Janez Salobir

Evaluation of different vitamin E recommendations and bioactivity of α-tocopherol isomers in broiler nutrition by measuring oxidative stress in vivo and the oxidative stability of meat

The aim of this study was to compare recommendations for vitamin E supplementation regarding high polyunsaturated fatty acid intake and to compare the bioactivity of RRR- and all-rac-α-tocopherol with respect to oxidative stress in vivo and the oxidative stability of broiler meat. Fifty male broilers were divided into 5 groups. All groups received diets with a high inclusion of fat (7.5%), one with palm fat and the others with linseed oil, which were either unsupplemented or supplemented with vitamin E to contain in total 85 or 200 IU of vitamin E as all-rac-α-tocopherol and 85 IU as RRR-α-tocopherol. Oxidative stress in vivo was studied by measuring the DNA damage; measuring malondialdehyde (MDA) in plasma, liver, and breast muscle; and analyzing the antioxidant capacity of the lipid-soluble compounds, total antioxidant status of plasma, and antioxidant enzyme assays. The tocopherols in plasma, liver, and breast muscle were also analyzed. In vitro oxidative stability was studied by measuring MDA in fresh, stored, and heat-treated breast meat. Linseed oil, as opposed to palm fat, induced DNA fragmentation and MDA formation. Both forms and concentrations of vitamin E reduced DNA damage and breast muscle MDA. The groups receiving 200 IU of all-rac-α-tocopherol and 85 IU of RRR-α-tocopherol had much higher values for antioxidant capacity of lipid-soluble compounds than did the controls. No differences were observed in the values of antioxidant enzymes. The α-tocopherol levels in tissues and plasma were significantly influenced by the level of α-tocopherol supplementation. Malondialdehyde formation in meat from the vitamin E-supplemented groups was decreased in comparison with that from the control linseed oil group. We conclude that both vitamin E concentrations were insufficient to prevent all harmful effects of lipid oxidation in vivo and that both were equally effective. On the contrary, to ensure good stability of meat lipids, higher vitamin E supplementation is needed, especially after heat treatment. The results of in vivo oxidative stress and meat lipid oxidation confirmed the currently accepted bioactivity of the RRR-α- to all-rac-α-tocopherol ratio of 1.39 in in vivo and in vitro systems.

In vivo antioxidant potential of Sweet chestnut (Castanea sativa Mill.) wood extract in young growing pigs exposed to n‐3 PUFA‐induced oxidative stress

BACKGROUND Farm animals in intensive farming systems are frequently exposed to oxidative stress, which demands adequate antioxidant protection. The aim of this study was to evaluate the antioxidant potential of different concentrations of Sweet chestnut wood extract (SCW; 0.75, 1.5 and 3 g kg⁻¹) in case of n-3 PUFA-induced oxidative stress in young pigs. RESULTS The highest concentration (3 g kg⁻¹) of SCW decreased malondialdehyde excretion in urine by 31.7%, but had no effect on plasma malondialdehyde. A linear trend towards decrease of urine isoprostanes iPF(2α)-VI was observed with the addition of SCW. All three concentrations of SCW efficiently protected blood lymphocytes from DNA damage and lowered plasma alanine aminotransferase levels. The antioxidative and antigenotoxic effect of 3 g SCW kg⁻¹ feed was comparable to the effect of 90.4 mg kg⁻¹ of added vitamin E. CONCLUSION The results from this study show that, besides being known as antihelmintic, antimicrobial and antiviral agent, Sweet chestnut wood extract could also be considered as a promising natural antioxidant in animal nutrition.

Oxidative stress markers in canine atopic dermatitis

There are no data in the veterinary literature relating to oxidative stress in canine atopic dermatitis (CAD). The study aimed to determine levels of oxidative stress markers, plasma malondialdehyde (MDA), total antioxidant capacity (TAC), whole blood glutathione peroxidase (GPX) and erythrocyte superoxide dismutase (SOD), in 15 CAD patients and 17 healthy dogs. A correlation between CADESI (Canine Atopic Dermatitis Extent and Severity Index) score and MDA was also determined. Significantly higher plasma MDA levels were found in patients than in healthy dogs. The significant, highly positive correlation determined between CADESI score and MDA in the patient group indicates an association between the severity of CAD and the extent of oxidative damage to membrane lipids. There were no significant differences in TAC, GPX and SOD between patients and healthy dogs. Our findings suggest that oxidative stress with increased lipid peroxidation could be involved in the pathogenesis of atopic dermatitis in dogs.

The comparison of black currant juice and vitamin E for the prevention of oxidative stress.

