Effect of high pressure treatment on the color of fresh and processed meats: A review.

High pressure (HP) treatment often results in discoloration of beef, lamb, pork, and poultry. The degree of color changes depends on the physical and chemical state of the meat, especially myoglobin, and the atmospheric conditions during and after pressurization. A decreased redness is attributed to a large degree to the oxidation of the bright red oxymyoglobin or the purplish deoxymyoglobin into the brownish metmyoglobin, as well as to the denaturation of myoglobin. Surely, the high myoglobin content makes beef more exposed to this discoloration compared to the white chicken meat. In addition, HP treatment causes denaturation of myofibrillar proteins followed by aggregation, consequently, changing the surface reflectance and increasing lightness. Other intrinsic and extrinsic factors may affect the pressure-induced color changes positively or negatively. In this review, the pressure-induced color changes in meat are discussed in relation to modification of the myoglobin molecule, changes in the meat microstructure, and the impact of the presence of different chemical compounds and physical conditions during processing.

The influence of feeding crimped kernel maize silage on broiler production, nutrient digestibility and meat quality.

Two experiments were carried out in parallel with male Ross 308 broilers over 37 d. An experiment with a total of 736 broilers was performed to study the effect of dietary inclusion of crimped kernel maize silage (CKMS) on broiler production and meat quality. Another study with 32 broilers was carried out from 21 to 25 d to investigate the inclusion of CKMS on nutrient digestibility. In both trials, 4 dietary treatments were used: wheat-based feed (WBF), maize-based feed (MBF), maize-based feed supplemented with 15% CKMS (CKMS-15) and maize-based feed supplemented with 30% CKMS (CKMS-30). Compared with MBF, the dry matter (DM) intakes of broilers receiving CKMS-15 and CKMS-30, respectively, were numerically 7.5 and 6.2% higher and feed conversion ratio 6 and 12% poorer (significant for 30% CKMS), although there were no significant differences in AME content between the three diets. At 37 d, the body weight of birds receiving 15% CKMS was similar to birds fed with MBF. However, the inclusion of 30% CKMS decreased broiler growth. Dietary supplementation with CKMS significantly reduced the apparent digestibility of phosphorus. The fat digestibility was significantly lower for CKMS-30 than for the other three diets. Broiler mortality decreased significantly when CKMS was added to the diet. The consumption of drinking water was significantly lower in all maize-based diets as compared to WBF and was lowest in broilers fed with CKMS-30. An improved litter quality in terms of DM content and a lower frequency of foot pad lesions was observed with broilers supplemented with both dietary levels of CKMS. The addition of CKMS to maize-based diets increased juiciness, tenderness and crumbliness of the meat. In conclusion, the dietary supplementation of 15% CKMS had no negative effect on broiler growth and positively influenced bird welfare in terms of mortality and foot pad health. Therefore, the addition of 15% CKMS to maize-based diets is considered an advantageous feeding strategy in broiler production.

Electrical stimulation affects metabolic enzyme phosphorylation, protease activation, and meat tenderization in beef.

The objective of this study was to investigate the response of sarcoplasmic proteins in bovine LM to low-voltage electrical stimulation (ES; 80 V, 35 s) after dressing and its contribution to meat tenderization at an early postmortem time. Proteome analysis showed that ES resulted in decreased (P < 0.05) phosphorylation of creatine kinase M chain, fructose bisphosphate aldolase C-A, ß-enolase, and pyruvate kinase at 3 h postmortem. Zymography indicated an earlier (P < 0.05) activation of µ-calpain in ES muscles. Free lysosomal cathepsin B and L activity increased faster (P < 0.05) in ES muscles up to 24 h. Immunohistochemistry and transmission electron microscopy further indicated that lysosomal enzymes were released at an early postmortem time. Electrical stimulation also induced ultrastructural disruption of sarcomeres. In addition, ES accelerated (P < 0.05) the depletion of ATP, creatine phosphate, and glycogen, as well as a pH decline and the more preferred pH/temperature decline mode. Finally, ES accelerated meat tenderization, resulting in lesser (P < 0.05) shear force values than the control over the testing time. A possible relationship was suggested between a change in the phosphorylation of energy metabolic enzymes and the postmortem tenderization of beef. Our results suggested the possible importance of the activation of µ-calpain, phosphorylation of sarcoplasmic proteins, and release of lysosomal enzymes for ES-induced tenderization of beef muscle.

PCR amplification of repetitive sequences as a possible approach in relative species quantification.

