Conventional freezing plus high pressure-low temperature treatment: Physical properties, microbial quality and storage stability of beef meat

Meat high-hydrostatic pressure treatment causes severe decolouration, preventing its commercialisation due to consumer rejection. Novel procedures involving product freezing plus low-temperature pressure processing are here investigated. Room temperature (20 °C) pressurisation (650 MPa/10 min) and air blast freezing (−30 °C) are compared to air blast freezing plus high pressure at subzero temperature (−35 °C) in terms of drip loss, expressible moisture, shear force, colour, microbial quality and storage stability of fresh and salt-added beef samples (Longissimus dorsi muscle). The latter treatment induced solid water transitions among ice phases. Fresh beef high pressure treatment (650 MPa/20 °C/10 min) increased significantly expressible moisture while it decreased in pressurised (650 MPa/−35 °C/10 min) frozen beef. Salt addition reduced high pressure-induced water loss. Treatments studied did not change fresh or salt-added samples shear force. Frozen beef pressurised at low temperature showed L, a and b values after thawing close to fresh samples. However, these samples in frozen state, presented chromatic parameters similar to unfrozen beef pressurised at room temperature. Apparently, freezing protects meat against pressure colour deterioration, fresh colour being recovered after thawing. High pressure processing (20 °C or −35 °C) was very effective reducing aerobic total (2-log₁₀ cycles) and lactic acid bacteria counts (2.4-log₁₀ cycles), in fresh and salt-added samples. Frozen + pressurised beef stored at −18 °C during 45 days recovered its original colour after thawing, similarly to just-treated samples while their counts remain below detection limits during storage.     

Fracture energy during slicing of frozen meat by a vibrating knife

The force, and hence fracture energy, required to cut horse quadriceps muscle, using a knife which was vibrated in the direction of material feed, were measured over a range of temperatures (−1.5 to −32.5 °C), vibration frequencies (no vibration and 1 to 1000 Hz), accelerations (0.073 to 75 m s⁻²), thicknesses (0 to 40 μm off-cut), and direction to the muscle grain. High vibration accelerations (75 m s⁻²) resulted in reduction of the cutting force; the reduction was independent of the off-cut thicknesses and resulted from lowering the coefficient of friction between the vibrating blade and the frozen meat. Rows of ice formed by the blade showed that pressure melting of the ice around the cutting region occurred during cutting and suggested that a hydrodynamic lubricating layer of water reduced the friction of the vibrating blade.     

The effects of antifreeze proteins on chilled and frozen meat

The effects of cryoprotectant proteins, trivially termed ‘antifreeze proteins’, from the Antarctic Cod and the Winter Flounder were assessed in meat during chilling and freezing. In light-microscopy studies, bovine muscle (Sternomandibularis) samples were soaked in phosphate buffered saline with and without 0·1 mg/ml antifreeze protein. Samples were then held frozen (−20°C) or chilled (2°C) for 3 days. Samples were freeze-substituted, embedded in resin and sectioned. With antifreeze protein present, transverse sections of frozen samples had many small intracellular spaces, probably representing ice crystals. Frozen controls had much larger intracellular single spaces. Antifreeze protein had no effect on chilled samples.

Similarly treated samples were examined by scanning electron microscopy using a cryostage attachment. Chilled ovine muscle samples (Peroneus longus) were soaked for various periods (0–7 days) in 0·9% saline containing various concentrations of antifreeze proteins (0–1 mg/ml). Samples were then held frozen (−20°C) or chilled (2°C) for 5 or 7 days. With frozen samples, antifreeze proteins reduced the size of ice crystals, compared to the control. This effect depended upon the concentration used and the period of soaking before the samples were frozen, but was independent of source. Antifreeze proteins had no effect on chilled samples.     

Definition of the optimal freezing rate-2. Investigation of the physico-chemical properties of beef M. longissimus dorsi frozen at different freezing rates

The influence of freezing rate on weight loss during the freezing, thawing and cooking, on water-binding capacity, on sensory and other physico-chemical properties of beef M. longissimus dorsi was investigated. The changes in myofibrillar proteins in muscle samples frozen at different freezing rates were also investigated.

