Carrick E. Devine

The effects of the ultimate pH of meat on tenderness changes during ageing

A range of ultimate pH (pH(u)) values, from 5.4 to 7.2, was produced in the m. longissimus thoracicum et lumborum (LD) of 55 sheep by subcutaneous injections of adrenaline, or by injection and subsequent exercise. The rate constant of ageing for each of 47 animals was calculated from shear force measurements, taken at intervals for up to 5 days from the LD held at 12 dgC. The relationship between the ageing rate and pH(u) could be adequately described by the equation rate constant (k) = 4.109 - 1.349(pH) + 0.1111(pH)(2); r(2) = 0.707, with a minimum rate constant at pH 6.07. The loin muscles from eight carcasses were selected to determine the effects of ultimate pH on the initial, unaged toughness value. Calpain activity was inhibited by intramuscular injection of ZnCl(2) dissolved in saline, and the muscles were aged at 10-12 °C on the carcass for up to 6 days. At all values of pH(u) (5.4-6.7), tenderness as measured by shear force were equivalent, from which it was concluded that the initial toughness of unaged lamb was not affected by pH(u). In the contralateral, untreated muscle, ageing was measured using both shear force measurement and changes in the myofibrillar fragmentation index (MFI). After 1 day, the intermediate pH range (5.8-6.2) has the highest shear force and the lowest fragmentation index value. However, in contrast to the shear force values, the ultimate MFI value in aged muscle was lowest in the high pH(u) range (6.4-6.7; minimum value at 6.4). The highest fragmentation values were found at the low pH(u) range. These observations suggest that pH(u)-related tenderness differences in meat result from different rates of ageing in the post rigor period, and therefore that these differences disappear following a sufficient ageing period. The slowest rate of tenderization measured on cooked meat was observed at pH(u) values around 6.07, but the least fragmentation of raw meat was observed at pH(u) values around 6.4, a difference of almost 0.33 pH unit.

Effect of rigor temperature on muscle shortening and tenderisation of restrained and unrestrained beef m. longissimus thoracicus et lumborum

Pairs of muscularis longissimus thoracicus et lumborum (LTL) from young bulls were removed within 1h of slaughter. Small portions of the muscles were placed in a rigormeter to continously follow the isometric tension and isotonic shortening developed, at constant temperatures of 15, 20, 25, 30 and 35°C, as the muscle went into rigor. The bulk LTL was placed in water baths at the same temperature. One of the bulk pairs was tightly restrained by wrapping, to reduce muscle shortening, the other was unrestrained free to shorten. For the bulk samples, shear values were measured using a Warner-Bratzler instrument (1, 7 and 14 days post mortem), and sensory attributes were measured using a sensory panel (7 and 14 days post mortem). Minimum tension and shortening occurred at 15°C. The activation energy for the muscle shortening process was larger than for the isometric tension process. This indicates that the isometric tension data, collected during rigor, does not solely reflect muscle shortening. Thus, a counteracting process that decreases the tension response, most likely ageing is simultaneously detected. Meat that went into rigor at 15°C had least shortening and was always more tender than meat going into rigor at higher temperatures. For meat entering rigor at temperatures higher than 15°C, restraining of the muscle by wrapping, significantly (p<0.05) decreased the amount of muscle shortening and resulted in an improved meat tenderness (p<0.001). It was also observed that at rigor temperatures higher than 15°C the meat tenderness is affected negatively by a reduced ageing capacity. It therefore appears that muscle shortening and enzyme activity both affect tenderness and that both are highly affected by rigor temperature and have the greatest beneficial effect at a rigor temperature of 15°C.

Electrical stimulation, muscle tension and glycolysis in bovine Sternomandibularis

Electrical parameters previously employed to hasten the onset of rigor mortis in carcasses were derived empirically. Various parameters have been studied to determine their relationship to the two-stage fall of muscle pH resulting from stimulation. At any given stimulation period pulse frequency had a considerable effect on the magnitude of the pH fall that occurs during stimulation, the greatest falls, ∼ 0·7 pH units in a 120 sec period, being achieved with 5 to 16·6 pulses. sec(-1). The same total number of pulses delivered at differing frequencies did not produce pH falls of equal magnitude. Variation in pulse shape and polarity caused minor changes. Increased energy levels per pulse increases the magnitude of the pH fall during stimulation. All stimulation parameters tested resulted in the rate of pH fall after stimulation being one and a half times to twice that of non-stimulated muscle. The tension-time curves obtained with lower frequencies (< 10 pulses . sec(-1)) show incomplete summation of pulses with sustained tension. At higher frequencies a complete tetanus develops, while tension falls rapidly soon after achieving the peak tension. The relationship between the mechanical and biochemical responses and the implications for stimulation as a practical processing method are discussed.

