Assessment of antioxidant capacity of bio-functional animal feed enriched with winery and oil mill products in broiler chickens and lambs

Grape pomace (GP) and Olive oil mill wastewater (OMW) are the main by-products generated during various stages of wine and olive oil production, respectively. While both of these derivatives are distinguished by their high organic load and their association to environmental pollution, they also display potential biological and clinical significance due to their high polyphenolic content, and thus antioxidant capacity. It is known that livestock are susceptible to diseases due to their low antioxidant defense system and their lack of adaptation to different dietary conditions. Additionally, their welfare and productivity are largely influenced by their farming environment, resulting in various pathological and stressful conditions. Recently, there is a great interest in the production and administration of animal feed with antioxidant activity and therapeutic properties. The aim of this thesis is to investigate the potential bioactivity of these animal feeds. To that end, 30 broilers and 24 lambs were fed with two bio-functional feeds enriched with grape pomace and olive oil mill wastewater respectively, which were previously evaluated for their antioxidant activity through in vitro techniques. Following feed administration, blood and tissues of vital organs from broilers and lambs were collected at two time points (30 & 50 days for broilers, 42 & 70 days for lambs post-birth). Biomarkers for antioxidant capacity, oxidative damage to lipids and proteins, as well as protein expression and antioxidant activity were assessed in the collected samples. In contrast to blood, tissue-specific results were observed in both species, but generally, bio-functional feed with GP and OMW reduced toxic effects from oxidative stress and enhanced animal redox status. Consequently, welfare and productivity of livestock were improved, while the harnessing of these by-products could contribute in diminishing environmental pollution due to their uncontrolled disposal to aquatic ecosystems.Τα στέμφυλα οινοποιίας και τα υγρά απόβλητα ελαιοτριβείων (ΥΑΕ) αποτελούν τα κύρια παραπροϊόντα που παράγονται κατά την επεξεργασία των σταφυλιών κατά την οινοποίηση και της ελιάς για την παραγωγή ελαιολάδου αντίστοιχα. Τα δυο αυτά παράγωγα χαρακτηρίζονται από υψηλό οργανικό φορτίο, με αποτέλεσμα να συνιστούν απόβλητα με σημαντικό περιβαλλοντικό αντίκτυπο. Όμως παράλληλα παρουσιάζουν αξιοσημείωτη βιοδραστκότητα και κλινικές ιδιότητες, κυρίως λόγω του υψηλού τους περιεχομένου σε πολυφαινολικές ενώσεις, οι οποίες είναι γνωστές για τις αντιοξειδωτικές τους ικανότητες. Έτσι παρουσιάζουν ένα σημαντικό δυναμικό για χρησιμοποίηση στη διατροφή των παραγωγικών ζώων τα οποία είναι ιδιαίτερα επιρρεπή σε εκδήλωση ασθενειών λόγω της χαμηλής αντιοξειδωτικής τους άμυνας και έλλειψης προσαρμογής στις διατροφικές αλλαγές. Η ευζωία και η παραγωγικότητα των ζώων αυτών επηρεάζεται σε μεγάλο βαθμό από το περιβάλλον εκτροφής και τις συνθήκες ενσταβλισμού τους, επάγοντας στρεσογόνα φαινόμενα που συχνά οδηγούν στην εκδήλωση ποικίλων παθολογικών καταστάσεων. Ως αποτέλεσμα, έχει αναπτυχθεί ένα σημαντικό ερευνητικό και επιχειρηματικό ενδιαφέρον για τη δημιουργία και χρησιμοποίηση ζωοτροφών που διαθέτουν με αντιοξειδωτική δράση και αποδεδειγμένες θεραπευτικές ιδιότητες. Στο πλαίσιο υλοποίησης της διατριβής, 30 κοτόπουλα (ηλικίας 2 ημερών) και 24 πρόβατα εκτράφηκαν με ζωοτροφές που ήταν εμπλουτισμένες με στέμφυλα οινοποιίας και ΥΑΕ αντίστοιχα, πρόσθετα με γνωστές αντιοξειδωτικές ιδιότητες, οι οποίες είχαν αποτιμηθεί με in vitro προσδιορισμούς. Η χορήγηση των ζωοτροφών πραγματοποιήθηκε αφ’ ενός για τα κοτόπουλα, από τις 2 μέχρι και τις 50 ημέρες μετά τη γέννησή τους, αφ’ ετέρου για τα πρόβατα από τη 15η μέχρι την 70η ημέρα μετά τη γέννησή τους. Μετά το πέρας της χορήγησης του κύκλου εκάστης ζωοτροφής, ελήφθησαν δείγματα αίματος και ιστών ζωτικών οργάνων τους σε δυο χρονικές στιγμές (μετά τη γέννησή τους 30 & 50 ημέρες για τα κοτόπουλα, 42 & 70 ημέρες για τα πρόβατα). Στα δείγματα μετρήθηκαν οι δείκτες που σχετίζονται με την αντιοξειδωτική τους ικανότητα, την οξειδωτική καταστροφή των λιπιδίων και πρωτεΐνών, καθώς και τη δραστικότητα και πρωτεϊνική έκφραση των αντιοξειδωτικών ενζύμων. Τα αποτελέσματα των αιματολογικών εξετάσεων έδειξαν σε όλες τις περιπτώσεις μια ιστο-ειδική επίδραση, με τη γενική εικόνα και για τις δυο εμπλουτισμένες ζωοτροφές (με στέμφυλα και ΥΑΕ) να υποδεικνύει μείωση των τοξικών επιδράσεων που προκαλούνται από το οξειδωτικό στρες και ενισχύουν την οξειδοαναγωγική κατάσταση των ζώων. Ως συνέπεια, η χρησιμοποίηση των προσθέτων αυτών αφενός βελτιώνει την ευζωία και παραγωγικότητα των εκτρεφόμενων ζώων και αφετέρου συμβάλει στη μείωση των περιβαλλοντικών προβλημάτων που προκαλείται από την ανεξέλεγκτη εναπόθεσή τους στο περιβάλλον

