Oxygen consumption is a muscle property that influences beef color. A greater amount of oxygen consumption results in less bloom and darker meat color while lower oxygen consumption produces a more desirable, bright, cherry red color. In postmortem muscles, mitochondria are greatly involved with the process of oxygen consumption. This research investigates the addition of Rotenone as a mitochondrial inhibitor to analyze the inhibition of ATP Synthesis in the mitochondria.Lew Wentz FoundationAnimal and Food Science

Denzer, M. L.

Kiyimba, F.

Lawson, M.

Mafi, G. G.

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• Oxygen consumption (OC) is a muscle property that influences fresh beef color (AMSA, 2012).• Greater OC results in less bloom and darker meat color, whereas lower OC can produce a bright-red color which is more desirable to the consumer for purchase and consumption. • In postmortem muscles, mitochondria and oxygen-consuming enzymes are primarily involved in OC (Ramanathan et al., 2020).• OC is important in characterizing beef color changes (Mancini and Hunt, 2005).• Although the role of mitochondria in beef color has been reported, limited research has validated the role of mitochondria in meat color. This research investigated the addition of rotenone as a mitochondrial inhibitor. Effect of Mitochondrial Inhibitor on Fresh Meat ColorM. Lawson1, M. L. Denzer1, F. Kiyimba1, G. G. Mafi1, R. Ramanathan11Department of Animal and Food Sciences, Oklahoma State University, Stillwater, OK 74078• The current research validates the role of mitochondria in beef color.• Meat color is determined by reciprocal interaction of biomolecules, mitochondria, and myoglobin.• In addition, lipid and myoglobin oxidation are inter-related. Primary and secondary oxidation products can enhance myoglobin oxidation. Secondary oxidation products can bind with myoglobin and promote heme release. • MRA occurs by three different pathways. In the current research, only complex I was blocked with rotenone. Decreased lipid oxidation and other MRA pathways may have increased MRA and redness. • Future research will determine the role of different mitochondrial inhibitors on beef color.  • Chuck Steaks were purchased from Homeland and were coarse ground, mixed with rotenone, vacuum packaged and stored for 24 h toreaction to occur, finely ground, and then packed into quarter pound patties. Control patties are packed on d0. Patties are PVC packaged and placed in retail display case for trials on d0 and d3.• Surface color, oxygen consumption, metmyoglobin reduction activity (MRA),  lipid oxidation, pH readings, and NADH levels weredetermined.• OC methodology was based on changes in oxygen myoglobin level when bloom patties were vacuum packaged. Briefly, patties were bloomed for one-hour, vacuum packaged (no continuous oxygen supply), and incubating samples at 30°C for 30 min to promote deoxymyoglobin formation. The HunterLab spectrophotometer readings provide the visible spectra to determine oxymyoglobin, which was calculated as 1 – (K/S 610 ÷ K/S 525). • MRA methodology using HunterLab spectrophotometer was used to evaluate metmyoglobin production.• Lipid oxidation methodology used to measure oxidative changes in meat. NADH was determined by enzyme recycling method. • The assays were replicated three times and the data were analyzed using the Mixed Proc of SAS.• Addition of rotenone improved redness of patties (Figure 2 & 4).• There was no effect of rotenone addition on pH (Figure 5). Addition of rotenone decreased oxygen consumption, probably due to inhibition of complex I activity and utilization of NADH. In support, NADH content was lower in rotenone treated samples. • Lipid oxidation was lower in rotenone treated samples compared with control (Figure 3). • Interestingly, MRA increased in rotenone-treated samples. The authors would like to thank the Wentz Scholar Program and the Office of Scholar Development and Undergraduate Research for their financial support to conduct this research. American, Meat, Science, and Association. 2012. Meat Color Measurement Guidelines. American Meat Science Association, Champaign, IL.Mancini, R. A., and M. C. Hunt. 2005. Current research in meat color. Meat Science 71(1):100-121. doi: https://doi.org/10.1016/j.meatsci.2005.03.003Ramanathan, R., S. P. Suman, and C. Faustman. 