BW: Body weight. CP : Crude protein ...... effective in terms of husbandry parameters and nutritional condition. 2.3.10. .... animals, air and water currents, and which can grow in various ..... C and stored in plastic bags which were kept in a refrigerator at. (-2 o .... tube with EDTA solution for measuring hemoglobin (Hb), and.
USING OF SPICES AS FEEDING STIMULANTS IN DIETS OF TILAPIA FISH BY Mohammed Abd Alla Eleryan Yousef B.Sc. Agricultural Science, Animal Production (Fish Production), El Azhar University, 2005
Thesis Submitted in Partial Fulfillment of the Requirements For the Degree of Master of Science
In
Agricultural Sciences - Animal production (Fish Nutrition) To Department of Animal Production Faculty of Agriculture Benha University
2012
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APPROVAL SHEET
USING OF SPICES AS FEEDING STIMULANTS IN DIETS OF TILAPIA FISH BY
Mohammed Abd Alla Eleryan Yousef B.Sc. Agricultural Science, Animal Production (Fish Production), El Azhar University, 2005 Thesis Submitted in Partial Fulfillment of the Requirements For the Degree of Master of Science This Thesis for M.Sc Degree has been approved by: Prof. Dr. Mohammed Khairy Ibrahim ………… Professor of Animal Breeding, Faculty of Agriculture, Moshtohor, Benha University Prof. Dr. Abdel-karim I. M. EL-Sayed ………… Professor of Animal Physiology, Faculty of Agriculture, Moshtohor, Benha University Prof. Dr. Deyab M. S. El-Saidy ………… Professor of Fish Nutrition, Faculty of Agriculture, Sheben El-Koum, Menofiya University Prof. Dr. Gaafar M. El-Gendi ………… Pofessor of poultry management,, Faculty of Agriculture, Moshtohor, Benha University Prof. Dr. Magdy A. Soltan (Supervisor) ……….. Professor of Fish Nutrition, Faculty of Agriculture, Moshtohor, Benha University Commttee in charge Date:
/
/ 2012
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SUPERVISION SHEET
USING OF SPICES AS FEEDING STIMULANTS IN DIETS OF TILAPIA FISH BY
Mohammed Abd Alla Eleryan Yousef B.Sc. Agricultural Science, Animal Production (Fish Production), El Azhar University, 2005 Thesis Submitted in Partial Fulfillment of the Requirements For the Degree of Master of Science
Under the supervision of : Prof. Dr. Mohammed Khairy Ibrahim ………… Professor of Animal Breeding, Faculty of Agriculture, Moshtohor, Benha University Prof. Dr. Abdel-karim I. M. EL-Sayed ………… Professor of Animal Physiology, Faculty of Agriculture, Moshtohor, Benha University Prof. Dr. Magdy A. Soltan (Supervisor) ……….. Professor of Fish Nutrition, Faculty of Agriculture, Moshtohor, Benha University
Dr. Hossam Mahmoud Agouz Abdel-ghany
………...
Assistant professor of Aquaculture, Central Laboratory for Aquaculture Research Abassa, Agriculture Research Center
2012
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CONTENTS 1.INTRODUCTION…………………………………………………….. 2.REVIEW OF LITERATURE ……………………………..………… 2.1. Spieces as a feed additive ............................................................ 2.2. Garlic as a feed additive in aquaculture ...................................... 2.3. Probiotic ……………………………………………………….. 2.3.1. Definition of probiotic .......................................................... 2.3.2. Developing probiotic for aquaculture ………….......... 2.3.3. Selection of good probiotic ................................................. 2.3.4. Mode of action of a probiotic …………………..…………. 2.3.5. Investogated mechanisms of probiotic ……………….…… 2.3.6. Selection criteria for potential probiotic …………..……… 2.3.7. Characteristics of good probiotic ......................................... 2.3.8. Probiotic effects ……………………………..…………… 2.3.8.1. Improvement of growth rate ……….………….…… 2.3.8.2. Improvement of feed utilization ……………….…… 2.3.8.3. Improvement of health and immune response……… 2.3.8.4. Improvement of reproduction performance ……….. 2.3.8.5. Production of polyunsaturated fatty acid vitamins And amino acid ……………….……….. 2.3.8.6. Proximate analysis …………………………………. 2.3.9.. How to provide probiotic ………………………………… 2.3.10. Propiotics evaluated for use in aquaculture …………………… 2.3.10.1. Lactic acid bactria as aprobiotic …………………. 2.3.10.2. Commercial probiotic …………………….………. 2.3.11. Probiotic protein interaction ……………………..……….. 2.4. Yeast as a probiotic ……………………………………………. 2.4.1. Effect of baker's yeast on fish growth ……………..……… 2.4.2. Effect of baker's yeast on feed utilization …………….……. 2.4.3. Effect of baker's yeast on the composition of Fish Body…. 2.4.4. Effect of baker's yeast on the physiological parameters….. 2.4.5. Effect of baker's yeast on fish immunity………………….. 2.4.6. Effect of baker's yeast on bacterial activity………………...