Both relative and absolute quantifications are possible in species quantification when single copy genomic DNA is used. However, amplification of single copy genomic DNA does not allow a limit of detection as low as one obtained from amplification of repetitive sequences. Amplification of repetitive sequences is therefore frequently used in absolute quantification but problems occur in relative quantification as the number of repetitive sequences is unknown. A promising approach was developed where data from amplification of repetitive sequences were used in relative quantification of species in binary mixtures. PCR LUX primers were designed that amplify repetitive and single copy sequences to establish the species dependent number (constants) (SDC) of amplified repetitive sequences per genome. The SDCs and data from amplification of repetitive sequences were tested for their applicability to relatively quantify the amount of chicken DNA in a binary mixture of chicken DNA and pig DNA. However, the designed PCR primers lack the specificity required for regulatory species control.

Investigation on CAST, CAPN1 and CAPN3 porcine gene polymorphisms and expression in relation to post-mortem calpain activity in muscle and meat quality.

This study aimed to detect variability in CAST, CAPN1 and CAPN3 porcine genes and to investigate the effect of CAST and CAPN1 polymorphisms on the activity of native and autolyzed µ-calpain and m-calpain, measured from 1 to 72 h post-mortem in Longissimus dorsi (LD) muscle of 30 pigs. Effects of polymorphisms on meat quality parameter such as pH, color and drip loss were also evaluated. Samples carrying CAST EU137105:g.76,872AA genotype showed higher autolyzed µ-calpain activity 24 and 72 h post-mortem, as well as lower drip loss values. Expression of CAST, CAPN1 and CAPN3 was assessed in LD muscles divergent for shear force. Higher CAST and CAPN3 expression was found in LD with high shear force (P<0.2), confirming a direct role for calpastatin but not for calpain 3 in meat tenderization. In conclusion, CAST gene affected post-mortem activation time of calpain and drip loss.

Effect of minor milk proteins in chymosin separated whey and casein fractions on cheese yield as determined by proteomics and multivariate data analysis.

The objective of this work was to find regressions between minor milk proteins or protein fragments in the casein or sweet whey fraction and cheese yield because the effect of major milk proteins was evaluated in a previous study. Proteomic methods involving 2-dimensional gel electrophoresis and mass spectrometry in combination with multivariate data analysis were used to study the effect of variations in milk protein composition in chymosin separated whey and casein fractions on cheese yield. By mass spectrometry, a range of proteins significant for the cheese yield was identified. Among others, a C-terminal fragment of beta-casein had a positive effect on the cheese yield expressed as grams of cheese per 100 g of milk, whereas several other minor fragments of beta-, alpha(s1)-, and alpha(s2)-casein had positive effects on the transfer of protein from milk to cheese. However, the individual effect of each identified protein was relatively low. Therefore, further studies of the relations between different proteins/peptides in the rennet casein or sweet whey fractions and cheese yield are needed for advanced understanding and prediction of cheese yield.

Effect of protein composition on the cheese-making properties of milk from individual dairy cows.

The objective of this study was to evaluate the effect of variations in milk protein composition on milk clotting properties and cheese yield. Milk was collected from 134 dairy cows of Swedish Red and White, Swedish Holstein, and Danish Holstein-Friesian breed at 3 sampling occasions. Concentrations of alphaS1-, beta-, and kappa-casein (CN), alpha-lactalbumin, and beta-lactoglobulin (LG) A and B were determined by reversed phase liquid chromatography. Cows of Swedish breeds were genotyped for genetic variants of beta- and kappa-CN. Model cheeses were produced from individual skimmed milk samples and the milk clotting properties were evaluated. More than 30% of the samples were poorly coagulating or noncoagulating, resulting in weak or no coagulum, respectively. Poorly and noncoagulating samples were associated with a low concentration of kappa-CN and a low proportion of kappa-CN in relation to total CN analyzed. Furthermore, the kappa-CN concentration was higher in milk from cows with the AB genotype than the AA genotype of kappa-CN. The concentrations of alphaS1-, beta-, and kappa-CN and of beta-LG B were found to be significant for the cheese yield, expressed as grams of cheese per one hundred grams of milk. The ratio of CN to total protein analyzed and the beta-LG B concentration positively affected cheese yield, expressed as grams of dry cheese solids per one hundred grams of milk protein, whereas beta-LG A had a negative effect. Cheese-making properties could be improved by selecting milk with high concentrations of alphaS1-, beta-, and kappa-CN, with high kappa-CN in relation to total CN and milk that contains beta-LG B.