2020. Biomolecular Interactions Governing Fresh Meat Color in Post-mortem Skeletal Muscle: A Review. Journal of Agricultural and Food Chemistry doi: 10.1021/acs.jafc.9b08098• The objective of the study was to determine the effect a mitochondrial inhibitor, rotenone, on fresh meat color.• The purpose of research was to validate the role of mitochondria on meat color (rotenone is a mitochondrial inhibitor and not intended for consumption).Objective Hypothesis ResultsDiscussion  and Conclusions References & AcknowledgementsIntroduction Methods and  Materials • We hypothesize that the addition of a mitochondrial inhibitor will inhibit electron flow at complex I and impact meat color. Novel probe-type oxygen sensor to measure oxygen consumption in beef steaks 199M. Lawson1, M. L. Denzer1, G. G. Mafi1, R. Ramanathan11Department of Animal and Food Sciences, Oklahoma State University, Stillwater, OK 74078Methods and MaterialsFigure 1. HunterLab MiniScan XE PlusEffect of Mitochondrial Inhibitor on Fresh Meat ColorM. Lawson1, M. L. Denzer1, F. Kiyimba1,G. G. Mafi1, R. Ramanathan11Department of Animal and Food Sciences, Oklahoma State University, Stillwater, OK 74078Patty Preparation                                           OCR                                                MRA Lipid OxidationSamples were removed from packaging and fresh meat was exposed for a one-hour bloom processVacuum packagedIncubated at 30 °C for 30 minHunterLab readings taken at 10, 20, and 30 minSamples submerged in .3% NaNO2 solution for 20 min with fresh cut portion displayedHunterLab readings taken One-hour bloom HunterLab readings Incubated at 30 °C for 2 hoursFinal HunterLab readings taken• Chuck Steaks coarsely ground on initial day and control patties formed (115g patties)• Rotenone added in 0%, .1%, and .2% for respective treatments (Control, Rotenone, 2Rotenone)*  and stored 24 hours to allow reaction time• Coarse ground beef with added rotenone finely ground and made into patties• Patties PVC packaged and put in retail case for trials to be run on d0 and d3*100mg of rotenone per 100g of ground beef Visible Spectra data is  then calculated as 1 – (K/S 610 ÷K/S 525). Measure 3g of meat sampleBlend 3g sample with 27mL of TCA in homogenizer for 10 secondsPour ground sample into funnel and let filter for 30 minutesCombine 1mL of filtrate with 1mL of TBA in test tube and vortexPlace glass test tube in 100°C  for 10 min and then cool for 5 minPlace 1mL of sample in Spectrometer and measure absorbance at 532nmNovel probe-type oxygen sensor to measure oxygen consumption in beef steaks 199M. Lawson1, M. L. Denzer1, G. G. Mafi1, R. Ramanathan11Department of Animal and Food Sciences, Oklahoma State University, Stillwater, OK 74078ResultsEffect of Mitochondrial Inhibitor on Fresh Meat ColorM. Lawson1, M. L. Denzer1, F. Kiyimba1,G. G. Mafi1, R. Ramanathan11Department of Animal and Food Sciences, Oklahoma State University, Stillwater, OK 74078Control, d0 Control, d3Rotenone (0.1%), d0 Rotenone (0.1%), d3Rotenone (0.2%), d0 Rotenone (0.2%), d300.10.20.30.40.50.6Control, day 0 Rotenone(0.1%), day 0Rotenone(0.2%), day 0Control, day 3 Rotenone(0.1%), day 3Rotenone(0.2%), day 3Lipid oxidation (Ab532 nm)a a abccFigure 3: Effects of rotenone and storage time on lipid oxidationabb bcda0510152025Control,InitialControl,d0Rotenone(0.1%), d0Rotenone(0.2%), d0Control,d3Rotenone(0.1%), d3Rotenone(0.2%), d0a* ValueFigure 4: Effects of rotenone on redness during three day storageFigure 5: Effects of rotenone addition on pH and NADH content. Least squares mean with different letters indicate difference (P < 0.05).  abb024681012Control Rotenone (0.1%) Rotenone (0.2%)%MRAMetmyoglobin reducing activity (MRA)abc0246810121416Control Rotenone (0.1%) Rotenone (0.2%)Oxygen Consumption (OC)Figure 6: Effects of rotenone addition on metmyoglobin reducing activity. Least squares mean with different letters indicate difference (P < 0.05).a a a4.55.05.56.0Control Rotenone(0.1%)Rotenone(0.2%)pHab b01020304050Control Rotenone(0.1%)Rotenone(0.2%)Nanomoles of NADHNADHFigure 7: Effects of rotenone addition on oxygen consumption. Least squares mean with different letters indicate difference (P < 0.05).Figure 2: Pictorial representation of the effects of rotenone on color• Oxygen consumption (OC) is a muscle property that influences fresh beef color (AMSA, 2012).