1 ºC increase

UV photons Reactive species

BACKGROUND

Operation of PCAPP Technology

Ø  For air, this process generates ozone, nitrogen oxides, peroxides, and other reactive gas species (RGS) inside the sealed package

Ø  Works with any gas (air, oxygen, nitrogen, helium, argon, carbon dioxide)

Ø  A variety of packaging materials – PP, LDPE, HDPE, PETE, glass, Tyvec, Tygon, multi-layered materials

Ø  In-package RGS generated have proven bactericidal effects Ø  These RGS species decay back to their original state (air,

modified atmosphere) after 24 h post-treatment Ø  Operate between 30 kV – 130 kV at 60Hz using less than

200 W power

PCAPP ADVANTAGES

Ø  Non-thermal treatment for raw products fruits and vegetables Ø  Elimination of spores and pathogens on products Ø  Dry process – no chemical residues Ø  Can be built into process or equipment – performed inside

containers or in a continuous process Ø  Extend shelf life of food products

< 1 ºC increase UV photons Reactive

species

PCAPP  Results:  Vine  Ripe  Tomatoes  (MA-­‐filled  Ziploctm  Bag,  30  s  treatment)  

Safebag

CANTALOUPE PROJECT OVERVIEW Project Objectives 1. Microbial profiling 2. PCAPP treatment of cantaloupe

Ø  Cut cantaloupe and Whole cantaloupe (Totes) Ø  Retail Grocery and Indiana Farms

3. Assessment of PCAPP treatment Ø  Plasma characterization Ø  Microbial assessment Ø  Quality measurement

4. Food safety training for Indiana cantaloupe growers

Variable   Levels  

1) Cantaloupe Product Origin  Retail Grocery (Spring Season: Honduras/Guatemala) Indiana Farms (Summer Season) Retail Grocery (Fall Season: California/Arizona)  

2) Product Sections   Cuts: Top (stem), bottom (blossom), side Whole  Cantaloupe  

3) Gas composition   Air (22% O₂ / 78% N₂) MA65 (65% O2 / 30% CO2 / 5% N2)  

4) Product Testing   Background flora (APC- Aerobic Plate Count), Quality: Color / Firmness  

5) Sample weight (g)  Cuts: 80-100 g (0.25 lb),

Whole: 3-5 lb  

6) Package Type   Cuts: PP Container with Cryovac™ B2630 film Whole: Tote  (30  Gallon)  

7) Plasma Treatment   Outside plasma field: 90 kV (PP container/cuts), 60kV (Tote/whole)  

8) HVACP Treatment times (min)   0 (Controls), 3 (PP/cuts), 30 (Tote/whole)  

9) Storage Temperature   Cuts: refrigeration (4° C/ 39° F), Whole: (21° C/ 70° F)  

Experimental Design and Measurements

Measurements 1. Microbial: Aerobic plate count (APC) 2. Color: l*/a*/b* values (Hunter Colorimeter) 3. Firmness: Compressive Strength (Texture Analyzer) 4. Reactive Gas Measurements: OES, NOx, O3, CO, H2O2 5. Microbial Characterization: BARDOT / 16S PCR Sequencing

 

Experimental Design: Measurements

Ø  1. MICROBIAL PROFILING

Microbial Profiling will be performed using BARDOT technology What is BARDOT?

Stands for: BActerial Rapid Detection using Optical Scattering Technology Inventor: Dr. Arun Bhunia, Purdue University (Food Science) What it does: It utilizes microbial profiling to generate information about the presence, types, and sheer numbers of organisms and bacterial populations naturally present

BARDOT Background Technology is based on imaging and identification of bacterial colonies from their formation of complex multi-organism structures Produces unique microbial fingerprints allowing species identification Automated and scans the petri plate producing an addressable map followed by pattern analysis Patterns are compared and isolates determined to be unique are isolated, cultured, and subsequently frozen for long term storage and identification by sequencing/further analysis

