EFFECT OF USING OMEGA-3 AND OMEGA-6 ON GROWTH AND HAEMATOLOGICAL PARAMETERS OF COMMON CARP (Cyprinus carpio L.)

A Thesis Submitted to the Council of the Faculty of Agricultural Sciences at the University of Sulaimani in Partial Fulfillment of the Requirements for the Degree of Master in Animal Production

(Fish Production) By

Avan Aladdeen Sadradeen B.Sc. Animal Production (2009), Faculty of Agricultural Science University of Sulaimani SUPERVISOR

Dr. Abdulmotalib Jassim Al-Rudainy Assistant Professor

2714 K

2014 A.D

‫بسم الرحمن الرحيم‬ ‫َو ﱠ‬ ‫سما ِء ماء‬ ‫ﷲُ أَن َز َل ِم َن ال ﱠ‬ ‫ض ب ْع َد َم ْوتِھا‬ ‫فَأ َ ْحيا بِ ِه ْاألَ ْر َ‬ ‫إِ ﱠن فِي ذلِ َك َآليَة لِقَ ْو ٍم‬ ‫س َم ُعون‬ ‫يَ ْ‬

‫‬ ‫   ‪65‬‬

‫‬

DEDICATION I dedicate this work to: My husband My smart father My lovely mother My honest brother and sisters sisters All my instructors and friends.

Linguistic Evaluation Certification I herby certify that this thesis prepared by (Avan Aladdeen Sadradeen), has been read and checked and after indicating all the grammatical and spelling mistakes; the thesis was given again to the candidate to make the adequate corrections. After the second reading, I found that the candidate has corrected the indicated mistakes. Therefore, I certify that this thesis is free from mistakes.

Dr. Nariman Salih Ahmad Lecturer Field crop Department Faculty of Agricultural Sciences / / 2014

ACKNOWLEGEMENTS First and foremost, I would like to thank God for his helping to complete this paper.I would like also to express my deepest thanks to my supervisor Dr. Abdulmotalib Jassim AlAl-Rudainy for the patience, guidance, instructive suggestion and continuing encouragement. I would like to express special thanks to my Teacher Dr. Nasreen M. Abdul - Rahman for help in fish Laboratory, and my friend Vian Muhammed for their support and Mr. Mr. Hemi Hemin nuradeen for help analysis data and all stuff in department of Animal production in Sulaimani University for them help and advice, I would like to thank thank my committee members for their very helpful insights, comments and suggestions for his input at my proposal meeting. Finally, I am very thankful to my wonderful family for their great assistance.

`SUMMARY This study was carried out to investigate the effect of linseed oil (LN) as a source of omega-3 and safflower oil (SFO) as a source of omega-6 on the growth performance of common carp. The experiment was conducted in the fish laboratory of Animal Production Department, Faculty of Agricultural Sciences,

Sulaimani

University,

Kurdistan

Region-Iraq.

The

actual

experimental feeding trials were durated for twelve weeks from 19th June till 11th September 2013; the trials lasted for three months after an adaptation period of 21 days. A total of a hundred and twenty six (126) common carp (C. carpio) were used in the experiment with an average weight of 35-40g and the range of 12-14 cm in length. A number of treatments with different concentrations of linseed oil and safflower oil were applied In control fish were fed a diet with 0% oils, fish were fed a diet 1.5 % linseed oil, fish were fed a diet 2% linseed oil, fish were fed a diet 2.5% linseed oil, fish were fed a diet 1.5% safflower oil, fish were fed a diet 2% safflower oil and fish were fed a diet 2.5% safflower oil. Each treatment was run in three replicates in which 6 common carp C. carpio

were stocked in each tank which fed the

experimental diets twice daily with 3% body weight. The present results of this study show that the effect of safflower oils on final biomass (g) revealed significant increase in SFO 2% and SFO2.5% compared with other treatments. Results indicated that treatments of SFO 2% and SFO2.5% were the in relative growth rate best compared with treatments (0.68, 0.64 g/day %), respectively. The highest specific growth rate (36) (g/ day %) was recorded for treatment SFO 2%, while the best feed efficiency rates (53.42 and 49.60 %) were obtained for the treatments of control and SFO1.5%, respectively. The traits of food conversion rate and protein efficiency rate had no significant differences at P>0.05 among all treatments. Blood parameters of tested fish were showed significant differences in RBC count (cells x 106µl) in SFO1.5%, SFO 2% and SFO 2.5% by 2.59, 2.27 and I

Summary ________________________________________________________________________________

2.14, respectively. While the white blood cell (cell x103µl) was differ significantly (17.93) for the seventh treatment and 17.23 for SFO 2%; and hemoglobin valued 9.66(Hb (g/dl) significantly in the LN 2.5%. The mean corpuscular hemoglobin (MCH (pg /l) was 54.83 in SFO 2.5%, mean corpuscular volume (MCV (fl)) was 160 in SFO 1.5%, packed cell volumes (PCV %) were 31.55% and 31.33% in SFO 2% and SFO 2.5%, respectively. Significant differences in granulocytes count 68.58% were recorded in SFO 2% and (68.00%) in the control. The lymphocytes were 16.86% in SFO 2.5%. Monocytes were differ significantly giving (36.20%) in SFO 2%. The percentage of Platelet values had no significant differences at P>0.05 between all treatments. Regards the Liver index, Gonadosomato index and blood cholesterol, no significant differences were observed at P>0.05 between experimental treatments. The results of statistical analysis showed no significant difference at P>0.05 in crude protein and ash values of common carp fed on diets with different type and levels of oils compared to the control. The dietary lipid content showed a significant increase at P≤ 0.05 between SFO 2.5% (37.82%), LN 2%, SFO 2.5% (37.69%, 37.12%) compared with other treatments

II

TABLE OF CONTENTS Subject No.

Subject Title

Page No.