The greatest weight losses during the freezing, thawing and cooking were registered at slow freezing procedures (freezing rate of 0·22 cm/h and 0·29 cm/h), when the meat was tougher and less soft. The solubility of myofibrillar proteins was least from those muscles frozen at such freezing rates.

The freezing of samples at freezing rates of 3·33 cm/h and 3·95 cm/h had less influence on their physico-chemical characteristics. The solubility of the myofibrillar proteins from such samples was greatest, and the cooked samples were the most tender.

From analysis of the results it was concluded that optimal conditions for meat freezing seem to be those when the average freezing rate is 2–5 cm/h.     

Serine peptidase inhibitors, the best predictor of beef ageing amongst a large set of quantitative variables

For consumers, tenderness is the most important sensory attribute of beef meat and, though to a lesser extent, of pork. Tenderness is therefore by far the most common cause of its unacceptability. The major challenge for the beef industry is to evaluate the toughness of the meat as soon as possible after death. In this context, the aim of the present work was to develop an equation to predict the myofibrillar ultimate resistance of raw meat.

The study was done on the Longissimus muscle from twenty three 19 months-old Charolais bulls grown in the same INRA farm. Muscles excised within 1 h post-mortem were vacuum packed and stored at 15 °C during 24 h and then transferred to 4 °C until used. The activities of lactate dehydrogenase, citrate synthase and myofibrillar Mg–Ca dependent ATPase, the levels of lactate dehydrogenase enzyme, myoglobin, myosin types I, IIa and IIb, cysteine and serine peptidase inhibitors, the pH, the osmolarity, the expressible juice, μ-calpain, m-calpain and calpastatin and meat toughness were measured. According to the physical method used here, the force measured on raw meat represents the resistance of the myofibillar structure.

Stepwise linear regression was used to determine the best equation (p < 0.05) for predicting toughness at 6 days post-mortem. A 6-variables predictive equation including serine peptidase inhibitors (partial R² = 0.4), the rate (partial R² = 0.25), and the extent of pH decline (partial R² = 0.03), the at death LDH activity (partial R² = 0.24), the extent of increase in osmotic pressure (partial R² = 0.13), and the rate of μ-calpain activity loss (partial R² = 0.09), explained 70% of the variability in meat toughness at 6 days post-mortem. This equation was developed from 20 animals and the other 3 animals, chosen randomly, were used to validate it. The absolute need for a predictive model of meat toughness and the nature of the serine peptidase inhibitors together with their potential target enzymes are discussed.     

Sorption of chloroanisole vapors by raisin packaging material

The behavior and concentration of 2,4,6-trichloroanisole (TCA) vapors migrating into low-density polyethylene film (PE) of 0.39 μm at various temperatures and desorption of TCA from PE were determined. After 12 h exposure, 1642 μg g⁻¹ TCA was sorbed at 30 °C compared with 675 μg g⁻¹ at 20 °C. For PE to reach equilibrium of 4200 μg g⁻¹ at 30 °C took 48 h, but 120 h at 20 °C. The transmission of TCA through PE occurred after 12 h at 30 °C (8.9 μg kg⁻¹ m⁻² h⁻¹) and after 36 h at 20 °C (5.0 μg kg⁻¹ m⁻² h⁻¹). Desorption of TCA from PE increased with temperature. At 80 °C, 99% TCA was desorbed in 1 h compared to 51% at 40 °C, 31% at 30 °C and 17% at 20 °C. The rate of sorption, desorption and transmission of TCA vapors by PE is highly temperature-dependent.     

Evaluation of the antioxidant potential of grape seed and bearberry extracts in raw and cooked pork