Determination of Redox Status in Different Tissues of Lambs and Kids and Their in-between Relationship

The objective of this study was to assess the resting values of the physiological oxidative stress exhibited by lambs and kids reared in Greece, and the potential correlations between redox biomarker levels in blood and other tissues (liver, diaphragm, quadriceps, psoas major muscle). For this purpose, lambs and kids at different developmental stages (d.s.) were used. The latter corresponded to four live weight categories (LWC), each representing 25%, 35%, 70% and 100% of mature body weight. In each of the above tissues, the levels of five common redox biomarkers were determined: glutathione (GSH), catalase (CAT), total antioxidant capacity (TAC), thiobarbituric reactive substances (TBARS), and protein carbonyls (CARBS). The results revealed that lambs and kids belonging to the 35% LWC had weaker endogenous antioxidant pools, while animals in the 70% and 100% LWC had elevated intrinsic antioxidant defense systems. Blood redox biomarkers were associated with the respective ones measured in the diaphragm, liver, quadriceps, and psoas major of both species. Importantly, TBARS levels in blood of animals in the 25% and 100% LWC are correlated with the TBARS levels in all other tissues tested. Blood antioxidant parameters might be used as potential biomarkers to predict the antioxidant status of tissues that affect meat quality. The latter would facilitate quality assessment prior to slaughter, allowing for timely nutritional interventions that can improve meat products

Tissue specific effects of feeds supplemented with grape pomace or olive oil mill wastewater on detoxification enzymes in sheep

The aim of the present study was to investigate the effects of livestock feed supplemented with grape pomace (GP) or olive oil mill wastewater (OMW) byproducts on the enzymatic activity and protein expression of antioxidants enzymes, in liver and spleen tissue of sheep. Thus, 36 male sheep of Chios breed were divided into 3 homogeneous groups, control group (n = 12), GP group (n = 12) and OMW group (n = 12), receiving standard or experimental feed. Liver and spleen tissues were collected at 42 and 70 days post-birth. The enzymatic activity of superoxide dismutase (SOD) and glutathione-s-transferase (GST) and also the protein expression of γ-synthase glutamyl custeine (γ-GCS) were determined in these tissues. The results showed GP group exhibited increased enzymatic activity of GST and protein expression of γ-GCS in liver compared to control group. In GP groupâs spleen, GST activity was increased compared to control but γ-GCS expression was not affected. In OMW groupâs liver, GST activity was increased and γ-GCS expression was reduced compared to control. In OMW groupâs spleen, GST activity was increased but GCS expression was not affected. SOD activity was not affected in both tissues either in GP or OMW group. Keywords: Grape pomace (GP), Olive oil mill wastewater (OMW), Glutathione-s-transferase (GST), Superoxide dismutase (SOD), γ-synthase glutamyl custeine (γ-GCS