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3. MATERIAL AND METHODS .......................................................... 3.1. The tested feed additives .............................................................. 3.2. Fish and culture technique ........................................................... 3.3. Experiment design ....................................................................... 3.4. Feed preparation ........................................................................... 3.5. Feeding system ........................................................................... 3.6. Water quality parameters ............................................................. 3.7. Proximate analysis diet and fish .................................................. 3.8. Growth performance and feed utilization parameters ………….. 3.9. Hematological parameters ……………………………………... 3.9.1. Determination of hemoglobin, Hb (gm%) ………………… 3.9.2. Liver enzyme ………………………………………….…… 3.10. Statistical analysis ….…………………………....…. 4. RESULTS AND DISCUSSION ……………………………………. 4.1. Rearing water quality probiotic …………………………………... 4.1.1. Water temperature …………………………………..………… 4.1.2. Dissolved oxygen (mg/1) ……………………………………… 4.1.3. PH ……………………………………………………………… 4.2. Effect of some feed additives and probiotic on growth performance of Nile tilapia O. niloticus………………………………………….. 4.2.1. Body weight (BW) ……………………………………………. 4.2.2. Weight gain (WG) ………………………..………………..…… 4.2.3. Specific growth rate (SGR) ………………………………….… 4.3. Feed intake and feed utilization ……………….…………………… 4.3.1. Feed intake …………………………………………………….. 4.3.2. Feed conversion ratio …………….………………………..…. 4.3.3. Protein efficiency ratio …………………………………..…… 4.4. Some haematological variables …………………………………… 4.5. Serum transaminases (AST&ALT) ………………….………… …. 4.6. Proximate analysis of whole fish ………………………………..….. 5. SUMMERY …………………………………………………..………. 6. REFERENCES……………………….....……………………………. ARABIC SUMMARY
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LIST OF TABLES No. Title of tables Page Table 1: The composition and chemical analysis of experimental diets……………………………………………………….. 40 Table 2: Effect of some feed additives and probiotics on some water quality parameters for aquaria stocked by Nile tilapia O. niloticus ……………………………………...… 46 Table 3: Analysis of variance for the effect of some feed additives and probiotics on some water quality parameters for aquaria stocked by Nile tilapia O. niloticus ……………… 46 Table 4: Effect of some feed additives and probiotics on growth performance of Nile tilapia O. niloticus………………… 48 Table 5: Analysis of variance for the effect of some feed additives and probiotics on growth performance of Nile tilapia O. niloticus 49 Table 6: Effect of some feed additives and probiotics on weight gain of Nile tilapia O. niloticus ………………………….. 52 Table 7: Analysis of variance for the effect of some feed additives and probiotics on weight gain of Nile tilapia O. niloticus .. 53 Table 8: Effect of some feed additives and probiotics on specific growth rate of Nile tilapia O. niloticus ……………………... 56 Table 9: Analysis of variance for the effect of some feed additives and probiotics on specific growth rate of Nile tilapia O. niloticus 57 Table 10: Effect of some feed additives and probiotics on feed intake of Nile tilapia O. niloticus……………………….… 65 Table 11: Analysis of variance for the effect of some feed additives and probiotics on feed intake of Nile tilapia O. niloticus. .. 66 Table 12: Effect of some feed additives and probiotics on feed conversion ratio of Nile tilapia O. niloticus …………..…. 68 Table 13: Analysis of variance for the effect of some feed additives and probiotics on feed conversion ratio of Nile tilapia O. niloticus 69 Table 14: Effect of some feed additives and probiotics on protein efficiency ratio of Nile tilapia O. niloticus 72 ……………………. Table 15: Analysis of variance for the effect of some feed additives and probiotics on protein efficiency ratio of Nile tilapia O.