Ø  1. MICROBIAL PROFILING

How BARDOT Technology was utilized for Cantaloupe: Identification of the number of unique isolates and their frequency on Indiana farm sourced cantaloupe both prior to and post PCAPP treatment Determine overall reduction of surface microbes and specific reduction in both population size and diversity (different microbial populations)

PRELIMINARY STUDY RESULTS: BARDOT– AEROBIC PLATE COUNT (APC) CHARACTERIZATION

Ø  74% reduction in diversity of organisms Ø  Retail Grocery (Spring Season: Honduran/Guatemala Imports)

Ø  The highest rate of survival were for Pseudomonas spp. bacteria. This is likely due to its excess exopolysacharide production as “barrier” to stress effects (e.g., PCAPP treatment).

PCAPP

PRELIMINARY STUDY RESULTS: 16S SEQUENCING– APC CHARACTERIZATION (BARDOT)

PCAPP

Ø  Microbacterium spp., Bacillus spp., Acinetobacter spp., Curtobacterium spp., Pantoea spp., and majority Enterobacter spp. were killed with PCAPP treatment

Ø  Treatment significantly reduced bacteria type except Pseudomonas spp.

Ø  Important note: BARDOT cannot quantify number of bacteria only type.

Ø  For Retail Grocery (Spring Season: Honduran/Guatemala Imports)

PRELIMINARY STUDY RESULTS: 16S SEQUENCING– APC CHARACTERIZATION (BARDOT)

For Indiana Farms (Summer Season 2014): Ø  How did these two Farms compare? Ø  Cantaloupe Varieties: Farm A: Athena, Atlantis, Aphrodite; Farm B: Aphrodite

   Combined  

Farm A   Farm B  

1   Pseudomonas   64.7%   Pseudomonas   50.5%  

2   Pantoea   15.4%   Pantoea   43.1%  

3   Arthobacter   4.6%   Erwinia   1.6%  

4   Massilia   4.6%   Paenibacillus   1.6%  

5   Stenotrophomonas   4.6%   Rahnella   0.8%  

6   Chryseobacter   3.1%   Chryseobacter   0.8%  

7   Kocuria   1.5%   Serratia   0.8%  

8   Serratia   1.5%   Uncultured   0.8%  

Ø  200 colonies were isolated from plates analyzed using BARDOT, then sequenced with percentage of aerobic classes/genera listed above

Ø  These bacteria can be found on plants or plant matter, in soil, or “rain” bacteria Note: Erwinia is known to cause plant disease and the Farm from which these strains

were isolated had complained about diseased cantaloupes.

PRELIMINARY STUDY RESULTS: 16S SEQUENCING– APC CHARACTERIZATION (BARDOT)

For Indiana Farms (Summer Season 2014): What is the effect of washing cantaloupe? Wash Methods: Ø  Farm A: Well water wash upon pick up (received “wet”) Ø  Farm B: Chlorine water wash, left to dry prior to pick up    

Combined  

Washed   Unwashed  1   Pseudomonas   48.7%   Pseudomonas   65%  2   Pantoea   40.2%   Pantoea   25%  3   Stenotrophomonas   2.8%   Massilia   3.7%  4   Chryseobacter   1.9%   Erwinia   2.4%  5   Paenibacillus   1.9%   Arthobacter   1.3%  6   Arthobacter   0.9%   Chryseobacter   1.3%  7   Kocuria   0.9%   Serratia   1.3%  8   Rahnella   0.9%          9   Serratia   0.9%        

10   Uncultured   0.9%        

Ø  Unwashed cantaloupe exhibited a lower diversity than washed cantaloupe, seven genera vs. 10+, respectively. Note: This could be due to contaminated wash water, or the fact that cantaloupes weren’t fully dried after, and the wet surfaces became excellent growing conditions for bacteria, or changes in competitive environment after washing

PRELIMINARY STUDY RESULTS: 16S SEQUENCING– APC CHARACTERIZATION (BARDOT)

For Indiana Farms (Summer Season 2014): Ø  What is the effect of PCAPP treatment?