The effect of grape seed extract (GSE) and bearberry (BB), on lipid oxidation (TBARS, mg malondialdehyde (MDA)/kg muscle), colour (CIE ‘a’ redness value), pH, microbial status (log₁₀CFU colony forming units/g pork) and sensorial properties of cooked pork patties was investigated. GSE (0–1000 μg/g muscle) and BB (0–1000 μg/g muscle) were added to raw pork (M. longissimus dorsi) patties which were stored in modified atmosphere packs (MAP) (75% O₂:25% CO₂) for up to 12 days at 4 °C. Cooked pork patties were stored in MAP (70% N₂:30% CO₂) for up to 4 days at 4 °C. Mesophilic plate counts and pork pH were unaffected by GSE and BB. GSE and BB addition decreased (P < 0.05) lipid oxidation (TBARS) in raw pork patties on days 9 and 12 of storage, relative to controls. Antioxidant activity of GSE and BB was observed in cooked pork patties demonstrating the thermal stability of GSE and BB. The ‘a’ redness values of raw and cooked pork patties marginally increased with increasing GSE concentration. The sensory properties of cooked pork patties were unaffected by GSE and BB addition. Results obtained demonstrate the potential for using health promoting nutraceuticals in meat and meat products.     

Identification of halothane genotypes by calcium accumulation and their meat quality using live pigs

The three halothane genotypes (NN, Nn, and nm) were identified by measuring the capacity for Ca²⁺ accumulation by sarcoplasmic reticulum in whole muscle homogenate preparations of M. longissimus dorsi with a Ca²⁺ specific electrode at 35°C. Significant differences (P < 0·001) in deterioration (%) of Ca²⁺ accumulation, 12% for NN, 35% for Nn, and 81% for nn pigs, were observed after ageing the whole muscle homogenate preparations for 24 h in ice.

Predictions of meat quality in live pigs (n = 34) based on the values for water-holding capacity, assessed as fluid (g/0·5 g wet wt LD), and pH (fluid) by using small biopsy LD samples (Cheah et al. 1993) were performed on all the halothane genotypes. The halothane genotype NN (n = 11) showed a fluid value of 0·37 ± 0·01 and a pH (fluid) value of 6·62 ± 0·03 as compared with 0·61 ± 0·02 and 5·84 ± 0·04, respectively, for the halothane genotype nn (n = 13). The Nn pigs (n = 10) showed fluid (0·49 ± 0·03) and pH (fluid) (6·19 ± 0·11) values between those values observed for the two homozygotes (NN and nn). Predictions of meat quality in live pigs from biopsy LD muscles were confirmed from assessments on post-mortem LD muscles based on pH₁ and fibre optic probe (FOP) measurements.

The extent of deterioration (%) in Ca²⁺ accumulation showed high correlations with fluid (r = −0·861) and pH (fluid) (r = −0·831) in the biopsy LD samples, and with pH₁ (r = 0·663), FOP (r = −0·812), and drip (%) loss (r = −0·777) in the post-mortem LD samples.     

Secondary sexual development (Masculinity) of bovine males: 2. Influence on certain meat quality characteristics

Differences in meat quality traits between bulls with secondary sexual development (bulls(+), n = 10), those without this development (bulls(−), n = 10) and steers (n = 10) were investigated. All animals had no permanent incisors (A-age group). Significant differences (P < 0·05) between bulls(+) and bulls(−) were found for the cooking loss percentage of the M. splenius (27·83% versus 31·11%, respectively), iron content of the M. splenius (56·02μg/g versus 49·43μg/g, respectively) and total collagen content of the M. splenius (3·74 versus 4·73 measured as Hyp N/Tot N x 1000, respectively). Drip loss of the wingrib cut (4·01% versus 5·18%, respectively) was also significantly different between bulls(+) and bulls(−). For the M. longissimus thoracis, no significant (P < 0·05) differences in any of the quality-indicating parameters investigated could be found. It is concluded that the M. splenius can be used as an indicator muscle for masculinity, based on meat quality attributes. This is supported by the correlation coefficients obtained between masculinity and the intramuscular collagen content of the M. splenius (r = −0·55) and the iron content of the M. splenius (r = 0·46). For all the other quality attributes investigated, non-significant (P > 0·05) differences between the three sex condition groups were found. It is concluded that the influence of masculinity on meat quality traits of young bulls is of little practical importance in a classification and grading system.     

Stability of catalase and its potential role in lipid oxidation in meat

The activity of catalase in microbial growth-controlled and uncontrolled ground beef muscle (semimembranosus, SM) did not change (P>0.05) during 6-day storage at 4°C. Likewise, catalase activity in ground, beef SM and longissimus dorsi (LD), pork LD, and chicken breast (B) and thigh (T) muscles was not affected (P>0.05) by 2-month storage at −20°C, with or without mid-month thawing/refreezing. When sodium azide (a catalase inhibitor) was added to ground beef SM, lipid oxidation (as measured by peroxide values) during 4-day refrigeration was higher (P<0.05) in treated samples — 43 and 55% higher at day 2 and day 4, respectively — than in the controls. It was concluded that catalase would be stable during meat storage/distribution and contribute significantly to the antioxidative process in raw meat products.     