Redox Status of Pregnant Ewes after Vaccination against Clostridial Diseases

The redox status shortly after the vaccination of pregnant ewes is rather unexploited. Thus, the present study was designed to evaluate the fluctuation of redox status after the administration of the annual booster dose of a polyvalent clostridial vaccine in pregnant ewes, 3 to 4 weeks before lambing, with or without a simultaneous injection of Vit E/Se. In total, 24 pregnant Lacaune ewes 3–4 weeks before lambing were randomly allocated into four equal groups: the V (vaccinated with a polyvalent clostridial vaccine), VE (vaccinated and injected IM with Vit E/Se), E (injected IM with Vit E and Se), and C (neither vaccinated nor injected with Vit E/Se). The study period lasted for 21 days, starting on the day of administration. Four redox biomarkers, the antioxidant capacity (TAC), the thiobarbituric acid reactive substances (TBARS), the reduced glutathione (GSH), and the catalase (CAT) were evaluated in blood samples collected from all ewes before the injections (0 h) and then at 12 (12 h), 24 (D1), and 48 h (D2), and thereafter on days 4 (D4), 6 (D6), 10 (D10), 14 (D14), and 21 (D21). The results reveal that the TAC was the only biomarker evaluated that was significantly affected by group and significantly lower in vaccinated animals (V and VE groups) compared to non-vaccinated (E and C groups). The absence of an increase in the TBARS values after vaccination in groups V and VE indicates the absence of significant oxidative stress. Overall, it can be assumed that annual booster immunizations against clostridial diseases do not impose acute oxidative stress on pregnant ewes in the last month of pregnancy

Assessment of Antioxidant and Antimutagenic Properties of Red and White Wine Extracts In Vitro

Wine is an alcoholic beverage of complex composition obtained through the fermentation of grape must. The consumption of wine has already been associated with a multitude of beneficial effects due to its high polyphenolic content. In this study, four Greek emblematic wines from two red (i.e., Xinomavro and Agiorgitiko) and two white (i.e., Assyrtiko and Malagouzia) varieties were analyzed for the estimation of their antioxidant profiles. To address this question, we assessed their ability to scavenge both synthetic and endogenous free radicals, such as DPPH•, ABTS+•, OH•, O2−, their potential reducing power, and their antimutagenic and antigenotoxic properties. All varieties exhibited potent antioxidant activity, as indicated by the results of methods above, with the red wines appearing more effective than the white ones regarding antioxidant capacity. Our small-scale study is the first to reveal that these wine varieties may have the ability to scavenge the most reactive endogenous radicals. In the future, this finding must be accompanied by larger studies to fill a knowledge gap in the scientific literature concerning a holistic approach of the in vitro antioxidant action of plant polyphenolic compounds. Conclusively, we believe that wines possess high bioactivity that allow them to settle in the industry of food additives and medicinal products

Assessment of Antioxidant and Antimutagenic Properties of Red and White Wine Extracts In Vitro

Wine is an alcoholic beverage of complex composition obtained through the fermentation of grape must. The consumption of wine has already been associated with a multitude of beneficial effects due to its high polyphenolic content. In this study, four Greek emblematic wines from two red (i.e., Xinomavro and Agiorgitiko) and two white (i.e., Assyrtiko and Malagouzia) varieties were analyzed for the estimation of their antioxidant profiles. To address this question, we assessed their ability to scavenge both synthetic and endogenous free radicals, such as DPPH center dot, ABTS(+center dot), OH center dot, O-2(-), their potential reducing power, and their antimutagenic and antigenotoxic properties. All varieties exhibited potent antioxidant activity, as indicated by the results of methods above, with the red wines appearing more effective than the white ones regarding antioxidant capacity. Our small-scale study is the first to reveal that these wine varieties may have the ability to scavenge the most reactive endogenous radicals. In the future, this finding must be accompanied by larger studies to fill a knowledge gap in the scientific literature concerning a holistic approach of the in vitro antioxidant action of plant polyphenolic compounds. Conclusively, we believe that wines possess high bioactivity that allow them to settle in the industry of food additives and medicinal products

Association of melatonin administration in pregnant ewes with growth, redox status and immunity of their offspring.

