NIR prediction of pork tenderness. Steven Shackelford, Andy King, and Tommy Wheeler USDA-ARS U.S. Meat Animal Research Center Clay Center, NE

NIR prediction of pork tenderness Steven Shackelford, Andy King, and Tommy Wheeler USDA-ARS U.S. Meat Animal Research Center Clay Center, NE

1. Slice shear force Background 2. Fresh (never frozen) vs Frozen (and thawed) 3. Noninvasive tenderness prediction system for beef longissimus using visible and near-infrared (VISNIR) spectroscopy 4. VISNIR for pork 5. NPB tenderness prediction RFP/Project

1. Slice shear force

1. Longissimus slice shear force A. Beef single measurement from 1 steak B. Pork duplicate measurement using a single slice from each of 2 chops C. Lamb single measurement (half slices from each of 2 chops combined)

Rep B slice shear force, kg 40 35 30 25 20 15 10 5 0 Repeatability = 0.90 n = 744 Pork 0 5 10 15 20 25 30 35 40 Rep A slice shear force, kg Mean = 15.3 SD = 5.0

Correlation of pork longissimus SSF with SPT

8 Sensory panel tenderness Sensory panel tenderness Correlation of WBSF and SSF with SPT 7 6 8 7 6 Tenderness 5 4 3 2 r = -.77 n = 479 Tenderness 5 4 3 2 r = -.82 n = 479 1 0 1 2 3 4 5 6 7 8 1 0 5 10 15 20 25 30 35 40 45 50 Warner-Bratzler shear force, kg Slice shear force, kg

Sensory panel tenderness Sensory panel tenderness SSF values are much greater than WBS values

2. Fresh (never frozen) vs Frozen (and thawed) A. Most loin chops/loins are sold fresh and most of those loin chops/loins are likely cooked fresh. B. If freezing and thawing alters tenderness, then we are likely biasing effects with freezing and thawing. C. The bias in most cases would be an underestimate of treatment differences.

Effect of freezing and thawing on beef longissimus slice shear force

Effect of freezing and thawing on pork longissimus slice shear force

Effect of freezing and thawing on beef longissimus slice shear force

8 Sensory panel tenderness Sensory panel tenderness Correlation of WBSF and SSF with SPT 7 If SSF has the same ability 8to predict SPT as WBS, why do we use SSF? 7 6 6 Tenderness 5 4 3 2 r = -.77 n = 479 Tenderness Throughput, which allows for large scale 4 (fresh) evaluations! 5 3 2 r = -.82 n = 479 1 0 1 2 3 4 5 6 7 8 1 0 5 10 15 20 25 30 35 40 45 50 Warner-Bratzler shear force, kg Slice shear force, kg

3. Prediction of beef longissimus tenderness with VISNIR spectroscopy

4. VISNIR for pork A. Began work on VISNIR for pork in 2001 as a part of pork quality genetics evaluation. i. We were unsuccessful, because there was very little tough pork!

4. VISNIR for pork B. In 2007, we were approached by the industry to adapt VISNIR for pork.

4. VISNIR for pork B. In 2007, we were approached by the industry to adapt VISNIR for pork. i. If there is very little tough pork, why should we do that?

Materials and Methods

4. VISNIR for pork B. In 2007, we were approached by the industry to adapt VISNIR for pork. i. If there is very little tough pork, why should we do that? ii. Given that pork is not ribbed, how can we do that? iii. Guess what happened? Initially, we were unsuccessful, because there was very little tough pork!

Materials and Methods Boneless pork loins (n = 901) were evaluated either on line on the loin boning and trimming line of large-scale commercial plants (n = 465) or at the U.S. Meat Animal Research Center abattoir (n = 436).

Materials and Methods Boneless loin sections were aged (2 C) until 14 days postmortem and two 2.54-cm thick chops were obtained from the 11 th rib region. Fresh (never frozen) chops were cooked (71 C) and longissimus slice shear force (SSF) was measured on each of the two chops. Those two values were averaged and that value was used for all analyses.

