LITHUANIAN UNIVERSITY OF HEALTH SCIENCES VETERINARY ACADEMY
Vilija Buckiūnienė
THE INFLUENCE OF DIFFERENT OILS
AND ANTIOXIDANTS ON LAYING
HENS EGGS AND BROILERS CHICKEN
MEAT QUALITY
Doctoral Dissertation Agricultural Sciences, Zootechnics (03A)
The dissertation is prepared between 2012 and 2016 at the Veterinary Academy of Lithuanian University of Health Sciences, Laboratory of Animal Productivity under Institute of Animal Rearing Technologies.
Dissertation is defended extramurally.
Scientific consultant –
Prof. Habil. Dr. Romas Gružauskas (Lithuanian University of Health Sciences, Agricultural Sciences, Zootechnics – 03A).
Dissertation is defended at the Zootechnics Research Council of Lithuanian University of Health Sciences
Chairperson –
Prof. Dr. Elena Bartkienė (Lithuanian University of Health Sciences, Agricultural Sciences, Zootechnics – 03A).
Members:
Dr. Jonas Jatkauskas (Lithuanian University of Health Sciences, Agri-cultural Sciences, Zootechnics – 03A);
Dr. Violeta Razmaitė (Lithuanian University of Health Sciences, Agri-cultural Sciences, Zootechnics – 03A);
Assoc. Prof. Dr. Antanas Šarkinas (Kaunas University of Technology, Technological Sciences, Chemical Engineering – 05T);
Prof. Dr. Qendrim Zebeli (University of Veterinary Medicine, Vienna, Agricultural Sciences, Veterinary – 02A).
Dissertation will be defended at the open session of the Zootechnics Research Council of the Lithuanian University of Health Sciences on the 19th of May, 2017 at 1:00 PM in Dr. S. Jankauskas Auditorium of the Vete-rinary Academy.
LIETUVOS SVEIKATOS MOKSLŲ UNIVERSITETAS VETERINARIJOS AKADEMIJA
Vilija Buckiūnienė
ĮVAIRIŲ ALIEJŲ BEI ANTIOKSIDANTŲ
ĮTAKA VIŠTŲ KIAUŠINIŲ IR VIŠČIUKŲ
BROILERIŲ MĖSOS KOKYBEI
Daktaro disertacijaŽemės ūkio mokslai, zootechnika (03A)
Disertacija parengta 2012–2016 metais Lietuvos sveikatos mokslų univer-sitete Veterinarijos akademijoje, Gyvūnų produktyvumo laboratorijoje prie Gyvūnų auginimo technologijų instituto.
Disertacija ginama eksternu.
Mokslinis konsultantas –
Prof. habil. dr. Romas Gružauskas (Lietuvos sveikatos mokslų universi-tetas, žemės ūkio mokslai, zootechnika – 03A).
Disertacija ginama Lietuvos sveikatos mokslų universiteto Veterinari-jos akademiVeterinari-jos Zootechnikos mokslo krypties taryboje:
Pirmininkė –
Prof. dr. Elena Bartkienė (Lietuvos sveikatos mokslų universitetas, žemės ūkio mokslai, zootechnika – 03A).
Nariai:
Dr. Jonas Jatkauskas (Lietuvos sveikatos mokslų universitetas, žemės ūkio mokslai, zootechnika – 03A);
Dr. Violeta Razmaitė (Lietuvos sveikatos mokslų universitetas, žemės ūkio mokslai, zootechnika – 03A);
Doc. dr. Antanas Šarkinas (Kauno technologijos universitetas, techno-logijos mokslai, chemijos inžinerija – 05T);
Prof. dr. Qendrim Zebeli (Vienos veterinarinės medicinos universitetas, žemės ūkio mokslai, veterinarija – 02A).
Disertacija ginama Lietuvos sveikatos mokslų universiteto Zootechnikos mokslo krypties taryboje 2017 m. gegužės 19 d. 13 val. dr. S. Jankausko auditorijoje.
TABLE OF CONTENT
ABBREVIATIONS ... 7
INTRODUCTION ... 8
1. REVIEW OF LITERATURE ... 13
1.1. Overview of the Physiological Roles and Corresponding Health Benefits of (n-3) PUFA ... 13
1.2. Benefits of oils in human nutrition ... 15
1.3. Digestion and metabolism of vitamin E ... 16
1.4. Antioxidants ... 18
1.5. Antioxidant in eggs ... 19
1.6. Selenium – an antioxidant with strong Pro-Oxidant properties ... 21
1.7. Health benefits of carotenoids ... 22
1.8. Beta-carotene ... 23
1.9. Lycopene ... 23
1.10. Lutein and Zeaxanthin ... 25
1.11. Lycopene in poultry products and its influence on human health ... 26
1.12. Demage of malondialdehydes (MDA) ... 28
1.13. Iron absorption ... 30
2. METHODICS ... 33
2.1. The experiments place, time and trial schemes ... 33
2.2. Feeding trials with laying hens and chicken broilers ... 34
2.3. Characteristics of in experiment used feed additives ... 37
2.4. The feed production and composition ... 38
2.5. In experiment used oils analysis methods ... 38
2.6. The evaluation methods of the laying hens ... 40
2.6.1. The methods of zootechnical analysis of laying hens ... 40
2.6.2. Methods of egg quality evaluation ... 40
2.6.3. Methods of egg sensory and texture properties evaluation ... 43
2.6.4. Physiological methods ... 44
2.7. The analysis methods of the trial with broiler chickens ... 45
2.7.1. The methods of zootechnical analysis of broiler chickens ... 45
2.7.2. Physiological analysis ... 46
2.7.3. The methods of broiler chickens meat quality analysis ... 46
2.7.4. Evaluation methods of sensory and technological properties for poultry meat ... 48
3. RESULTS ... 50
3.1. Usage of different oils, selenium and vitamin E in laying hens’ feeding ... 50
3.2. Usage of different oils, antioxidants and lycopene on laying hens feeding .... 79
3.3. Influence of iron sulphate and iron glycinate on laying hens’ productivity, digestive process, physiology status and egg quality parameters ... 111
3.4. Influence of iron sulphate and iron glycinate on productivity, digestive process, physiology status and meat quality parameters of broilers chicken ... 134
4. DISCUSSION ... 156
4.1. Effect of plant oil ... 162
4.2. Effect of lycopene ... 162 4.3. Effect of antioxidant ... 163 CONCLUSIONS ... 164 PRACTICAL RECOMMENDATION ... 168 REFERENCES ... 169 PUBLICATIONS ... 190 SANTRAUKA ... 221 ANNEX ... 245 CURRICULUM VITAE ... 259 ACKNOWLEDGEMENTS ... 260
ABBREVIATIONS
SCFA – short chain fatty acidTBARS – Thiobarbituric acid reactive substances (lipid oxidation degree) GPT – alaninaminotransferase
GOT – aspartataminotransferase GGT – gama-gliutamiltranspeptidase SFA – saturated fatty acid
MUFA – monounsaturated fatty acid PUFA – polyunsaturated fatty acid AI – aterogenic index TI – trombogenic index h/H – hipocholesterolemic/Hipercholesterolemic index IP – peroxidabity index L* – lightness a* – redness b* – yellowness WBC – leucocytes RBC – erythrocytes HGB – hemoglobin MCV – mean cell volume
MCH – mean corpuscular hemoglobin MCHC – mean cell hemoglobin concentration PLT – platelets
MPV – mean platelet volume MDA – malondialdehyde n-3 – omega 3 fatty acid n-6 – omega 6 fatty acid CVD – cardiovascular disease GRx – glutathione peroxidase TRxR – thioredoxin reductase DIO – iodothyronine deiodinase ROS – reactive oxigen species ID – iron deficiency FCR – feed conversion ratio LC – long chain DHA – docosahexaenoic EPA – eicosapentaenoic SO – sunflower oil RO – rapeseed oil LO – linseed oil
INTRODUCTION
In recent years, consumers are becoming increasingly aware of the nutritional quality and healthy benefits of the food they consume. It is widely acknowledged that cholesterol content and fatty acid composition in poultry products are closely related to the occurrence of cardiovascular heart diseases (Sacks, 2002). Therefore, much attention has been paid toward the modulation of poultry products to decrease the risk of cardiovascular disease by improving the properties of carcasses and meat quality (Kamboh and Zhu, 2013).
