Lamiaa Fathy Asal

3. BIOLOGICAL ROLE OF NATURAL ANTIOXIDANT FOR DECREASING THE DAMAGE IMPACT OF FREE RADICALS UNDER AGING AND HEAT STRESS CONDITION IN RABBITS.

4. LamiaaFathyAsal Animal and Fish Production Department, Faculty of Agriculture, Saba Basha ALEXANDRIA UNIVERESITY

5. Advisor's Committee Prof. Dr. Hassan Saber Zeweil (Supervisor) Professor of Poultry Nutrition, Animal and Fish Production Department, Faculty of Agriculture (Saba Basha), ALEXANDRIA UNIVERESITY • Prof. Dr. Mohamed Hassan Ahmed • Professor of Animal Nutrition, Animal and Fish Production Department, • Faculty of Agriculture (Saba Basha), • ALEXANDRIA UNIVERESITY • Prof. Dr. Mokhtar Ibrahim Yousef • Professor of Environmental Animal Physiology and Reproductive Toxicology, Department of Environmental Studies. Dean of Institute of Graduate Studies and Research (IGSR) Alexandria University, Egypt Prof. Dr. Kamel Ibrahim Kamel Chairman of Research, Animal Physiology, Animal Production Res. Institute, Agric. Research Center, Ministry of Agriculture

6. Examiners Committee

7. Introduction

8. What are Free Radicals? Free radicals are molecules that contain one or more unpaired electrons and are, consequently, very reactive, particularly with respect to lipids. Free radicals damage healthy cells and in turn impair the body’s normal functions.

9. Superoxide anion Hydrogen peroxide Hydroxyl radical Organic hydroperoxide Alkoxy radical Hydroperoxy radical O2 H2O2 _ . OH. ROOH RO. ROO. Reactive Oxygen Species

10. How are Free Radicals Produced? Aging causes oxidative stress (Liu, 2002) and decreases in levels of antioxidants and antioxidant enzymes in the heart (Somaniet al., 1995) and blood (Ji, 1993) and accumulation of oxidative damage to deoxyribonucleic acid (DNA), lipids, and protein (Beckman and Ames 1998).

11. Hot weather Cold weather High humidity STRESS environmental stress, including high temperature, causes oxidative stress with increased production of MDA and decreased concentrations of antioxidant vitamins and minerals under stress conditions (Sahin et al., 2002a, b).

12. Oxidative Stress = Imbalance between pro-oxidants (free radicals, reactive oxygen species) and anti-oxidants

13. Sperm function is undisturbed when the levels of ROS and antioxidants are balanced, as this ensures that no significant damage will occur. In the case of metabolic oxidative stress, however, provoked by excessive ROS production or low antioxidant status or both, impaired sperm function may occur (Agarwal et al., 2003; Aitken and Baker, 2006; Makker et al., 2009). In addition, DNA fragmentation in both nuclear and mitochondrial genomes is a consequence of oxidative stress (Agarwal et al., 2003; Aitken and Baker, 2006).

14. MEASUREMENT OF FREE RADICALS When a fatty acid is peroxidized it is broken down into aldehydes, which are excreted. Aldehydes such as thiobarbituric acid reacting substances (TBARS) have been widely accepted as a general marker of free radical production. The most commonly measured TBARS is malondialdehyde (MDA) .

15. ANTIOXIDANT DEFENSES Antioxidant means "against oxidation." Antioxidants work to protect lipids from peroxidation by radicals. Antioxidants are effective because they are willing to give up their own electrons to free radicals. When a free radical gains the electron from an antioxidant it no longer needs to attack the cell and the chain reaction of oxidation is broken. After donating an electron an antioxidant becomes a free radical by definition. Antioxidants in this state are not harmful because they have the ability to accommodate the change in electrons without becoming reactive.

16. TOMATOES • Dried tomatoes present a special interest, because some bioactive antioxidants are found in their composition at concentrated state. Lycopene has the highest value, with antioxidant and curative specific features on some diseases (George et al., 2004). In addition, tomato and tomato products are excellent source of vitamins A, C, and E. They also contain many other bioactive compounds, such as carotenoids (α-r , β-, γ-carotene, and lutein), and othecarotenes including phytoene and phytoflune. Moreover, tomato and tomato products also contain flavonoids. Many of these nutrients and phytochemicals have antioxidant properties and, in combination with lycopene, may contribute to the protection against peroxidation( Alshatwi et al.,2010) .

17. Lycopene Lycopene Content of common fruits and vegetables Lycopene, the pigment of red tomatoes, determines the biological and curative value of dried tomatoes. It is a carotenoid belonging to the same group of ß-carotene and gives the red colour to tomatoes, rosé grapefruits, apricots, red oranges and water melons. This compound is not only a pigment but also a strong antioxidant, which neutralizes the free radicals, and, especially, the oxygen derived ones. Its ability to inhibit the oxidative activity of the active oxygen is twice higher than in case of ß-carotene and 10 times higher than in case of α- tocopherol (Shi et al., 2000). Source :Rao et al. (2006)

18. Objective of the study The present work aimed to study the effect of dietary supplementation with 1% tomato powder on semen quality and blood and seminal plasma biochemical and antioxidant and digestibility coefficients of young and old adults V- line rabbit bucks during winter and summer seasons.

