INTEGRATED STRATEGIES FOR IMPROVING NEW YORK STATE DAIRY PRODUCTS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 0208331
• Project Start Date: Oct 1, 2006
• Project End Date: Sep 30, 2009
• Program Code: [(N/A)]- (N/A)

Recipient Organization
CORNELL UNIVERSITY
(N/A)
ITHACA,NY 14853

Performing Department
FOOD SCIENCE

Non Technical Summary
Milk is New York's leading agricultural product, accounting for more than one-half of the state's total agricultural receipts. Milk's relative loss of market share is a specific problem for NYS, as our state is 3rd in the nation in milk production, and the dairy industry comprises the largest sector of our state's agricultural economy. The long-term economic viability of the state's dairy industry is at least partially dependent on the quality of products provided to consumers. This project integrates research and extension programming to apply biotechnological tools to identify and track bacterial contaminants that are responsible for reduced product quality to their point(s) of origin and develop effective strategies for eliminating these organisms from milk production systems. The purpose of this project is threefold: 1. Enhance the production of high quality processed milk products with extended shelf-life; 2. Increase geographical marketability for the processed products, which will contribute to the financial stability of the NYS milk processing industry; and 3. Improve the quality of the fluid milk in NYS to help ensure a market for our state's dairy farmers.

Animal Health Component

50%

Research Effort Categories

Basic

40%

Applied

50%

Developmental

10%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
503	3450	1040	40%
503	3450	1060	20%
503	3450	1100	40%

Knowledge Area
503 - Quality Maintenance in Storing and Marketing Food Products;

Subject Of Investigation
3450 - Milk;

Field Of Science
1040 - Molecular biology; 1060 - Biology (whole systems); 1100 - Bacteriology;

Keywords

milk

shelf life

bacterial tracking

molecular tracking

dairy production systems

Goals / Objectives
The long-term objective of this project is to improve the economic position of the NY State dairy industry through development and application of strategies that will reliably extend product shelf-lives and thus improve product quality. We will apply biotechnology-based diagnostic tools to identify, and ultimately eliminate, reservoirs of spoilage organisms limiting shelf-life. Construction of a comprehensive microbial database will enable us to predict and model bacterial transmission routes, and hence, facilitate development of effective recommendations for eliminating spoilage microorganisms from milk production systems. Our specific objectives are to: 1. Identify sources of spoilage organisms in fluid milk production systems. 2. Generate a WWW-based, searchable database of spoilage organisms and their characteristics. 3. Disseminate findings to processing plants in NYS through Processing Plant Superintendent updates, plant visits by Extension field staff, and at professional meetings. 4. Monitor efficacy of information transfer and implementation of intervention approaches through plant visits and product testing.

Project Methods
We will identify critical points of spoilage organism entry into dairy products by sampling milk and environmental sites at various points in the processing system. Because the Cornell dairy manufactures milk with the targeted spoilage profile and because its milk is supplied from only one farm, it represents an ideal model system for our initial study. Bulk tank milk samples (4) and environmental samples (at 10 pre-determined milk contact sites) will be collected at the Cornell Teaching and Research Center. Environmental samples (at 10 pre-determined milk contact sites) and 2 percent fat processed milk samples (4) will be collected monthly over the period of one year. Up to 5 representative colonies from each sample will be characterized by Gram stain, spore stain, and for the following reactions: KOH, oxidase, and catalase. Bacillus spp. and related genera are difficult to reliably identify to the species level through the strategies described above, therefore, we will characterize up to 200 microbes by 16S rDNA and rpoB sequencing. A single colony from a plate will be used to inoculate Brain Heart Infusion (BHI) broth, which will be incubated overnight at 30C with shaking (250 rpm). DNA lysates for PCR will be prepared from overnight cultures as described by Furrer et al. (1991) with slight modification. Briefly, an overnight culture will be centrifuged at 13,000 rpm for 10 minutes and pellets will be resuspended in 95 ml of 1X PCR buffer (Promega, Madison, WI). Lysozyme will be added to achieve a final concentration of 2.5 mg/ml. After 15 min of incubation at room temperature, proteinase K (20 mg/ml) will be added and the mixture will be incubated at 58C for 1 hr. Enzymes will be subsequently inactivated by heating at 95C for 8 minutes. 16S rDNA PCR will be performed using primers PEU7 and DG74; subsequent DNA sequencing of these PCR products will be performed using primers PEU7 and P3SH. rpoB sequencing will be performed. Bacterial identities and characteristics will be entered into a WWW-based searchable database. We hypothesize that the targeted spoilage organisms exist in similar reservoirs among milk production systems. This hypothesis will be tested by collecting targeted samples in up to 10 additional commercial milk processing plants. The emerging 'bacterial reservoir model' will be continuously modified and refined as data are compiled from all processing operations. Identification of specific bacterial niches will allow implementation of systematic strategies for reduction and elimination of the organisms.