Summary………………………………………………………………………I List of contents …………………………...……….……………………….….II List of table…………………………………………………………………...VI List of figures…………………………………....……………………….…...VI List of abbreviations……………………………….. ……………………… III

CHAPTER ONE: INTRODUCTION Introduction………………………………………………………………………..1 CHAPTER TOW: LITERATURE REVIEW Literature Review………………………………………………………………….4 2- Overview………………………………………………………………………..4 2-1- The use of omega-3 and omega-6 in human diets…………………………....7 2-2- The use of omega-3 and omega-6 in animal diets…………………………....9 2-3- The use of omega-3 and omega-6 in common carp diets…………………....10 2-4-Nutritient requirement for common carp ………………..…….……….…….12 2-4-1-Energy requirements………………………………………………...……..12 2-4-2-Protein and amino acid requirements……………………………………....12 2-4-3-Lipid and fat …………………………………………………………….....13 2-4-5-Vitamins requirements…………………………………………..…………15 2-4-6-Mineral requirements.............................................................................…...16 2-5-Fatty acids structure………………………………………………………… 16 2-5-1-Omega-3 fatty acids………………………………………………………. 17 2-5-2 Omega-6 fatty acids………………………………………………………. 18 2-6-Safflower oil…………………………………………………………………..19

III

2-7-Linseed oil…………………………………………………………………….20

CHAPTER THREE: MATERIALS AND METHODS Materials and methods…………………………………………….………………24 3-1- Experimental fish………………...….……………….………..……………..24 3-2- Experimental system ………………………….……………………...……...25 3-3 Water quality …………………….……………………..…………………….26 3-4- Design of the experimental study……...…………………………………….26 3-5- Diet formulation ………….……………………………………..………..….27 3-6- Growth parameters………….…………………………………….……….…29 3-7- Biological parameters……………….………………………………………..30 3-8- Haematological examination…………………………………………..……..30 3-8-1- Red Blood Cells Counting (RBC)……………………….………………....30 3-8-2: Hemoglobin Test (Hb)…………………………………….…….……….....31 3.8.3 Packed Cell Volume (PCV %)……………………………………………....31 3.8.4 White Blood Cells Counting (WBC)…………….………...………………..32 3.8.5 Measuring the concentration of cholesterol…………………...…………….32 3-9 Chemical composition………………………………….……………………..33 3-9-1 Crude protein………….………………………………………………....…33 3-9-2 Ether extract ………….…………………………………………….….…...33 3-9-3 Ash……………………………………………………………………….....33 3-9 Statistical analysis………………………………….………...…….................34

IV

CHAPTER FOUR: RESULTS AND DISCUSSION Results and discussion……………………..……………………………………...36 4.1. Water quality ………………………………………….…………….………...36 4.2. Growth Performances…………………………...………………………….…37 4.2.1. Weight Gain………………………………………………...…………….…37 4.2.2. Food Conversion Ratio …………………………………….…………..…...46 4.3. Biological parameters………………………………………..………………..51 4.4. Hematological parameters………………………………………………….....52 4.4.1 Hematology……………....……………….…………………………............52 4.4.2 White blood cell counting...…………………………………………............56 4.5 Body composition……….…………………………………………………….58 Conclusions and recommendation……………………………………..................62 Conclusions……………………………………………………..…………............62 Recommendation………………………………………………...………………...63 References…………………………………………………………...………….....64 Summary in Arabic……...…………………………………………………….…..i-ii Summary in Kurdish…….…………………………………………………….…A-B

V

LIST OF TABLES Table 1: Fatty acid composition for linseed oil and safflower oil as a source of on selected fatty acids (i.e; omega- 3 and omega- 6) (g/100g of total fatty acids) (Ruyter et al. 2000)…………….………..………………………...22 Table 2: formulation of experimental diets…………………………………….....28 Table 3: Chemical composition (%) of with diets……………………………… 29 Table 4: Water parameter during the experimental trails………………………... 36 Table 5: Effect of safflower oil and linseed oil on growth performance of common carp 12 weeks fed on experimental diets (mean ±SE)…………………… 40 Table 6: Feed utilization; FCR, PER and FER of common carp after 12 weeks feeding Experimental diets (±SE) in C. carpio L………………………….47 Table 7: Effect of feeding trail of linseed oil and safflower oil on liver somatic index and gonado somatic index(± SE) in C. carpio……………….…… 51 Table 8: Haematological parameter of common carp fed on Experimental diets. (Cont. LN 1.5%, LN 2%, LN 2.5%, SFO 1.5%, SFO 2% and SFO 2.5%)…54 Table 9: Effect of linseed oil and safflower oil (± SE) Wight blood cell………....57 Table 10: Effect of linseed oil and safflower oil on whole body composition at the end of experiment on dry matter basis (mean±SE)…………………..… 59

LIST OF FIGURES Figure1: The reared fish common carp Cyprinus carpio …………………….….24 Figure 2: Experimental design Control, LN1.5, LN2, LN2.5, SFO1.5, SFO2 and SFO2.5 represent the seven treatments in this trial (A and B)………….25 Figure3: Shows the experimental design of the study …………...………...……..26

VI

LIST OF ABBREVIATION AA ALA AOAC DGLA DHA EDTA EFA EPA FAO FCR FEC G GLA GRAN GSI H HB HMG-COA

ARACHIDONIC ACID ALPHA-LINOLENIC ACID ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS DIHOMO-GAMMA-LINOLENIC ACID DOCOSAHEXAENOIC ACID ETHYLENEDIAMINETETRAACETIC ACID ESSENTIAL FATTY ACID EICOSAPENTAENOIC ACID FOOD AND AGRICULTURE ORGANIZATION FOOD CONVERSION RATE FOOD EFFICIENCY RATE GRAM GAMMA –LINOLENIC ACID GRANULOCYTE GONADOSOMATIC INDEX HOURS HEMOGLOBIN HEPATIC 3-HYDROXYL-3- METHYLGLUTARYL COENZYME

HPUFA

HIGHLY POLYUNSATURATED FATTY ACID

KRG

KURDISTAN REGIONAL GOVERNMENT

KJ

KILO JOULES

L

LITRE

LA

LINOLEIC ACID

LDL

LOW DENSITY LIPOPROTEIN

LN LSD LYMPH MCH MCV MG ML MON MUFA

LINSEED OIL LEAST SIGNIFICANT DIFFERENCE LYMPHOCYTE MEAN CORPUSCULAR HEMOGLOBIN MEAN CORPUSCULAR VOLUME MILLE GRAM MILLE LITTER MONOCYTE MONOUNSATURATED FATTY ACIDS

VII

N-3 N-6 NIH NRC PCV PER PG PLT PUFA RBC RGR SD SE SFO SGR T TG U.S. W0 W1 WBC

OMEGA-3 OMEGA-6 NATIONAL INSTITUTES OF HEALTH NATIONAL RESEARCH COUNCIL PACKED CELL VOLUME PROTEIN EFFICIENCY RATE PICO GRAM PLATELET POLYUNSATURATED FATTY ACIDS RED BLOOD CELL RELATIVE GROWTH RATE STANDARD DIVISION STANDARD ERROR SAFFLOWER OIL SPECIFIC GROWTH RATE TIME TRIGLYCERIDE UNITED STEAD INITIAL WEIGHT FINAL WEIGHT WHITE BLOOD CELL

