Effect of grazing pastures of different botanical composition on antioxidant enzyme activities and oxidative stability of lamb meat

The aim of this work was to study the influence of different pastures (Intensive ryegrass, Botanically diverse and Leguminosa rich pastures) on the antioxidant status and oxidative stability of meat from lambs that had been exclusively grazing for three months. Lipid, colour and protein oxidation, -tocopherol content and activity of antioxidant enzymes (superoxide dismutase (SOD), catalase (Cat) and glutathione peroxidase (GSH-Px)) were measured in Longisimus thoracis et lumborum muscle samples taken 1 day after slaughter. Pasture type significantly affected protein oxidation and the activity of GSH-Px, but no significant differences were found for the -tocopherol content, colour and lipid oxidation, and the activities of SOD and Cat. Grazing a Botanically diverse pasture induced significantly higher protein oxidation in meat, as measured by the free thiol and carbonyl contents, compared to a Leguminosa rich or Intensive ryegrass pasture (P < 0.05). The GSH-Px activity was significantly higher in meat from lambs on the Leguminosa rich pasture compared to the other pasture groups (P < 0.01).     

Chloride salt type/ionic strength, muscle site and refrigeration effects on antioxidant enzymes and lipid oxidation in pork

The effects of NaCl and KCl at varying ionic strengths on catalase and glutathione peroxidase (GSH-Px) activities and lipid oxidation in refrigerated ground pork muscles from different anatomical locations were studied. Catalase and GSH-Px activities were higher in boston butt (BB) than in longissimus dorsi (LD), whereas lipid oxidation measured by 2-thiobarbituric acid substances (TBARS) content was higher in LD. Catalase activity was stable in both BB and LD during 4-day storage; GSH-Px activity decreased in LD. GSH-Px activity decreased more with NaCl than KCl, whereas salt type had no consistent effect on catalase activity. TBARS content, however, increased more with NaCl than with KCl. NaCl at the highest ionic strength decreased GSH-Px activity by 19.2 and 18% in LD and BB, respectively, and increased TBARS content by 8- and 3.6-fold. Results indicated that pork samples with higher catalase and GSH-Px activities would undergo less lipid oxidation, and the accelerated lipid oxidation in salted pork may be partly related to a decrease in GSH-Px activity.     

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.     

Green tea catechins upregulate superoxide dismutase and catalase in fruit flies

Chinese Longjing green tea is an excellent source of polyphenol antioxidants. HPLC analysis revealed that Longjing green tea catechin extract (GTC) contained 62% epigallocatechin gallate (EGCG), 19% epigallocatechin (EGC), 9% epicatechin gallate (ECG), and 7% epicatechin (EC). Investigating the effect of GTC on the lifespan of Drosophila melanogaster, we observed that a 10 mg GTC/mL diet could prolong its 50% survival time by 36% and mean lifespan by 16%. This was consistent with 17% reduction in total body lipid hydroperoxide (LPO) level in GTC-treated flies compared to the control group. Supplementation of 10 mg GTC/mL diet increased the survival time only in wild type Oregon-R-C (OR) but not in two mutant fly lines, SOD(n108)/TM3 (gene for superoxide dismutase (SOD) was knocked out) and Cat(n1)/TM3 (gene for catalase was knocked out), when the flies were challenged with paraquat or hydrogen peroxide. Accordingly, SOD and catalase activities in OR wild type increased by 40 and 19%, respectively. RT-PCR analysis indicated that the genes for copper-zinc containing SOD (CuZnSOD), manganese containing SOD (MnSOD), and catalase were upregulated. It was concluded that prolonging lifespan by GTC in D. melanogaster was influenced, among others, by upregulation of endogenous antioxidant enzymes.     

Chloride salt type/ionic strength and refrigeration effects on antioxidant enzymes and lipid oxidation in cattle,camel and chicken meat

The effects of NaCl and KCl at varying ionic strengths on catalase and glutathione peroxidase (GSH-Px) activities and lipid oxidation in ground Longissimus dorsi (LD) of cattle and camel and breast muscle of chicken during refrigerated storage were studied. NaCl and KCl significantly increased 2-thiobarbituric reactive substances (TBARS) and peroxide values. TBARS and peroxide values increased and GSH-Px activity decreased during 4 day storage in the 4 °C, but catalase activity was stable. Salt type had no consistent effect on GSH-Px and catalase activities. Chicken samples had lower enzyme activities and TBARS content than cattle and camel. Their peroxide values were lower than camel samples. Camel meat showed higher catalase activity and TBARS content than cattle meat. Results indicated that negative correlation between lipid oxidation and GSH-Px activity and the accelerated lipid oxidation in salted meat may be partly related to a decrease in GSH-Px activity.     