The description of active compounds in food is an indispensable step in the validation process of such nutritional strategies and it is essential for the exact definition of “functional food” (Jezewska-Zychowicz, 2009). The discovery of active compounds contained in food daily consumed, is an essential step in the realization of a nutraceutical food having the capacity to prevent chronic disorders (Leoncini et al., 2012). The term nutraceutical (a combination of nutrition and pharmaceutical) indicates a food component that provides health benefits, including the prevention of diseases. Consu-mers are progressively more interested in functional foods, therefore in present scenario they are extremely careful on their wellbeing as a result demands of beneficial foods worldwide growing. In spite of slight variations in overall nutritional content of eggs from country to country (mostly related to the variations in diet fed to the hens), eggs are widely recognized as naturally rich in proteins and a number of vitamins and minerals. The egg is often referred to as one of the original and natural functional foods. Nutritional claims allowed for regular eggs (standard diet fed to the hens) vary from country to country mostly due to regulations (Guyonnet, 2011).
Nowadays, however, it is known that the response of cholesterol in human serum levels to dietary cholesterol consumption depends on several factors, such as ethnicity, genetic makeup, hormonal factors and the nutritional status of the consumer. Additionally, in recent decades, there has been an increasing demand for functional foods, which is expected to continue to increase in the future, owing to their capacity to decrease the risks of some diseases and socio-demographic factors such as the increase in life expectancy.
Eggs provide a completely packaged, highly nutritious food including essential nutritional components (Laudadio et al., 2014). However, the uncertainties are related to the cholesterol level in eggs. It was reported by Gilbert (2000) that eggs result more healthier and valuable food when cholesterol content is reduced. Nevertheless, the cholesterol level derived
from egg is much lesser than the cholesterol concentration synthesized by humans (Singh and Sachan, 2010). The interior components of egg can easily be transformed by poultry feeding practices (Laudadio and Tufarelli, 2011; Khan et al., 2012a). The eggs can also be improved thanks to addi-tional nutrients such as selenium, vitamins B-group, vitamin E and parti-cularly antioxidant compounds. Nevertheless, the fortification of different nutrients in egg is exclusively dependent on nutritional changes of laying hen diet (Ahmed and Abdelati, 2009; Singh and Sachan, 2010). This egg is moreover able to attract the market attention by varying their health status and interesting to a kind of the consumer who are available to spend for these changes in egg. Therefore, as need and production of enriched eggs increases, research is needed to establish their solidity and adequacy. Egg is a nutritive food rich in proteins and it is a cheap source of protein (Hasler, 2000). Altering the nutritive contents of the egg can thus help alleviating the problem of nutritional deficiencies in target population (Kassis et al., 2010). World bodies recommend for the intake of 3 servings of vegetables and 2 servings of fruits a day but this is not practically feasible in many countries which are developing. So there is a definite need for an alternative that can sort these problems (Dike, 2014). One should have a food item that is accepted by most of the people in the community, egg is one such food item that can be fortified with various nutrients. Egg is superior to animal meat in case of nutrients because animal meat contains more fat that can cause various health problems to human which include heart attack (Simopoulos, 1991).
Poultry meat is an accepted valuable source of nutrients for consumers. In general, consumers are interested in good tasting and healthy food with relevance to nutritional physiology. At the same time, they are afraid of potentially harmful ingredients such as drug residues, intoxicants, allergenic components, and microbial contamination, which may contribute to health problems. On the other hand, consumers are more and more interested in products enriched with beneficial components (for example, probiotics in yogurt), which will improve their wellbeing. Therefore, consumers decide to buy food with special “healthy” contents and, what is also important, they are willing to pay more money for healthy food. One restriction for this kind of improved food is that it has to be produced in a natural way. One way to fulfill these demands is the production of “functional foods”.
This definition can be extended to cover the fact that functional foods may also assure the recommended daily intake (RDI) of relevant substances, which are often lacking in a daily diet. Therefore, functional foods can improve the health and well-being of humans and may reduce the risk of metabolic disease. Functional foods are produced in a natural way by
enri-ching existing components and thus correspond to human expectations of this type of product. Functional poultry meat may be capable of enhancing the status of poultry meat and in this way further increase poultry meat consumption. It can therefore be confirmed that enriching poultry meat with health-promoting substances is an interesting future issue for poultry meat production (Uijttenboogaart, 2000), as was the case for eggs (Pritchard, 2003).
Poultry meat is an important source of high-quality protein, minerals, and vitamins to balance the human diet (Regmi, 2007) and, in 2010, was consu-med more than any other meat at an average of 12.5 kg per person worldwide (Brainsma, 2015). Demand for poultry meat is projected to double by 2030 (Brainsma, 2015).
Fat is a high energy source, improves the diet palatability and fat–vitamin absorption, increases the intestinal absorption of all nutrients reducing the passage rate of digesta, modulates immunity, modifies meat composition and quality, and can be a relatively economic feed (sub)product from crop, livestock and fishery industries.
The broiler chicken needs high-energy diet to support its rapid growth and metabolism. Thus, approximately 2 to 5 per cent oil is recommended in broiler chicken diets for optimal growth performance. Vegetable oils are mainly triacylglycerols which account for more than 95 per cent of total oil. They also contain small quantities of diacylglycerols, phospholipids, toco-pherols, free fatty acids, etc.
Nowadays, the positive effects of increasing n-3 PUFA (polyunsaturated fatty acid) and balanced intake of n-3 and n-6 fatty acids have been linked to coronary artery disease, hypertension, diabetes, arthritis, other inflammatory and autoimmune disorders, cancer and mental illnesses (Griffin, 2008; Rie-diger et al., 2009).
Improvement in the n-3 fatty acid composition of animal products for human consumption could be achieved by increasing the intake of the feed of animal origin (marine sources) that is rich in n-3 long chain polyunsatu-rated fatty acids (n-3 LC PUFA, including eicosapentaenoic, docosapenta-enoic and docosahexadocosapenta-enoic fatty acid), mainly docosahexadocosapenta-enoic (DHA) and eicosapentaenoic acid (EPA). Another possibility is to provide feed with a high level of a-linolenic acid, which could be bioconverted to longer and more unsaturated n-3 LC PUFA. Linseed oil is one of the best vegetable sources of indispensible PUFA, with a-linolenic acid content above 50 per cent of total fatty acid content.
According to Poureslami et al. (2010), bioconversion of a-linolenic acid into LC PUFA is inversely related to animal’s place on evolutionary scale (rainbow trout > broiler chicken > human).
Taking into account the very low bioconversion of alinolenic fatty acid into n-3 LC PUFA in humans and low fish consumption, it seems that the most beneficial way to increase n-3 fatty acid levels in human food is the consumption of linseed-fed animals (Kartikasari et al., 2012).
The fatty acid composition of the egg may be influenced by the dietary composition offered to the laying hens (Bavelaar and Beynen 2004). Howe-ver, attempts to manipulate the total amount of fats in the egg through diet change have only marginally succeeded. Polyunsaturated fatty acids (PUFA) are considered healthy fatty acids and are classified as α-linolenic acid, eicosapentaenoic acid, and docosahexaenoic acid and have been clinically proven to minimize the incidence of heart-associated diseases and certain mental disorders (Simopoulos, 1999; Riediger et al., 2009).
A number of different seeds and their oils and some nuts contain considerable amounts of a-linolenic acid. Linseeds (flaxseeds) and their oil typically contain 45-55 per cent of fatty acids as a-linolenic acid, whereas soybean oil, rapeseed oil, and walnuts all typically contain ~10 per cent of fatty acids as a-linolenic acid. There is little a-linolenic acid in corn oil, sunflower oil, or safflower oil, which are all rich in linoleic acid (Burdge and Calder, 2006; Blasbalg et al., 2011).