VIII

IX

CHAPTER ONE INTRODUCTION Aquaculture has been defined in many ways to denote all forms of culturing aquatic animals and plants in fresh, brackish water and the marine environment (Pillay and Kutty, 2005). The most current accepted definition has been given by the food and Agriculture Organization as the farming of aquatic organisms including fish (FAO, 2010). The importance of aquaculture as one of the primary sources that can be relied upon to protect food security in the countries, especially securing animal protein with high nutritional value, as well as to protect and strengthen the stocks of natural and preservation of the environment (FAO, 2010). Many countries depend on fish to cover 50% of their daily demand on animal proteins, the world yearly consuming of fish reached to 18kg/individual/year, whereas Iraqi population consuming of fish is not more than 1.5kg/individual/year during the few last years (Al-Rudainy, 2007). Fish has easy digested meat, which has a good percentage of protein, oil and vitamins, as well as unsaturated fatty acid especially omega- 3 and omega- 6 which is very important for human body and the growth for children (bureau et al., 2008). The fish is the most important sources of animal protein contained the proportion of protein ranged from 18-23%, while the beef and egg contain 16.8%and eggs13.6% of protein, respectively. proteins of contained fish contain essential amino acids for human body especially lysine, fish proteins are important for growth and development of the body, maintenance and repairing of damage tissues, they are required for production of enzymes and hormones, they also necessary for many body processes containing all the eight essential amino acids

1

Chapter One

Introduction

including the sculpture (containing lysine, methionine and cysteine) (bureau et al., 2008). Fish is contains iodine which is essential for the growth of the thyroid gland and some receiver in children, in addition to the above mentioned it contains a reasonable ratio of vitamins (A, B and D) and minerals (calcium, phosphorus, iron) (FAO, 1996). Fish is a rich source of vitamins, particularly vitamins a and d from fatty species as well as vitamins B1, B2 and B3, vitamin is required for normal vision and for bon growth, vitamin d is crucial for bone growth, while B1, B2 and B3 are important for energy metabolism, the minerals present in fish include iron, calcium, zinc, iodine, phosphorus, selenium and fluorine, these minerals are highly ‘’bioavailable’’ being absorbed easily by body (kris-etherton et al., 2002). Fats are the most intense in the production of energy for the fish and give almost 38.5kJ/g fat, compared with the energy of protein 23.6 kJ/g protein and carbohydrates are the least giving almost 17.3 kJ/g carbohydrates, fats and nutrients are important for fish because they contain soluble vitamins (A, K, E and D) and minerals and they are the source of essential fatty acids (Bureau et al., 2008). Many of the fatty acids can be synthesized by humans, but there is a group of polyunsaturated fatty acids that the human body cannot produce: omega-3 and omega-6 fatty acids depending on the species, animals may require certain types of that are found in three families that have been called omega fatty acids: Linolenic acid is the smallest molecular weight fatty acid in that family, Other families of polyunsaturated fatty acids are the oleic acid and linoleic families, with the lowest molecular weight fatty acids in the omega-3 and omega-6 families being oleic and linoleic acids (kris-etherton et al., 2002).

2

Chapter One

Introduction

The present study aims to investigate the effect of adding different level of linseed oil and safflower oil on growth performance and feed utilization, carcass composition and blood parameters of fingerlings common carp.

3

CHAPTER TWO LITERATURE REVIE 2- Overview The common carp cyprinuscarpio belongs to the family cyprinidae. in nature, common carp inhabits in rivers, reservoirs and marshes, they are omnivorous in nature as they feed on a wide variety of plants and animals such as; zooplankton, insects, worms, aquatic plants, algae and seeds, the common carp can tolerate a very wide range of conditions, common carp can live in a wide range of temperatures between 1-40 ºc, the best growth occurs at a temperature of above 13 ºC, and they reproduce at temperatures up to 20 oC, they originally live in fresh or brackish water with a 7.0-7.5 pH (Swift, 1993; Billard, 1995; Takeuchi et al., 2002). Common carp are distributed all over Asia, most parts of Europe and in some African countries. Also, they have been introduced in the USA and Australia (Pillay and Kutty, 2005). The required protein levels in the diets of common carp are between 30-38% crude proteins. Carbohydrates and lipids are easily utilized by common carp as a dietary energy sources. Common carp has to be feed more frequently (4 times a day) due to absence of a stomach (Swift, 1993; Billard, 1995; Takeuchi et al., 2002). Aquaculture continues to be the fastest growing food producing sector worldwide. Aquaculture production in 2009 stood at 55.1 million tons contributing to 38% of the total world production of sea food (145.1 million tons) with a total value of USD 106 billion. The contribution of aquaculture will account for over 50% of the global fish supply by 2020 if this sector continues to expand at the present rate(FAO, 2010).

4

Chapter Two

Literature Review

The Euphrates and Tigris rivers are the main sources of water in Iraq, with freshwater bodies covering between 6 -7 million ha, from this 39% are natural lakes, 13.3% dams and reservoirs, 3.7% rivers and their branches and 44% marshes. The total area with employed for aquaculture consists of about 7500 ha at around 2000 farms (Kitto and Tabish, 2004). In Iraq, fisheries and aquaculture output was estimated to be: 49,623 tones (captured) 3,622 tones (cultured) in 1980. Afterward the production was declined substantially to less than half (22,629) tones capture, 1,600 tones cultured due to the conflict and awareness crises in the region,however recently the fish production has rapidly increased whereby capture fish was estimated at 34, 472 tones and cultured fish production at 19,246 tons in 2008 (FAO, 2010). The average annual production of common carp for 1998 was 7,500 tones and decreased to 2,183 tons in 1999. This declination was due to the consistency drainage of the majority of Iraqi wetlands. In 2008 the production of common carp was recorded to elevate significantly to approximately 10,700 tones. The aquaculture sector in Iraq is totally based on freshwater production. The rearing system of aquaculture is mainly earthen ponds, with some cage farming being used for culture as a common way for commercial production (Fan, 1996). In Kurdistan region, aquaculture is purely based on freshwater production. The sources of freshwater are the Tigris and its tributaries (Khabur River, Greater Zab River, Lesser Zab River, Sheena River, RawandozRiver, Awa Spi River and Sirwan River), and water reservoirs in the form of dams (Dukan dam, Darbandikhan dam, Dohuk dam and Mosul dam). The water from these tributaries feed into the Tigris annually and are estimated to be at 16 billion m3 which equals ~ 33.45% of the Tigris water (Tigris water is estimated at 48 billion m3 per year) (Ismael, 2004).