Materials and Methods Carcasses were blocked by plant (n = 3), production day (n = 24), and observed SSF Mean = 13.9 kg; SD= 3.7; CV = 26.8%; Range 6.4 to 32.4 kg)

Materials and Methods One-half of the carcasses were assigned to a calibration data set (CDS), which was used to develop regression equations, and one-half of the carcasses were assigned to a prediction data set (PDS), which was used to validate the regression equations.

Frequency, % 70 65 60 55 50 45 40 35 30 25 20 15 10 Results - CDS Calibration data set n = 451 Comparison of means; SEM = 0.3 kg; P < 10-3 Comparison of % > 20 kg; P = 0.06 VISNIR predicted tender Mean SSF = 13.4 kg SD = 3.2 kg Range 8.1-28.2 kg 3.6% > 20 kg n = 279 VISNIR not predicted tender Mean SSF = 14.5 kg SD = 4.1 kg Range 6.4-30.0 kg 8.1% > 20 kg n = 172 5 0 5 to 10 10 to 15 15 to 20 20 to 25 Slice shear force at 14 d postmortem, kg 25 to 30 30 to 35

Frequency, % 70 65 60 55 50 45 40 35 30 25 20 15 10 Results - PDS Prediction data set n = 450 Comparison of means; SEM = 0.3 kg; P < 10-7 Comparison of % > 20 kg; P < 10-5 VISNIR predicted tender Mean SSF = 13.3 kg SD = 2.8 kg Range 7.9-25.5 kg 1.8% > 20 kg n = 274 VISNIR not predicted tender Mean SSF = 15.2 kg SD = 4.9 kg Range 6.8-32.4 kg 13.6% > 20 kg n = 176 5 0 5 to 10 10 to 15 15 to 20 20 to 25 Slice shear force at 14 d postmortem, kg 25 to 30 30 to 35

Observed longissimus intramuscular fat percentage 8 7 6 5 4 Results IMF prediction VISNIR conducted on ventral side of longissimus Calibration data set R^2 = 0.62 RSD = 0.62% n = 451 Prediction data set R^2 = 0.63 RSD = 0.63% n = 450 R² = 0.62 R² = 0.63 3 2 1 0 0 1 2 3 4 5 6 7 8 Predicted longissimus intramuscular fat percentage

5. National Pork Board RFP/Project A. In response to the results of the NPB Consumer Preference Study, NPB issued a RFP for development of systems to predict pork tenderness in 2009 B. USMARC submitted a proposal to field test our VISNIR system C. TAMU submitted a proposal to test VISNIR and some less well-developed technologies. D. NPB staff encouraged us to combine efforts E. Collaborative effort was developed to maximize resources

5. National Pork Board RFP/Project F. USMARC established collaboration with four large-scale commercial packing plants i. To maximize the likelihood that we sampled ample variation in tenderness for testing the tenderness prediction systems, packing plants were selected to represent a diversity of processing systems and hog ii. iii. sources Two plants had conventional chilling systems with spray chill and two plants had blast-chilling systems All four plants had similar CO2 stunning systems

5. National Pork Board RFP/Project F. iv. A total of 1,208 loins were sampled. a. Plant 1 -- 300 loins were sampled on Nov 17, 2009 b. Plant 2 -- 300 loins were sampled on Nov 19, 2009 c. Plant 3 -- 304 loins were sampled on Jan 12, 2010 d. Plant 4 -- 304 loins were sampled on Jan 14, 2010

5. National Pork Board RFP/Project VISNIR predicted SSF class Plant 1 Plant 2 Plant 3 Plant 4 1_< 13.4 kg 2_13.4 to 13.8 kg 3_13.8 to 14.2 kg 4_> 14.2 kg Grand Total

5. National Pork Board RFP/Project F. iv. A total of 1,208 loins were sampled. a. Plant 1 -- 300 loins were sampled on Nov 17, 2009