The aim of the study
To investigate the impact of different oils and antioxidants on producti-vity of broilers chicken and laying hens, physiological status and quality of production.
Objectives of the study
1. To investigate the impact of different oils, natural and synthetics antioxidants on chicken broiler and laying hens productivity and livability.
2. To ivestigate the impact of different oils, natural and synthetics antioxidants on broiler chicken and laying hens physiological status.
3. To investigate the impact of different oils, natural and synthetics antioxidants on broiler chicken and laying hens quality of produc-tion.
4. To investigate the impact of of different oils, natural and synthetics antioxidants on broiler chicken and laying hens production of textural and sensory characteristics.
Actuality and novelty of the work
Functional foods are those that provide a specific health benefit to the consumer over and above their nutritional value. Functional foods are relatively recent developments that meet a strengthening consumer demand for foods that enhance health and wellbeing. Eggs are of particular interest from a functionality point of view, because they are relatively rich in fatty acids and the associated fat-soluble compounds. The type and ratio of fatty acids are important determinants of human health. The idea of egg enrichment with n-3 fatty acids simultaneously with antioxidants and other vitamins.
The impact of compound feed supplement with the different oils (sunflower, rapeseed and flaxseed) and antioxidants (selenium, vitamin E, lycopene and santoquin) on laying hens and broiler chickens’ productivity, quality of production, PUFA, MUFA, n-6, n-3 concentration and their ratio, atherogenic, thrombogenicity, peroxidation, hipocholesteremic/hypercho-lesteremic indices, MDA concentrations, selenium, iron, vitamin E and pigment accumulation in eggs and broiler meat was analysed.
Practical value of the dissertation is characterized by the created com-pound feed formulations and prepared specific recommendations for increa-sed quality of broiler chickens’ meat and eggs. Optimal insertion of feed additives into compound feed creating the optimal n-6 and n-3 ratio and maximum Se and vitamin E accumulation in eggs and broiler chickens’ meat to ensure optimal lipid oxidation processes was analysed.
The novelty of the research is characterized by the fact that using com-pound feed enriched with different oils and antioxidants, fatty acid profile, atherogenic, thrombogenicity, peroxidation, hipocholesteremic/hypercholes-teremic indices, content of pigment substances, tocopherols, selenium and iron concentration, malondialdehyde content of sensory and textual proper-ties of eggs and poultry meat (fresh and stored for a longer period of time) have been extensively analysed. Effects of compound feed additives on broiler chickens and laying hens’ physiological status – as short-chain fatty acids and ammonia nitrogen concentration in the caecum, blood parame-ters – as alkaline phosphatase, bilirubin (total and direct), gamaglutamin-transferase, alanine aminotransferase were also studied.
1. REVIEW OF LITERATURE
1.1. Overview of the Physiological Roles and Corresponding Health Benefits of (n-3) PUFAVery long-chain (n-3) fatty acids have been demonstrated to have a wide range of physiological roles, as summarized in Table 1, where these roles are linked to certain health or clinical benefits. They have been demonstrated to beneficially modify a number of risk factors for cardiovascular disease. Risk factors improved include blood pressure (Geleijnse et al., 2002), platelet reactivity and thrombosis (Geleijnse et al., 2002), plasma TG concentrations (Harris, 1996), vascular function (Nestel et al., 2002), cardiac arrhythmias (von Schacky, 2008), heart rate variability (von Schacky, 2008), and inflame-mation (Calder, 2006). Due to these effects, increased intake of very long-chain (n-3) fatty acids is associated with a reduced risk of cardiovascular morbidity and mortality (Calder, 2004; Calder and Yaqoob, 2010). Supple-mentation of at-risk patients with very long-chain (n-3) fatty acids reduced mortality (Marchioli et al., 2002; Bucker et al., 2002; Studer et al., 2005; Yakoyama et al., 2007). In addition, several noncardiovascular actions of these fatty acids have been described (Table 1.1.1), suggesting that increasing their intake could reduce the risk of (i.e., protect against) a number of condi-tions and may even be used as a treatment. As an example, very longchain (n-3) PUFA have been used successfully in rheumatoid arthritis (Calder, 2008a) and, to a lesser extent, in inflammatory bowel diseases (Calder, 2008b) and asthma (Calder, 2006). DHA has an important structural role in the eye and brain. Consequently, ensuring a good DHA supply early in life when the brain and eye are developing is vitally important to optimize visual and neurological development (San Giovanni et al., 2000a; San Giovanni et
al., 2000b). Very long-chain (n-3) fatty acids may contribute to enhanced
mental development (Helland et al., 2003) and improved childhood learning and behavior (Richardson, 2004) and may lower the burden of psychiatric illnesses in adults (Freeman et al., 2006). There also appears to be a role for very long-chain (n-3) PUFA, especially DHA, in preventing neurodegenera-tive disease of ageing (Solfrizzi et al., 2010). The effects of very long-chain (n-3) PUFA on health outcomes are likely to be dose dependent, but clear dose response data have not been identified in most cases.
During the last 2-3 decades, a substantial growth in poultry industry has been observed, largely secluded to large and small scale organized poultry farming. This is mainly due to exploitation of various modern growth pro-moting strategies and appropriate disease preventive and control measures.
Table 1.1.1. Summary of the physiological roles and potential health benefits of very long-chain (n-3) fatty
Physiological role of very long-chain (n-3) fatty acids
Potential health benefit Disease target Regulation of blood
pressure
Decreased blood pressure Hypertension, CVD1 Regulation of platelet function Decreased likelihood of thrombosis Thrombosis, CVD Regulation of blood coagulation Decreased likelihood of thrombosis Hypertriglyceridemia, CVD Regulation of plasma TG concentrations Decreased plasma TG concentrations Hypertriglyceridemia, CVD Regulation of vascular function Improved vascular reactivity CVD Regulation of cardiac rhythm Decreased cardiac arrhythmias CVD Regulation of heart rate Increased heart rate
variability
CVD
Regulation of inflammation Decreased inflammation Inflammatory diseases (arthritis, inflammatory bowel diseases, psoriasis, lupus, asthma, cystic fibrosis, dermatitis, neurodege-neration, etc.), CVD Regulation of immune function Improved immune function Compromised immunity Regulation of fatty acid and
TG metabolism
Decreased TG synthesis and storage
Weight gain, weight loss, obesity Regulation of bone
turnover
Maintained bone mass Osteoporosis Regulation of insulin
sensitivity
Improved insulin sensitivity
Type-2 diabetes Regulation of tumor cell
growth
Decreased tumor cell growth and survival
Some cancers Regulation of visual
signaling (via rhodopsin)
Optimized visual signaling
Poor infant visual development (especially preterm)
Structural component of brain and central nervous system
Optimized brain development leading to better cognitive and learning processes
Poor infant and childhood cognitive processes, learning, and behavior
Lipids rich in PUFAs, used as food additives in order to fortify and/or amend FA composition of specific foods (e.g. infant foods), are highly demanded in the food industry. The food fortification in PUFAs can be achieved in several ways such as direct addition of PUFAs or PUFA-produ-cing edible microorganisms in foods, or through the utilization of feed rich in PUFAs that leads to the production of PUFA-rich animal products (e.g. eggs, meat) (Bellou et al., 2016).
For optimal health, many government and public health authorities recommend increasing ω-3 fatty acids in diet.
1.2. Benefits of oils in human nutrition
Sunflower oil is a high-quality edible oil. It is used in cooking, frying, and in the manufacture of margarine and shortening and considered by some as desirable as olive oil. Sunflower oil was selected in this study due to its high use in food as it is a rich source of linoleic acid. Furthermore, it is light in taste and appearance and has a high vitamin E content compared to other vegetable oils (Shahidi et al., 1992).