Water-holding capacity in chicken breast muscle is enhanced by pyruvate and reduced by creatine supplements.

In commercial production, chickens are subjected to feed withdrawal prior to slaughter and exposed to stress during transport and handling of the animals at the slaughterhouse; this causes plasma glucose and glycogen stores in liver and muscle to decrease, which has a negative impact on meat quality. The aim of the present study was to investigate how supplementation of the energy complements creatine and pyruvate during the fasting period would affect postmortem pH decrease, water-holding capacity, and color of the meat. Female Ross 208 broilers were supplemented with glucose combined with either pyruvate or creatine via the drinking water for 18 or 42 h prior to slaughter, i.e., before and throughout the fasting period. Chickens were slaughtered at 42 or 43 d of age. Temperature and pH were measured at 1, 10, 30, and 45 min and 1, 2, 4, 8, and 12 h postmortem. The results showed that the pyruvate and glucose supplementation increased the pH at 45 min postmortem by 0.25 units and decreased drip loss of musculus pectoralis major (PM) by 50 to 65% in chickens supplemented for 42 h. The creatine and glucose supplementation reduced pH at 3 and 4 h postmortem by 0.32 to 0.42 units, increased the lightness (L*) by 2.3 to 5.6 units, and increased drip loss by 51 to 137% in the PM of chickens supplemented for 18 and 42 h. Pyruvate and glucose supplementation thus appear beneficial but whether this is concomitant with an overall improvement in meat quality remains to be determined.

Ascorbic acid, alpha-tocopherol, and oregano supplements reduce stress-induced deterioration of chicken meat quality.

In order to ameliorate a negative effect of stress on meat quality characteristics, chickens were fed a diet supplemented with a combination of ascorbic acid (1,000 ppm) and alpha-tocopherol (200 ppm) or oregano (3%), which has a high content of antioxidants. Chickens were slaughtered by cervical dislocation in the stable (no stress) or after transport and electrical stunning at the slaughter plant (stress). Activities of antioxidative enzymes (catalase, superoxide dismutase, and glutathion peroxidase) in pectoralis major (PM), iliotibialis (IL), and liver were unaffected by supplementation. However, erythrocyte stability, which is a more complex model system for determining oxidative status, increased with ascorbic acid-alpha-tocopherol supplementation and tended to increase after oregano supplementation. In nonstressed birds, this improved antioxidative status was reflected in decreased TBA-reactive substances (TBARS) in PM and liver of ascorbic acid-alpha-tocopherol-supplemented chickens and likewise in liver from oregano-supplemented chickens compared to that of nonstressed control birds. However, postmortem temperature, pH, and water-holding capacity were not affected by supplementation. Drip loss from oregano-supplemented chickens showed increased protein oxidation in specific bands, but this did not relate to water-holding capacity or antioxidative status. When exposed to stress, the concentration of TBARS in the control animals increased in PM and IL. Ascorbic acid-alpha-tocopherol supplementation protected IL, and oregano supplementation protected PM from stress-induced increases in TBARS. This differential effect between muscles may indicate differences in protection mechanisms. In conclusion, ascorbic acid-alpha-tocopherol and oregano supplements to chickens protect against stress-induced increase in TBARS, in different muscles.

Manipulation of critical quality indicators and attributes in pork through vitamin E supplementation, muscle glycogen reducing finishing feeding and pre-slaughter stress.

The combination of a muscle glycogen reducing diet or a standard diet (control group) with normal (80 mg/kg) and high vitamin E levels (500 mg/kg) and exercise immediately prior to slaughter was used on 56 pigs to investigate the influence on meat quality indicators (pH and temperature) and attributes (drip loss, colour and Warner-Bratzler shear force). The drip loss was reduced in M. longissimus dorsi, M. biceps femoris and M. semimembranosus in pigs given the muscle glycogen reducing diet compared with the control groups, the greatest effect was seen in exercised pigs. These results can be explained by an early post mortem reduction in glycometabolism in pigs fed muscle glycogen reducing diets rather than by an increase in ultimate pH. Noticeably, high dietary vitamin E level increased muscle glycogen stores by about 10% on the day prior to slaughter but not on the day of slaughter in both dietary groups compared with the low dietary vitamin E level, which in fact reduced rather than improved the water-holding capacity, especially in pigs fed the standard diet.