5. National Pork Board RFP/Project VISNIR predicted SSF class Plant 1 Plant 2 Plant 3 Plant 4 1_< 13.4 kg 139 2_13.4 to 13.8 kg 80 3_13.8 to 14.2 kg 47 4_> 14.2 kg 34 Grand Total 300

5. National Pork Board RFP/Project F. iv. A total of 1,208 loins were sampled. a. Plant 1 -- 300 loins were sampled on Nov 17, 2009 b. Plant 2 -- 300 loins were sampled on Nov 19, 2009

5. National Pork Board RFP/Project VISNIR predicted SSF class Plant 1 Plant 2 Plant 3 Plant 4 1_< 13.4 kg 139 26 2_13.4 to 13.8 kg 80 39 3_13.8 to 14.2 kg 47 62 4_> 14.2 kg 34 173 Grand Total 300 300

5. National Pork Board RFP/Project G. Loin processing i. Center-cut boneless loins were identified and USMARC conducted VISNIR on-line during or immediately ii. iii. iv. following boneless loin production Loins were captured in combo bins Meat quality measurements and a second VISNIR measurement was obtained approximately 1 hour later to help facilitate development of a robust model Center-cut boneless loins were vacuum-packaged, boxed, and transported (-2.8ºC) to USMARC v. Loins arrived at USMARC within 12 hours of production and were immediately placed in a holding cooler, unboxed, sorted, inventoried, and placed on carts for aging (1.5ºC) vi. The following day, vacuum-packaged loins were weighed for subsequent purge loss determination.

5. National Pork Board RFP/Project G. Loin processing vii. 14 days postmortem, loins were unpackaged viii. VISNIR spectra collected by USMARC

5. National Pork Board RFP/Project G. Loin processing vii. 14 days postmortem, loins were unpackaged viii. VISNIR spectra collected by USMARC ix. Impedance evaluated by TAMU

5. National Pork Board RFP/Project G. Loin processing vii. 14 days postmortem, loins were unpackaged viii. VISNIR spectra collected by USMARC ix. Impedance evaluated by TAMU x. VISNIR spectra collected by TAMU xi. Loins weighed for purge loss xii. Chops cut a. 1 Chop overwrapped and placed in simulated retail display b. 2 Chops for SSF (next day 15 d postmortem) c. 2 Chops for WBSF (frozen 14 d postmortem)

Measurements made with two instruments

Results

Frequency, % Plant differences in slice shear force 70 NPB Project 60 50 40 30 Plant 1 Mean = 13.6 kg 1.3% > 25 kg n = 300 Plant 2 Mean = 18.8 kg 15.7% > 25 kg n = 300 20 10 0 < 10 10 to 15 15 to 20 20 to 25 25 to 30 30 to 35 35 to 40 40 to 45 > 45 Pork longissimus slice shear force at 15 d postmortem, kg

5. National Pork Board RFP/Project VISNIR predicted SSF class Plant 1 Plant 2 Plant 3 Plant 4 1_< 13.4 kg 139 26 41 2_13.4 to 13.8 kg 80 39 45 3_13.8 to 14.2 kg 47 62 93 4_> 14.2 kg 34 173 125 Grand Total 300 300 304

5. National Pork Board RFP/Project VISNIR predicted SSF class Plant 1 Plant 2 Plant 3 Plant 4 1_< 13.4 kg 139 26 41 60 2_13.4 to 13.8 kg 80 39 45 52 3_13.8 to 14.2 kg 47 62 93 80 4_> 14.2 kg 34 173 125 112 Grand Total 300 300 304 304

Frequency, % Plant differences in slice shear force 70 NPB Project 60 50 Plant 1 Mean = 13.6 kg 1.3% > 25 kg n = 300 Plant 4 Mean = 14.3 kg 1.6% > 25 kg n = 304 40 30 20 Plant 2 Mean = 18.8 kg 15.7% > 25 kg n = 300 Plant 3 Mean = 20.7 kg 24.7% > 25 kg n = 304 10 0 < 10 10 to 15 15 to 20 20 to 25 25 to 30 30 to 35 35 to 40 40 to 45 > 45 Pork longissimus slice shear force at 15 d postmortem, kg