   Combined  

Treated   Untreated  

1   Pseudomonas   48.8%   Pseudomonas   69.4%  

2   Pantoea   40.8%   Pantoea   19.4%  

3   Massilia   2.4%   Chryseobacter   3.2%  

4   Stenotrophomonas   2.4%   Serratia   3.2%  

5   Arthobacter   1.6%   Kocuria   1.6%  

6   Erwinia   1.6%   Rahnella   1.6%  

7   Paenibacillus   1.6%   Uncultured   1.6%  

8   Chryseobacter   0.8%        

Ø  Plasma Treated samples indicated eight unique genera of bacteria Ø  Untreated or control samples indicated seven unique genera Ø  Post treatment, % of Pantoea increased dramatically, while Pseudomonas decreased

Note: Pantoeas and Pseudomonas bacteria, exhibit mucoid colony morphologies, where this mucoid or “slime” layer acts as a protective barrier to plasma treatment.

PRELIMINARY STUDY RESULTS: 16S SEQUENCING– APC CHARACTERIZATION (BARDOT)

For Indiana Farms (Summer Season 2014): Ø  How did each part of cantaloupe (top/bottom/side) compare?

   Combined  

Top (stem)   Bottom (blossom)   Side  1   Pseudomonas   64.1%   Pseudomonas   50%   Pseudomonas   61.1%  2   Pantoea   20%   Pantoea   46.7%   Pantoea   22.2%  

3  Stenotrophomo

nas   4%   Chryseobacteria   1.1%   Paenibacillus   11.1%  

4   Massilia   4%   Kocuria   1.1%   Chryseobacteria   5.6%  5   Arthobacter   2.7%   Serratia   1.1%          6   Erwinia   2.7%                  7   Chryseobacter   1.3%              8   Rahnella   1.3%                  9   Serratia   1.3%              

10   Uncultured   1.3%              

Ø  The cantaloupe stems had the highest diversity with 10+ unique genera of bacteria Ø  The cantaloupe blossom ends indicated five unique genera of aerobic bacteria Ø  The sides of the cantaloupe indicated four unique genera of aerobic bacteria Note: The cantaloupe stems may contain the highest diversity and/or greatest populations due to higher water activity (nutrient potential) at the stem area.

Ø  2. PCAPP TREATMENT OF CANTALOUPES Ø  Indiana cantaloupe (Aphrodite/Ariel/Minerva varieties) grown in

different locations and under different growing practices were acquired and treated with PCAPP for microbiological reduction.

Ø  PCAPP Treatment was performed on 0.25 lb cantaloupe sections (stem, blossom, and side), or whole cantaloupe (tote container).

Ø  PCAPP treatments were carried out in polypropylene (PP) storage containers (cuts) or totes (whole) using air or MA65 to achieve significant bacterial reductions without significant quality changes.

Ø  Treated samples were stored under refrigeration (cuts) or room temperature (whole) for 24 hours and then evaluated for microbial reductions.

Ø  2. PCAPP TREATMENT OF WHOLE CANTALOUPE

Ø  A PCAPP prototype treatment system was built to treat 16-20 whole cantaloupe inside a 30 gallon container.

Ø  The prototype treatment system was evaluated for resultant quality and its ability to reduce bacteria on whole cantaloupe.

Inside tote

tote

Ø  3. ASSESSMENT OF PCAPP TREATMENT RESULTS: MICROBIAL – CUT

CANTALOUPE Retail Grocery (Spring and Fall Seasons)

Control

Control

Control

Treated

Treated

Treated

Stem

Blossom

Side

0h

0h

24h

24h

24h

Ø  3. ASSESSMENT OF PCAPP TREATMENT RESULTS: MICROBIAL – CUT

CANTALOUPE

Control

Control

Control

Treated

Treated Treated

3 Log = 1,000 CFU 2 Log = 100 CFU

1 Log = 10 CFU

Ø  The higher the Log number the greater the population of bacterial colonies present

Ø  Log10 expresses scientific notation in short form

Ø  CFU = Colony Forming Unit (bacterial colony) Ø  1 Log = 1 x 10^1 = 10 CFU Ø  2 Log = 1 x 10^2 = 100 CFU Ø  3 Log = 1 x 10^3 = 1,000 CFU Ø  4 Log = 1 x 10^4 = 10,000 CFU Ø  5 Log = 1 x 10^5 = 100,000 CFU Ø  6 Log = 1 x 10^6 = 1,000,000 CFU

Bacterial populations expressed in Log10. What does this mean?