5

Chapter Two

Literature Review

Since the early 1960ʼs, common carp has been placed into the Dukan and Darbandikhan lakes. It aims to improve fish production in the region but this strategy has not successfully elevated the production to the economic value in these two lakes. Subsequently in 1998, the Food and Agriculture Organization (FAO) established two stations; Erbil (Enkawa) and Sulaimnia (Dukan) for fish production. After that, in 2001, the FAO offered the stations to the Ministry of Agriculture and Water Resources-Kurdistan Region-Iraq (Gowdet, 2007). In Kurdistan small-scale fish farms are operated by individual farmers in rural lake and river areas.Commoncarp is considered as the main important fish species due to the efficient production, wide range of tolerance, high resistance for diseases, ease to hatch naturally or artificially and their ability to be cultured in different systems (El-Sayed, 1994;El-Khalil, 2005).The General Directorate of Fish Production and Development (GDFPD) presently owns Enkawa station it comprises 20 fish ponds of different sizes with an annual capacity of over 2,000 tones and a hatchery to produce common silver and grass carp fingerlings for distribution to farmers to practice and develop fish farming as a profitable food source. In Erbil Enkawa station, a fish feed factory with a capacity of 3 tonnes per hour was established by (MOAWR) in 2007 to produce the diets for carp feeds. These diets are produced as a pellet and distributed to famers in the region. Indeed, the number of fish farms increased rapidly to 260 by 2008. The average size of most of these farmers is comparatively small, about 1-3 ha. The priority plan for the Kurdistan Regional Government (KRG) is to regenerate agriculture production in the State (Omar, 2011).

6

Chapter Two

Literature Review

2-1- The use of omega-3 and omega-6 in human diets: The most important dietary are omega-3 and omega-6 fatty acids include alpha-linolenic acid (ALA), linoleic acid (AL), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) in human nutrition. For most people, the primary sources of omega-3 fatty acids include oils , cold-water fish such as salmon, halibut, and sardine, which contain EPA and DHA, and oils from plants such as linseed oil, safflower oil, canola, soybean, and walnut, which contain ALA, AL. ). Safflower oil meal is mainly used as animal feed. Safflower cake has the potential to be used asa human food if the bitter principles are removed (Nagaraj, 1995). The U.S.A. Food and Drug Administration have not published recommended daily allowances (also referred to as Dietary Reference Intakes) for omega-3, omega-6 fatty acids, but other groups have. For example, the Institute of Medicine and National Academy of Sciences suggest including between 1.3 g and 2.7 g of omega-3, omega-6 fatty acids in a 2000 kcal daily diet (National Academy of Sciences, 2002) World Health Organization and North Atlantic Treaty Organization have made formal, population-based recommendations: for EPA plus DHA, daily recommended amounts range from 300 to 500 mg/day, and for ALA, the range is 800–1100 mg/day (Kris-Ethertonet al., 2002), a cumulative amount comparable to that recommended by the National Academy of Sciences (2002) above. The EPA plus DHA can be acquired by consuming two to three servings of fatty fish per week or by taking fish oil supplements. Since ALA becomes converted internally to EPA and DHA, people on diets low in plant oils (such as the traditional diet of Inupiat and other Alaska Natives).Some omega-3, omega-6 fatty acids have been suggested or shown to be beneficial in the prevention or treatment of a number of disease conditions, including high cholesterol, highblood pressure, heart disease, stroke, diabetes, arthritis depression, schizophrenia,

7

Chapter Two

Literature Review

attention deficit/hyperactivity disorder, inflammatory bowel disease, asthma, macular degeneration, and colon, breast, and prostate cancer (Storlienet al., 1991; Egelandet al., 1998; Gurr, 1999; Kris-Ethertonet al., 2002; Li et al., 2003). Appropriate levels of omega-3, omega-6ftatty acids in the diet are also important for healthypregnancy and proper neonatal growth and development(Lantinget al., 1994). Omega-3 fatty acids correct imbalances in modern diets that lead to health problems. Eating foods rich inomega-3 fatty acids can help lower the risk of chronic diseases such as heart disease, stroke, and cancer aswell as lower LDL or “bad” cholesterol.ALA – Studies show a diet high in ALA helps reduce heart disease and stroke by reducing cholesterol and triglyceride levels, enhancing the elasticity of blood vessels, and preventing the build-up of harmful fat deposits in the arteries. In fact, the National Institutes of Health (NIH) has reported the majority of U.S. diets no longer contain the amount of omega-3 fatty acids needed by our bodies for overall health and wellness.EPA/DHA – Studies show that diets high in amounts of EPA and DHA help with brain and eye development, prevents cardiovascular disease, and can help to prevent Alzheimer’s disease. For example, diets notably high in DHA have been known to protect against degenerative processes within the retina of the eye to increasing the problem solving skills in nine month old infants. 10-year study correlated increased intakes of DHA/EPA as consumed by various population sectors with relative risk of heart related deaths. Those who increased consumption of DHA/EPA up to 664 mg/day were associated with an approximate 40 percent reduction in cardiovascular disease and a significant reduction in all-cause mortalit, all infant formula is now supplemented with DHA Hibbeln and Joseph (2006).Most omega-6 fatty acids are consumed in the diet from vegetable oils such as linoleic acid. Excessive amounts of linolenic acid can contribute to inflammation and result in heart disease, cancer, asthma, arthritis,and depressionLands and William (2005).