Antioxidant enzyme activities and antioxidant capacity in longissimus muscle from bulls fed diets rich in polyunsaturated fatty acids

The effect of a treatment diet composed of grass silage and concentrate including rapeseed (with/without feeding restriction) was compared with a control diet of maize silage/grass silage (70:30) and concentrate including soybean, on the antioxidant enzyme activities of fresh longissimus muscle from German Simmental bulls. Additionally, the effect of diet on antioxidant capacity (AOC) of hydrophilic and lipophilic antioxidants was evaluated in fresh and stored beef muscle using the FRAP-ferric reducing ability and TEAC – Trolox-equivalent antioxidant capacity assays at different reaction times. Catalase and superoxide dismutase activities were significantly higher in the treatment diet groups, and glutathione peroxidase activity was not different. AOC was not affected by the diet. However, storage affected the values of FRAP and TEAC assays, and the results were time-depending. 30 min were found like a minimum reaction time for both assays. Generally, AOC values of the hydrophilic antioxidants were significantly higher than lipophilic values.     

Nutritional value and digestion rate of rhea meat proteins in association with storage and cooking processes

The nutritional value of proteins was investigated after the storage and cooking of rhea M. Gastrocnemius pars interna. Oxidation of basic and aromatic amino acids, surface hydrophobicity and aggregation state of proteins, were determined in raw and cooked meat. In addition, myofibrillar proteins were exposed in vitro to proteases of the digestive tract. Cooking markedly affected the protein surface hydrophobicity. The BBP bound content was three times greater in cooked than in fresh rhea meat. A small increment in tryptophan content after cooking was observed. Storage influenced Schiff bases formation indicating the presence of protein-aldehyde adducts after cooking. High content of Schiff bases was found after cooking of samples stored for 5 days, demonstrating a probable implication of free amino groups, most likely from lysine. Cooking decreased the myofibrillar protein susceptibility to pepsin activity. After cooking, the proteolysis rate by pancreatic enzymes increased. Our findings support the importance of protein aggregation in the nutritional value of meat proteins.     

First evaluation of unfermented and fermented rooibos (Aspalathus linearis) in preventing lipid oxidation in meat products

This study consisted of two trials aiming to evaluate, for the first time, the antioxidant potential of rooibos in meat products. With this purpose, the first trial evaluated three unfermented (green) rooibos forms (dried leaves, water extract, freeze-dried extract) added at 2% inclusion level to ostrich meat patties on an 8-day shelf-life trial. A Control group without green rooibos inclusion was also considered. The second trial evaluated the addition of different concentrations (0%, 0.25%, 0.5% and 1%) of a fermented rooibos extract to nitrite-free ostrich salami. The 2% green rooibos inclusion considerably lowered the TBARS content of ostrich patties, in this way extending their shelf-life. The fermented form (0.5% and 1%) was also effective in delaying lipid oxidation in ostrich salami until 15 days of ripening. The antioxidant potential of both green and fermented forms of rooibos in meat products was confirmed, even if its effect on lipid oxidation requires further study and long-term effects are not yet fully understood.     

Antioxidant activities of malt extract from barley (Hordeum vulgare L.) toward various oxidative stress in vitro and in vivo

The antioxidant activities of malt extract from barley were evaluated by various methods in vitro and in vivo. Scavenging effects on the hydroxyl and superoxide radicals, and protection against reactive oxygen species induced lipid, protein and DNA damage were evaluated. The d-galactose induced mouse aging model was used to evaluate ability of malt extract to behave as an antioxidant in vivo. The extract exhibited high antioxidant activities both in vitro and in vivo, evidenced by its ability to scavenge hydroxyl- and superoxide-radicals, high reducing power, and protection against biological macromolecular oxidative damage. Furthermore, malt extract prevented the decrease of antioxidant enzyme activities, decreased liver and brain malondialodehyde levels and carbonyl content, and improved total antioxidant capability in d-galactose-treated mice. In conclusion, these results demonstrate potential antioxidant activities and antiaging effect of malt, providing scientific support for the empirical use of malt as an antioxidant for diseases caused by reactive oxygen species.     