Ø  3. ASSESSMENT OF PCAPP TREATMENT RESULTS: MICROBIAL – CUT CANTALOUPE

Retail Grocery (Spring and Fall Seasons)

0.00  

0.50  

1.00  

1.50  

2.00  

2.50  

3.00  

3.50  

4.00  

Top   BoHom   Side  

Popu

laKo

n  (lo

g10  C

FU/g)  

Cantaloupe  SecKon  

Average  Microbial  Load:  Air  

Untreated   Treated  

0.00  

0.50  

1.00  

1.50  

2.00  

2.50  

3.00  

3.50  

4.00  

Top   BoHom   Side  

Popu

laKo

n  (lo

g10  C

FU/g)  

Cantaloupe  SecKon  

Average  Microbial  Load:  MA65  

Untreated   Treated  

Stem   Stem   Blossom   Blossom   Side   Side  

Microorganisms   Control   Surviving   Control   Surviving   Control   Surviving  

Aerobic Plate Count (APC)   population   population   population   population   population   population  

(log10 CFU/g)   (log10 CFU/g)   (log10 CFU/g)   (log10 CFU/g)   (log10 CFU/g)   (log10 CFU/g)  

Untreated   Treated   Untreated   Treated   Untreated   Treated  

Aerobic Plate Count (AIR)   3.30 ± 0.15   1.29 ± 0.11   3.02 ± 0.26   1.59 ± 0.35   2.76 ± 0.18   1.28 ± 0.25  

Aerobic Plate Count (MA65)   3.12 ± 0.16   0.33 ± 0.15   3.03 ± 0.22   1.65 ± 0.50   2.64 ± 0.20   0.29 ± 0.11  

3. ASSESSMENT OF PCAPP TREATMENT RESULTS: MICROBIAL – WHOLE CANTALOUPE Retail Grocery (California) vs. Local Farms (Indiana)* Note: Indiana cantaloupe 50% larger than CA variety Average weights: 3 - 4.5 lbs (CA); 4.5 - 6+ lbs (IN)

Stem   Stem   Blossom   Blossom   Side   Side  

Microorganisms   Control   Surviving   Control   Surviving   Control   Surviving  Aerobic Plate Count

(APC)   population   population   population   population   population   population  

(log10 CFU/g)   (log10 CFU/g)   (log10 CFU/g)   (log10 CFU/g)   (log10 CFU/g)   (log10 CFU/g)  

Untreated   Treated   Untreated   Treated   Untreated   Treated  Aerobic Plate Count

(CA)   4.88  ±  0.15   2.64  ±  0.61   4.79  ±  0.36   2.57  ±  0.80   4.32  ±  0.70   2.13  ±  0.13  Aerobic Plate Count

(IN)   5.95  ±  0.14   4.57  ±  0.13   5.09  ±  0.20   3.86  ±  0.20   4.99  ±  0.19   4.11  ±  0.20  

0.00  

1.00  

2.00  

3.00  

4.00  

5.00  

6.00  

7.00  

Top   BoHom   Side  

Popu

laKo

n  (lo

g10  C

FU/g)  

Cantaloupe  SecKon  

Average  Microbial  Load:  Farms  (CA)  

Untreated   Treated  

0.00  

1.00  

2.00  

3.00  

4.00  

5.00  

6.00  

7.00  

Top   BoHom   Side  

Popu

laKo

n  (lo

g10  C

FU/g)  

Cantaloupe  SecKon  

Average  Microbial  Load:  Farms  (IN)  

