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Fırat Üniversitesi Sağlık Bilimleri Veteriner Dergisi
2009, Cilt 23, Sayı 1, Sayfa(lar) 015-019
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Yüksek Enerjili Diyetler ve Soğuk Çevre Şartlarında (15 ºC) Yetiştirilen Broylerlerde Selenyum ve Vitamin C Katkısının Lipit Peroksidasyon Üzerine Etkileri
Pınar TATLI SEVEN1, İsmail SEVEN2, Seval YILMAZ3, Bestami DALKILIÇ1
1Fırat Üniversitesi, Veteriner Fakültesi, Hayvan Besleme ve Beslenme Hastalıkları Anabilim Dalı, Elazığ, TÜRKİYE
22 Fırat Üniversitesi, Sivrice Meslek Yüksekokulu, Elazığ, TÜRKİYE
33 Fırat Üniversitesi, Veteriner Fakültesi, Biyokimya Anabilim Dalı, Elazığ, TÜRKİYE
Anahtar Kelimeler: Yüksek enerji, selenyum, vitamin C, antioksidan enzimler, kan parametreleri, soğuk şartlar, broyler
Özet
120 broyler (1 günlük), her biri 30 hayvan içeren 1 kontrol ve 3 deneme grubuna ayrıldı. Deneme grupları şöyledir; temel diyet yedirilen Grup I (kontrol), yüksek enerji diyeti yedirilen (başlangıç diyeti olarak 3250 kcal/ kg; büyütme diyeti olarak 3300 kcal/kg) Grup II, yüksek enerjili diyete sodyum selenit olarak 1 mg/kg Se katılan Grup III, yüksek enerjili diyete askorbik asit olarak 250 mg /kg Vitamin C katılan Grup IV olarak belirlendi. Plazma triiodotiroidin ve trigliserit düzeyleri selenyum ve vitamin C katılan gruplarla karşılaştırıldığında grup II'de önemli oranda arttı. Karaciğer ve abdominal yağ malandialdehit aktiviteleri kontrol ve grup II'de daha yüksek (P<0.01) bulundu. En yüksek karaciğer katalaz (CAT) düzeyi kontrol ve grup II de bulundu. Abdominal yağın CAT düzeyi kontrol grubunda grup III' den önemli oranda daha yüksekti. Karaciğer, abdominal yağ ve kalp redükte glutatyon düzeyleri istatistiksel olarak önemli bulunmadı (P>0.05). Sonuçlar gösterdi ki soğuğa maruz kalma ve yüksek enerjili diyetler dokularda oksidatif hasarı indükledi, fakat bu hasar diyete selenyum ve vitamin C katkısıyla kısmen azaldı.
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    Stress factors reduce the production along with increase the death rate in poultry1,2. Cold conditioning, a physical environmental stressor, has been shown to have variable modulator effects on cells of the immune system in animals3. Despite the general awareness that energy demands are increased by cold and that the magnitude of those demands is moderated by total body insulation, few quantitative data exist relating environment, nutrient need, and productive efficiency4. Low ambient temperature causes to increased feed intake and decreased performance in poultry5,6. A study indicated that animals acclimated to cold could be able to achieve a higher maximum sustained energy intake rate than non acclimated ones7. Besides, the negative effects of cold conditioning might be lowered by antioxidants such as dietary selenium (Se) and vitamin C (Vit C )8,9.

    Se is a type of trace mineral which supports healthy activity within your immune system, functions as an important part of the potent antioxidant glutathione. Se supplementation could be of great interest in protecting cells against oxidative stress10. Vit C has been supplemented to diets of poultry reared under stress. It is known that Vit C are used for reducing the negative effects of environmental stress in the diets of poultry because of the reported benefits of Vit C supplementation on poultry reared under heat or cold stress3,11.

    This study was planned to determine the effects of Se and Vit C supplemented on high energy diet and low enviromental temperature on blood parameters and antioxidant enzymes in broilers.

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    In this study, 120 broiler chicks (Ross 308) at oneday- old were used. Chicks were randomized into 3 experimental and a control group containing 30 birds in each. Treatment groups were comprised of 3 replicates of 10 birds. Corn and soybean meal-based diets were formulated according to the requirements of the National Research Council12. Diets were formulated as starter and finisher (Table 1). The experimental groups were as follow; Group I (control) was fed with basal diet, Group II was fed with high energy diet (3250 kcal/kg as starter diet; 3300 kcal/kg as finisher diet), Group III was fed with high energy diet supplemented with 1mg/kg Se as sodium selenite, Group IV was fed with high energy diet supplemented with 250 mg/kg Vit C as ascorbic acid. The birds were fed with a starter diet until 28 d of age, then fed with finisher diet until 40 d. Diets and water were offered ad libitum. Chicks were warm-room reared at 33.20±2.50 ºC during the first week and 26.20±1.85 ºC for the second week. Starting at day 14 and continuing through week 6, birds were cold-stressed at an average room temperature of 15.10 ±1.98 ºC. On days 40, 10 birds from each group were killed by cervical dislocation. Blood samples were collected from brachial vein. Liver, abdominal fat and heart tissue samples were taken immediately.