Progress 10/01/06 to 09/30/09

Outputs
OUTPUTS: Project staff organized, conducted or participated in over 40 workshops/ conferences/in-service events during 2009, attracting more than 2,500 participants. Among the off-site programs were: five regional seminars for dairy laboratory personnel, five regional workshops for processing plant superintendents, four regional refresher courses for experienced Certified Milk Inspectors (CMI), a plant HACCP training, and one training program on the sensory evaluation of milk. Programs offered at Cornell included: a Hands-On Laboratory Workshop, the Certified Milk Inspectors (CMI) School for New Inspectors, an FDA Milk Plant Sanitation and Inspection Course, a Vat Pasteurizer Workshop, a Cheese Making Workshop, two High Temperature Short Time (HTST) Pasteurizer Operator Workshops and one Rural Youth Program presentation. In addition to the off-site training program, MQIP Sensory Analysis kits were provided to several dairy plants for use in training and provided for use in the CMI School, the Cheese Making Workshop, the FFA State Fair Judging and a practice session for the FFA national competition. Personnel actively participated as speakers, organizers and committee members in many county, state and national programs and related associations including the Finger Lakes Association for Food Protection, the NYS Association for Food Protection, the International Association for Food Protection, the NY State Cheese Manufacturers Association, the Pennsylvania Association for Milk and Food Sanitarians, the Metropolitan Association for Food Protection, the Oregon Dairy Industry, the National Conference on Interstate Milk Shipments, the NCIMS HACCP Auditors Training, the Dairy Practices Council, the Seneca Castle Farm Field Days, and the NE FDA Regional Seminar. PARTICIPANTS: Nancy Carey, Research Support Specialist, Food Science Department, 412 Stocking Hall, Cornell University, Ithaca, NY 14853; 607-254-7079 (phone); 607-254-4868 (fax); [e-mail]; Shelley Laue, Technician, Food Science Department, 200 Stocking Hall, Ithaca, NY 14853, 607-255-7643 (phone), 607-254-4868 (fax), [e-mail]; Tanya Pierik, Technician, Food Science Department, 200 Stocking Hall, Ithaca, NY 14853, 607-255-7643 (phone), 607-254-4868 (fax), [e-mail]; Patricia Wood, Technician, Food Science Department, 213 Stocking Hall, Ithaca, NY 14853, 607-255-2894 (phone); 607-254-4868(fax), [e-mail]; Nicole Woodcock, Technician, Food Science Department, 200 Stocking Hall, Cornell University, Ithaca, NY 14853; 607-255-7643 (phone); 607-254-4868 (fax), [e-mail]; Janene Lucia, Extension Support Specialist, Food Science Department, 172 Stocking Hall, Cornell University, Ithaca, NY, 607-255-2892 (phone); 607-255-7619 (fax), [e-mail]; Steven Murphy, Sr. Extension Associate, Food Science Department, 171 Stocking Hall, Cornell University, Ithaca, NY, 607-255-2893 (phone); 607-255-7619 (fax), [e-mail]; Robert Ralyea, Sr. Extension Associate, Food Science Department, 200 Stocking Hall, Cornell University, Ithaca, NY 14853; 607-255-7643 (phone); 607-254-4868 (fax); Jason Huck, Dairy Plant Manager, Food Science Department, 255D Stocking Hall, Cornell University, Ithaca, NY 14853; 607-255-7900 (phone); 607-254-4868 (fax). TARGET AUDIENCES: This project targeted the New York State Dairy Processing Industry. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
In 2009, project staff worked with 30 fluid milk processing plants in NY/PA/NJ/ME/NH/MA/VT. Each plant was visited two times, and 315 raw and processed product samples were collected. Over 4,200 individual analyses were conducted on samples, and rapid feedback on relative plant ranking and processed product quality throughout shelf-life for all products was provided to each plant. In summary, through coordinated research and outreach strategies, the investigators monitored and contributed to measurable improvements in fluid milk quality and product shelf life extension in NYS.