19. Materials and methods

20. EXPERIMENTAL design V-line rabbit bucks SUMMER 20 bucks WINTER 20 bucks Without TP (NTP) With TP (TP) Without TP (NTP) With TP (TP) • 7-9 Months • 22-24 Months • 7-9 Months • 22-24 Months • 7-9 Months • 22-24 Months • 7-9 Months • 22-24 months

21. Studied parameters • Changes in body weight, feed,water intake and rectal temperature • Body weight (BW) • Feed intake (FI) • Water intake (WI) • Rectal temperature (RI) • Reproductive performance • Ejaculate volume (EV) • Initial hydrogen (PH) • Sperm motility (SM) • Packed sperm volume (PSV) • Sperm concentration (CON) • Total sperm output (TSO) • Total motile sperm (TMS) • Total functional sperm fraction (TFSF) • Live sperm • abnormal sperm • Initial Fructose • Seminal plasma analysis • Alanineaminotransferase (ALT) • Aspartateaminotransferase (AST) • Alkaline phosphatase (ALP) • Acidephosphatase (ACP) • Lactate dehydrogenase (LDH) • Thiobarbaturic reactive substances (TBARS) • Superoxide dismutase (SOD) • Glutathion S-transferase (GST) • Blood plasma analysis • Alanineaminotransferase (ALT) • Aspartateaminotransferase (AST) • Alkaline phosphatase (ALP) • Acidephosphatase (ACP) • Lactate dehydrogenase (LDH) • Total protein • Albumin (Alb) • Globulin (Glb) • Glucose • Total lipid (TL) • Triglceride (TG) • Urea • Thiobarbaturic reactive substances (TBARS) • Superoxide dismutase (SOD) • Glutathion S-transferase (GST) • Testosteron

22. Studied parameters • Apparent digestibility coefficients of nutrients • Dry matter (DM) • Organic matter (OM) • Crude protein (CP) • Crude fiber (CF) • Ether extracts (EE) • Nitrogen free extract (NFE)

23. Results

24. Table 1:Overall means (Mean ±SE) and the interaction of live body weight (BW), feed intake (FI), water intake (WI) and rectal temperature(RT) of young and old V-line bucks rabbit as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

25. Table 1:continued

26. Table 2:Overall means (Mean ±SE) and the interaction of ejaculate value (EV), initial hydrogen ion concentration (pH), sperm motility (SM) and packed sperm volume (PSV) of young and old V-line bucks rabbit as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

27. Table 2:continued

28. Table (3): Overall means (Mean ±SE) and the interaction of sperm concentration (CON), total sperm output (TSO) total motile sperm (TMS) and the total functional sperm fraction (TFSF) of young and old V-line bucks rabbit as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

29. Table 3:continued

30. Table (4): Overall means (Mean ±SE) and the interaction of live and abnormal sperm and initial fructose of young and old V-line rabbit bucks as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column not sharing similar superscripts are significantly different (P<0.05).

31. Table 4:continued

32. Table (5): Overall means (Mean ±SE) and the interaction of alanineaminotransferase (ALT), aspartateaminotransferase (AST) and alkaline phosphatase (ALP) of young and old V-line bucks rabbit as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

33. Table 5:continued

34. Table (6 ): Overall means (Mean ±SE) and the interaction of seminal plasma acid phosphatase (ACP) and lactic dihydrogenase (LDH) of young and old V-line bucks rabbit as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

35. Table 6:continued

36. Table (7): Overall means (Mean ±SE) and the interaction of seminal plasma thiobarbituric acid-reactive substances (TBARS), superoxide dismutase (SOD) and Glutathione S-transferase (GST) of young and old V-line bucks rabbit as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

37. Table 7:continued

38. Table 8: Overall means (Mean ±SE) and the interaction of blood plasma total protein, albumin (Alb) globulin (Glb) and glucose of young and old V-line rabbit bucks as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

39. Table 8:continued

40. Table 9: Overall means (Mean ±SE) and the interaction for blood plasma enzymes as affected by tomato powder dietary supplementation of aging male rabbit aged during winter and summer condition. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

41. Table 9:continued

42. Table 10: Overall means (Mean ±SE) and the interaction of blood plasma total lipids (TL), triglyceride (TG), urea and lactic dihydrogenase (LDH) for young and old VL rabbit bucks as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

43. Table 10:continued

44. Table 11: Overall means (Mean±SE) and the interaction for blood plasma thiobarbituric acid-reactive substances (TBARS), superoxide dismutase (SOD) and Glutathione S-transferase (GST) for young and old VL rabbit bucks as affected by tomato powder dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

45. Table 11:continued

46. Table 12: Overall means (Mean ±SE) and the interaction for blood plasma testesterone as affected by tomato powder dietary supplementation of aging male rabbit aged during winter and summer condition. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).

47. Table 12:continued

48. Table 13: Overall means (Mean ±SE) and the interaction for digestibility coefficients of dry matter (DM), organic matter (OM), crude protein (CP) and crude fiber (CF)of young (Y) and old (O) V-line rabbit bucks as affected by tomato powder (TP) dietary supplementation during winter and summer seasons. a,b Means within a column not sharing similar superscripts are significantly different (P<0.05).

49. Table 13:continued

50. Table 14: Overall means (Mean ±SE) and the interaction for digestibility coefficients of ether extract (EE) and nitrogen-free extract (NFE) of young (Y) and old (O) V-line rabbit bucks as affected by tomato powder (TP) dietary supplementation during winter and summer seasons. a,b Means within a column for each factor not sharing similar superscripts are significantly different (P<0.05).