Post mortem energy metabolism and pH development in porcine M. longissimus dorsi as affected by two different cooling regimes. A (31)P-NMR spectroscopic study.

(31)P-NMR spectroscopy was carried out on M. longissimus dorsi samples chilled by two different cooling profiles corresponding to commercial batch and tunnel chilling. The half-life of post mortem phosphocreatine (PCr) degradation was found to be significantly less in muscle samples exposed to tunnel chilling (rapid) compared with muscle samples exposed to batch chilling (soft) conditions, while no difference in the post mortem ATP degradation was found. Moreover, the post mortem pH development in the muscle samples differed considerably between the two cooling regimes. A maximum difference of approx. 0.25 pH units between the two cooling profiles was observed around 150 min post mortem. Theoretical calculations of the registered pH difference between rapid and soft chilling of muscle samples revealed that the temperature effect on the buffer capacity of muscle is the major determining factor in the detected difference in intracellular pH between the two cooling profiles, while any contribution from a temperature-induced delayed progress in the lactate formation post mortem seems negligible. Moreover, calculations on the effect of the registered pH difference between rapid and soft chilling of muscle samples resemble a 2.5 times greater denaturation of myosin in samples which were chilled softly compared with samples chilled more rapidly. Finally, the relationship to the functionality of meats from soft and rapid chilled pork carcasses is discussed.

Origin of multiexponential T(2) relaxation in muscle myowater.

To obtain a further understanding of the nature of the multiexponential T(2) relaxation seen in muscle tissue water (myowater), relaxation measurements were carried out on whole, minced, and homogenized pork of three different qualities with regard to water-holding capacity (normal, red soft exudative, and dark firm dry). Whole, minced, and homogenized pork all resulted in multiexponential T(2) relaxation (three components) independently of the quality, even though microscopic studies on homogenized meat revealed considerable disruption of the macroscopic structure. This states that the relaxation behavior in meat cannot be explained by intra-/extracellular compartmentalization of the water as suggested in earlier studies. Subsequent studies of T(2) relaxation in either whole meat, where the structure integrity was changed by the introduction of dimethyl sulfoxide (membrane disruption) or urea (protein denaturation), or minced meat with added NaCl (inter-/intraprotein interactions) lead to the suggestion that in whole meat (i) the fastest relaxation component reflects water tightly associated with macromolecules, (ii) the intermediate relaxation component reflects water located within highly organized protein structures, for example, water in tertiary and/or quaternary protein structures and spatials with high myofibrillar protein densities including actin and myosin filament structures, and (iii) the slowest relaxation component reflects the extra-myofibrillar water containing the sarcoplasmatic protein fraction. Finally, relaxation patterns in heat-set gels of superprecipitated actomyosin and bovine serum albumin similar to that identified in whole meat support the proposed nature of T(2) relaxation in muscle myowater.

Muscle glycogen stores and meat quality as affected by strategic finishing feeding of slaughter pigs.

The aim of the present study was to investigate whether muscle glycogen stores in slaughter pigs could be decreased through strategic finishing feeding before slaughter. Moreover, preliminary meat quality traits were measured to see whether such a regulation of muscle glycogen stores affected ultimate pH, color, and tenderness in the meat. The strategic finishing feeding was carried out the last 3 wk prior to slaughter. Seven experimental groups with eight animals per group were fed diets low in digestible carbohydrates. A control group with four animals was fed a traditional grower-finishing diet. The muscle glycogen stores were reduced in longissimus muscle (LM) 11 to 26% at the time of slaughter in pigs that were fed the experimental diets compared with the control group. Meat quality measured as ultimate pH and color on LM muscle in half the pigs 24 h postmortem showed that ultimate pH in LM was not affected by the reduction in glycogen stores in the muscles from pigs fed any of the experimental diets. However, the meat from pigs fed the experimental diets was darker than the meat from pigs that were fed the control diet with two of the experimental diets, resulting in significantly lower L* values. Activities of key enzymes in the glycolytic pathway, glycogen phoshorylase a and b, phosphofructokinase, and the fatty acid oxidative pathway, beta-hydrozyacyl-CoA-dehydrogenase, were not affected by the strategic feeding. In contrast, the activity of the proteolytic enzyme calpain as well as its inhibitor calpastatin was influenced by the strategic feeding. Lower activity of mu-calpain and greater activity of calpastatin in the muscle samples from the strategically fed pigs indicate a lesser muscle protein degradation in the muscles compared with muscles of control animals. The present study showed that the muscle glycogen stores in slaughter pigs can be reduced at the time of slaughter through strategic finishing feeding with diets low in digestible carbohydrate without compromising growth rate.