Publications

• Latorre, A. A., J. A. S. van Kessel, J. S. Karns, M. J. Zurakowski, A. K. Pradhan, R. N. Zadoks, K. J. Boor, and Y. H. Schukken. 2009. Molecular ecology of Listeria monocytogenes: evidence for a reservoir in milking equipment on a dairy farm. Applied and Env. Microb. 75 (5):1315-1323.
• Oliver, S. P., K. J. Boor, S. C. Murphy and S. E. Murinda. 2009. Food safety hazards associated with consumption of raw milk. Foodborne Pathogens and Disease 6(7):793-806.
• Woodcock N. H., Hammond B. H., Ralyea R. D., Boor K. J. 2009. Short communication: N alpha-lauroyl-L-arginine ethylester monohydrochloride reduces bacterial growth in pasteurized milk. J Dairy Sci. 92(9):4207-4210
• Ranieri, M. L., J. R. Huck, M. Sonnen, D. M. Barbano, and K. J. Boor. 2009. High temperature, short time pasteurization temperatures inversely affect bacterial numbers during refrigerated storage of pasteurized fluid milk. J. Dairy Sci. 92:4823-4832.
• Ranieri, M. L. and K. J. Boor. 2009. Short communication: bacterial ecology of high-temperature, short-time pasteurized milk processed in the United States. J. Dairy Sci. 92:4833-4840.

Progress 10/01/07 to 09/30/08

Outputs
OUTPUTS: We have generated a WWW-based, searchable database of spoilage organisms and their characteristics (www.pathogentracker.com). We have also disseminated findings to processing plants in NYS through Processing Plant Superintendent updates, plant visits by Extension field staff, and at professional meetings. PARTICIPANTS: Cornell Teaching and Research Center (Tom Eddy), source of raw milk and environmenatal samples for microbiological analyses. Cornell Dairy Plant (Dan Sykes and Jason Huck), source of raw and processed milk for microbiological analyses. New York State Milk Promotion Advisory Board, provides matching funds for this project. Steven Murphy, Sr. Extension Associate, Cornell University, outreach, extension and field services. Matthew Ranieri, Graduate Research Assistant primarily responsible for labwork associated with this project. Nicole Woodcock, Laboratory Technician, Milk Quality Improvement Program, Cornell University. Crowley Foods, Binghamton (Stephen Lally, Mike Hesch), dairy processing plant that has provided raw and processed milk and environmental samples for microbiological analyses. Stewarts Processing, Saratoga, NY (Marguerite Petit), dairy processing plant that has provided milk and environmental samples for analysis. Upstate Farms, Rochester (Jim Tomczak, Bob Kuhn), dairy processing plant that has provided milk and environmental samples for analyses. TARGET AUDIENCES: New York State, national and international dairy processing and farming industries PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
We have generated a WWW-based, searchable database of spoilage organisms and their characteristics (www.pathogentracker.com). This database allows any interested party in the world with the capability of using our data, or adding to the database, which is a collection and storage warehouse for microbial milk spoilage information. To date, we have isolated 93 dairy farm isolates, 57 raw milk tank truck isolates, 167 dairy plant storage silo isolates and 1099 pasteurized milk isolates (1416 total) which yielded a total of 257 rpoB allelic types, representing an extensive diversity of Bacillus and Paenibacillus spp. Our molecular subtype data clearly show that certain endospore-forming bacterial subtypes are present in the dairy farm environment as well as in the processing plant. The potential for entry of these ubiquitous heat-resistant spoilage organisms into milk production and processing systems, from the dairy farm to the processing plant, represents a considerable challenge that will require a comprehensive farm-to-table approach to fluid milk quality. Finally, we disseminated these findings to processing plants in NYS through Processing Plant Superintendent updates, plant visits by Extension field staff, and at professional meetings, keeping the New York State dairy industry personnel, regulatory personnel, and dairy farming community informed of current research and findings as it relates to dairy product quality.