8

Chapter Two

Literature Review

2-2- The use of omega-3 and omega-6 in animal diets: All vertebrate species have absolute dietary requirements for certain PUFA. When a dietary deficiency occurs, the animal stops growing and reproducing, develops various pathologies, and eventually dies. The various deficiency symptoms and pathologies were summarized in the previous edition of this book (Sargent et al., 1989). The poly unsaturated fatty acid (PUFA) in question are termed essential fatty acids (EFA) and they include members of both the omega6and the omega-3 series typified by omega-6 and omega-3. All vertebrate species probably require both omega-6 and omega-3 poly unsaturated fatty acid i.e., both omega-6 and omega-3 poly unsaturated fatty acidare essential fatty acids for vertebrates, and the biologicallyactive forms of essential fatty acids are generally the poly unsaturated fatty acid and highly unsaturated fatty acids metabolites of omega-6and omega-3, which are oftentermed HUFA (highly unsaturated fatty acids). Some, but not all, vertebratespecies can convert omega-3 and poly unsaturated fatty acid to the higher omega-6 andomega-3highly unsaturated fatty acids through a series of alternating desaturation and chain elongation reactions mediatedby microsomal systems containing elongases and (∆6 and ∆5) enzyme fatty acid desaturases (Cook, 1996). Several studies have been published on fatty acids and skin health in dogs. In dogs with atop, the importance of LA in skin health was also observed when this fatty acid was significantly reduced compared to normal animals(Gurr, et al., 2002).Another study found that coetaneous fatty acids in dogs with seborrhea also have decreased LA content. With respect to the omega-3 fatty acids, one study found that increased amounts of a dietary α-linolenic acid fed to normal dogs for 56 days significantly lowered epidermal water loss with notable hair coat improvement. Because flaxseed oil provided the source of α-linolenic acid in the

9

Chapter Two

Literature Review

diet, it was suggested that this omega-3 fatty acid may also function as does LA due to its incorporation into skin creaminess(Gurr, et al., 2002). However, it appears from studies John (2011)laboratory that the α-linolenic acid (omega-3) effect may be due, in part, to the sparing of linoleic acid because of the known metabolic competition between these two fatty acid types.When inflammation exists, omega-3 fatty acids and some omega-6 fatty acidsmay be needed to help control the inflammatory response. This effect is mediated via the production of less pro-inflammatory mediators from precursors Research into the benefits of omega-3 fatty acidsin equine diets is increasing. Fat as a sourceof dietary energy for horses is now widely accepted,and its nutritional advantages are irrefutable.Substitution of starch with fat can helprelieve painful muscle conditions such as tyingup, modify behavior, and control metabolicconditions such as insulin resistance. Scientistsare further exploring how certain types of fatshelf horses (Fritschet al., 2010). 2-3- The use of omega-3 and omega-6 in common carp diets: The vegetable source of oil is used as “The replacement”(Turchiniet al., 2009),The vegetable oil can replace substantial amount of fish oil in the diets of many fish species without affecting growth and feed efficiency. However, the drawback of these alternatives is that they lack the omega-3, omega-6 and therefore are compromising the nutritive value of farmed fish for consumers. Several alternative oil sources, derived from unicellular algae, pelagic organisms or benthic invertebrates containing high amounts of omega-3 have been identified and tested in aqua feeds. Nevertheless, their prices are still too high to be commonly used in aqua feeds (Turchiniet al.,2009).Freshwater microalgae unlike marine microalgae generally have omega-3 linseed oil rather than omega-6 and

10

Chapter Two

Literature Review

omega-3fish oilas their principal poly unsaturated fatty acid (Ahlgrenet al., 1992). In addition,omega-6 is not prominent in marine microalgae but it can be abundant in freshwatermicroalgae (Ahlgrenet al., 1992). The major poly unsaturated fatty acid in the green leaves ofterrestrial and freshwater plants is omega-3 andomega-6 is abundant in theseed oils of plants. Freshwater insects can have substantial amounts of bothomega-3 and omega-6 in their lipids, butomega-3is usually very low or absentand omega-6and omega-3 are frequently the major PUFA in this invertebrategroup (Stanley–Samuelson et al., 1988; Ogget al., 1993). Bell et al. (1994b)showed that other freshwater invertebrates contained mainlyomega-3 andomega-6, and omega-3 as their principal and Henderson et al. (1996) showedthat insect larvae (bloodworms) used commercially as fish feed have omeg6as their major PUFA, withomega-3, omega-6, and omega-3 being present only in low amounts. Therefore, although lipids in freshwater organisms are generallyless well defined than their marine counterparts, it is clear that omega-6 and omega3are at least as well represented as omega-3 at the base of the freshwaterfood webs and that both omega-3 andomega-6are present. This correlateswith the widespread ability of freshwater fish to convertomega-3 and omega-6to thebiologically active omega-3 and omega-6 and with freshwater fish having substantialdietary requirements for both omega-6 and omega-3. It was reported that commoncarp, in contrast to marine fish, are be able to bio-convert omega-3 source of linseed oil to omega-3 andomega-6 source of fish oil (Zhenget al., 2004; Tocher, 2003; Olsen et al., 1990; Farkas, 1984). Itis therefore of interest to understand and maximize the ability of carp tosynthesize omega-3 andomega-6 source of fish oilfromomega-3 source of linseed oil in order to preserve the lipid quality ofthe fish as human food and for sustainable utilization of feed resources. Commoncarpculture might be capable of becoming net producer of EPA and DHA byselecting fish with high enzyme activities in fatty acid elongation anddesaturation.The common carp have

11

Chapter Two

Literature Review

also relatively low requirementsboth for omega-3 and omega-6fatty acids (0.5-1%) which can be fulfilled by plant 18 carbon fatty acids(Takeuchi, 1996). 2-4-Nutritient requirement for common carp: Nutrition has a major impact on the lipid content and composition in fish. Thus good pond management to maintain a sufficient amount and appropriate structure of the planktonic and benthic community in ponds is of great importance when seeking to improve common carp fatty acid composition.Cereals are usually used as a supplemental feeding for carp. Since they arerich in carbohydrates and have very low levels of omega-3, the flesh of thefarmed common carp generally contains a high level of oleic acid and a low level offavorable omega-3 HUFA

2-4-1-Energy requirements: Energy is released during the metabolic oxidation of carbohydrate, protein and lipid; energy does not strictly account as a nutrient component but is a property of nutrients (Parker 2002; Lim and Webster 2006; NRC 2011). However, almost of all dietary energy is derived from protein and lipid (Takeuchi et al., 2002). Parker (2002) reported that energy requirements for common carp (expressed as concentration of diet) at 13.4 MJ kg-1 DM agrees with the study by Takeuchi et al., (2002) which states dietary energy requirement values for the common carp of 12.97-15.06 MJ kg-1. The growth performance of carp required for optimum growth based on the total gross energy intake (100%), the maintenance and activity, productive and heat increment energy as well as, faecal and non-faecal energy values was; 12.6, 24, 31.9, 30 and 1.5, respectively (Ohta and Watanabe 1996).