5. National Pork Board RFP/Project Source of plant differences in tenderness will be discussed this afternoon during the reciprocation session entitled The effect of chilling rate on tenderness But, implications for VISNIR will be discussed here

Frequency, % Previous tenderness prediction model 55 50 Experiment 2 - predicted with model from Experinent 1 Comparison of means; SEM = 0.24 kg; P < 10-11 Comparison of % > 25 kg; P < 10-9 45 40 35 30 VISNIR predicted tender Mean SSF = 15.7 kg SD = 5.1 kg Range 7.9 to 47.1 kg 5.5% > 25 kg n = 604 25 20 15 10 VISNIR not predicted tender Mean SSF = 18.1 kg SD = 6.5 kg Range 7.7 to 40.6 kg 16.2% > 25 kg n = 604 5 0 < 10 10 to 15 15 to 20 20 to 25 25 to 30 30 to 35 35 to 40 40 to 45 > 45kg Pork longissimus slice shear force at 15 days postmortem, kg

Split-overall Plant Predicted tender Not predicted tender 1 247 53 2 94 206 3 121 183 4 142 162 This VISNIR model was not completely reflective of tenderness differences among plants

Reflectance Average spectra for each plant 0.40 0.35 0.30 0.25 0.20 0.15 Plant 1 Plant 2 Plant 3 Plant 4 0.10 0.05 0.00 400 450 500 550 600 650 700 750 800 850 900 950 1000 Wavelength, nm

Correlation analysis To develop a robust model that would properly reflect tenderness variation among and within packing plants, correlation analysis was conducted for each plant to identify the wavelength range at which reflectance was most highly related to SSF. For each plant, the strongest correlation was found at or near 822 nm.

Correlation analysis Also, variation in the plant means for reflectance at 822 nm accounted for virtually all of the variation in plant means for SSF (r = -0.99). Thus, reflectance at 822 nm was indicative of variation in tenderness both among and within plants.

Frequency, % Single variable model 60 55 Experiment 2 - predicted with reflectance at 822 nm Comparison of means; SEM = 0.23 kg; P < 10-26 Comparison of % > 25 kg; P < 10-16 50 45 40 35 30 25 20 15 10 VISNIR predicted tender Mean SSF = 15.1 kg SD = 4.5 kg Range 7.7 to 32.4 kg 3.6% > 25 kg n = 604 VISNIR not predicted tender Mean SSF = 18.7 kg SD = 6.6 kg Range 7.9 to 47.1 kg 18.0% > 25 kg n = 604 5 0 < 10 10 to 15 15 to 20 20 to 25 25 to 30 30 to 35 35 to 40 40 to 45 > 45kg Pork longissimus slice shear force at 15 days postmortem, kg

Frequency, % Follow-up project single variable model 50 45 Experiment 3 - predicted with reflectance at 822 nm Comparison of means; SEM = 0.28 kg; P < 10-9 Comparison of % > 25 kg; P < 10-5 40 35 30 25 VISNIR predicted tender Mean SSF = 16.1 kg SD = 4.1 kg Range 8.4 to 37.8 kg 3.0% > 25 kg n = 300 20 15 10 VISNIR not predicted tender Mean SSF = 18.6 kg SD = 5.3 kg Range 8.5 to 37.9 kg 12.4% > 25 kg n = 299 5 0 < 10 10 to 15 15 to 20 20 to 25 25 to 30 30 to 35 35 to 40 40 to 45 > 45kg Pork longissimus slice shear force at 15 days postmortem, kg

Conclusion It works So why hasn t it been implemented Not for a lack of interest by the industry Instrument supplier

Conclusion It works So why hasn t it been implemented Not for a lack of interest by the industry Instrument supplier We have sought other instruments capable of being used for beef and pork.

Thank you