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    Table 1: Composition of the experimental diets, %

    The basal diet and high energy diet was analysed for Se13. Triiodothyronine (T3) concentration was determined using commercially available radioimmunoassay kit (Byk-Sangtec Diagnostica, Dietzenbach-Germany; Immulite 2000, DPC, LA). Plasma biochemical parameters were measured using an auto analyzer (Olympus AU 600, Japan).

    Plasma14 and tissue15 MDA concentration was measured. Catalase (CAT) level was estimated by measuring the breakdown of H2O2 at 240 nm16. Tissue reduced glutation (GSH) concentration was measured by using the dithionitrobenzoic acid recycling17. Hemoglobin concentration and tissue protein contents were determined according to Drabkin and Austin18 and Lowry et al.19, respectively.

    Data collected were subjected to analysis of variance, and where significant differences were observed, means were further subjected to Duncan's multiple range test20. The results were considered as significant when p values were less than 0.05.

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    According to our results, T3 level in group supplemented with Se was the lowest among groups (P<0.01) (Table 2). Tona et al.21 reported that T3 hormone levels in standart broiler breeder lines were 3.84, 2.80 and 1.55 nmol/l at 14, 28 and 41 days, respectively. T3 level (1.72 nmol/l) in group supplemented Se at 40 days was in an aggrement with Tona et al.21. Significant reduction in T3 levels of Se and Vit C groups in compared to control and high energy groups may be result from antioxidant effects of Se and Vit C22,11.


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    Table 2: Plasma triiodothyronine hormone (T3) levels and some biochemical parameters of in the study groups (n=10)

    In our study, we found that plasma glucose, total protein, albumin and total cholesterol levels were not different among groups. However, plasma triglyceride in group supplemented with Se were significantly found higher than control and high energy group. A study indicated that the acute cold stress caused an increase in plasma triglyceride because of lipid peroxidation23. The control and high energy group without Se and Vit C supplement may be increased lipid peroxidation, and this following, lipid peroxidation may be increased plasma triglyceride level23. Cold conditioning induced a significant increase in MDA levels in liver and abdominal fat, thus, suggesting cold stress during this treatment (Table 3). Besides, in liver and abdominal fat, MDA level of the high energy group in compared to that of control group was similar, while its MDA level was increased in compared to those of Se and Vit C supplement groups. Alptekin et al.24 showed that stress induces a significant increase in lipid peroxide levels in the liver and small intestine. McArdle and Jackson25 have demonstrated a significant increase in free radical production together with an increase in the expression of antioxidant enzymes during a period of non-damaging exercise. These increases in antioxidant enzyme activities have been considered as a protective response against oxidative stress26. Hydrogen peroxide, a precursor of more potent radical species, is scavenged at higher concentrations by CAT. In the present study, the CAT activity was increased significantly in liver and abdominal fat of broilers exposed to cold (Table 3), while it was similar in the heart. Kaushic and Kaur et al.27 reported that the activities of antioxidant enzymes such as CAT and glutation peroxidase may change according to different tissues. They reported that the CAT activity was increased significantly in the kidney of stressed animals, while it was decreased in the heart, liver and small intestine. In the liver and abdominal fat, hydrogen peroxide produced in response to stress is taken care of primarily by CAT, which is shown to be present in higher concentrations in these tissues. The decreased activities of CAT in the livers of groups supplemented with Se and Vit C and in the abdominal fat of group supplemented with Se indicate the highly reduced capacity to hydrogen peroxide produced in these tissues in response to cold conditioning28. Factors such as housing in low temperature, rapid growth rates, high energy rations are known to influence the occurence of ascites in broilers29. Previous studies reported that cold temperature is one of the most effective factors ascites. In this study, control and high energy group may have induced to ascites29. The MDA levels of liver and abdominal fat, and CAT activities of liver in control group and high energy group without supplement significantly increased (P<0.05). This may be due to rising tend to ascites in these groups. Likewise, the previous study reported that lipid peroxidation were elevated in birds with ascites8. Moreover, it was found that the MDA and CAT activities of groups supplemented with Se (group III) and Vit C (group IV) decreased. In this study, especially, Se supplemention on account of decreasing of lipid peroxidation found the more effective than Vit C supplemention as agreement with study of Stanley et al.28. Because, Se levels in diet could be also be critical since Se has long been recognised to have antioxidant proporties due to its importance for glutathione peroxidase activity9. In this study, GSH activities of liver, abdomial fat, lung and heart were not found statistically significant between all groups (Table 4). In a previous study indicated that GSH activities may be not significant statistically as related to factors caused and exposed period to oxidatif damage11.


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    Table 3: MDA (nmol/mg protein) and CAT levels (k/g protein) of liver and abdominal fat in groups (n=10) (mean ± SE)


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    Table 4: Reduced glutation (GSH) levels (k/g protein) of some tissues in the groups (n=10) (mean ± SE)

    It was found similar effects on lipit peroxidation and antioxidant enzymes of control and diet with high energy. Lipid peroxidation and antioxidant enzyme activities (CAT) were significantly affected negatively to diets of control and group II in broilers exposed to cold conditioning. The supplementation to diet of Se and Vit C decreased significant these negative effects.