12

Chapter Two

Literature Review

2-4-2-Protein and amino acid requirements: Protein is the most expensive dietary nutrient in fish diets (Jauncy 2000; Lim and Webster 2006) and essential for maintenance, growth and reproduction as well as energy (Lim and Webster 2006). A deficiency of protein will cause impairment and retardation or cessation of growth or loss of weight because of withdrawal of protein from less important tissues to retain the function of more vital organs. Ogino and Chen (1973) indicated that the common carp needs 1g kg-1 body weight of protein to meet their daily requirement and for 12g kg-1 body weight for maximum protein retention. The protein requirements of C. carpiohave been demonstrated with crude protein levels ranging from 30-42% as being adequate for fish to grow well. Siddiquiet al. (1988) showed that tilapia fingerlings (50 g mean weight) required only 30% proteins in the diet compared to fry (0.8 g) requiring 40%. Protein requirement therefore depends on the fish age, size, type of rearing, and protein quality presence of natural feed.

2-4-3-Lipid and fat: Each gram of lipid contains 2.5 fold the energy content compared to a gram of carbohydrate or protein (Jauncy 2000; Parker 2002).Dietary lipids have two primary functions: the first of which is that they are a source of metabolic energy. Secondly, as phospholipids they maintain biological structure and normal functions of cell membranes (Jauncey,1982; Sargent et al., 1999; NRC, 2011). The consumption of fish and fish lipids can provide poly unsaturated fatty acid especially omega-3 (Kmínkováetal. 2001), moreover, proteins, unsaturated essential fatty acids, minerals, and vitamins can also be provided by them (Ackman 2000). Polyunsaturated fatty acids are known to diminish the level of the blood

13

Chapter Two

Literature Review

cholesterol, therefore, they can prevent cardiovascular diseases (Kmínkováet al. 2001). These fatty acids can be further converted to highly unsaturated fatty acids (HUFA), and carp is able toprolongthe process ofremoving the saturation of carbon chain (Zhenget al., 2004). Csengeri (1996) studied the effects of starvation on lipid content andcomposition in common carp and observed a consistent decrease in the oleicacid levels in both muscle and liver, whereas PUFA were partly protected. Healso concluded that the effect of starvation was dependent on the previousfeeding regime. A supplementaryfeedstuff which is rich in ALA could be an alternative way to increase the omega-3 HUFA content in carp flesh. Feedstuffs with a high level of ALA that arecheap and easily available include safflower oil, linseed and hempseed.Safflower oil and linseed oilhave a moderate / high level of ALA (13% and 60%, respectively) and afavorable omega-3/omega-6 ratio (around 1:2 and 5:1, respectively) (Pickova and Morkore, 2007). Dietary lipids have two primary functions: the first of which is that they are a source of metabolic energy, while the Second(as phospholipids) renew the structural integrity of cellular membranes (Jauncey and Ross 1982). Lipids, and specifically fatty acids, are the favored source of metabolic energy in fish, especiallymarine fish, as evidenced by the very high oil levels (more than 20% of the wet weight) that can be achieved by fish such as capelin and herring. Fatty acids not only are the major source of metabolic energy in fish for growth from the egg to the adult fish (Tocheret al., 1985a), but also arethe major source of metabolic energy for reproduction (Henderson et al., 1984b; Sargent et al., 1989). Indeed, as pointed out elsewhere (Sargentet al., 1995a). It has been known that common carp can utilize lipids as an effective dietary energy source,fats present a double edged sword: those which are saturated, often

14

Chapter Two

Literature Review

coming from animals, have a lower digestibility, whereas unsaturated fats may be easy to digest, but present the danger of oxidation and ultimately feed spoilageto counteract this spoilage, antioxidants are frequently added to fish diets in storage.Fats carry out many important functions in the body,they help the body absorb certain vitamins, produce hormones and build body tissues,fats are important for the development of the brain and the central nervous system,adequate fat is an important part of a healthy diet that meets individual energy and nutrient needs and takes into account appropriate levels of physical activity, most of the fat in the diet should come from unsaturated fats, especially from seeds, nuts and fatty fish that provide omega-3 fatty acids,small amounts should come from saturated fatty acids (less than 10%) ofcalories in the diet for adults and 8% for children), trans fats and foods containing trans fats (partially hydrogenated oil) should be avoided or consumed as little as possible (less than 1% of calories)(Takeuchi et al., 2002).

2-4-5-Vitamins requirements: Other dietary nutrient requirement that are essential in the diets, which are required in small quantities for normal growth, reproduction and good health are vitamins (Jauncey 2000; Lim and Webster 2006; NRC 2011). In general, vitamins are divided into two distinct groups, which are water soluble vitamins (B-complex vitamins, inositol, Choline and vitamin C) and fat soluble vitamins (vitamin A, D, E and K) ( Lim and Webster 2006; NRC 2011). Vitamin deprivation in diets will cause morphological changes and functional alteration as reported in numerous fish species (Lim and Webster 2006; NRC 2011). They concluded that many factors may impact vitamin requirements of finfish including carp and tilapia, such as fish size, water temperature and diet composition. Takeuchi et al. (2002) stated

15

Chapter Two

Literature Review

that juvenile and adult carp do not require vitamin C because they can synthesize ascorbic acid from D-glucose. 2-4-6-Mineral requirements There is small amount of information are available on the mineral requirement of fish and crustaceans. Minerals play a vital role in the body of aquatic animals for tissue formation and basic metabolic functions including osmoregulation, proper function of muscles and nerves and acid-base balance (Lim and Webster 2006; NRC 2011). Fish are able to directly absorb minerals through their gills and skin which are required in normal metabolism from their external aquatic environment, thus not all elements that are used in metabolism are required in the fish diet (Lovell 1989; De Silva and Anderson 1995; Pillay and Kutty 2005; NRC 2011). 2-5-Fatty acids structure The importance of fatty acidsincludes the risk reduction of self-activated reactions in the body (the interactions of immune cells attack healthy tissues of the body).The human body needs the essential fatty acids to from the cell walls and decides the extent of human health and immunity. Without such acids the process of cell renewal and protect the body from disease cannot be happened (NRC, 2011). The lack of essential fatty acids includes deficiency appearance at any place and in any form as long as it is related to weak immune, disorder and regulates inflammation and its effects on the gastrointestinal includes constipation, swelling, indigestion and inflammation of the gastrointestinal tract or food allergies (Swift, 1993; Billard, 1995; Takeuchi et al., 2002).