Predicting variability of ageing and toughness in beef M. Longissimus lumborum et thoracis

The object of this study was to determine muscle characteristics which might predict meat toughness. Eleven Charolais cattle were slaughtered at approximately 26 months of age and the Longissimus lumborum et thoracis muscle was taken 1 hr post mortem and stored at 12 °C for 24 hr and then at 4 °C.

The average half-life for ageing in these raw muscles was 4.6 days but the toughness varied widely between the animals. Toughness varied 3-fold and the rate of ageing varied 20-fold between animals.

Correlations were done to determine which characteristics might explain this variability. Toughness was correlated positively with increase in oxidative status of muscle and the initial levels of calpastatin. Toughness was correlated negatively with the initial levels of μ- and m-calpains and cysteine and serine proteinase inhibitors, the initial pH values and the rates of their decline. The rates of ageing were highly correlated positively with the initial levels of proteinase inhibitors and the rates of decline of calpastatin and negatively with the ultimate amounts of expressible juice.

There was a wide variability in tenderness in M. Longissimus lumborum et thoracis from similar animals. Variations in metabolism and enzyme activity controlled by inhibitors and calpains appear to be largely responsible for this variability.     

The effects of residual oxygen concentration and temperature on the degradation of the colour of beef packaged under oxygen-depleted atmospheres

Samples of beef longissimus dorsi (LD), approximately 5 × 5 × 1 cm, were packaged in pairs under 10 litre volumes of N₂ or CO₂ containing O₂ at concentrations between 100 and 1000 ppm. The packaged samples were stored at temperatures of 5, 1, 0 or −1·5°C, for times between 4 and 48 h. Samples of beef psoas major (PM) were packaged under N₂ or CO₂ containing O₂ at between 100 and 600 ppm, and stored at −1·5°C for 24 or 48 h. After storage, each sample was assessed for colour deterioration and discoloration, and for the fraction of metmyoglobin in the surface pigment.

The results obtained with N₂ and CO₂ atmospheres were similar. The colours of all LD samples had deteriorated after 4 h storage at 5 or 1°C, although the degree of deterioration increased with increasing O₂ concentration. All LD samples stored for 12 h at 5 or 1°C were extensively discoloured, with metmyoglobin fractions generally exceeding 60%, but those stored at −1·5°C for 48 h or less, under O₂ concentrations ≤ 400 ppm had undergraded colours. The colours of some LD samples stored at −1·5°C under about 600 ppm of O₂ were also undergraded, but the colours of samples stored under 800 or 1000 ppm had deteriorated by 24 h. The colours of LD samples stored at 0°C under > 200 ppm had deteriorated after 24 h storage, and the colours of samples stored under 100 ppm O₂ had deteriorated after 48 h storage. All PM samples were wholly discoloured after storage at −1·5°C. Evidently, the colour of beef muscle of high colour stability is resistant to degradation by atmospheres containing < 600 ppm of O₂ when the meat is stored at sub-zero temperatures, but not when the storage temperature is at or above 0°C. Beef muscle of low colour stability, such as the PM, will discolour at all low concentrations of O₂ irrespective of the storage temperature.     

The effects of spray and blast-chilling on carcass shrinkage and pork muscle quality

Combinations of blast- and spray-chilling of pork carcasses were compared to spray-chilling at conventional chilling temperatures with regard to carcass shrinkage during chilling and pork muscle quality. In experiment 1, pork sides were spray-chilled at 1°C for the first 10 h (40 spray cycles of 60-s duration every 15 min) of cooling or blast-chilled at −20°C for 1, 2 or 3 h followed by spray-chilling for 9, 8 or 7 h duration, respectively. All pork sides were then chilled to 24 h post mortem at 1°C. Experiment 2 followed the same procedures as experiment 1, except that −40°C was used as the blast-chill temperature.