Black currant is known as a fruit with a very strong in vitro antioxidative capacity, but its in vivo antioxidant efficacy has not yet been characterized. The aim of the experiment was to determine the potency of black currant juice in comparison to vitamin E, for decreasing oxidative stress. Oxidative stress was induced by high intake of polyunsaturated fatty acids (PUFAs) in pigs as a model for humans. Twenty-four growing pigs were divided into four groups. All groups received isocaloric daily rations composed of an equal amount of basal diet that was supplemented with starch (CONT), linseed oil (OIL), linseed oil and black currant juice (OIL+BCJ), or linseed oil and vitamin E (OIL+VIT E). The experiment confirmed that the high proportion of PUFAs in the OIL group increased oxidative stress. In comparison with the OIL group, vitamin E supplementation significantly lowered plasma malondiadehyde (MDA) and the 24-hour urine MDA excretion rate, and reduced the degree of DNA damage in leukocytes to the level of the CONT group. The black currant juice intake failed to significantly decrease plasma MDA and 24-hour urine MDA excretion rate, but did reduce the degree of DNA damage in leukocytes to the level of the CONT group, as well as increase plasma beta+gamma-tocopherol concentrations. Although black currant juice did not reduce the formation of MDA, it efficiently prevented DNA damage induced by the high intake of PUFAs. It could be concluded that under these experimental conditions vitamin E was more efficient as an antioxidant that black currant juice.

Effect of Ganoderma lucidum (Reishi mushroom) or Olea europaea (olive) leaves on oxidative stability of rabbit meat fortified with n-3 fatty acids

The objective of the present study was to evaluate the effect of Ganoderma lucidum (Reishi mushroom) or Olea europaea (olive tree) leaves on oxidative stability of rabbit meat fortified with n-3 fatty acids. Forty-eight slovenska kunka (SIKA) rabbits were divided into four homogeneous groups. The control group (CONT-) received diet with 6% palm fat; other groups received diet with 6% linseed oil and were either unsupplemented (CONT+) or supplemented with 1% of G. lucidum (REISHI) or O. europaea leaves (OLIVE). Rabbits were slaughtered and fatty acid composition, concentration of vitamin E and malondialdehyde (MDA) in back muscle were analyzed. The results showed that linseed oil addition improved fatty acid composition by increasing polyunsaturated fatty acid (PUFA) proportion, decreasing proportion of saturated fatty acid (SFA) and reducing n-6/n-3 ratio in rabbit meat. Groups that were supplemented with linseed oil had lower content of α-tocopherol and higher content of γ-tocopherol, compared to the CONT- group. The addition of potential antioxidants did not effectively prevent oxidation of rabbit meat.

The effect of α-tocopherol, sweet chestnut wood extract and their combination on oxidative stress in vivo and the oxidative stability of meat in broilers

1. This study examined the effect of α-tocopherol (α-T), sweet chestnut wood extract (SCW) and their combination on oxidative stress in vivo and oxidative stability of meat in broilers given diets rich in PUFA. 2. A total of 60 male broilers were individually caged and divided into 6 groups of 10. The C-PALM group received a diet with 7·5% palm fat and the other 5 groups with 7·5% linseed oil. The linseed oil groups were either un-supplemented (C-LIN) or supplemented with α-T or/and SCW as follows: αT-85 (C-LIN diet + 68 IU vit E as all-rac-α-T/kg), αT-200 (C-LIN diet + 183 IU vit E as all-rac-α-T/kg), SCW (C-LIN diet + 3 g SCW/kg) and αT-SCW (C-LIN diet + 68 IU vit E as all-rac-α-T/kg + 3 g SCW/kg). Different parameters of oxidative stress were measured. 3. Linseed oil induced DNA fragmentation and malondialdehyde (MDA) formation, while α-T reduced both parameters, and SCW reduced the DNA damage. A combination (αT-SCW) also reduced plasma MDA. Larger antioxidant capacity of lipid soluble compounds were recorded in groups αT-85, αT-200 and αT-SCW than in the controls but there were no differences between these groups in antioxidant enzymes and total antioxidant status. A combination (αT-SCW) increased tocopherol concentrations in breast muscle and in comparison to the C-LIN group MDA concentrations were reduced in groups αT-85, αT-200 and αT-SCW. 4. It can be concluded that neither of the α-T concentrations were able to prevent all the negative effects of lipid oxidation in vivo and only high concentrations of α-T improved the stability of meat. With the exception of DNA damage, SCW had no impact on in vivo and in vitro measured markers of oxidative stress but may have a sparing or regenerating effect on α-T.

Vitamin E supplementation in canine atopic dermatitis: improvement of clinical signs and effects on oxidative stress markers

Low levels of plasma vitamin E concentrations were found in canine atopic dermatitis (CAD). The present study was aimed at determining the effect of an eight-week vitamin E supplementation on clinical response (Canine Atopic Dermatitis Extent and Severity Index (CADESI-03) scores and pruritus intensity) in dogs with atopic dermatitis. Levels of oxidative stress markers (plasma malondialdehyde and total antioxidant capacity (TAC), blood glutathione peroxidase and erythrocyte superoxide dismutase, plasma and skin vitamin E concentrations) were also determined. Twenty-nine dogs with CAD were included in the study. Fourteen received vitamin E (8.1 IU/kg once daily, orally) and 15 received mineral oil as placebo (orally). All dogs were treated with antihistamine fexofenadine. Levels of oxidative stress markers (with the exception of skin vitamin E), CADESI-03 and pruritus intensity were determined at the beginning, then every two weeks. Skin vitamin E was determined at the beginning and at the end of the treatment. Significantly higher plasma levels of vitamin E and TAC were observed in the vitamin E group than in the placebo group. CADESI-03 scores determined throughout the treatment in the vitamin E group were significantly lower than in the placebo group. The findings of this study support the supplementation of vitamin E in dogs with atopic dermatitis.