Untreated   Treated  

*Results across 3 different Farms in California and 3 different Farms in Indiana

Ø  3. ASSESSMENT OF PCAPP TREATMENT RESULTS: RIND COLOR (AIR VS MA65)- CUT CANTALOUPE

0.00  

10.00  

20.00  

30.00  

40.00  

50.00  

60.00  

70.00  

L*  AVG   a*  AVG   b*  AVG  

Color  U

nits  

24h  Color  (MA65):  Cantaloupe  Tops  (stem)  

Control   Treated  

0.00  

10.00  

20.00  

30.00  

40.00  

50.00  

60.00  

70.00  

L*  AVG   a*  AVG   b*  AVG  

Color  U

nits  

24h  Color  (AIR):  Cantaloupe  Tops  (stem)  

Control   Treated  

Cantaloupe Tops (stem):

0h 24h

Retail Grocery 2015 (Spring and Fall Seasons)

Ø  3. ASSESSMENT OF PCAPP TREATMENT RESULTS: RIND COLOR (AIR)- WHOLE CANTALOUPE Indiana (Summer 2015)- Results across 3 different Indiana Farms

0.00  

20.00  

40.00  

60.00  

80.00  

L*  AVG   a*  AVG   b*  AVG  

Color  U

nits  

24h  Color  (AIR):  Whole  Cantaloupe  (IN)    Tops  (stem)  

Control   Treated  

0.00  

20.00  

40.00  

60.00  

80.00  

L*  AVG   a*  AVG   b*  AVG  

Color  U

nits  

24h  Color  (AIR):  Whole  Cantaloupe  (IN)    BoHoms  (blossom)  

Control   Treated  

0.00  

20.00  

40.00  

60.00  

80.00  

L*  AVG   a*  AVG   b*  AVG  

Color  U

nits  

24h  Color  (AIR):  Whole  Cantaloupe  (IN)    Sides  

Control   Treated  

Ø  3. ASSESSMENT OF PCAPP TREATMENT RESULTS: FIRMNESS

Texture  Analysis  Instrument  (lower  le^)  and  Program  (right)  used  in  compressive  strength  tesKng  of  Cantaloupe  secKons

TA Setting   Parameter  

Test Mode:   Compression  

Pre Test Speed:   1.5 mm/sec  

Test Speed:   1.0 mm/sec  

Post Test Speed:   10.0 mm/sec  

Target Mode:   Distance  

Distance:   10.0 mm  

Trigger Type:   Auto (Force)  

Trigger Force:   Auto – 5.0 g  

Break Mode:   Off  

Stop Plot at:   Start Position  

Tare Mode:   Auto  

0.00  

5.00  

10.00  

15.00  

20.00  

25.00  

30.00  

35.00  

40.00  

Top   BoHom     Side  

Firm

enss  (N

ewtons)  

Cantaloupe  SecKon  

24h  Firmness:  Cantaloupe  (MA65)  

Control   Treated  

Ø  3. ASSESSMENT OF PCAPP TREATMENT RESULTS: FIRMNESS (AIR VS MA65)- CUT CANTALOUPE

0.00  

5.00  

10.00  

15.00  

20.00  

25.00  

30.00  

35.00  

40.00  

Top   BoHom     Side  

Firm

ess  (New

tons)  

Cantaloupe  SecKon  

24h  Firmness:  Cantaloupe  (AIR)  

Control   Treated  

Retail Grocery 2015 (Spring and Fall Seasons)

0.00  1.00  2.00  3.00  4.00  5.00  6.00  7.00  8.00  9.00  

10.00  11.00  12.00  13.00  

Top   BoHom     Side  

Firm

ess  (New

tons)  

Cantaloupe  SecKon  

24h  Firmness:  Whole  Cantaloupe  (CA)  

Control   Treated  

Ø  3. ASSESSMENT OF PCAPP TREATMENT RESULTS: FIRMNESS (AIR)- WHOLE CANTALOUPE

Retail Grocery (California) vs. Local Farms (Indiana)* Note: Indiana cantaloupe 50% larger than CA.