Today rapeseed is one of the top five oilseed crops cultivated world-wide (Miri, 2007; Khattab et al., 2012, Liersch et al., 2013). It is mainly utilized for its oil, which is one of the most common edible and healthy cooking oils due to its low content of saturated fatty acids (SFA, 7 per cent), high content of the monounsaturated fatty acids (MUFA, 60 per cent), and adequate content of polyunsaturated fatty acids (PUFA, 8-12 per cent). In addition, rapeseed seed is rich in tocols and phenolic compounds (Ackman, 1994; Naczk et al., 1998; Oraby and Ramadan, 2015).
Flaxseed oil mainly originated as triacylglycerol (98 per cent) with lower contents of phospholipids (0.9 per cent) and free fatty acids (0.1 per cent) (Mueller et al., 2010). The concentration of α-linolenic acid (ω-3 PUFA), a potent anti-carcinogenic compound in flaxseed oil ranges approxi-mately from 40-60 per cent. Other bioactive components, such as linoleic acid and oleic acid are also present, each at 15 per cent level (Williams
et al., 2008). Flaxseed (Linum usitatissimum L.) is an oilseed crop
conside-red a functional food (Shima et al., 2014) that has a high concentration of a-tocopherol (Oomah and Sitter, 2009) and an extremely high concentration of a-linolenic acid (Khattab and Zeitoun, 2013) and plant lignans, such as Secoisolariciresinol diglucoside 3699 lg/g as reported by Patel et al. (2012), considered anticancer effects compounds (Daleprane et al., 2010). Several studies have pointed to benefits of including extra virgin flaxseed oil (EFO) in the human diet (Pilar et al., 2014). Previous studies showed that inclusion
of EFO in the diet can inhibit cancer cell growth and spontaneous metastasis (Chen et al., 2006; Dabrosin et al., 2002) and, thus, is a great ally in the treatment of breast cancer (Mason et al., 2010) and ovarian cancer (Eilati
et al., 2013).
Good results were demonstrated in controlling the rates of LDL and total cholesterol concentrations (Patade et al., 2008) and treating cardiovas-cular diseases (Dodin et al., 2008; Leyva et al., 2011) related to the con-sumption of EFO. Due to their nutraceutical properties and high cost, EFO can be adulterated by the addition of edible oils with lower nutritional and commercial value, generating not only economical losses, but also a decrease in health benefits. This fraud is difficult to detect since refined soybean oil (SO) and refined sunflower oil (SFO), the most used adulterant oils, show similar physicochemical characteristics to EFO. Thus, it is necessary to establish efficient and feasible methods for the identification and quantification of adulteration in EFO (Yang et al., 2005).
1.3. Digestion and metabolism of vitamin E
Vitamin E is the most potent lipid-soluble antioxidant in human plasma. Dietary components with antioxidant activity of vitamin E include α-, β-, γ-and δ-tocopherols γ-and α-, β-, γ- γ-and δ-tocotrienols. These compounds containing a chromanol ring attached to a saturated phytyl (tocopherols) unsaturated (tocotrienols) and vary as to the number of methyl groups on the chromanol ring (Traber, 2012). Among the tocopherol homologs, the alpha-tocopherol has the highest biological potency (Ortíz et al., 2006).
Digestion of tocopherols and tocotrienols is similar to lipids, so that the emulsification and incorporation into mixed micelles takes place. Then, these compounds are absorbed by the enterocytes and transported primarily in the proximal intestine. In these cells, the tocopherols are incorporated into chylomicrons and transported by the lymphatic system (Nagy et al., 2013).
Os chylomicrons reach the blood circulation, where its triglycerides are hydrolyzed by the lipoprotein lipase (Asakura, 2006). Tocopherols reach the liver via the remnant chylomicrons and alpha-tocopherol is preferably incorporated into VLDLs by having greater affinity for the transport protein alpha-tocopherol (α-tocopherol transport protein or α-TTP). Therefore, alpha-tocopherol is the isomer that is in higher concentrations in plasma and tissues (Fu et al., 2014).
Several reactive oxygen/nitrogen species induce oxidative stress. The removal of these compounds is the main function of antioxidants, vitamin E being the most important and abundant fat-soluble antioxidant in vivo. The
peroxyl radicals are removed by vitamin E in order to disrupt and inhibit lipid peroxidation reactions (Niki, 2014). The alpha-tocopherol presents anti-inflammatory activity and modulates the expression of proteins invol-ved in cholesterol metabolism (Wallert et al., 2014).
The richest dietary sources of vitamin E are vegetable oils. Other important sources of this vitamin are whole grains and nuts (Traber, 2012). Triacylglycerols are the predominant components of vegetable fats and oils. Minor compounds include: mono-and diglycerides, free fatty acids, sterols, tocopherols, among others. Tocopherols play an important role for inhibi-ting the lipid oxidation in vegetable oils. The result of the autoxidation of fats and oils is the development of objectionable flavors and odors characte-ristic of the condition known as “oxidative rancidity” (Asif, 2011).
Vitamin E (α-tocopherol acetate) is a fat-soluble vitamin as well as an effective antioxidant that can protect cells from oxidative damage (Bautista-Ortega and Ruiz-Feria, 2010; Xiao et al., 2011). Numerous nutritional and physiological studies have shown that the vitamin E supplementation is beneficial for growth performance in various animal models (Colnago et al., 1984; Gao et al., 2010). Lin and Chang (2006) reported that the oral supple-mentation of vitamin E increased body weight gain in laying hens during the peak-laying period. Vitamin E supplementation of the diets effectively elevates pig meat quality such as stabilised meat colour (Buckley et al., 1995) and improved tenderness of muscles (Li et al., 2009) in meat animals. It is well known that vitamin E increase antioxidant capacity (El-Demer-dash, 2004) and react directly with reactive oxygen species (ROS) (Wissam
et al., 2000) to reduce lipid peroxidation damage in biological systems
(Packer and Landvik, 1990). Poultry diets are normally supplemented with vitamin E as a feed additive. However, vitamin E is easily oxidised in nature and loose its activity due to its chemical instability during processing, transportation and storage (Li et al., 2005). Other challenges associated with the application of vitamin E are poor water-solubility (Sagalowicz and Leser, 2010) and variation in its bio-availability (Yang and McClements, 2013) in the feed industry. These issues ensure the necessity of research for the development of new products to overcome these shortages of vitamin E.
The dietary requirement for vitamin E in poultry feed is highly variable and depends on the concentration and type of fat in the diet, the concent-ration of selenium, and the presence of pro-oxidants and antioxidants (NRC, 1994). Vitamin E is one of the essential nutrients in poultry feed, and its deficiency causes a wide variety of disorders in poultry species.
Fig. 1.3.1. Commonly known vitamin E deficiency diseases and most vulnerable tissues in poultry species (Rengaraj and Hong, 2015)
1.4. Antioxidants
Antioxidants are one of the essential components which improve food quality, especially if they reduce lipid oxidation. Tocopherols are natural phenolic antioxidants which play important role as lipid oxidation inhibitors in food and biological systems (Tasan and Demirici, 2005).
Antioxidants play a vital role against various diseases like cancer, cardiovascular diseases, inflammation, ageing process, rheumatoid arthritis, diabetes, as well as disease associated with cartilage and Alzheimer’s di-sease (Chauhan and Chauhan, 2006). Another field that is strongly affected by lipid peroxidation is the food sector, where the free radicals not only affect the quality of lipid in raw or processed food but also lead to loss of nutritional value. In the past decade search for natural antioxidant comp-ounds has gained considerable attention and a number of publications on antioxidants from natural sources appeared (Huang et al., 2004). Antioxi-dants from natural sources are preferred by consumers (Kranl et al., 2005), due to concern on the toxic and carcinogenesis effects of synthetic antioxi-dants (Ito et al., 1986; Safer and Al-Nughamish, 1999). Natural antioxiantioxi-dants have a low capacity to reduce oxidative reactions, but their price is relati-vely high (Haliwell et al., 1988). Therefore, there is a need to develop potent, cheaper and safer natural antioxidants.