High and low rigor temperature effects on sheep meat tenderness and ageing

Immediately after electrical stimulation, the paired m. longissimus thoracis et lumborum (LT) of 40 sheep were boned out and wrapped tightly with a polyethylene cling film. One of the paired LTs was chilled in 15°C air to reach a rigor mortis (rigor) temperature of 18°C and the other side was placed in a water bath at 35°C and achieved rigor at this temperature. Wrapping reduced rigor shortening and mimicked meat left on the carcass. After rigor, the meat was aged at 15°C for 0, 8, 26 and 72 h and then frozen. The frozen meat was cooked to 75°C in an 85°C water bath and shear force values obtained from a 1×1 cm cross-section. The shear force values of meat for 18 and 35°C rigor were similar at zero ageing, but as ageing progressed, the 18 rigor meat aged faster and became more tender than meat that went into rigor at 35°C (P<0.001). The mean sarcomere length values of meat samples for 18 and 35°C rigor at each ageing time were significantly different (P<0.001), the samples at 35°C being shorter. When the short sarcomere length values and corresponding shear force values were removed for further data analysis, the shear force values for the 35°C rigor were still significantly greater. Thus the toughness of 35°C meat was not a consequence of muscle shortening and appears to be due to both a faster rate of tenderisation and the meat tenderising to a greater extent at the lower temperature. The cook loss at 35°C rigor (30.5%) was greater than that at 18°C rigor (28.4%) (P<0.01) and the colour Hunter L values were higher at 35°C (P<0.01) compared with 18°C, but there were no significant differences in a or b values.

The protective effect of electrical stimulation and wrapping on beef tenderness at high pre rigor temperatures.

A three factorial experimental design involving electrical stimulation (ES/NES), wrapping (wrapped/unwrapped) and pre rigor temperature (15°C or 35°C) was applied to 70 beef M. longissimus lumborum muscles to obtain a wide variation in shear force and drip loss. The shear force of all treatment groups decreased during ageing. As anticipated, wrapping and electrical stimulation had positive effects on shear force. However, high pre rigor temperature (35°C) did not result in higher shear force values if the muscles were electrically stimulated, wrapped or both. The results suggested that electrical stimulation protects against the negative effects of high pre rigor temperatures. The drip loss of all treatment groups increased during ageing in a manner that was unrelated to treatment but was correlated to tenderness (r(2)=0.70; p<0.0001). It was concluded that the application of electrical stimulation, whatever the pre rigor temperature, protects beef from toughening through the prevention of rigor shortening and the avoidance of inhibition of ageing enzymes.

The effects of non-vacuum packaging systems on drip loss from chilled beef

This study investigated drip loss in chilled beef (hot-boned m. longissimus lumborum (LL)) under conventional packaging systems, in which a vacuum is applied, and non-vacuum packaging systems. The use of vacuum during the packaging process (vacuum packaging, CO(2) atmosphere packaging) was associated with increased drip. Drip was lower in heat-shrunk vacuum packaging than in non-shrunk vacuum packaging. A slow vacuum onset had no effect on drip formation, or may have increased it. The extent of pressure reduction (vacuum; range 0-1 atm) did not significantly affect drip formation, although the standard vacuum pressure (0 atm) tended to cause more drip loss than higher pressures. The three non-vacuum anaerobic packaging systems tested were: flushed with carbon dioxide without the pressure being reduced below atmospheric (Flush), or the same system with no CO(2) flush but a proprietary oxygen absorber added (Scavenger), or a combination of both (Flush/Scavenger). Storage was at -1.5 °C for up to 20 weeks. The Flush and Flush/Scavenger systems had considerably lower drip loss than the CAP standard system (6.4%, 6.5%, and 9.1% respectively); the Scavenger system had the lowest drip loss (5.2%). Drip losses generally increased with storage period, irrespective of packaging system. All non-vacuum packaging systems except the Flush system had very low oxygen levels (<0.l% v v ). The Flush system had considerably higher levels of oxygen (0.9%) with associated browning of meat samples. All packaging systems gave a hygienic shelf-life of at least 16 weeks. At 16 weeks, microbial numbers were highest (5 × 10(6)) in the Scavenger system. The meat from all packaging systems was acceptable to taste panels even after 16 weeks of storage at -1.5 °C. There were no significant differences between any of the packaging systems for any of the sensory attributes tested. The packaging systems with the best all round performance were the Flush/Scavenger and the Scavenger systems, depending on the storage-life required.

Pre-slaughter stress arising from on-farm handling and its interactions with electrical stimulation on tenderness of lambs.

