Safety based shelf life for ready to eat pre-packaged refrigerated foods Cold Chain Management III QuickTime™ and a TIFF (Uncompressed) decompressor are needed to see this picture. QuickTime™ and a TIFF (Uncompressed) decompressor are needed to see this picture. Bonn Germany June 2-3, 2008 QuickTime™ and a TIFF (Uncompressed) decompressor are needed to see this picture. Dr. Ted Labuza & Dr. Francisco Diez & Dr. Amit Pal University of Minnesota St Paul MN 55108 Slide # 1 Greetings from Minnesota, USA We are 4.4 million 15,129 lakes Two ocean going seaport ports State bird is a mosquito Rated one of best places to live in USA but climate goes from -35°C to +37°C Slide # 3 What do we want to know ? • Get the location of a case-lot of food in the cold distribution chain in case of an adverse event (eg. recall) – ISO 9000-2000 Clause 3.5.4 Traceability is the ability to trace the history or location of what is under consideration QuickTime™ and a TIFF (Uncompressed) decompressor are needed to see this picture. • What is the shelf life left of the product at each point in the distribution? “Quality” • Is the food safe when we eat it, determined either by sensors or by modeling the time-temperature history in the cold chain? Slide # 4 Expiration Dating of foods & Safety The last package of of hotdogs you sold me was no good !!! Did the date expire ? Mike’s Hotdog Stand No, but the dog I gave it to did ! Mike’s Hotdog Stand Question of safety vs food quality US regulatory Value of life = $6.5 MM Slide # 5 U.S. Regulatory Stance on Shelf Life • Federal Laws – Required for drugs, OTC and infant formula – Drugs 10% loss below label value on lower 95% CL line – All other food products voluntary • State laws – 30 states regulate some dates (dairy, meat) – Minnesota ≤ 90 days – None based on safety , more for commerce • GMA v Mass. Dept. Public Health – 393 NE2nd 881 (1981) – Court rules state can require dating based on public health concerns re US Constitution Slide # 6 EU Food Dating Rules Directive 97/4/EEC Article 9 of 79/112/EEC 1.The date of minimum durability of a foodstuff shall be the date until which the foodstuff retains its specific properties when properly stored. It shall be indicated in accordance with the provisions of this article. 2.The date shall be preceded by the words: --“Best before…” when the date includes an indication of the day, --“Best before end…” in other cases 3. In the case of foodstuffs which, from the microbiological point of view, are highly perishable and are therefore likely after a short period to constitute an immediate danger to human health, the date of minimum durability shall be replaced by the “use by” date. Slide # 7 Types of food dating Code date Born on date Sell by date Better if used by Freeze by Best when purchased by Best if used by - minimum durability Death date - use by (expiration) Slide # 8 Temperature Abuse • Szybist and Labuza 2001 – 100 homes 42°F = 5.6°C Slide # 9 Taoukis study Greece 20 20 9 11 temperature ( C) 0 3 7 5 3 0 7 5 17 5 15 10 13 10 11 15 9 15 5 % of cases te m pe r atur e ( oC) Left: Temperature distribution in commercial chilled storage. (Measurements in 150 supermarkets in the metropolitan area of Athens). Right: Temperature distribution in domestic refrigerators. (Based on measurements at 40 households).