Differential response of protein metabolism in splanchnic organs and muscle to pectin feeding

The aim of the present study was to determine whether the addition of soluble fibre in the diet affected protein metabolism in the intestinal tissues, some visceral organs and in skeletal muscle. A diet supplemented with pectin (80 g/kg) was fed to young growing rats and the effect on organ mass and protein metabolism in liver, spleen, small and large intestines and gastrocnemius muscle was monitored and compared with the control group. Protein synthesis rates were determined by measuring [13C]valine incorporation in tissue protein. In the pectin-fed rats compared with the controls, DM intake and body weight gain were reduced (9 and 20 %, respectively) as well as gastrocnemius muscle, liver and spleen weights (6, 14 and 11 %, respectively), but the intestinal tissues were increased (64 %). In the intestinal tissues all protein metabolism parameters (protein and RNA content, protein synthesis rate and translational efficiency) were increased in the pectin group. In liver the translational efficiency was also increased, whereas its protein and RNA contents were reduced in the pectin group. In gastrocnemius muscle, protein content, fractional and absolute protein synthesis rates and translational efficiency were lower in the pectin group. The stimulation of protein turnover in intestines and liver by soluble fibre such as pectins could be one of the factors that explain the decrease in muscle turnover and whole-body growth rate.

Relevance of Meat Fat Content and Fruit and Vegetable Intake for the Oxidative Status of Pigs

Background: The aim of the present study was to evaluate the effect of substituting lean meat with fat meat on oxidative stress in a diet with or without fruit and vegetables. Methods: Thirty-two pigs were divided into groups and fed isocaloric daily rations: LM+FV (balanced diet with lean meat and fruit and vegetables); FM+FV (as LM+FV, but lean meat was substituted with fat meat); LM–FV (as LM+FV, but without fruit and vegetables), and FM–FV (as FM+FV, but without fruit and vegetables). Oxidative stress was evaluated by measuring the 24-hour urine malondialdehyde excretion rate, the degree of leukocyte nuclear DNA damage, the concentration of tocopherols in blood plasma, erythrocyte glutathione peroxidase activity and the total antioxidant status of plasma. Results: The substitution of lean meat with fat meat modestly increased the rate of leukocyte DNA damage only in the diet with fruit and vegetables but had no effect in the group deprived of fruit and vegetables. Regardless of the fruit and vegetable content of the diet, the substitution of lean meat with fat meat did not affect any other parameters measured. In comparison to both fruit- and vegetable-containing diets, the deprivation of fruits and vegetables in the LM–FV and FM–FV groups significantly increased the rate of leukocyte DNA damage and reduced the plasma α-tocopherol level (significant only for FM+FV). Conclusion: The substitution of fat meat with lean meat in a diet with or without fruit and vegetables has only a marginal or no effect on oxidative stress. But fruit and vegetable exclusion markedly increased the level of oxidative stress.

Effects of supplementation with α-tocopherol, ascorbic acid, selenium, or their combination in linseed oil-enriched diets on the oxidative status in broilers

&NA; A trial was conducted to investigate whether, and if so to what extent, the combined supplementation of vitamin E, vitamin C, and selenium was superior to their sole supplementation concerning the oxidative stress induced by a high n‐3 dietary polyunsaturated fatty acids (PUFA) intake in broilers. Four hundred 21‐day‐old Ross 308 male broilers were allocated to 5 experimental groups fed the following linseed oil (5%)‐enriched finisher diets: Cont (no supplement), +E (200 IU vitamin E/kg feed), +C (250 mg vitamin C/kg feed), +Se (0.2 mg selenium/kg feed), or +ECSe (concentrations as in the sole supplementation, combined). Analyses of malondialdehyde (MDA), vitamin C, and &agr;‐ and &ggr;‐tocopherols in plasma, antioxidant capacity of water‐ (ACW) and lipid‐ (ACL) soluble compounds in serum, and glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities in whole blood were performed. In breast muscle, analyses of MDA, &agr;‐ and &ggr;‐tocopherols, ACW, selenium, and fatty acid (FA) composition were carried out. Birds fed the combination of antioxidants showed reduced average daily gain (ADG) and average daily feed intake (ADFI) and, as +E, lower MDA and &ggr;‐tocopherol, together with raised &agr;‐ tocopherol levels in plasma and lower MDA and raised &agr;‐ tocopherol levels in breast muscle compared to the control. The combination of antioxidants in the +ECSe group raised GPx activity in whole blood compared to the control. In conclusion, results indicated that vitamin E is the most effective antioxidant to alleviate oxidative stress caused by high dietary PUFA and that the supplementation with additional vitamin C and selenium did not have clear synergistic effect.