Oxidation is one of the primary reasons for quality deterioration in meat products. During this process, the muscle haeminic pigment changes from red oxymyoglobin to brown metmyoglobin, turning the meat an undesirable
brownish colour (Jose et al., 2008). Moreover, lipid oxidation results in the production of free radicals, which are linked to the formation of off-flavours and odours, a reduction in polyunsaturated fatty acids and the production of undesirable compounds such as potentially toxic peroxides and aldehydes (Morrissey et al., 1994). All these modifications cause a decrease in the freshness of meat and lower consumer acceptance, resulting in an economic loss to the meat industry.
1.5. Antioxidant in eggs
Egg is considered a perfect natural food that has been consumed for centuries worldwide. Although egg contains all the indispensable nutrients for life, its consumption in several areas has reduces due to the public insight on its high cholesterol level (Nimalaratne et al., 2011). The antioxi-dant content of egg is almost nill and a fraction is found in egg yolk. So fortifying egg with antioxidant can enhance its quality which can be achie-ved by altering the ration in poultry feed (Carlsen et al., 2010). On the other hand, present data report that there was no full correlation between egg intake and plasma cholesterol concentrations as stated by Qureshi et al. (2007). Egg-yolk is a valuable source of many compounds significant for humans as well as animal wellbeing. It was well demonstrated that the diet supplied to laying hens has direct influence to yolk qualitative traits. By changes in diet, specific molecules having positive health effects can be incorporated in the egg-yolk (Kuhnle et al., 2008). To date, different investi-gations stated that active molecules in feed can be shifted from hens diet to egg-yolk. Among compounds, both lutein and zeaxanthin resulted the most widely studied molecules in eggs. These help to reduce macular degenera-tion related to the age either filtering harmful light blue and/or as antioxi-dants (Chung et al., 2004). Isoflavones from soybean was found to contain phytoestrogens having possible health effects and they were detected in yolks (Kuhnle et al., 2008). Phenolic compounds usually contained in cereals, fruit and many vegetables are significant antioxidants, suggesting to occupy a defensive function in the various disorders of chronic nature (Liu, 2007). All the natural phenolic compounds derived from a common biosyn-thetic pathway, incorporating precursors from the shikimate and/or the acetate-malonate pathways (Colombo et al., 2014). Up till now, few data is existing on the possible polyphenols in poultry eggs. Hens are well known to be able to ‘bioconvert’ health-related molecules from diet to egg. Both wheat and maize are valuable food/feed ingredients in diet containing anti-oxidants.
(a) General assumptions of the model based on observations of carotenoid distribution in
feed and different tissues.
(b) The routes through the nexus (black) and mechanisms that may be involved (red) based
on this study and previous reports.
Fig. 1.5.1. The nexus model, showing the fate of carotenoids provided to poultry as biofortified diets (Moreno et al., 2016)
Table 1.5.1. Natural sources in enriched eggs and role for human and animal well being
Herbs* Active ingredient
in herbs Benefits in relation to human health
Turmeric powder Flavonoids compounds
Antimicrobial, antioxidant Garlic, onion and their leaves Allicin, Allyl sulfide Reduce LDL cholesterol,
anticarcinogenic properties Sufar beer, grape pulp Betaine Decrease plasma homocysteine
which ruptures arterial walls Alfaalfa, marigold petals,
red pepper, spirulina
Carotenoid pigments Antioxidant, anticarcinogenic Basil leaves Eugenic acid, Eugenol Immunomodulatory properties Marigold petals, bay Lutein Antioxidants, improves vision Tomato pomace, grape pulp Lycopene Decrease LDL cholesterol,
antioxidant, anticarcinogenic Citrus pulp Nirangenin Reduce LDL cholesterol Flaxseed, canola, fish, oils,
insects, worms
n-3 PUFA Decrease LDL, hypertension, angina, atherosclerosis Seeds, legumes, weeds, yest,
fermented products
Phytosterols Increase HDL, decrease blood sugar
Fenugreek, spices Quercitin, Lutein, Citogenin
Induce insulin secretion, antimicrobial and tonic activity Brewery waste, yest,
fermented products
Statin Reduce LDL cholesterol Broccoli, cauliflower,
cabbage, radish leaves, waste
Sulphoraphane Anticarcinogenic and antioxidant properties
Bran Tocotrienols Decrease LDL cholesterol Milk, eggs, meat products Taurine Prevent atherosclerotic *Adapted from Narahari et al. (2004) and Singh et al. (2012)
1.6. Selenium – an antioxidant with strong Pro-Oxidant properties
Trace element deficiency in many countries is a common problem in farming animals, among which selenium (Se) deficiency is of major impor-tance. The Se deficiency in livestock species have been described to be involved in the different pathogenesis syndromes (Naziroglu et al., 2012; Zhou et al., 2013), myopathies (Alehagen and Aaseth, 2015) and fertility problems (Guyot et al., 2007).
Many trace elements, including those essential for life, such as sele-nium, are naturally accumulated by yeast (Kieliszek and Błażejak 2013;
Schrauzer 2006). Yeast cells are widely used in the production of food and fodder as well as in the biotechnology and pharmaceutical industries. In addition, they can be employed as a eukaryotic cell culture model in research (Brozmanová et al., 2010).
Selenium may be present in inorganic or organic form in the diet. The main dietary factors affecting availability of Se include methionine, thiols, heavy metals, and vitamin E. Se resorption from the gastrointestinal tract, its retention and metabolism in the body all depend on the quantity of Se intake and its chemical form.
Selenium is a member of a group of trace elements, which are essential to proper functioning of an organism. It is an integral part of selenoproteins and several antioxidant enzymes such as glutathione peroxidase (GPx), thioredoxin reductase (TRxR), and iodothyronine deiodinase (DIO), which protect cells from the harmful effects of free radicals that are generated during the oxidation process (Drutel et al. 2013).Oxidation-reduction reac-tions are critical for the maintenance of the physiological homeostasis both in unicellular and multicellular organisms. The term “oxidative stress” is defined as the perturbations of the physiological redox homeostasis when the rate of cellular reduction is overwhelmed by the rate of cellular oxide-tion (Halliwell. 2007). Both the enzymatic and non-enzymatic antioxidants can reverse this balance when the cellular damage is not beyond repair. In the context of selenium, the antioxidant properties are predominantly exer-ted by its incorporation into selenoproteins that can catalyze the reduction of disulfide bonds in proteins and peptides (Labunskyy et al., 2014; Maghadas-zadeh and Beggs, 2006). Such reductive properties are important for protec-ting against indiscriminate oxidation of intracellular and extracellular cons-tituents by intrinsic and extrinsic oxidants, spanning the spectrum of disease conditions to harmful chemicals. Therefore, deciphering the physiological roles of selenium as an antioxidant is one of the key research areas that have been investigated and reinvestigated ever since the discovery of selenium as an essential trace element by Schwarz and Foltz in 1957 (Schwartz and Foltz, 1957).
1.7. Health benefits of carotenoids
Several carotenoids commonly found in foods are thought to play a role in maintaining bodily functions and preventing disease. Beta-carotene, lycopene, lutein, and zeaxanthin are some of the most well known caro-tenoids considered to have health benefits. Each of these compounds and their proposed benefits are discussed below.
1.8. Beta-carotene
Beta-carotene (C40H56) (Fig. 1.9.1.) is one of the major carotenoids present in the diet (Johnson, 2002). It is found in a variety of orange, yellow, and green fruits and vegetables. Particularly good sources of beta-carotene include various greens (collard, turnip, spinach, lettuce), mangos, cantaloupe melons, peppers, pumpkin, carrots, and sweet potatoes (Holden et al., 1999). Beta-carotene, along with alpha-carotene and betacryptoxanthin, are sources of provitamin A. Once converted to vitamin A, health benefits derived from these compounds include maintenance of normal eye health, epithelial function, embryonic development, and immune system function (NAS, 2001). Epidemiological studies have found that diets high in beta-carotene or in fruits and vegetable intake are associated with decreased cancer risk (Vanpoppel and Goldbohm, 1995). These findings led to the belief that beta-carotene may help reduce the risk of lung cancer (Johnson, 2002). However, several human intervention studies in which beta-carotene supplements were used actually found that beta-carotene increased the risk for lung cancer in high risk populations (smokers and asbestos-exposed workers) (Omenn et
al.,1996). These results have led to questions as to whether these effects were
due to the high betacarotene dosage levels, the breakdown of beta-carotene into oxidation products, or if other components or a combination of compo-nents is responsible for the anticarcinogenic effects of fruit and vegetable consumption (Johnson, 2002; Palozza, 1998). In addition to the debated anti-cancer benefits of beta-carotene, this carotenoid has also been proposed to decrease risk factors for cardiovascular disease.