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    2) Cerci IH, Tatli P, Azman MA, et al. The effect of restricted feed on feed intake, egg production and feed conversion in pullets. Indian Veterinary J 2003; 80: 1153-1157.

    3) Tatli Seven P, Seven I. Effects of triiodothyronine hormone and iodine supplements on the performance of broilers in cold environment (15 oC). Indian Veterinary Journal 2008; in press.

    4) National Research Council. Subcomitee on Environmental stress, ISBN: 0-309-03181. http://www.nap.edu/catalog/4963.html. 1981. 16.09.2008

    5) Spinu M, Degen AA. Effect of cold stress on performance and immune responses of stress and white leghorn hens. British Poultry Science 1993; 34: 177-181.

    6) Stillman MJ, Shukitt-Hale B, Levy A, et al. Spatial memory under acute cold and restraint stress. Physiologic Behavior 1998; 64: 605-609.

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    8) Enkvetchakul B. Antioxidants, lipid peroxides, and pathophysiology of male broiler chickens with ascites. Ph.D. Thesis, University of Arkansas, Fayetteville AR, 1994.

    9) Bottje W, Enkvetchakul B, Moore R, et al. Effect of alphatocopherol on antioxidants, lipid peroxidation, and the incidence of pulmonary hypertension syndrome (ascites) in broilers. Poultry Sci 1995; 74: 1356-1369.

    10) Benefit of Glutathione, Selenium: Activates Antioxidant Glutathione for Protection of cell. http://benefitsofglutathione.com/?p=22 16.09.2008

    11) Tatli Seven P, Yilmaz S, Seven I, et al. Responses of broilers under cold conditioning (15 oC) to dietary triiodothyronine and iodine combined to antioxidants (Se and vitamin C). Indian Veterinary Journal 2008; in press.

    12) National Research Council. Nutrient requirements of poultry. 9th Rev., Washington DC: National Academy Pres, 1994.

    13) Brown MW, Watkinson JH. An automated fluorimetric method stress determination of nanogram quantities of selenium. Analytica Chimica Acta 1977; 89: 29-35.

    14) Satoh K. Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clin Chim Acta 1978; 90: 37-43.

    15) Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979; 95: 351.

    16) Aebi H. Catalase. Methods of Enzymatic Analysis. 2nd Edition, Wenheim: Verlag Chemie 1974: 673-678.

    17) Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys 1959; 82: 70-77.

    18) Drabkin DL, Austin JM. Spectrophotometric studies, spectrophotometric constants for common haemoglobin derivatives in human, dog and rabbit blood. Journal of Biological Chemistry 1932; 98: 719-733.

    19) Lowry OH, Rosebrough NJ, Farr AL, et al. Protein measurements with the folin phenol reagent. J Biol Chem 1951; 193: 265-275.

    20) Spss for Windows. Relased 11.5 version, Spss Inc., USA, 2002.

    21) Tona K, Onagbesan OM, Bruggeman V, et al. Comparison of feed intake, blood metabolic parameters, body and organ weights of growing broilers originating from dwarf and standard broiler breeder lines. International Journal of Poultry Science 2004; 3: 422-426.

    22) Gerloff BJ. Effect of Se supplementation on dairy cattle. Journal Animal Science 1992; 70: 3934-3940.

    23) Martinello F, Soares SM, Franco JJ, et al. Hypolipemic and antioxidant activities from Tamarindus indica L. Pulp fruit extract in hypercholesterolemic hamsters. Food and Chemical Toxicology 2006; 44: 810-818.

    24) Alptekin N, Seçkin F, Yelkenci N, et al. Lipid peroxides, glutathione, γ-glutamylcysteine synthetase and γ- glutamyltranspeptidase activities in several tissues of rats following water-immersion stress. Pharmacol. Res 1996; 34: 167-169.

    25) McArdle A, Jackson MJ. Exercise, oxidative stress and ageing. Journal of Anatomy 2000; 197: 539-541.

    26) Altan O, Pabuccuoglu A, Altan A, ve ark. Effect of heat stress on oxidative stress, lipid peroxidation and some stress parameters in broilers. British Poultry Science 2003; 44: 545-550.

    27) Kaushic S, Kaur J. Chronic cold exposure affects the antioxidant defense system in various rat tissues. Clinica Chimica Acta 2003; 333: 69-77.

    28) Stanley G, Chukwu H, Thompson D, et al. Singly and combined effects of organic selenium (Se-yeast) and vitamin E on ascites reduction in broilers. Association of Research Directors Eleventh Biennial Research Symposium. (1997).

    29) Bolukbasi SC, Aktas MS, Guzel M. The effect of feed regimen on ascites induced by cold temperatures and growth performance in male broilers. International Journal of Poultry Science 2005; 4: 326-329.

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