Carcass shrinkage was similar for all treatments in experiment 1 at 24 h ranging from 0·5–0·7 g 100 g⁻¹. Blast/spray-chilling increased the rate of chilling and reduced the rate of post-mortem pH decline in two muscles (longissimus thoracis, LT and semimembranosus, SM) compared to the combined conventional/spray-chill treatment. Carcasses that were blast-chilled for 3 h had LT muscles that were darker with a higher protein solubility, less drip loss, shorter lengths and higher shear values compared to those from carcasses in the conventional/spray-chill treatment. In experiment 2, carcasses blast-chilled for 3 h at −40°C recorded a weight gain at 24 h of 0·4 g 100 g⁻¹, compared to a weight loss in all other treatments (0·2–0·4 g 100 g⁻¹). Muscle colour was darker in both the LT and SM of carcasses blast-chilled for 3 h at −40°C compared to carcasses from the conventional/spray-chill treatment, but most other measurements of muscle quality showed an inconsistent response to chilling treatment.     

The storage life of chilled pork packaged under carbon dioxide

Pork cuts of longissimus dorsi muscle with overlaying fat and skin were packed under vacuum in film of low oxygen transmission rate, or under CO₂ in gas impermeable aluminium foil laminate. Cuts were stored at +3 or −1·5°C. Vacuum packaged cuts were grossly spoiled by Brochothrix thermosphacta after 2 weeks' storage at 3°C and after 5 weeks at −1·5°C. Cuts packaged under CO₂ were grossly spoiled by B. thermosphacta after 5·5 weeks' storage at 3°C. Growth of B. thermosphacta was suppressed when CO₂ packaged cuts were stored at −1·5°C. At that temperature, slow growth of enterobacteria was detected after a lag of about 18 weeks. The enterobacteria caused gross spoilage of an increasing proportion of cuts between 18 and 26 weeks. Muscle tissue with pale, soft, exudative (PSE) characteristics tended to lose colour after long storage periods, apparently because of loss of myogglobin with exudate. Until spoilage, the eating qualities of pork appeared little affected by prolonged storage.     

Relationship between heat transfer parameters and the characteristic damage variables for the freezing of beef

One of the most suitable parameters for relating the freezing rate to the volume of drip produced during the thawing of meat is the characteristic time, defined as the time necessary to reduce the temperature of the sample from −1·1°C (initial freezing point in beef) to −7°C (80% of the water frozen).

However, as the freezing of beef in factories takes place with important temperature gradients, distributions of these characteristic times must be expected along the pieces of frozen meat.

In order to relate these characteristic time distributions to heat transfer parameters under industrial freezing conditions, a mathematical model which simulates the freezing of beef is developed in this paper.

The model establishes the heat transfer equations with simultaneous change of phase, taking into account the dependence of the thermal properties with the ice content and considering the anisotropy of the thermal conductivity according to the direction of the fibres.

Boundary conditions include the possibility of thermal resistances in the refrigerated interphase.

The model developed was compared with laboratory experiments performed under factory freezing conditions and showed a satisfactory agreement between theory and experiment.     

Effects of freezing, thawing and storage on some quality factors for portion-size beef cuts

Portion-size beef cuts packaged in oxygen impermeable plastic bags were used to study the effects of rates of freezing and thawing, and storage time and temperature on drip and cooking losses, shear force, destruction of glutathione and accumulation of protein-breakdown products in meat. Portions weighing 150 g or over and frozen in an air-blast at −30°C gave lower losses of drip and lower amounts of nitrogenous constituents in drip than samples weighing less than 150 g or samples frozen in cardboard boxes in still air at −18°C. Freezing and thawing or frozen storage had no significant effect on shear force of meat frozen after ageing. During frozen storage, the destruction of glutathione and accumulation of protein-breakdown products increased, depending directly on storage temperature and time. The results show that a test based on these two biochemical changes would be suitable for assessing the quality of frozen beef.     