16

Chapter Two

Literature Review

When hydrogen added to change their characteristics the unsaturated fatty acid is divided into two groups: Montounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA), monounsaturated fatty acids with double bond between one pair of atoms carbon, or fatty acids polyunsaturated with double bond between more than one pair of carbon atoms (Chou and Shiau, 1996). 2-5-1-Omega-3 fatty acids Omega-3 polyunsaturated fatty acids (PUFAs) are long chain PUFAs found in plants and marine sources such as fish, mussel, oyster, shrimp but primarily cold water fish (Friedman & Moe, 2006) but also exist in a wide range of plant products such as nuts, especially English walnuts, seeds, namely sesame (Namiki, 2007), linseed oil and vegetable oils such as soybean and canola besides olive (Whelan & Rust, 2006). omega-3 fatty acids, unlike saturated fatty acids, have been associated with various health benefits relating to treatment of rheumatoid arthritis (Rennieetal., 2003)and coronary artery disease (Freeman, 2000) whilst improving blood pressure control and preserve renal function even in hypertensive heart transplant recipients(Holm et al., 2001). The effects of omega-3 on various cancers and also on other clinical disorders including oedema, rheumatoid arthritis, cardiovascular disease and others, is closely related to their metabolism. Although most studies define omega-3 as lowering cholesterol and LDL as reducers Schackyet al., (1999). In another experiment conducted by Leeet al., (1967) on the group omega-3 to test its impact on the growth rate of rainbow trout larvae Salmogairdneri,using five different diets in a content of group omega-3, it is realized that the diets containing corn oil was supported the growth rates in the fish with the advent of the proportion of mortality compared with diets containing salmon oil.

17

Chapter Two

Literature Review

2-5-2 Omega-6 fatty acids Omega-6 fatty acid is also a polyunsaturated fatty acid, Omega-6 containing fats and oils are essential for life and recently their necessity for health has been reconfirmed (Willett, 2007 and Lands, 2008). Consumption of certain foods such as vegetable oils, nuts and some fish are encouraged because of their healthful qualities, including fat content, and their potential to reduce the risk of disease, including heart disease and cancer. Some scientists believe that diets high in omega-6 fats relative to omega-3 fats may be associated with increased prevalence of chronic diseases, including heart disease and certain cancers (Wijendran and Hayes, 2006). Omega-6 dietary factors can play a major role in the risk of heart disease, numerous studies have shown how consumption of certain foods may impact total blood cholesterol, including low-density lipoprotein (LDL) or “bad cholesterol,” high-density lipoprotein (HDL) or “good cholesterol,” as well as triglycerides Clinical trials help to clarify the mechanisms related to omega-6 fats and cholesterol, reduction, and new studies continue to add insights (Lands, 2008). However, omega-6 fats also may play a role in breast and pancreatic cancer incidence, dietary consumption of omega-6 fats was correlated with lower incidence of breast cancer (Rissanen, et al.,2003).

18

Chapter Two

Literature Review

2-6-Safflower oil: Traditionally, safflower has been grown for centuries from China to the Mediterranean region and along the Nile valley up to Ethiopia (Weiss, 1971). Presently it is grown commercially in India, the U.S.A, Mexico, Ethiopia, Kazakhstan, Australia, Argentina, Uzbekistan, China, and the Russian Federation. Pakistan, Spain, Turkey, Canada, Iran, and Israel also grow safflower to a limited extent. Safflower acreage and production around the world have witnessed wide fluctuations in the past. However, recently interest in safflower flowersas a source of color for use in food is gaining importance owing to a recent ban on the use ofsynthetic colors in food in the European countries and elsewhere. Also, flowers are reported tohave medicinal properties to cure several chronic diseases, like hypertension, cardiovasculardiseases, arthritis, spondylosis, and sterility in both men and women. Detailed information aboutclinical uses of safflower flowers has been given in the monograph on safflower written by Li andMundel (1996). Safflower oil is nutritionally similar to olive oil, is containing high levels of linoleic and oleic acid. The monounsaturated fatty acid like oleic acid is alsoknown to reduce low-density lipoprotein (LDL; bad cholesterol) without affecting highdensity lipoprotein (HDL; good cholesterol) in blood (Smith, 1996). Safflower oil is highly stable, and its consistency remains the same at low temperatures, thereby making it suitable for application in frozen/chilled foods (Weiss, 1971). Safflower oil is also better suited to hydrogenation for margarine reduction than soy or canola oils (Kleingarten, 1993). Safflower oil is nonallergenic, and therefore suitable in inject able medications (Smith, 1996). Safflower oil is a preferred for the paint and varnish industry owing to its specific properties of absence of linolenic acid, presenceof high linoleic acid, low color values, low free fatty acids, low unsaponifiables, and no wax, which make the quality in paints, alkyd resins, and

19

Chapter Two

Literature Review

connection beyond comparison (Smith, 1996). Table (1) show the fatty acid composition of safflower oil. 2-7-Linseed oil Since the 15th century, linseed oil has been extensivelyused in varnishes and oilbased house paints (LazzariandChiantore, 1999). An herbaceous plant, Linumusitatissimum, and linseedalso called flax(Linumusitatissimum) produces seeds which are oval and flattened in shape, 4–6 cm long, pale to dark brown and shiny. This occurs as a consequence of high content of glycerol esters (also known as glycerides or triacylglycerols) of linolenic acid in linseed oil. There are four varieties of linseed oil that are available in the market, including raw, boiled, stand and refined linseed oils.Characteristics of linseed oil composition and its structure of linseed oil,as linseed is grown in several geographical areas, includes Europe, North and South America (especially Argentina) and Asia (especially India).The linseed oil pressed from the seeds displays natural variation in composition that reflects growing, agronomic and environmental conditions (Gunstone, 1996). In particular, climate affects the abundance of the unsaturated fatty acid in the oil, the colder the climate, the higher the iodine value of oil or the degree of its unsaturation (Fjällströmet al., 2002). Iodine value is normally expressed in terms of grams of iodine added per 100 grams of oil. The oil consists almost exclusively of the esters of glycerol (C3 alcohol with three hydroxyl groups, one on each carbon atom) and five fatty acids, two of them are saturated, palmitic and stearic, and three are unsaturated,oleic, linoleic and linolenic, exhibiting one, two andthree double

bonds,

respectively.