• Greater OC results in less bloom and darker meat color, whereas lower OC can produce a bright-red color which is more desirable to the consumer for purchase and consumption. • In postmortem muscles, mitochondria and oxygen-consuming enzymes are primarily involved in OC (Ramanathan et al., 2020).• OC is important in characterizing beef color changes (Mancini and Hunt, 2005).• Although the role of mitochondria in beef color has been reported, limited research has validated the role of mitochondria in meat color. This research investigated the addition of rotenone as a mitochondrial inhibitor. Effect of Mitochondrial Inhibitor on Fresh Meat ColorM. Lawson1, M. L. Denzer1, F. Kiyimba1, G. G. Mafi1, R. Ramanathan11Department of Animal and Food Sciences, Oklahoma State University, Stillwater, OK 74078• The current research validates the role of mitochondria in beef color.• Meat color is determined by reciprocal interaction of biomolecules, mitochondria, and myoglobin.• In addition, lipid and myoglobin oxidation are inter-related. Primary and secondary oxidation products can enhance myoglobin oxidation. Secondary oxidation products can bind with myoglobin and promote heme release. • MRA occurs by three different pathways. In the current research, only complex I was blocked with rotenone. Decreased lipid oxidation and other MRA pathways may have increased MRA and redness. • Future research will determine the role of different mitochondrial inhibitors on beef color.  • Chuck Steaks were purchased from Homeland and were coarse ground, mixed with rotenone, vacuum packaged and stored for 24 h toreaction to occur, finely ground, and then packed into quarter pound patties. Control patties are packed on d0. Patties are PVC packaged and placed in retail display case for trials on d0 and d3.• Surface color, oxygen consumption, metmyoglobin reduction activity (MRA),  lipid oxidation, pH readings, and NADH levels weredetermined.• OC methodology was based on changes in oxygen myoglobin level when bloom patties were vacuum packaged. Briefly, patties were bloomed for one-hour, vacuum packaged (no continuous oxygen supply), and incubating samples at 30°C for 30 min to promote deoxymyoglobin formation. The HunterLab spectrophotometer readings provide the visible spectra to determine oxymyoglobin, which was calculated as 1 – (K/S 610 ÷ K/S 525). • MRA methodology using HunterLab spectrophotometer was used to evaluate metmyoglobin production.• Lipid oxidation methodology used to measure oxidative changes in meat. NADH was determined by enzyme recycling method. • The assays were replicated three times and the data were analyzed using the Mixed Proc of SAS.• Addition of rotenone improved redness of patties (Figure 2 & 4).• There was no effect of rotenone addition on pH (Figure 5). Addition of rotenone decreased oxygen consumption, probably due to inhibition of complex I activity and utilization of NADH. In support, NADH content was lower in rotenone treated samples. • Lipid oxidation was lower in rotenone treated samples compared with control (Figure 3). • Interestingly, MRA increased in rotenone-treated samples. The authors would like to thank the Wentz Scholar Program and the Office of Scholar Development and Undergraduate Research for their financial support to conduct this research. American, Meat, Science, and Association. 2012. Meat Color Measurement Guidelines. American Meat Science Association, Champaign, IL.Mancini, R. A., and M. C. Hunt. 2005. Current research in meat color. Meat Science 71(1):100-121. doi: https://doi.org/10.1016/j.meatsci.2005.03.003Ramanathan, R., S. P. Suman, and C. Faustman. 2020. Biomolecular Interactions Governing Fresh Meat Color in Post-mortem Skeletal Muscle: A Review. Journal of Agricultural and Food Chemistry doi: 10.1021/acs.jafc.9b08098• The objective of the study was to determine the effect a mitochondrial inhibitor, rotenone, on fresh meat color.• The purpose of research was to validate the role of mitochondria on meat color (rotenone is a mitochondrial inhibitor and not intended for consumption).Objective Hypothesis ResultsDiscussion  and Conclusions References & AcknowledgementsIntroduction Methods and  Materials • We hypothesize that the addition of a mitochondrial inhibitor will inhibit electron flow at complex I and impact meat color. 

Effect of mitochondrial inhibitor on fresh meat color

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Effect of mitochondrial inhibitor on fresh meat color

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