(Adapted from Taoukis et al., 1998.) Slide # 10 So dates by themselves ignore t-T history in refrigerated distribution Thus to ensure safety need t-T integration Slide # 11 Degradation Kinetic parameters • Rate of degradation as f(T) • Arrhenius function ln rate vs 1/T in K --> Ea • simple Ln time to X (or rate) vs temperature --> Q10 • Olley square root of rate vs T 100 ARRHENIUS PLOT TTI I −E a Ea k = ko e ln k = ln ko − RT logτ = τ r − bT RT k = A + bT k Or 10 TTD 1 3.2 3.3 3.4 3.5 3.6 3.7 3 1/Tx10 (K -1 ) Slide # 12 Sensory Shelf Life Plot of Skim Milk 500 marker problem 2x104 2x102 200 CFU/mL CFU/mL 3x102 CFU/mL 4x107 CFU/mL 100 Hours 8x104 CFU/mL 50 Q10 ~ 5 10 0 2 4 6 8 10 Temperature °C 12 14 MN-SD Dairy Research Center Slide # 13 Area under T vs t curve Shelf life depends on the rate and temperature sensitivity Ea or Q10, ie how much faster for a 10 °C increase in T Slide # 14 Using kinetics Hot dog Q10 = 3.3 ts 0°C = 90 days %fconsumed =100 (t/ ts0°C ) x Q10 ΔT/10 % fcon=100(1/90) x 3.320/10 =10.7% fcon =100 (20/90) x 3.31/10 = 25% Slide # 15 Sensory of Skim Milk Amount of Change for Q10 = 5 t 0°C=28 days • for 1 day @ 20°C • %fconsumed =100 (1/ 28 )x 6 20/10 =128% (long dead) • for 20 days at 1°C • %fconsumed = 100 (20/28) x 6 1/10 = 85% or 4 days left @1°C Slide # 16 Time Temperature Integration • Combine T vs t, k and Ea or Q10 functions in algorithm – Temperature vs time measurement – Algorithm for Reaction extent as f(t,T) – Use an integrating tag TTI T °C Δt time Slide # 17 USDA -FSIS 1998 Guidance for Beef Grinders to Better Protect Public Health Guidance for Minimizing Impact Associated with a Food Safety Hazard in Raw Ground Meat and Other FSIS Regulated Products Install a time-temperature indicator on the package to indicate adequate temperature of storage, distribution, and display (in grocery and other retail establishments). Slide # 18 Shelf Life plots food or drug vs tag shelf life shelf life Log time TTI TTI Temperature Temperature shelf life Log time TTI TTI shelf life Temperature Temperature Illustration of proper and improper TTI design Slide # 19 Shelf Life Dating Warnings • August 1998 Prevention Magazine - NBC survey – 61% feel sell by is last date to safely sell – 34% feel use by is last date to safely use • 1999 US IFT document to RCs related to safety through label date • 1999 National Enquirer – Use by date is a stern warning on meats, poultry, fish and other perishables. Pay close attention and do not use once date is passed • Food Technology July 1999 “Playing the Open Dating Game” Ted Labuza and Lynn Szybist Slide # 20 • FIFO vs LSFO system Taoukis et al – ≥15% savings (EU programs including SMAS) – Taoukis, P.S., Bili M., Giannakourou M. (1998). “Application of shelf life modelling of chilled salad products to a TTI based distribution and stock rotation system.“ Proceedings of the International Symposium on Applications of Modelling as an Innovative Technology in the Agri-Food-Chain Ed. L.M.M. Tijskens, Wageningen, Netherlands, p. 131-140. – Case study with fish in Greece to Italy chain store – Basis for formation of SMAS • • http://www.vitsab.com/htdocs/default.htm Contact [email protected] Slide # 21 % Life Consumed TTI Center Box Time (h) TTI Top Box out Inside box Center of box out Inside box Center of box 60% 40% 20% 70% 45% 25% 85% 50% 25% >100% 75% 40% >100% 90% 45% >100% >100% 60% 48 78 120 Field Test : Monitoring seabream exported from Greece to Italy Note test showed if use LSFO increase profit by 15% Slide # 22 Theoretical Probability of Reduction in illness using LSFO 0.50 FIFO Probability Probability 0.40 SMAS 0.30 0.20 0.10 0.00 -16 -14 -12 -10 -8 -6 -4 -2 0 -0.10 Log probability of illness To prove it need data for growth of pathogens or toxin production Slide # 23 Slide # 24 1. What are the scientific parameters for establishing safetybased ‘‘use-by’’ date labels for refrigerated RTE foods? 