1.9. Lycopene
Lycopene (C40H56) is a hydrophobic, acyclic carotenoid containing eleven conjugated double bonds (Fig. 1.9.1) (Wilcox et al., 2003; Shi, 2000). Tomatoes, watermelon, guava, and grapefruit are the main sources of lycopene in the diet (Holden et al., 1999). In these raw plant products, approximately 95 per cent of lycopene is found in the all-trans form (Shi, 2000; Nguyen and Schwartz, 1999), and is located in the photosynthetic pigmentprotein complex of the thylakoid membrane (Shi and Le Maguer, 2000). In some thermally processed foods and in human serum and tissues, higher quantities of cis isomers are found (Nguyen and Schwartz, 1999; Clinton, 1998). The popularity of lycopene as a bioactive food component has stemmed from a number of epidemiological studies that have concluded that diets rich in high lycopene foods are associated with a reduced risk of several diseases. In some cases, cell culture and dietary intervention studies
have provided additional evidence that lycopene may play a positive role in health. Consumption of high lycopene foods has been proposed to reduce the risk of diseases including cardiovascular disease (Wilcox et al., 2003), and cancers of the prostate (Stahl and Sies, 1996; Hadley et al., 2002; Miller
et al., 1996), breast (Shi, 2000), cervix, colon, esophagus, skin, pancreas,
bladder, and stomach (Wilcox et al., 2003; Nguyen and Schwartz, 1999; Clinton, 1998). Of these conditions, the most evidence exists for lycopene and a reduced risk of prostate cancer.
1.10. Lutein and Zeaxanthin
Lutein and zeaxanthin (C40H56O2) (Fig. 1.9.1) are oxygenated carote-noids, making them members of the xanthophyll group of carotenoids (Alves-Rodrigues and Shao, 2004). Lutein and zeaxanthin are stereo isomers, which complicates analytical techniques for determining the quantities of each stereo isomer present, and likewise, has created difficulties in determining the influence of the individual isomers on human health (Stacewicz-Sapuntzakis
et al., 1993; Johnson, 2000). Particularly good sources of these carotenoids
are leafy greens like spinach, collard greens, kale, corn, persimmons, and broccoli (Holden et al., 1999). Commercially produced lutein is derived from marigolds and is used in the poultry industry to impart yellow color to the yolks of eggs and the skin of broilers (Sowbhagya et al., 2004). Some evidence suggests that lutein and zeaxanthin are associated with a reduced incidence of age-related macular degeneration and cataracts as determined by epidemiological and intervention studies (Alves-Rodrigues and Shao, 2004; Sowbhagya et al., 2004; Richer, 2004). The mechanism by which these carotenoids are thought to decrease macular degeneration and cataract formation, is by increasing the macular pigmentation of ocular tissues, which helps to filter damaging blue light, thus preventing oxidation damage that eventually leads to tissue damage (Alves-Rodrigues and Shao, 2004; String-ham and Hammond, 2005; Berendschot et al., 2000). However, other studies have not found a relationship between lutein and zeaxanthin and eye health (Mozaffarieh et al., 2003), indicating that more work is needed in this area of nutrition (Stringham and Hammond, 2005). While eye health is the predo-minant health benefit associated with lutein and zeaxanthin, beneficial effects of these carotenoids have been proposed for other health conditions as well. Lutein has been proposed to reduce risk factors for coronary heart disease and stroke, breast cancer, and improving skin health, although research in these areas is still limited (Alves-Rodrigues and Shao, 2004; Johnson, 2000).
Fig. 1.10.1. Lycopene from poultry products for human nutrition (Laudadio et al., 2015)
1.11. Lycopene in poultry products and its influence on human health
Some vitamins (A, D, E) and carotenoids are liposoluble compounds that are naturally present in the food, or are used as excipients in different industrial products, such as pharmaceuticals, cosmetics, or foods (Gonnet
et al., 2010). Carotenoids are among the most investigated lipophilic
bioactive compounds due to their numerous beneficial roles in human health because of their potential as prophylatic and therapeutic agents for the diseases such as hypertension, diabetes, cardiovascular diseases and some types of cancer (Fraser and Bramley, 2004; Ratnam et al., 2006). Carote-noids are also one of the most important groups of natural pigments because they are widely distributed in the plants (Yuan et al., 2008; Papaioannou and Liakopoulou-Kyriakides, 2010). There are several types of carotenes, but one of the most important is β-carotene due to its high provitamin A activity and antioxidant capacity (Hentschel et al., 2008; Cao-Hoang et al., 2011).
Lycopene is a natural dye manufactured by plants and microorganisms during the process of photosynthesis to protect them from the activity light
and increase light sensitivity (Rao, 2000; Rao and Rao, 2003; Rao, 2004) is found in vegetables and some types of fruit with red dye (such as pineapple, orange, tomatoes, grapefruit, strawberries and sweet peppers) and is tomato main source of his in the human diet, and quantity of existing depend on the type and maturity of tomato (Sies and Stahl, 1995; Stahl and Sies, 1996; Gerster, 1997; Rao and Agarwal, 1999; Markovic et al., 2006) which is a powerful antioxidant that provides protection against damage to the cells of the body due to free radicals, and dietary antioxidants such as carotenoids role in health and disease, which has increased the interest in them is largely to make sure the benefits of these compounds in the diet as is of great importance in the fight against the free radicals generated as a result of oxidative stress and protect cells from damage (Nierenberg et al., 1997; Leal
et al., 1999). There are more than 700 kinds of carotenoids have been
iden-tified, but only six forms of which are present in food and in the blood and tissues of the body and this carotenoids are α-and β-lycopene and β-crypto-xanthin, lutein, zeaβ-crypto-xanthin, carotene (Borel et al., 2007) and lycopene effectiveness is very high in the fight against diseases, a preventive measure against heart disease, cardiovascular and cancer of the prostate gland and the gastrointestinal tract, skin, pancreas, uterus, and there are many studies indicate the importance of lycopene to humans in health and disease (Ševčíková et al., 2008) also, a diet rich in tomato increases the levels of high-density lipoproteins high Density Lipoprotein and reduces the level of lipoproteins and low-lying density Low Density Lipoprotein (Napolitano
et al., 2007), either vitamin E is one of the most powerful antioxidants and
has played a major role in many vital functions within the body and use extensively as additives to animal feed to improve the performance and enhance the immune status and improve the quality of meat, eggs and increase vitamin E in animal products increases the content within the human body during eating these products (Sunder et al., 1997; Flachowsky, 2000) has pointed (Chan and Decker, 1994) the inability of poultry to manufacture vitamin E, therefore, should be affixed to the feed as one of the basic requirements fodder has been observed that added to the diets of birds improves growth and productive performance and improves the quality of the meat against oxidative deterioration as well as the role of the powerful antioxidant scavenging ability on free radicals (Skrivan et al., 2010; Guo
et al., 2001). Given the importance of antioxidants as additives fodder and
for its role in improving the qualities of productivity as the researcher found (Ševčíková et al., 2008) that lycopene is important in the fight against free radicals and this importance be useful for poultry as consisting of free radicals in the body of the chicken at higher temperatures and in cases of stress, when the rapid growth in cases of higher production and metabolism.