Effects of freezing temperature and duration of frozen storage on lipid and protein oxidation in chicken meat

This study examined the effects of freezing temperature and duration of frozen storage on lipid and protein oxidation in chicken leg and breast meat. The meat was frozen at three different temperatures (−7, −12 and −18 °C) and then stored at −18 °C for up to 6 months. A significant effect of frozen storage duration on lipid oxidation was detected in leg and breast meat, whereas freezing temperature had no significant effect. In leg meat, freezing at −7 °C had a significant impact on protein oxidation, measured as the increase in carbonyl groups and the decrease in total sulphydryl groups, after 3 months of frozen storage. Lipid and protein oxidation appeared to occur simultaneously in chicken meat during frozen storage and was more intense in leg meat than in breast meat.     

Inhibition of protein and lipid oxidation by rapeseed, camelina and soy meal in cooked pork meat patties

In this study protein-containing by-products of deoiling processes rich in phenolics were applied to meat to be used as potential food ingredients in developing meat products with antioxidant effect. The effect of rapeseed meal (Brassica rapa L.), camelina meal (Camelina sativa), soy meal and soy flour (from soybean, Glycene max L.), in inhibiting oxidation of lipids and proteins was tested in cooked pork meat patties. A commercial CO₂ extract from rosemary (Rosmarinus officinalis) was used as a reference material alone and in combination with the other plant materials. The cooked pork meat with added plant materials was oxidized for 10 days at 5 °C under light. The oxidation was followed by measuring the formation of hexanal, pentanal and propanal by headspace gas chromatography and the formation of protein carbonyls by converting them to 2,4-dinitrophenylhydrazones (DNPH). Rapeseed meal (0.5 and 0.7 g/100 g meat) and camelina meal (0.7 g/100 g meat) as such and their combination (addition of 0.5 g/100 g) with rosemary extract (0.04 g/100 g) were effective antioxidants toward both protein and lipid oxidation while soy meal and flour were effective only in combination with rosemary extract.     

Effect of multiple freeze-thaw cycles on protein and lipid oxidation in rabbit meat

Frozen storage is chosen to be the preferred method in meat preservation. However, multiple freeze-thaw (F-T) cycles will occur during the transfers or distribution processes under poor cold-chain conditions. This study investigated the effects of multiple freeze-thaw (F-T) cycles on water mobility, microstructure damage, protein oxidation and lipid oxidation in rabbit meat during seven F-T cycles through 56 days. Scanning electron microscopy results indicated that the increase of F-T cycles induced the rupture of endomysia and the increase of gaps between fibres. Low-field nuclear magnetic resonance (LF-NMR) results showed that the peak area of T₂₁ and T₂₂ decreased with increasing number of repeated F-T cycles, indicating that immobile water shifted to free water, and free water mobility increased. During repeated freeze-thaw process, the peroxide value, thiobarbituric acid-reactive substances, non-haem iron content and carbonyl groups of rabbit meat gradually increased, whereas haem content, haem iron content and sulfhydryl content of rabbit meat gradually decreased. In addition, the results of Fourier transform infrared, sodium dodecyl sulphate polyacrylamide gel electrophoresis, and intrinsic fluorescence spectroscopy indirectly proved that multiple F-T cycles could cause α-helix structure disruption, protein aggregation and degradation, and tryptophan oxidation of rabbit meat myofibrillar protein (MP). Overall, multiple repeated F-T cycles changed moisture migration, damaged microstructure and promoted lipid and protein oxidation of rabbit meat.     

Early post-mortem sarcoplasmic proteome of porcine muscle related to lipid oxidation in aged and cooked meat

In order to identify specific markers of lipid oxidation generated in meat during refrigerated storage and cooking an analysis was conducted to investigate the relationships between the early post-mortem sarcoplasmic proteome, which contains the majority of enzymes involved in the oxidative process, and the level of lipid oxidation. This study was performed in Longissimus lumborum pig muscle. Proteome was analysed by 2-D electrophoresis in combination with liquid chromatography–tandem mass spectrometry (LC–MS/MS) and lipid oxidation was estimated by the TBA reactive substances (TBA-RS) measurement. Many markers of lipid oxidation were identified, but no single marker covered the oxidative process in its entirety. The role of five protein groups (albumin, redoxins, annexins, lipid transporters and enzymes of aerobic respiration), from which a link with lipid oxidation can be established, is discussed. This study, which completes a precedent work focused on protein oxidation, clearly demonstrates that a combination of several markers is needed to assess the sensitivity of meat to oxidation during both ageing and cooking.     