2. What effect do the multiple factors that influence the growth and survival of L. monocytogenes, i.e., strain differences, food matrices, production and distribution systems, consumer susceptibility, etc., have on the establishment of safety based ‘‘use-by’’ date labels for refrigerated RTE foods? 3. What data need to be acquired to scientifically validate and verify the adequacy of a proposed safety-based ‘‘use-by’’ date label for a refrigerated RTE food? Note sushi/sashimi are RTE consumed as raw products as are oysters Slide # 25 4. Should safety-based ‘‘use-by’’ date labels for refrigerated RTE foods be established using mathematical modeling techniques? If so, what modeling approaches are best suited to the development of labels for refrigerated RTE foods? 5. What impact would safety-based ‘‘use-by’’ date labels created for one psychrotrophic pathogen, e.g., L. monocytogenes, likely have on the control of other foodborne pathogens in refrigerated RTE foods? Note that in US under FDA, Listeria is an adulterant so if you detect it (1 CFU/25 g) then the food is considered adulterated while EU & Canada rules allow up to 100 CFU/g or 2500 per 25 g in a food as long as it does not allow growth above that. Slide # 26 Time-to-Detect (TTD) Concept Growth from initial counts below detection since if present is adulterated (TTD) or growth to 100 CFU/g Or time to + toxin ¾ Vital to satisfy FSO (food safety objective) Limit time = TTD (conservative) or Limit time = TTD + λ Dispatch Shelf life → TTD + [ln(N/No)]/μ Or just assume growth rate below detection is same as above starting at some probable level for No Baranyi and Pin, 1999 Slide # 27 Days to detect botulinum toxin in CAP/MAP partially cooked fish Days Baker & Genigeorgis 1990 100 Safety Line 10 Q10 ~ 4 Ea ~ 21.3 Kcal/mole 1.0 0.1 0 10 20 30 Temperature °C 40 Slide # 28 Shelf Life and Storage Conditions Shelf life of frankfurters and other deli meats: ≤ 90 days at X°C At 5 °C, mean generation time of Lm from math modeling program ¾ 1.77 days in frankfurters or 12 days to reach 100 CFU/g @ 5°C ¾ 2.45 days in deli meats or 16 days (US FDA/CFSAN, USDA/FSIS & CDC, 2003) The risks from Lm could be considered minimal if Initial contamination No with Lm is at very low levels (≤10-5 CFU/g) No temperature/time abuse during storage If assume same growth rate at below detection then at No = 10-5 TTD = [ ln (1/No)] x G/0.693 or for hotdogs TTD is 29 days @ 5°C and at No = 10-6 it is 35 days ¾ Seems too short or likely growth rate much less REALITY ¾ ¾ ¾ ¾ 69-78% of consumers store opened packages of deli meats for a week ¾ 10-13% for 1-3 weeks in their refrigerators so vacuum lost (USDA-FSIS, 2006) ¾ USDA-FSIS noted in BilMar outbreak that contaminated hotdogs were consumed near or beyond end of shelf life labeled on package Slide # 29 Shelf Life Evaluation of Ready-to-Eat Meat and Poultry Products based on Listeria monocytogenes growth Dan Belina MS & Amit Pal Ph.D. Slide # 30 RTE Meat Processing Trimming Cooler & Peeler Blender Stuffer Smoking/ Cooking Weighing & Sorting Vacuum Packing CONSUMER Source: http://www.hotdogcartsdirect.com/how_hot_dogs_are_made.htm Slide # 31 Safety-Based Shelf life Dating (SBDL) • Strain differences • Food matrices • Competing microflora and packaging • Production, distribution, and handling practices • Consumer susceptibility • Initial level • Growth kinetics Lm contamination levels in RTE meats are mostly <10 CFU/g (Gombas et al., 2003; Draughon et al., 2006) Slide # 32 So what temperature should date be based on? Audits International, 1999 Lunch Meat Temperature (°F) Home (%) Retail (%) < 32 9 4 33 – 35 10 3 36 – 38 25 10 39 – 41 29 23 42 – 44 18 22 45 – 47 5 12 48 – 50 3 15 51 – 53 0.4 4 54 – 56 0.5 3 57 – 59 0.4 2 60 – 63 0.1 1.3 13% door and 4% bottom of the household refrigerators >45°F Godwin et al, 2007 (>7.2 °C) Slide # 33 Inoculum Size and TTD TTD (hours) Robinson et al., 2001 TSB 1.2 M NaCl in TSB Inoculum size Log (cells/mL) Fuqua et al., 1994; Robinson et al., 2001 Note Ln(1/No) = [0.693/G] x time This data suggests the model works with constant G and thus could model to level at below detection. Slide # 34 PRELIMINARY STUDY Francisco Diez-Gonzalez, Daniel Belina, Theodore P. Labuza and Amit Pal. Modeling the Growth of Listeria monocytogenes Based on the Time-to-Detect in Culture Media and Frankfurters Intl. J. Food Microbiology 113:277-283; 2007 Listeria monocytogenes H7776 Implicated in hotdog outbreak Growth part No = 5 CFU/g TTD part No= 0.01 CFU/g or 0.25 CFU/25 g ie less than detection Six temperatures 4 to 36°C Two media ¾ TSB broth ¾ HPP processed hotdogs Slide # 35 Phase 1 preliminary evaluation in TSB broth shows can work L. mono Time to Detection 2 R = 0.9324 1000 Q10 ~ 3.5 Hours 100 10 1 0 10 20 30 40 Temp (C) Slide # 36 Phase 1 TTD in hot dogs Ln Q10 =(Ea x 10)/[R T1T2] Q10 = 3.45 Slide # 37 Arrhenius plots of TTD TSB R2 ~ 0.91 hotdogs R2 ~ 0.96 Slide # 38 18°C Growth Curve TSB so time to 100 ~1 day Stationary phase 10 9 Log Log CFU/m L CFU /mL 8 7 6 5 Log phase 7.9 hours per log k =0.293 hr-1 Lag = 13 hr 4 3 time to 102 = 25 hr 2 1 0 0 20 40 60 80 100 120 Hours Slide # 39 Summary of growth data 9 days 29 days Note at 4°C (39.2°F) G is ~ 33 hr much longer than USDA model of 1.77 hr Slide # 40 Summary of temperature effect Q10 4.39 3.35 Q10 3.32 7.2 4.01 9.68 Note that this means just can’t use one Q10 or use largest one which would mean throwing away good food Slide # 41 Phase 2 Finding fastest growers Total 19 Lm ribotypes (ID by DUP-XXXX) with 2 reps 109 CFU/ml + 1 part frank OR turkey breast ×2 3 part PW 103-4 CFU/ml With or without PL/SD Plate Counting on PALCAM 4 °C Stomached to slurry 8 °C 12 °C 30 ml slurry × 2 Slide # 42 Growth Models Transformation → y = (N/No) [ μ ( t -λ )] Linear y = exp t>λ y =1 t<λ Growth curve y = µt + c No Lag time Time λ = [Ln(No) – c] / µ Gompertz Logistic y = A. exp [-exp μ .e A ( λ -t)+1] ] A y= 1 + exp[ Baranyi [ 4μ (λ − t ) + 2] A e μA(t) + 1 y = yo + μA(t)- ln(1+ (ymax − yo ) ) e A(t ) = t + 1 μ ln(eμt + e−μλ − e−μ (t +λ ) ) Slide # 43 Listeria monocytogenes Strains • Three strains used − Provided by Dr. M. Weidman’s ILSI Listeria database, Cornell University − Selected based on their manifesting the fastest growth characteristics on culture media and a frankfurter slurry • DUP-1044A − 1998-99, multistate outbreak, frankfurter, 4b • DUP-1042B − 2000, epidemic, Mexican style cheese, 4b • DUP-1039C − 1998, sporadic, human, 3a Slide # 44 Phase 2 Findings No significant difference between model performances (P > 0.05) So use the simplest ln N/No vs time Variability in lag times and maximum growth rates was not similar among strains – No single strain consistently had the fastest growth at all growth conditions on broth or meat slurries ¾ fastest strains selected were: DUP-1044A, 1039C, 1030A, and 