1.12. Demage of malondialdehydes (MDA)
Oxidation is a result of natural metabolic processes, but excessive formation of reactive species, such as free radicals, can damage important biomolecules (i.e., lipids, proteins, and nucleic acids) in the body of humans and animals alike. The rate of oxidation increases in result of the following: high intake of oxidized lipids and prooxidants; deterioration of sensitive polyunsaturated fatty acids (PUFA); and low intake of antioxidative nutrients (Morrissey et al., 1998; Smet et al., 2008). In muscle foods, oxidetive reactions continue postmortem and are a leading cause of quality deteriora-tion during processing and storage. With a relatively high propordeteriora-tion of PUFA, poultry meat is more susceptible to oxidative processes, specifically lipid oxidation, than beef or pork. Therefore, incorporation of dietary anti-oxidants, such as vitamin E and Se in poultry feed, has been supplemented to achieve optimal growth performance, reproduction, and meat quality.
However, oxidation (oxidative rancidity) is the main deterioration pro-cess in fats and oils and lipid based feed stuffs. The oxidation propro-cess is related to a decrease in nutritional value, feed palatability and broiler perfor-mance, and can also cause health problems (Baiao & Lara, 2005; Tavarez
et al., 2011).
The reactive oxygen species (ROS), are continuously formed as a result of normal metabolic processes; however, when not effectively and safely removed by an endogenous antioxidant system, they can oxidize and dama-ge cellular macromole-cules, possibly leading to oxidative stress (Kaden-bach et al., 2009). Oxidative stress is believed to play an important role in the regulation of the metabolic activity of some organs and productivity in farm animals. It may impair health both directly, by peroxidative damage to lipids and macromolecules, and indirectly by changes in cellular membranes or modifying some metabolic pathways, resulting in altered physiology and possibly pathology (Celi et al., 2014). Exogenous antioxidants are important because they have a twofold function: to prevent food oxidation, in particular lipid oxidation, and at the same time to increase the amount of antioxidant agents present in the organism, protecting against metabolic disorders.
One of the major causes of chemical deterioration of foods, especially those containing polyunsaturated fatty acids (PUFA), is lipid oxidation (Belitz
et al., 2009). As a result of this process, unsaturated fatty acids form
odour-less and tasteodour-less hydroperoxides, which are further decomposed to secondary oxidation products mainly aldehydes. Malondialdehyde (MDA) and hydroxy-lated a,bunsaturated aldehydes such as 4-Hydroxy-2-(E)-Nonenal (HNE) and
4-Hydroxy-2-(E)-Hexenal (HHE) are formed among other saturated or unsa-turated aldehydes. MDA is a three carbon dialdehyde with carbonyl groups at the C-1 and C-3 positions and is known to be produced from the decompo-sition of hydroperoxides derived from the oxidation of both n-3 and n-6 PUFAs (Frankel, 2005). HNE and HHE are unsaturated in the a,b position with a hydroxyl group attached on the forth carbon atom. HNE is formed during the oxidation of n-6 PUFA, while HHE is mainly related to the oxidation of n-3 PUFA (Long and Picklo, 2010; Surh et al., 2010; Han and Csallany, 2009; Pryor and Porter, 1990; Guillen and Uriarte, 2012).
MDA, HNE and HHE have attracted the attention in biological systems due to their potential toxicity to humans which is attributed to their high reactivity with proteins and DNA, consequently leading to structural dama-ge and alteration of their functionality (Esterbauer, 1982; Esterbauer and Cheeseman, 1990; Esterbauer et al., 1991; StAngelo, 1996; Uchida, 2003; Guillen and Goicoechea, 2008; Voulgaridou et al., 2011). More specifically, it has been confirmed that MDA can modify double-stranded DNA by formation of amino-imino-propene crosslinks between the NH2 groups of a guanosine base and the NH2 group of the complementary cytosine base (Esterbauer et al., 1991). Furthermore, it has been reported that MDA can react with NH2 containing amino acids and it is noteworthy that compared to amino acids, proteins have been found to be more readily modified by MDA in physiological conditions (Nair et al., 1986). Concerning hydroxyla-ted a,b-unsaturahydroxyla-ted aldehydes, similar involvement in protein and DNA modification has been reported (Uchida and Stadtman, 1992; Uchida, 2003; Wakita et al., 2011).
Feed additives are used in animal nutrition to improve feed quality and the performance and health of animals. The European Union introduced the complete ban of growth promoters for broilers in January 2006, allowing only 4 antibiotics that are not associated with human treatment (Hernández
et al., 2004). This has had significant consequences on the growth
perfor-mance of animals, especially for the poultry industry (Attia et al., 2011). The dietary strategies adopted to improve the nutritional value, oxide-tive stability and sensory properties of poultry products have been reviewed by Bou et al. (2009), and these include plant derived or phytogenic addi-tives.
These compounds, including polyphenols, are characterized by low-molecular-weight reactive oxygen species-scavenging activity (Pastorelli
et al., 2012), and are thought to be potential growth promoters in animal
diets (Hashemi and Davoodi, 2011).
Lipid oxidation is a main factor affecting the quality of the poultry meat, and oxidation results in characteristic off-flavors due to volatile
secondary lipid oxidation products (Bragagnolo et al., 2006). The mineral content of meat is highly linked to oxidative capacity of muscles, with redu-ced oxidative capacity being associated with reduredu-ced levels of iron, zinc, and selenium (Kelman et al., 2014). In this regard, particular interest has focused on selenium which is important for improving health and perfor-mance of the birds and improving meat quality for human consumption (Haugh et al., 2007; Ševčikova et al., 2006).
The products of lipid oxidation can decrease the nutrient content of the feed by reacting with proteins, lipids, and fat-soluble vitamins, which may even form toxic products that can adversely affect broiler performance and health.
Adding antioxidants to poultry feed is one of the most efficient strate-gies to protect the sensitive nutrients in feedstuffs. Some natural antioxi-dants and synthetic or commercial antioxidant blend (CAB) compounds for feed inclusion were recently studied and available in order to use in the poultry industry (Tavarez et al., 2011; Delles et al., 2014; Lu et al., 2014).
1.13. Iron absorption
Food fortification strategies involving the addition of micronutrients are designed to reduce deficiencies within defined populations. Identifying which food to fortify (known as the vehicle) is a multidisciplinary task, as there are many requirements of the candidate vehicle that need to be fulfilled (Allen et al., 2006).
Anemia affects 33% of the world population and accounts for 8.8% of global disability (Kassebaum et al., 2014). Although the etiology of anemia is multifactorial, iron deficiency (ID) is considered to be themost prevalent cause globally. Iron deficiency and low iron status are common all around the globe (WHO, 2008), and women of reproductive age are a vulnerable population because of their high iron requirements (Hallberg et al., 1995; Hallberg and Rassander-Hulten, 1991). Besides inadequate iron intake, low iron bioavailability is the predominant reason for iron deficiencies in populations subsisting on plant-based diets, independent of sex (Hoppe
et al., 2008). Strategies to increase the intake of foods rich in iron, as well as
dietary factors with enhancing effect on iron absorption, are therefore important.
Iron is an essential element in all living organism, which has several vital functions in the body, such as a major role as an oxygen carrier in blood hemoglobin and muscle myoglobin, a component of many enzymes,
and a required factor for a number of metabolic processes (Quintana et al., 2006)
A number of dietary factors affect the absorption of nonhaem iron. Intake of ascorbic acid and meat stimulates absorption, whereas calcium, polyphenols (e.g. in tea, coffee, vegetables) and phytates (e.g. whole grain cereals) inhibit absorption (Hallberg and Hulthen, 2000). Lactic acid-fermented foods may also improve the non-haem iron absorption. A number of single-meal studies with fermented vegetables and cereals have shown a significant increase in iron absorption in humans (Scheers et al., 2015; Brune et al., 1992; Gillooly et al., 1983). Lactic acidfermented foods can increase iron absorption in humans, possibly by lowering pH, activating phytases, producing organic acids or by the viable lactic acid bacteria.