SIMPLE SUMMARY: Melatonin is a known antioxidant and anti-inflammatory regime, while in sheep it is broadly used to accelerate the onset of the breeding season. Our recent study showed that melatonin administration during pregnancy in heat-stressed ewes improved fertility rate and number of lambs born per ewe, the redox status of the maternal organism and the produced milk quantity until weaning. In this study, we present the impact of melatonin administration in stressed ewes during pregnancy considering: (a) humoral response of both maternal organism and offspring during the first two days after parturition, (b) chemical composition and antioxidant parameters of colostrum and milk until weaning and (c) redox status of the offspring until weaning. The results indicated that melatonin improved the redox status of the offspring and the quality of colostrum. Moreover, melatonin could be administered as immune-modulatory regime, apart from antioxidant, in prenatally stressed offspring in order to cope with the crucial first days of their life, as the humoral response results suggested. ABSTRACT: In this study, the effects of melatonin treatment on growth, redox status and immunity in prenatally stressed newborn lambs were evaluated. Thirty-seven newborn lambs were allocated into two groups (melatonin-MEL and control-CON), based on whether their mothers were treated with melatonin implants or not, respectively. All pregnant ewes were exposed to heat stress. The body weight of lambs was recorded at birth (L0), and then on days 15 (L15) and 40 (L40). Redox biomarkers [total antioxidant capacity (TAC), glutathione (GSH), thiobarbituric acid reactive substances (TBARS)] were assayed in blood samples collected from lambs on days L0, L1, L2, L5, L10 and L40. Chemical analysis and antioxidant capacity were evaluated in colostrum and milk samples collected at the same time points with blood samples. Cytokines (IL-1β, IL-6, IL-10, IFN-γ) and immunoglobulin (IgG) were assayed in blood and colostrum samples collected from ewes on days L0 and L1, and in lambs’ blood on days L0, L1 and L2. The results revealed that body weight gain of newborn lambs did not differ between the two groups (p > 0.05). Better redox status was found in MEL lambs until L2, as well as higher antioxidant capacity in the colostrum of MEL ewes compared to CON ones on day L0 (p < 0.05). In MEL ewes’ colostrum, higher protein content was measured on day L0 and higher fat content on L1 compared to CON group (p < 0.05). The highest level of IL-6 was found in MEL ewes on L1, with a concomitant increase of IL-10 level in MEL lambs in comparison to CON lambs on L2. Moreover, CON colostrum resulted in a higher level of IL-10 within time, coupled with an increased level of IgG found in lambs’ plasma on L2 (p = 0.04). This study indicated that melatonin could be administered as antioxidant and immune-modulatory regime in prenatally stressed offspring in order to cope with the crucial first days of their life. This effect of melatonin was also amplified by crosstalk between IL-6, IL-10 and IgG production, resulting in an improved quality of produced milk

Association of Melatonin Administration in Pregnant Ewes with Growth, Redox Status and Immunity of Their Offspring

In this study, the effects of melatonin treatment on growth, redox status and immunity in prenatally stressed newborn lambs were evaluated. Thirty-seven newborn lambs were allocated into two groups (melatonin-MEL and control-CON), based on whether their mothers were treated with melatonin implants or not, respectively. All pregnant ewes were exposed to heat stress. The body weight of lambs was recorded at birth (L0), and then on days 15 (L15) and 40 (L40). Redox biomarkers [total antioxidant capacity (TAC), glutathione (GSH), thiobarbituric acid reactive substances (TBARS)] were assayed in blood samples collected from lambs on days L0, L1, L2, L5, L10 and L40. Chemical analysis and antioxidant capacity were evaluated in colostrum and milk samples collected at the same time points with blood samples. Cytokines (IL-1β, IL-6, IL-10, IFN-γ) and immunoglobulin (IgG) were assayed in blood and colostrum samples collected from ewes on days L0 and L1, and in lambs’ blood on days L0, L1 and L2. The results revealed that body weight gain of newborn lambs did not differ between the two groups (p &gt; 0.05). Better redox status was found in MEL lambs until L2, as well as higher antioxidant capacity in the colostrum of MEL ewes compared to CON ones on day L0 (p &lt; 0.05). In MEL ewes’ colostrum, higher protein content was measured on day L0 and higher fat content on L1 compared to CON group (p &lt; 0.05). The highest level of IL-6 was found in MEL ewes on L1, with a concomitant increase of IL-10 level in MEL lambs in comparison to CON lambs on L2. Moreover, CON colostrum resulted in a higher level of IL-10 within time, coupled with an increased level of IgG found in lambs’ plasma on L2 (p = 0.04). This study indicated that melatonin could be administered as antioxidant and immune-modulatory regime in prenatally stressed offspring in order to cope with the crucial first days of their life. This effect of melatonin was also amplified by crosstalk between IL-6, IL-10 and IgG production, resulting in an improved quality of produced milk

Redox biomarker baseline levels in cattle tissues and their relationships with meat quality

Cattle breeds or crossbreds with high productivity traits have been developed to meet a growing demand for food. When intensive farming practices are followed, animals face several challenges which can result in poor performance, compromised welfare and the reduced quality of their products. Our study aims to highlight the resting values of the physiological oxidative stress that three cattle breeds exhibit, and their potential relationship with meat quality. For this purpose, we determined the levels of five common redox biomarkers (glutathione (GSH), catalase (CAT), total antioxidant capacity (TAC), thiobarbituric reactive substances (TBARS) and protein carbonyls (CARBS)) in the tissues of three commonly used beef cattle breeds (Charolais (CHA), Limousin (LIM) and Simmental (SIM)) and their association with specific meat quality traits that depend on color, pH and texture. The results revealed that LIM cattle breed animals have elevated intrinsic antioxidant defense systems in comparison to CHA and SIM cattle breed animals. In addition, the meat quality parameters were associated with the redox biomarkers. We propose that the determination of specific antioxidant parameters in the blood might be used as potential biomarkers to predict meat quality. This would allow farmers to nutritionally intervene to improve the quality of their products