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niloticus………………………………………………….. Table 16: Effect of some feed additives and probiotics some feed additives and probiotics in Nile tilapia diets on blood parameters………………………………………………. Table 17: Analysis of variance for the effect of effect of some feed additives and probiotics in Nile tilapia diets on blood parameters………………………………………………. Table 18: Effect of some feed additives and probiotics on serum transaminases (AST&ALT) of Nile tilapia O. niloticus..… Table 19: Analysis of variance for the effect of some feed additives and probiotics on serum transaminases (AST&ALT) of Nile tilapia O. niloticus…………………………………... Table 20: Average of chemical analysis whole body of Nile Tilapia O. niloticus as affected with different feeding diets……... Table 21: Analysis of variance for the effect of some feed additives and probiotics on chemical analysis of whole body of Nile tilapia O. niloticus. …………………………………..….
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LIST OF ABBREVIATIONS BL : Body length BW: Body weight CP : Crude protein DO : Dissolved oxygen FCR: Feed conversion ratio FI : Feed intake Kcal : Kilo calorie KJ: Kilo joule O : Oreochromis PER : Protein efficiency ratio PPT: Part per thousand PPM: Part per million SGR :Specific growth rate T : Tilapia WG: Weight gain AST: Aspartate amino transferase ALT: Alanine amino transferase
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ACKNOWLEDGMENT First thanks to ALLAH who made me able to accomplish this work. It is a pleasure to express the sincere gratitude and thankfulness to Prof. Dr. Mohammed Khairy Ibrahim, Professor of Animal Breeding, Faculty of Agriculture, Moshtohor, Benha University for his valuable advice, continuous help, encouragement guidance, supervision and constructive criticism throughout this work. The auther also efforts his gratitude to Prof. Dr. Abdel-karim I. M. EL-Sayed Professor of Animal Physiology, Faculty of Agriculture, Moshtohor, Benha Universit, for his sincere supervision, and constructive criticism throughout this work . The deepest appreciation is to Prof. Dr. Magdy A. Soltan Professor of Fish Nutrition, Faculty of Agriculture, Moshtohor, Benha University for his supervision, encouragement, guidance and constructive criticism throughout carrying out this work in addition to his great effort in follow up all fobricating stages till the final practical results I wish to express my sciencere appreciation to Dr. Hossam Mahmoud Agouz Abdel-ghany Assistant professor of Aquaculture, Central Laboratory for Aquaculture Research (CLAR), Abassa, Agriculture Research Center for his supervision, and constructive criticism throughout carrying out this work. Sincere thanks and appreciation is expressed to all members of the Department of Animal production, Faculty of Agriculture, Moshtohor, for their help and cooperation.
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1. INTRODUCTION The demand for animal protein for human consumption is currently on the rise and is largely supplied from terrestrial farm animals. Aquaculture, however, is an increasingly important option in animal protein production. This activity requires high-quality feeds with a high protein content, which should contain not only the necessary nutrients but also complementary additives to keep organisms healthy and promote favourable growth. Some of the most utilized growth-promoting additives include hormones, antibiotics and some salts (Gongora, 1998 and Klaenhammer and Kullen 1999). Although these do promote growth, their improper use can result in adverse effects in the animal and the final consumer and can lead to resistance of pathogenic bacteria in case of antibiotics (El-Haroun et al., 2006). An alternative to the antibiotics is the use of beneficial bacteria (probiotic) to fight the pathogenic bacteria by competitive exclusion, which is an acceptable practice in animal husbandry (Sissons, 1989). In aquaculture industry, several probiotic species were used, including Saccharomyces spp. (Surawicz et al., 1989), Lactobacillus acidopilus (Venkat et al., 2004), B. subtilis (Kumar et al., 2006, Ghosh et al., 2007 and Keysami et al., 2007) and mixed cultures (Lessard and Brisson, 1987). There has been increasing interest in the possible use of probiotics in aquaculture, including application in black tiger shrimp, Penaeus monodon (Rengpipat et al., 2000), salmonids Oncorhynchus nerka and rainbow trout, Salmo gairdneri (Irianto and Austin 2002), white shrimp, Litopenaeus vannamei Boone (Venkat et al., 2004), Indian major carp, Labeo rohita Ham. (Kumar et al., 2006) and Penaeus japonicus and P. semisulcatus (Saleh, 2007). Most probiotics are supplied as live supplements in the