The main advantage of chelates is the greater physical stability. The separation of trace elements and vitamins in feed oxidation is reduced and the absorption is increase (Stanacev et al., 2013). Iron is the most abundant metal found in the earth crust, in the water and as well as naturally in different food stuffs. The average adult human body contains 3 to 4 gram of iron. About 60 to 70 per cent of total iron is present as a part of haemo-globin in the red blood corpuscles (RBC) of our body. Circulating iron plays a vital role in the transportation of oxygen from the lungs to the various tissues in the body. The remaining 30 to 40 per cent of iron (1 to 1.5 g) is stored in liver, kidney, spleen and bone marrow (Park, 2011).
Iron deficiency is a major cause of anaemia, which concerns nearly one-quarter of the world’s population (McLean et al., 2009). Iron deficiency anaemia is associated with poor pregnancy outcome (Allen, 2000), increa-sed susceptibility to infection (De Silva et al., 2003) and decreaincrea-sed work capacity (Haas and Brownlie, 2001). The health consequences of iron defi-ciency without anaemia are more controversial, but there is evidence that this is associated with reduced work performance (Haas and Brownlie, 2001) and impaired cognitive function (Murray-Kolb and Beard, 2007).
Low iron intake and/or bioavailability is responsible for most anaemia in industrialized countries, but this accounts only for about half of anaemia cases in developing countries (Allen et al., 2006). The daily nutritional requirement of iron for adult males and females is given as 17 mg and 21 mg, respectively (ICMR, 2010). The higher level of iron needed for women is due to iron losses by regular menstrual blood.
Most studies investigating associations between dietary intake and iron status have focused on individual nutrients (e.g., iron) and foods (e.g., meat) (Heath et al., 2001; Pynaert et al., 2009; Harvey et al., 2005; Ramakrishnan
et al., 2002; Rangang, 1997; Galan et al., 1998; Asakura et al., 2009; Cade et al., 2005; Brussaard et al., 1997) which has several limitations. People do
not eat foods and nutrients alone but as meals consisting of a variety of foods and nutrients that may impact on one another (Newby and Tucker, 2004; Hu, 2002). By example, phytic acid (e.g., from whole grain breads and cereals) decreases absorption of nonhaem iron (e.g., from vegetables) (Hurrell et al., 2003). Thus, the analysis of dietary patterns can help to overcome this problem by considering the whole diet and describing how foods are consumed in combination. In recent years, empirically derived dietary patterns have been used to assess the association between dietary intake and anaemia (Shi et al., 2006) and dietary intake and iron status (Broderstad et al., 2010; Beck et al., 2013). The studies in a mouse model of iron overload showed that iron deposition enhances fatty acid oxidation and decrease glucose oxidation in skeletal muscle (Huang et al., 2011), whereas, comparable results could not be found in birds in the same study.
Fig. 1.13.1. Hormonal control of iron homeostasis by hepcidin. The major flows of iron into plasma and extracellular fluid are controlled by the effects of hepcidin on the iron exporter ferroportin (FPN). Hepcidin regulates the absorption of iron from the diet, including oral supplements. It also controls the release of stored iron in the liver and the release of
iron recycled from senescenterythrocytes, mostly in the spleen (Broderstad et al., 2010)
2. METHODICS
2.1. The experiments place, time and trial schemes
The experiments were carred out in 2012-2016. At LUHS VA Animal productivity laboratory by Animal raring technology institute and JST “Vilniaus paukstynas”.
Scientific research were carried out in accordance with the new version of the 2013-01-01 1997-11-06 Republic of Lithuania, animal care, storage and use of the law (Republic of Lithuania, animal welfare and protection of the law), (Valstybės žinios, 2012, Nr. 122-6126) and an executive act – Minister of State Food and Veterinary “Service of the order on animals used for experimental and other scientific research, storage, maintenance and operation of approval” (Valstybės žinios, 2009-01-22, Nr. 8-287). It is also according with 2010 of 22 September, European Parliament and Council Directive 2010/63/EU and EC recommendations 2007/526 EC Animal Use and storage for experimental and other purposes.
Activities which aid has been implemented work objectives shown in this work principal Figure 2.1.1.
2.2. Feeding trials with laying hens and chicken broilers 1 trial
The feeding trial was carred out with 60 laying hens of Lohman Brown cross at the age of 30 weeks. The laying hens were divided into 6 groups, 10 birds in each group. The I control group compound feed was supplemented sunflower oil 4.5% + 0.5 mg Na2SeO3 + 40 mg vit. E/kg, II control group was added rapeseed oil 4.5% + 0.5 mg Na2SeO3 + 40 mg vit. E/kg, III cont-rol group was added linseed oil 4.5% + 0.5 mg Na2SeO3 + 40 mg vit. E/kg, I experimental group was added sunflower oil 4.5% + 0.5 mg Alkosel®R397 + 40 mg vit. E/kg, II experimental group was added rapeseed oil 4.5% + 0.5 mg Alkosel®R397 + 40 mg vit. E/kg and the last one III experimental group which was added linseed oil + 0.5 mg Alkosel®R397 + 40 mg vit. E/kg.
Table 2.2.1. Usage of different oils, selenium and vitamin E on laying hens feeding
Parameter
Groups
Control Experimental I II III I II III
Standart compound feed+ Sunflower oil +
0.5 mg Na2SeO3 + 40 mg vit. E/kg + – – – – –
Standart compound feed + Rapeseed oil +
0.5 mg Na2SeO3 + 40 mg vit. E/kg – + – – – –
Standart compound feed + Linseed oil +
0.5 mg Na2SeO3 + 40 mg vit. E/kg – – + – – –
Standart compound feed + Sunflower oil +
Alkosel®R397 0.5 mg + vit. E (40 mg/kg) – – – + – –
Standart compound feed + Rapeseed oil +
0.5 mg Alkosel®R397 + vit. E (40 mg/kg) – – – – + –
Standart compound feed + Linseed oil +
0.5 mg Alkosel®R397 + vit. E (40 mg/kg) – – – – – + During the feeding trial, the laying hens were held in the individual cages (0.36 m × 0.44 m) with stationary drinking-bowl and feed box under the same feeding and holding conditions. The birds were fed with compound feed 125 g per day and matched Lohman brown cross hens recommendations (www.isapoultry.com).
2 trial
Table 2.2.2. Usage of different oils, lycopene and antioxidants on laying hens feeding
Parameter
Groups
Control Experimental
I II III I II III IV V VI
Standart compound feed +
sunflower oil (4.5%) + – – – – – – – – Standart compound feed +
rapeseed oil (4.5%) – + – – – – – – – Standart compound feed +
linseed oil (4.5%) – – + – – – – – –
Standart compound feed + sunflower oil (4%) +
lycopene (25 g/kg feed) – – – + – – – – – Standart compound feed +
rapeseed oil (4%) + lycopene
(25 g/kg feed) – – – – + – – – –
Standart compound feed + linseed oil (4%) + lycopene
(25 g/kg feed) – – – – – + – – –
Standart compound feed + sunflower oil (4.5%) +
antioxidant (0.15 g/kg feed) – – – – – – + – – Standart compound feed +
rapeseed oil (4.5%) +
antioxidant (0.15 g/kg feed) – – – – – – – + – Standart compound feed +
linseed oil (4.5%) +
antioxidant (0.15 g/kg feed) – – – – – – – – +
The feeding trial was carred out with 63 laying hens of Lohman Brown cross at the age of 30 weeks. The laying hens were divided into 9 groups, 7 birds in each group. The I control group compound feed was supple-mented sunflower oil 4.5%, II control group was added rapeseed oil 4.5%, III control group was added linseed oil 4.5%, I experimental group was added sunflower oil 4.5% + lycopene 25 g/kg, II experimental group was added rapeseed oil 4.5% + lycopene 25 g/kg, III experimental group which was added linseed oil + lycopene 25 g/kg, IV experimental group compound feed was supplemented 4.5% sunflower oil + 0.15 g/kg antioxidant, V expe-rimental group was added 4.5% rapeseed oil + 0.15 g/kg antioxidant and the