Effects of rigor temperature and electrical stimulation on venison quality

The effects of rigor temperature and electrical stimulation on venison quality were assessed using venison longissimus dorsi muscle. In the first trial, effect of rigor temperature (0, 15, 25, 30, 35 and 42 °C) and time post-mortem (at rigor, 3, 7 and 14 days) on drip and cooking losses, % expressible water (water holding capacity, WHC), sarcomere length, protein solubility, meat tenderness and colour were investigated. In the second trial, the effects of rigor temperature (15 and 35 °C), electric stimulation (stimulated or not stimulated) and time (at rigor, 3 and 6 weeks post-mortem) on tenderness and colour were further investigated. Results of the first trial showed no clearly established trends of the effect of rigor temperature and time on the cooking and drip losses and protein solubility except venison muscles that went into rigor at 42 °C tended to have higher drip loss and lower protein solubilities compared to muscles that went into rigor at the other temperatures. Venison water holding capacity (WHC) decreased with the increase in rigor temperature (P < 0.001) and venison became more tender with time post-mortem. Venison colour improved with increasing rigor temperature. During display, samples that went into rigor at 15, 25 and 35 °C had the lowest and those at 0 and 42 °C had the highest rate of change of redness (a^*) value with time. In the second trial, tenderness was improved by stimulation (P = 0.01). Redness (a^*) values were affected by rigor temperature (P < 0.01) and post-mortem time (P < 0.001) but not by electrical stimulation. It is concluded that venison tenderness can be improved via the manipulation of rigor temperature to obtain acceptable level of tenderness early post-mortem with less damaging effect on colour stability.     

Definition of the optimum freezing rate-1. Investigation of structure and ultrastructure of beef M. longissimus dorsi frozen at different freezing rates

The influence of freezing rate on location, shape and size of ice crystals formed during freezing of beef M. longissimus dorsi, as well as its influence on ultrastructure, were investigated. Muscle samples were frozen at different rates: 0·22 cm/h and 0·39 cm/h (cooling agent was chilled air), and 3·33 cm/h, 3.95 cm/h, 4·92 cm/h and 5·66 cm/h (cooling agent was liquid carbon dioxide which expanded in the sucking-pipe of the tunnel freezer).

It was found that by slow freezing (freezing rates 0·22 cm/h and 0·39 cm/h) 30·00 μm). An increase in the freezing rate was followed by a change in ice crystal location. In this case they had also been formed intracellularly. The number of crystals increased while their size decreased.

The most intensive fibre damage was found in samples frozen at a rate of 0·22 cm/h, and the least in samples frozen at a rate of 3·95 cm/h with a freezing temperature of −50°C.     

Development of backfat and individual fat layers in the pig and its relationship with carcass lean

The development of backfat (total and individual layers) was monitored in 140 Yorkshire pigs (71 castrates and 69 gilts) during a growing period extending from 14·5 kg to 137·0 kg live weight using a serial slaughter procedure. The allometric coefficient (b) for fat thickness was calculated at several locations extending from the shoulder to the M. gluteus medius at the mid-line and lateral to the mid-line. The relationships between backfat (total and individual layers) and the yield of trimmed boneless meat were also obtained from 80 carcasses.

Total backfat was the thickest at the shoulder, decreased gradually to the last rib, increased at the maximum loin, decreased to the middle of the M. gluteus medius and then increased posterior to the M. gluteus medius. The development of total backfat relative to weight at slaughter was generally slowest at the shoulder (b = 0·555−0·767), most rapid in the region extending from the 5/6 last rib to the last rib b = 0·729−0·810) and intermediate at the loin and in the region of the M. gluteus medius (b = 0·609−0·834). The development of the middle fat layer (b = 0·609−1·107) was more rapid than that of the outer layer (b = 0·500−0·817), the inner layer being intermediate (b = 0·515−0·966). Consistent with the rate of development observed for the individual fat layers, at light weights, the outer layer was usually more predominant, particularly at positions above the M. longissimus, whereas, at heavier weights, the middle layer became predominant.

It was observed that backfat (total or individual layers) measurements, which were the most precise predictors of yield of trimmed boneless meat, that is, measurements from the 5/6 last rib to the last rib (RSE = 4·6−3·1), also had the most rapid rate of development. Furthermore, the middle layer of backfat which exhibited the fastest rate of development of all three individual layers (b = 0·609−1·107), also contributed the most to the observed precision (RSE = 4·6−3·1).