0.00  1.00  2.00  3.00  4.00  5.00  6.00  7.00  8.00  9.00  

10.00  11.00  12.00  13.00  

Top   BoHom     Side  

Firm

ess  (New

tons)  

Cantaloupe  SecKon  

24h  Firmness:  Whole  Cantaloupe  (IN)  

Control   Treated  

*Results across 3 different Farms in California and 3 in Indiana

•  PCAPP treatment samples •  90kV/5min/air •  Not significant changes compared

to the untreated samples. •  Firm tissue, no color changes

•  Controls •  No treatment •  Significant mold growth •  Deteriorated surface and tissue

Physical Quality Observation – PCAPP Treatment

Stem (top) Blossom (bottom) Side

Ø  Three samples were cut from fresh cantaloupes, top-bottom-side Ø  All samples were stored at room temperature for 8 days

Ø  4. FOOD SAFETY TRAINING – CANTALOUPE GROWERS

Ø  Food safety risks from growing cantaloupe is high and growers need to be informed of latest science and technology options available.

Ø  Series of educational factsheets and modules are being planned to deliver GAP’s training and share latest research on cantaloupe food safety.

OPERATIONAL COST: Ø  What does HVACP Technology Cost?

Ø  Electricity: ($0.11/kw-hr) Ø  Container (bin) volume: 500 L (90 cantaloupe), Package Gas: AIR Ø  Time: 600s, Voltage: 70 kV, power/package, 1000 W package Ø  Equipment Cost: HV - $40,000 per system, treats 1,000 cantaloupe in 11 bins at the same time.

Electrodes in each bin would be added when bin constructed - $10 per bin Ø  A single HV unit could treat 50 bins per hour. Ø  Treat 4 million cantaloupe over 40 days equals 100,000 (1111 bins) per day. Ø  Equipment operated 400 hr/year = $500 in electricity cost. Ø  Electrode costs - $10 per bin x 44,444 bins = $444,440

Ø  Estimate based on breakdown above (5 year payback): Ø  Capital equipment cost per cantaloupe/year (CEPC) = $0.002 per melon Ø  Total operating costs per year = $444,940 (replacement electrodes, gas safety

monitoring, electricity) Ø  Operational cost per cantaloupe (OCPC) = $0.11 Ø  Total cost per cantaloupe (CEPC + OCPC) = $0.112 per cantaloupe Ø  Alternative electrode designs or electrode reuse may provide much lower per unit cost.

CANTALOUPE PROJECT SUMMARY 1. Are there human pathogens present on Indiana or retail cantaloupe? Ø  We did not find any. However, they may be present below LOD of our methods. Ø  Note: Study was not designed to directly look for foodborne pathogens 2. Are there plant pathogen/spoilage organisms present on Indiana or retail cantaloupe? Ø  Yes 3. Do different farms have different microbes present? Ø  Yes (confirmed by the results of Farm data) 4. Are there areas of the cantaloupe that support more microbes than others? Ø  Yes (confirmed by the results of Stem vs. Blossom vs. Side) 5. Does PCAPP treatment reduce numbers of microbes present? Ø  Yes 6. Does PCAPP treatment change diversity or types of microbes present? Ø  Yes (confirmed by BARDOT and 16S Sequencing results) 7. Does PCAPP treatment affect quality of the cantaloupe? Ø  No (confirmed by color and firmness results) 8. Is PCAPP treatment affordable in comparison to other farm methods in controlling

microbes and maintaining product quality?

PROJECT TEAM

Ø  Bruce Applegate

Ø  Liz Maynard

Ø  Jean Jensen

Ø  Ximena Yepez

Ø  Kevin Keener

Ø  Ellen Phillips

Ø  Trevor Lim

Ø  Phil Myer

THANK YOU … QUESTIONS ???

Kevin M. Keener, Ph.D., P.E. Kevin M. Keener, Ph.D., P.E. Professor and Director Center for Crops Utilization Research and BioCentury Research Farm Iowa State University 1041 Food Sciences Building Ames, Iowa 50011-1061 Office: 515-294-0160 Cell: 515-735-2427 F: 515-294-6261 Email: kkeener@iastate.edu CCUR Internet: www.ccur.iastate.edu BCRF Internet: www.biocenturyresearchfarm.iastate.edu