1042B No definitive link between serotype and fastest strains Average lag Q10 = 7.6 and Average Log phase Q10 = 7 Time to 102 Q10 = 7.4 Pal, A, Labuza, T.P. and Diez-Gonzalez. F. Comparison of Primary Predictive Models Study the Growth of Listeria monocytogenes at Low-Temperatures in Liquid Cultures and Selection of Fastest Growing Ribotypes in Meat and Turkey Product Slurries J. Food Microbiology 25:460-470; 2008 Slide # 45 Phase 3 Growth on Frankfurters ~52.44 g and 121.9 cm2 109 CFU/ml 0.1 ml rinse or 1 ml in 4 plates 4 °C 8 °C 102 CFU 12 °C ×2 ×2 20 ml PW 40 sec rinse ×2 Lm Psychrotrophs Counting Counting on PALCAM on PCA Using the 3 fastest growers in air vs vacuum, w/wo antimicrobial, & each w/wo competition with psychrotrops at 39.2, 46.4 & 53.6 °F Initial No at < 2 CFU/g (0.3 Log) ie less than 100 CFU/gSlide # 46 DUP-1044A at 4 °C (Vacuum packaged) Control (V) HPP (V) PL/SD + HPP (V) PL/SD (V) 8 2 log10(CFU/cm ) 10 6 Sig. Diff. between HPP and control 4 2 Survived but no growth 0 -2 0 20 40 60 Time (days) 80 100 V vacuum packed HPP slices treated in package at 400 MPa (15 min) for 106 reduction, then inoculated PL/SD 2%Potassium acetate + 0.2% Na-Lactate in meat formula Slide # 47 DUP-1044A at 12 °C (Vacuum packaged) Control (V) HPP (V) PL/SD + HPP (V) PL/SD (V) 2 log10(CFU/cm ) 10 8 6 >3 log(CFU/cm2) 4 2 0 25 days vs label of 90 days but started at above detection -2 0 20 Time (days) 40 60 Slide # 48 Lm vs PPC – Comparison at 8°C 10 8°C-vacuum 8 (□) HPP (○) Control Listeria monocytogenes growth - Safety Indicator 6 4 2 (♦) PL/SD + HPP 0 (▲) PL/SD -2 0 20 40 60 80 10 (□) HPP 8°C-vacuum 8 Psychotrophs growth Spoilage Indicator Q10 ~ 6 (○) Control 6 (▲) PL/SD 4 (♦) PL/SD + HPP 2 0 -2 (♦) PL/SD + HPP (▲) PL/SD (□) HPP (○) Control 0 20 40 60 80 Slide # 49 Phase 3 Results Summary Time to 100-fold Listeria population With Without ‘P Lact. + Sod. Diac.’ ‘P Lact. + Sod. Diac.’ Franks oC 4 8 12 4 Psy 45 day 8 Psy 18 day 12 Psy 6 day Q10 = 6 DUP1044A NG NG 18-25 22-44* 3-5* 2-3* DUP1042B NG DUP1039C NG Strain Q10 = 4.7 NG-65 (air packaging) NG-21 (air packaging) 19-49* NG NG 18-45* 5-12* 3-6* Q10 = 4.7 8-10* 4-7* Q10 = 4.7 NG: No Growth ≤ 1 log growth in >90 days psy = 106psychrotrops Slide # 50 Phase 3 Findings Lm growth strain dependent so which one do we use for standard ? Even with PL/SD, Lm DUP-1044 was able to grow to 102 CFU/g but longer than quality-based shelf life of 6 days at 12 °C Without the PL/SD, pathogen level to 100 CFU/g occurs before spoilageat all temps whereas in fish@ < 10°C, bot toxin slower than spoilage Reddy et al 1999 Results could be used to create a safety based tag but would need to account for initial levels below the detection limit Pal, A, Labuza, T.P. and Diez-Gonzalez. F. Evaluating the growth of Listeria monocytogenes in refrigerated ready-to-eat frankfurters – Influence of strain, temperature, packaging, lactate and diacetate, and background microflora. (J. Food Protection accepted – in press) 2008 Slide # 51 Phase 4 TTD Modeling Assumption 100 lb = 1000 g 25 g = 0.55 g ~10 CFU (DUP-1044A) Size (g) 0.55 g × 3 No (CFU/25g) 100 2.5 250 1 1000 0.25 1500 0.18 37 °C for 48 h + + 5 ml PDX-LIB MOX agar True positive = when at least two out of the three replicates showed confirmed presence Sampling frequency: 3 days (4 °C), 2 days (8 °C), and 1 day (12 °C) TTD = first out of three consecutive positive samples Slide # 52 TTD (days) vs. Inoculum size (Ln No) 60.0 a a TTD = -3.95 ln(No) + 32.09 2 R = 0.97 50.0 b 40.0 10.0 a a 8.0 TTD = -1.24 ln(No) + 1.92 2 R = 0.89 a 6.0 b 4 °C 4.0 2.0 30.0 b 8 °C 0.0 -5.50 -4.50 0.007 0.01 -3.50 -2.50 0.04 -5.50 -4.50 -3.50 -2.50 0.1 CFU/g 6.0 Observed TTD Expected TTD (Phase 2 data) Safe growth limit TTDs with common letter are not significantly different (P > 0.05) from pair-wise t-test 4.0 a TTD = -0.62 ln(No) + 0.56 2 R = 0.95 a ab 2.0 b 12 °C 0.0 -5.50 -4.50 -3.50 -2.50 Slide # 53 Shelf Life Model (TTD vs Temp.) ShelfLife T = ShelfLife o exp( − bT ) 0.1 CFU/g 0.04 CFU/g 0.01 CFU/g Guadagni, 1968; Labuza, 1972 0.007 CFU/g 100.0 47 days TTD (days) 23 days TTD1 = 185.17e-0.34T R2 = 0.96 10.0 6 days Q10= 31 TTD2 = 151.84e-0.38T 3.4 days R2 = 0.93 Q10= 24 1.0 2 4 6 8 10 12 14 Temperature (°C) Q10 = exp(10bT ) Slide # 54 Phase 5 Findings Significant difference between TTDs existed when the inoculum sizes differed by at least 2-log (P < 0.05) but followed expected pattern of log No vs time The Q10 values for the TTD of Listeria shelf life plot ranged from 24 to 31 while lag and growth phase was about 7 in slurry and 4.6 on hotdog Q10 values change with process, composition, matrix and packaging We cannot design the proper safety tag without agreement on the right input data and would need a dual tag for safety & shelf life! Slide # 55 Micro-electronic TTI tag • Infratab (US) (www.infratab.com) – Micro-electronic TTI integrator • RFID capability for traceability • US Patent # 5,442,669 • ePC global compatible – Possible to program for all 3 growth phases which one cannot do with chemical tag Slide # 56 References • Audits International/FDA. 2006. U.S. food temperature evaluation. Available at: http://www.foodrisk.org/Audits-FDA_temp_study.htm. Accessed 12 June 2007. • Center for Food Safety and Applied Nutrition, Food Safety and Inspection Service, Centers for Disease Control and Prevention. 2003. Quantitative assessment of the relative risk to public health from foodborne Listeria monocytogenes among selected categories of ready-to-eat foods. Washington, D.C.: U.S. Department of Health and Human Services and U.S. Department of Agriculture. Available at: http://www.foodsafety.gov/~dms/lmr2-toc.html. Accessed 12 June 2007. • Mead, P. S., L. Slutsker, V. Dietz, L. F. McCaig, J. S. Bresee, C. Shapiro, P. M. Griffin, and R. V. Tauxe. 1999. Food-related illness and death in the United States. Emerg. Infect. Dis. 5:607-625. • NACMSF. 2005. Considerations for establishing safety-based consume-by date labels for refrigerated ready-to-eat foods. J. Food Prot. 68:1761-1775. • Pleasant, A.B., Soboleva, T.K., Dykes, G.A., Jones, R.J., and Filippov, A.E. 2001. Modelling of the growth of Listeria monocytogenes and a bacteriocin-producing strain of Lactobacillus in pure and mixed cultures. Food Microbiol. 18:605-615. • Reddy, N. R., H. M. Solomon, and E. J. Rhodehamel. 1999. Comparison of margin of safety between sensory spoilage and onset of Clostridium botulinum toxin development during storage of modified atmosphere(MA)-packaged fresh marine cod fillets with MA-packaged aquacultured fish fillets. J. Food Saf. 19:171-183. • USDA-FSIS. 2006. Consumer attitudes and behaviors regarding ready-to-eat foods. Available at: http://www.fsis.usda.gov/OPPDE/rdad/FRPubs/02-041N/conley_lm.htm. Accessed 12 June 2007. Slide # 57 Contact Dr. Theodore Labuza Department of Food Science and Nutrition University of Minnesota [email protected] 612-624-9701 fax 651-483-3302 cell 651-307-2985 http://www.ardilla.umn.edu/Ted_Labuza Slide # 58
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