Glutathione peroxidase activity, and content of total and soluble selenium in five bovine and porcine organs used in meat production

Glutathione peroxidase (GSHPx) activity, and total and soluble selenium content were compared in five bovine and porcine organs. The highest GSHPx activity in porcine tissues was found in the liver (35.0 U/g), spleen (29.3 U/g) and kidney (27.3 U/g) with much lower values in the heart (1.8 U/g) and diaphragm (0.8 U/g). A different pattern with lower inter-organ variation in GSHPx activity was observed in cattle: kidney (8.5 U/g), spleen (8.0 U/g), heart (5.8 U/g), liver (4.0 U/g) and diaphragm (2.1 U/g). The total selenium content was similar in both species with the highest content in the kidney (1764 and 1665 ng/g; pig/bovine), followed by liver (533 and 307 ng/g), spleen (370 and 284 ng/g), heart (201 and 205 ng/g) and diaphragm (144 and 116 ng/g). The percentage of soluble selenium varied more among the pig organs (46–94%) than among bovine organs (61–75%). The results show a marked variation in the activity of the selenium-containing GSHPx among organs and species in spite of a similar rank order of selenium content in the two species. Since GSHPx has a role in food stability and the intake of selenium is marginal in many European countries, the results add to the background information concerning the use of selenium rich organs as human foods.     

Effect of sage and garlic on lipid oxidation in high-pressure processed chicken meat

Sage was found to protect minced chicken breast processed with high hydrostatic pressure up to 800 MPa for 10 min against lipid oxidation during subsequent chilled storage for 2 weeks. Garlic showed prooxidative effects especially at moderate high pressure around 300 MPa, an effect partly counteracted by simultaneous addition of sage. From the rate of lipid oxidation, measured as thiobarbituric acid reactive substances, the apparent volume of activation for pressure-induced lipid oxidation during subsequent chilled storage was estimated, which showed that the prooxidative effect of garlic and pressure-induced lipid oxidation were additive. Based on electron spin resonance (ESR) spectroscopy radical formation was measured in pressurized lipid and aqueous phases of minced chicken thighs, and a high radical scavenging capacity of sage in the lipid phase was identified as most important for the protective effect of sage.     

Synthesis of superoxide dismutase,catalase and other enzymes and oxygen and superoxide toxicity during changes in oxygen concentration in cultures of brewing yeast

The physiological effects on brewing yeast, growing in semi-defined wort medium, of a sudden transition from aerobiosis to anaerobiosis were studied. Two yeast strains were examined, used for ale and lager fermentations respectively. The reverse transition (from anaerobiosis to aerobiosis) was also examined. Transitions were applied by changing the sparging gas during growth or in stationary phase, and the effects on the specific activities of certain key enzymes and on the viability of the cultures were examined. Neither type of transition led to significant changes in growth rate, the rate of ethanol production or the specific activities of alcohol dehydrogenase and pyruvate decarboxylase. The most significant change was in the specific activity of CuZn-superoxide dismutase, which showed a rapid increase in activity on transition from anaerobiosis to aerobiosis, and a decrease in activity on the reverse transition. Catalase activity in the ale yeast generally followed that of CuZn-superoxide dismutase, whereas in the lager yeast it remained unchanged by the transitions. The transition from anaerobiosis to aerobiosis caused increases in citrate synthase and Mn-superoxide dismutase, though only after a significant lag period. Aerobic to anaerobic transitions caused a decrease in Mn-superoxide dismutase activity, while citrate synthase remained unchanged. Anaerobically grown cells showed a rapid loss in viability on exposure to oxygen (5–7% in the first hour), while aerobically grown cells were unaffected. When anaerobically grown cells were exposed to 0·25 mM -potassium superoxide, there was an 8% loss of viability within 10 min, whereas aerobic cells were not affected. It is concluded that the toxic effect of oxygen is due to superoxide (or a species derived from it) and that the CuZn-superoxide dismutase (but not the Mn-isoenzyme) plays a role in protecting the cells. The de novo synthesis of the CuZn-enzyme is not always rapid enough to confer full protection.     