Publications

• Huck, J. R., M. Sonnen, and K. J. Boor. 2008. Tracking heat-resistant, cold-thriving fluid milk spoilage bacteria from farm to packaged product. J. Dairy Sci. 91:1218-1228.

Progress 10/01/06 to 09/30/07

Outputs
OUTPUTS: We have generated a WWW-based, searchable database of spoilage organisms and their characteristics (www.pathogentracker.com). We have also disseminated findings to processing plants in NYS through Processing Plant Superintendent updates, plant visits by Extension field staff, and at professional meetings. PARTICIPANTS: Cornell Teaching and Research Center (Tom Eddy), source of raw milk and environmental samples for microbiological analyses. Cornell Dairy Plant (Dan Sykes), source of raw and processed milk for microbiological analyses. New York State Milk Promotion Advisory Board, provides matching funds for this project. Jason Huck, Graduate Research Assistant, laboratory support and outreach. Steven Murphy, Sr. Extension Associate, Cornell University, outreach, extension and field services. Matthew Ranieri, Graduate Research Assistant, primarily responsible for labwork associated with this project. Nicole Woodcock, Laboratory Technician, Milk Quality Improvement Program, Cornell University. Crowley Foods, Binghamton/Albany (Stephen Lally, Tim Barnard, Mike Hesch), 2 dairy processing plants that will provide raw and processed milk and environmental samples for microbiological analyses. HP Hood, Maine/Massachusetts/Vermont (Rebecca Piston) Quality Assurance Manager for 5 dairy processing plants that will provide raw and processed milk and environmental samples for microbiological analyses. Stewarts Processing, Saratoga, NY (Marguerite Petit), dairy processing plant that will provide raw and processed milk samples and environmental samples for microbiological analyses. Upstate Farms, Buffalo/Rochester (Jim Tomczak, Bob Kuhn), 2 dairy processing plants that will provide raw and processed milk samples and environmental samples for microbiological analyses. TARGET AUDIENCES: New York State, national, and international dairy processing and farming industries.

Impacts
We have generated a WWW-based, searchable database of spoilage organisms and their characteristics (www.pathogentracker.com). This allows any interested party in the world the capability of using our data, or adding to the database, which is a collection and storage warehouse for microbial milk spoilage information. During the study we isolated 93 dairy farm isolates, 57 raw milk tank truck isolates, 138 dairy plant storage silo isolates and 336 pasteurized milk isolates (624 total) which identified a total of 154 rpoB allelic types, representing an extensive diversity of Bacillus and Paenibacillus spp. Our molecular subtype data clearly show that certain endospore-forming bacterial subtypes are present in the dairy farm environment as well as in the processing plant. The potential for entry of these ubiquitous heat-resistant spoilage organisms into milk production and processing systems, from the dairy farm to the processing plant, represents a considerable challenge that will require a comprehensive farm-to-table approach to fluid milk quality. Finally, we disseminated these findings to processing plants in NYS through Processing Plant Superintendent updates, plant visits by Extension field staff, and at professional meetings, keeping the New York State dairy industry personnel, regulatory personnel, and dairy farming community informed of current research and findings as it relates to dairy product quality.

Publications

• Huck, J. R., B. H. Hammond, S. C. Murphy, N. H. Woodcock and K. J. Boor. 2007. Tracking spore-forming bacterial contaminants in fluid milk processing systems. J. Dairy Sci. 90:4872-4883.
• Huck, J. R., N. H. Woodcock, R. D. Ralyea and K. J. Boor. 2007. Molecular subtyping and characterization of psychrotolerant endospore-forming bacteria in two NY State fluid milk processing systems. J. Food Prot. 70:2354-2364.