Comparative study of low-field NMR relaxation measurements and two traditional methods in the determination of water holding capacity of pork.

The correlation between transverse relaxation, T(2,) and water holding capacity (WHC) determined either by Honikels bag method (Honikel, 1998) or centrifugation has been investigated in meat samples from m. longissimus dorsi (LD) 24 h post mortem from 74 pigs. Bi-exponential analysis of the transverse relaxation, T(2), showed highly significant correlations between both the two NMR time constants (T(21),T(22)) and water holding capacity determined by both Honikels bag method (r(T(21))=-0.72 and r(T(22))=0.77) and centrifugation (r(T(21))-0.50 and r(T(22))=0.75). This shows that transverse relaxation measurement is an efficient method for determination of water holding capacity in pork. Significant correlations were also found between T(21) and T(22) measured 24 h post mortem and pH measured at various time post mortem. This indicates that transverse relaxation, T(2), reflects pH-induced structural changes occurring in muscles post mortem.

Strategic finishing feeding as a tool in the control of pork quality.

A standard diet and two finishing feeding strategies known to reduce muscle glycogen stores were investigated in combination with exercise immediately prior to slaughter in pigs. The objective was to determine the influence of muscle glycogen at slaughter on temperature and pH in post-mortem muscle, the colour, drip loss and Warner-Bratzler shear force of the meat. The muscle glycogen stores were reduced by strategic finishing feeding. In general, pH(45 min) was higher in muscles from strategically fed pigs compared with control pigs. Exercise also resulted in higher pH(45 min) in control pigs compared to non-exercised control pigs, while the opposite was seen in muscles from strategically fed pigs. Exercise resulted in higher muscle temperatures in the carcasses irrespective of feeding strategy. pH(24 h) were higher in M. biceps femoris and M. semimembranosus from exercised, strategically fed pigs compared with the corresponding controls. In contrast, irrespective of feeding strategy no difference in pH(24 h) was registered in the meat of non-exercised pigs. Drip loss was lower in meat of strategically fed pigs compared with meat of control pigs. Moreover the drip loss was lowest in the meat of non-exercised pigs. The present study shows that strategic finishing feeding has high potential for the control of pork quality.

Comparison of sample preparation methods for the high-performance liquid chromatographic analysis of cell culture extracts for triphosphate ribonucleosides and deoxyribonucleosides.

We compared four different procedures for the purification and concentration of nucleoside triphosphates in cell extracts prior to HPLC analysis. Two methods involved precipitation, with either acetonitrile or calcium fluoride. The acetonitrile procedure yielded reasonable recovery and sufficient purity for the subsequent HPLC analysis. The calcium fluoride coprecipitation procedure gave both good recovery and purity; but the recovery was shown to be dependent on the concentration of the nucleoside triphosphates. The other two methods involved small Sep-Pak cartridges. The silica cartridge procedure yielded unfavorable recoveries in periodate-treated cell extracts, apparently due to poor solubility of nucleoside triphosphates in the requisite solvents. The strong anion exchange cartridge procedure yielded both good recovery and purity. This procedure was found to be fast, efficient, and reliable for purifying and concentrating nucleotides in cell extracts.

Influence of acyclovir and bucyclovir on nucleotide pools in cells infected with herpes simplex virus type 1.

The effects of the acyclic guanosine analogs acyclovir (ACV) and (R)-9-(3,4-dihydroxybutyl)guanine (bucyclovir, BCV) on the deoxyribonucleoside triphosphate (dNTP) pools of herpes simplex type 1 (HSV-1)-infected African green monkey kidney (GMK) and human embryonic lung fibroblast (HL) cells were investigated. HSV-1 infection increased the dNTP pools in both cell types compared with those in uninfected cells. Mock-infected GMK cells showed a 10-fold-higher dTTP concentration than comparable HL cells. ACV or BCV treatment of HSV-1-infected cells yielded further increases of the dNTP pools. ACV- or BCV-treated, HSV-1-infected HL cells showed 20- to 50-fold-higher concentrations of ACV triphosphate and BCV triphosphate, respectively, than similarly treated GMK cells. This is in accord with previous results, which showed that ACV and BCV are less active in GMK cells than in HL cells. This difference in activity is attributed to the substantial deoxythymidine pools previously found in GMK cells. The results are discussed in relation to known metabolic and kinetic parameters.