Influence of thermal treatment on formation of volatile compounds,cooking loss and lipid oxidation in foal meat

The influence of four different cooking methods (roasting, grilling, microwaving baking and frying with olive oil) on volatile compounds profile (using solid phase microextraction coupled to GC–MS), lipid oxidation (by TBARs measurement) and cooking loss of foal meat was studied. Cooking losses were significantly (P < 0.001) affected by thermal treatment, being higher (29.9%) after microwaving and lower after grilling (19.1%) treatments. As expected, all the cooking methods increased TBARs content, since high temperature during cooking seems to cause an increase of the oxidation processes in foal steaks, being this increment significantly (P < 0.001) higher when foal steaks were roasted or microwaved.     

Effect of different cooking methods on lipid oxidation and formation of volatile compounds in foal meat

The influence of four different cooking methods (roasting, grilling, microwaving and frying) on cooking loss, lipid oxidation and volatile profile of foal meat was studied. Cooking loss were significantly (P < 0.001) affected by thermal treatment, being higher (32.5%) after microwaving and lower after grilling (22.5%) and frying (23.8%). As expected, all the cooking methods increased TBARs content, since high temperature during cooking causes increased oxidation in foal steaks, this increase was significantly (P < 0.001) higher when foal steaks were microwaved or roasted.     

Colour,lipid and protein stability of Rhea americana meat during air- and vacuum-packaged storage: Influence of muscle on oxidative processes

Physicochemical characteristics and oxidative stability during storage were determined in Gastrocnemius pars interna (GN) and Iliofiburalis (IF) muscles of Rhea americana. Glycolytic potential (GP) and pH decline of muscles were measured within the first 24 h post mortem. Colour, lipid and protein stability were determined during storage of meat, i.e. 5 days under air-packaging at 4 °C, or 28 days under vacuum-packaging at 4 °C. In parallel, anti-oxidant status of muscles was estimated by measuring α-tocopherol content and anti-oxidant enzyme activities (superoxide dismutase and catalase), while pro-oxidant status was evaluated by determining haeminic iron and long chain fatty acids (especially polyunsaturated fatty acids). The ultimate pH was similar in both muscles, but the GP value was significantly higher in IF than in GN muscle. Haeminic iron and alpha-tocopherol content differed between muscles, with 30% more haeminic iron (p < 0.05) and 134% more alpha-tocopherol (p < 0.001) in IF than GN muscle. The IF muscle presented higher lipid content and lower PUFA/SFA ratio (polyunsaturated fatty acids/saturated fatty acids) than GN muscle. With storage under air-packaging, lipid and protein oxidation of rhea muscles increased up to 275% and 30%, respectively. This increase was more rapidly and marked in IF muscle. The IF also showed high level of metmyoglobin accumulation after 3 days of storage (47%) and was rejected by 1 consumer out of 2 in sensorial analysis. Under vacuum-packaging, both muscles showed a high stability of colour and no oxidation of lipids and proteins.     

Influence of dietary conjugated linoleic acid on volatile profiles, color and lipid oxidation of irradiated raw chicken meat

Forty-eight, 27-week-old White Leghorn hens were fed a diet containing 0, 1.25, 2.5 or 5.0% conjugated linoleic acid (CLA) for 12 weeks. At the end of the 12-week feeding trial, hens were slaughtered, and boneless, skinless breast and leg meats were separated from carcasses. Meats were ground through 9 and 3-mm plates, and patties were prepared. Patties prepared from each dietary treatment were divided into two groups and either vacuum- or aerobic-packaged. Patties were irradiated at 0 or 3.0 kGy using a linear accelerator and stored at 4°C. Samples were analyzed for thiobarbituric acid reactive substances, volatile profiles, color and odor characteristics at 0 and 7 days of storage. Dietary CLA reduced the degree of lipid oxidation in raw chicken meat during storage. The content of hexanal and pentanal in raw chicken meat significantly decreased as dietary CLA level increased. Irradiation accelerated lipid oxidation in meat with aerobic packaging, but irradiation effect was not as significant as that of the packaging. Dietary CLA treatment improved the color stability of chicken patties. Color a*-value of irradiated raw chicken meat was higher than that of the nonirradiated meat. Dietary CLA decreased the content of polyunsaturated fatty acid and increased CLA in chicken muscles, which improved lipid and color stability and reduced volatile production in irradiated and nonirradiated raw chicken meat during storage.