The effect of electrical stimulation of lamb carcasses (n=269) or its absence (n=257) on shear force of m. longissimus thoracis et lumborum (LT) was monitored during ageing in pasture-fed merino lambs (n=526). The lambs were slaughtered on four different days allowing durations of between one to 10 days of recovery from pre-slaughter handling (yarding, weighing and crutching) that affected ultimate pH (pH(u)). The right LT was removed 20-40min post-slaughter, tightly-wrapped in cling film (prevents the muscle cross-section increasing and thus minimising shortening) and rapidly cooled to 15°C to enter rigor mortis and age. At 0, 4, 24 and 72h post-slaughter, pH measurements and samples for shear force measurement were taken. Pre-slaughter handling had a significant negative effect on pH(u) and several days recovery were required for pH(u) to reach values associated with optimal meat quality as reflected by pH(u). Lambs with one and three days recovery (no significant difference between them) had a pH(u)>5.7 in 50% of the muscles and 19.4%>pH(u) 5.8. Whereas, in lambs with 8-10 days recovery (no significant difference between them), only 8% had a pH(u)>5.7 and 3.1%>pH(u) 5.8. Within each slaughter day electrically stimulated lambs were always more tender than non-stimulated lambs. For non-stimulated muscles at 72h, shear force values >40N occurred for 11.2% of the muscles: for electrically stimulated muscles at 72h, shear force values >40N occurred for 1.9% of the muscles. The rates of tenderisation were slower for intermediate pH(u) values resulting in higher shear force values at all ageing durations. With ageing at 72h for intermediate pH(u), non-stimulated muscles (n=38) 17.64% were >40N and for stimulated muscles (n=34), 7.9% were >40N.

Effect of meat ultimate pH on rate of titin and nebulin degradation.

The processes involved in the tenderisation of meat were studied on muscles with a range of ultimate pH values (5.4-7.0), produced by subcutaneous injection of various doses of adrenaline and exercise. The m. longissimus thoracicum et lumborum (LD) was removed from carcasses stored at 10 °C and held for 1, 3 or 6 days after slaughter, then frozen until tenderness assessment. The tenderness of meat cooked from the frozen state was determined as the force to shear samples of 10 mm × 10 mm cross-section using a MIRINZ tenderometer. The maximum toughness of 15 kgF occurred at an ultimate pH (pH(u)) of about 6.0, resulting in a curvilinear relationship between tenderness and pH(u) at 1 day post-slaughter. By 6 days post-slaughter, all meat had reached the same low shear value of approximately 3 kgF. SDS-PAGE patterns obtained from samples at 12, 24 and 48 hr post-slaughter showed increasing titin and nebulin degradation over time, with the slowest rate of degradation occurring at pH(u) values 6.0-6.3. Titin and nebulin are known to play an important role in the stabilisation of myofibril structure, and it is suggested that the curvilinear relationship results when pH-dependent titin and nebulin degradation occurs.

Contribution of amino acid transmitters to epileptiform activity and reflex suppression in electrically head stunned sheep

In sheep, administration of a combination of zolazepam and tiletamine hydrochloride resulted in a dose dependent reduction in the duration of epileptic activity induced by an electric stun applied to the head. The compound also lengthened the normal period of reflex suppression that occurs after a stun. Excitatory amino acid receptor antagonists (2-amino-7-phosphonoheptanoic and 2-amino-5-phosphonovaleric acids) also reduced the duration of epileptic activity following an electric stun. These drugs did not alter the time of pedal and ear pinch reflex suppression. Administration of bicuculline (a gamma amino-4-butyric acid [GABA] receptor antagonist) reduced the period of stun induced reflex suppression and increased seizure duration. Administration of a GABA receptor agonist, baclofen, increased the duration of reflex suppression. The results suggest that the development of epileptiform-like activity following application of an electric current to the head is dependent upon excitatory amino acid receptors. The reflex suppression that also arises following an electric stun is contributed to by the activation of GABA receptor mechanisms.

Studies in electrical stimulation: Post-mortem decline in nervous response in lambs.

A study has been made of the effects of electrical stimulation, either indirectly through the nerves or directly through the muscles, on glycolysis in lamb hind leg muscles. Responses were affected by delay before application and the stimulating voltage. At 5 min post mortem the same fall in pH during stimulation (ΔpH) and subsequent increased rate of pH fall (dp H/dt) occurred in muscles stimulated both directly and indirectly with 12 V. With 200V, ΔpH was 50% greater in muscles stimulated directly and 30% less if stimulated indirectly. At 30 min post mortem there was very little effect of stimulation at either voltage on glycolysis when muscles were indirectly stimulated, indicating that the nerves had lost their capacity to trigger muscles to contract. In contrast, if the legs were stimulated directly, maximum ΔpH and dpH/dt values were achieved with 200 V pulses. The significance of these findings to the application of electrical stimulation to commercial meat processing is discussed.