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diet, which has the ability to survive the passage through the intestinal tract (Verschuere et al,. 2000). The direct nutritional benefits of probiotic bacteria include vitamin production, availability of minerals and trace elements and production of important digestive enzymes. Probiotics increases growth, improve efficiency, prevent intestinal disorders and stimulate pre-digestion of anti-nutritional factors present in the ingredients (Holzapfel et al., 1998). Biogen® contains Bacillus licheniformis and B. subtilis. The advantage of these spore-forming bacteria is that they are able to survive the pelletization process. After transit passage through the stomach, they germinate in the intestine and use a large number of sugars (carbohydrates) for their growth and produce a range of relevant digestive enzymes, amylase, protease and lipase (ElHaroun et al., 2006). Garlic (Allium sativum L.) is widely cultivated in Egypt and used as flavoring agent, a common feed and popular remedy. Because the thyroxin like activity of garlic, it is suggested that it has growth stimulating effect. Garlic stimulated growth by increasing the inflow of glucose into tissues and thyroid like activity (El-Nawawy, 1991). Sulfur compounds in garlic are considered as active antimicrobial agents and improve immunity and therefore stimulate growth (El-Afify, 1997) and have a mode of action similar to antibiotics (Ibrahim et al., 2004). The present study was designed to investigate the effect of some probiotics (B. subtilis and Biogen®); some spices (garlic and fennel) or the combination of B. subtilis and each of garlic or fennel on viability, growth performance, feed utilization, liver function, feeding and proximate composition of O. niloticus.
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2. REVIEW OF LITERATURES 2.1. Spices as a feed additive: Many attempts have been undertaken in order to improve the utilization of diet nutrients using several growth promoters feed additives from different sources. The use dietary supplementation has greatly increased although feed additives contain chemical components, hence the accumulative effect of these components induced deterrent effects on human health (Salem and El-Mahdy, 2001). Therefore, the use of natural feed additives is important to minimize these adverse effects. Some vegetables, herbs, edible plants and seeds are used at tonics and restoratives (Abdel-Latif et al., 2004). Several studies in animal nutrition showed that adding some spices or medical and aromatic plants to diets of sheep, goats, rabbits, chicken, cow and buffaloes had favorable effects on live weight gain, feed intake, feed efficiency and nutrient digestibility (Hanfy 1995; Gaber et al., 1996 and Allam et al., 1999). The most recent studies showed successful use of spices and natural herbs in fish nutrition including marjoram, basil, licorice roots, black seeds, peppermint leaves, fenugreek seeds and caraway seeds (Abdel-Maksoud et al., 2002; Abd- Elmonem et al., 2002; Sakr 2003; Shalaby et al., 2003, El-Dakar 2004; El-Dakar et al., 2004 a and b and Shalaby 2004). In this context, Lawerance et al., (2001) recommended to use environmental-friendly feeds in shrimp diets in order to reduce the amount of pollutants in the effluent water. Incorporation of some natural plants may be caused a reduction of uneaten feed percent by aquatic culture animals (Sakr, 2003; ElDakar 2004 and El-Dakar et al., 2004 a & b) about third of feed introduced to aquatic cultured animals in wasted into water (Axler et
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al., 1996) causing adversely effects on water quality. Reduction of the feed waste is very important especially in culture systems depends on the reuse rearing water, Marjoran, basil and peppermint might decrease feed waste to 3-16% (Sakr, 2003 and El-Dakar et al., 2004 a & b). El-Dakar et al., (2005) evaluated fennel seeds, Marjoram leaves and ginger roots meals as feed additives for shrimp, Penaeus japonicus juvenile. A commercial shrimp diet containing 45% protein were used to prepare four experimental diets with 0.5% of each tested spices and they found that, shrimp fed the tested diets were exhibited higher body weight, gain and specific growth rate than those fed the control diet. Growth performance, feed conversion and nutrient utilization of shrimp juveniles fed fennel diet were significantly higher (P