The

two

and

three

double

bonds

are

nonconjugated,being separated from each other by CH2 groups. The minor components that may also be present are monoacylglycerols (monoesters of glycerol), diacylglycerols (diesters of glycerol) and free fatty acids (Gunstone,

20

Chapter Two

Literature Review

1996). Although linseed oil contains around 60% of linolenic acid, the most unsaturated fatty acid, this acid occurs only in small amounts, usually below 1% in other oil types. The notable exceptions are soybean and rape oil, with 8% and 7% linolenic content, respectively (Gunstone, 1996). This high level of linolenic acid in linseed oil affects the drying property of the oil, making it particularly suitable in formulations of drying alkyd paints. The natural unsaturated fatty acids exhibit one or more carbon double bonds with configuration starting at the location of the ninth carbon atom (Roberfroid and Calderon, 1995). Table (1) show the fatty acid composition of linseed oil.

21

Chapter Two

Literature Review

Table 1: Fatty acid composition for linseed oil and safflower oil as a source of on selected fatty acids (i.e; omega- 3 and omega- 6) (g/100g of total fatty acids) (Ruyter et al. 2000)

Fatty acid

Linseed oil as a source Safflower oil as a of Omega-3

source of Omega-6

%

%

Myristic acid

0.142

2.1

Palmitic acid

6.28

7.0

Stearic acid

2.72

3.9

Oleic acid

21.58

20.5

Linoleic acid

20.06

66.2

α-Linolenic acid

48.57

0.4

S.F.A

9.14

9

MUFA

21.58

12

PUFA

68.63

75

22

CHAPTER THREE MATERIALS AND METHOD 3.1. Experimental fish The study was conducted at the Faculty of Agricultural Sciences, Sulaimani University. A total of 126 fingerlings of common carp C. carpio L. (Figure 1) were obtained a local fish supplier located in Babel. Fish total length varied ranged between12-14 and weight ranged between 35-40 g, with no visible signs of disease or morbidity. The fish were supplied with continuous oxygen. The fish were acclimated to laboratory conditions for 21 days before starting the experimental trials.

Figure 1: The reared fish common carp Cyprinus carpio

24

Chapter Three

Materials and Methods

3.2. Experimental system The system was contains of 21tanks (100L tap water/ tank) were used in the current study (Figure 2). Each tank was provided with a proper continuous aeration. Seven different treatments were used in the experiment with three replicates; each treatment consisted of six fish. Water was replaced every 24 h after feeding in order to maintain a healthy environment with enough oxygen. Fish were fed twice daily with formulated fish feed (3% of initial body weight per a day having 30% of crude protein).

A

B

Figure 2: Experimental design Control, LN1.5, LN2, LN2.5, SFO1.5, SFO2 and SFO2.5 represent the seven treatments in this trial (A and B).

25

Chapter Three

Materials and Methods

3-3- Water quality Water quality parameters of different treatments in different replicates were measured during the study. The water temperature was measured ranged between 27-33˚C. Dissolved oxygen was measured ranged between 6.75-7.57 mg L-1. The pH value was measured ranged between 6.01 - 7.22, used HQ40d Dual-Input pH and DO meter. 3-4- Design of the experimental study 126 common carp

T1

T2

T3

T4

T5

T6

T7

0% oil

1.5% Linseed oil

2% Linseed oil

2.5% Linseed oil

1.5% Safflower

2% Safflower

2.5% Safflower

oil

oil

oil

Parameters

Chemical composition(%) For experimental fish

-Crude protein - Ether extract -Ash

Blood parameters -R.B.C. -W.B.C. -P.C.V. -Hb.

Biological parameters

-Liver index -Gonadosomatic index

- MCH , MCV , GRAN Lymph,, Mid and PLT

- Cholesterol Figure 3: Shows the experimental design of the study.

26

Growth - Weight increase -R.G.W. -S.G.R. -S.R. -F.C.R. -F.E.C -P.E.R

Chapter Three

Materials and Methods

3-5- Diet formulation Experimental diets were formulated with fishmeal, yellow corn, barley, wheat bran, soybean, salt and vitamins, fat and plant oil (linseed oil and safflower oil) as a source of omega-3 and omega-6 (Table 2), the chemical composition of the experimental diets (Table 3). The ingredients were mixed with water to obtain dough. Then, the dough was passed through an electrical mincer for pelleting by using Kenwood Multi-processors. The pellets were dried at room temperature for a few days and crushed to yield fine particles. The fish were fed 2 times a day, at 9:00 am and at 2:00 pm. Feeding rate started with 5% of biomass then the accurate feeding rate was determined to be 3% by second week depending on satiation level. The feeding amount was then recalculated according to weekly weights. The feeding trial continued for 12 weeks.

27

Chapter Three

Materials and Methods

Table 2: formulation of experimental diets Treatment

T1(con)

T2 1.5%

T3 2%

T4 2.5%

T5 1.5%

T6 2%

T7 2.5%

0% oil

linseed oil

linseed oil

linseed oil

Safflower

Safflower

Safflower

oil

oil

oil

Pellet content

Fishmeal

25

25

25

25

25

25

25

Yellow corn

6.5

5.5

5.5

5.5

5.5

5.5

5.5

Barley

10

10

10

10

10

10

10

Wheat bran

20

20

20

20

20

20

20

Salt& Vitamin

2

2

2

2

2

2

2

0

1.5

2

2.5

0

0

0

0

0

0

0

1.5

2

2.5

(Omega-6 source) Soybean

35

35

35

35

35

35

35

Fat

1.5

1

0.5

0

1

0.5

0

Linseed oil (Omega-3 source) Safflower oil

28

Chapter Three

Materials and Methods

Table 3: Chemical composition (%) of with diets Treatments

Control

LN 1.5

LN 2

LN 2.5

SFO 1.5

SFO 2

SFO 2.5

Crud protein%

31.59±0.51

32.29±0.33

32.36±0.31

31.96±0.45

31.32±0.66

31.40±0.35

30.72±0.63

Fiber%

3.89±0.05

4.04±0.02

3.90±0.04

4.33±0.02

3.87±0.01

3.76±0.04

3.81±0.07

Ether extract%

4.95±0.05

4.84±0.1

4.88±0.06

4.21±0.02

4.46±0.05

5.05±0.04

5.08±0.03

Carbohydrate%

51.31±0.76

50.61±0.67

50.34±0.77

51.02±0.65

51.58±0.72

51.33±0.74

52.16±0.78

Ash%

8.26±0.03

8.22±0.04

8.52±0.03

8.48±0.03

8.76±0.05

8.46±0.02

8.28±0.03

%

3-6- Growth parameters The individual body weight (g) and total body length (cm) of fish for all treatments (126 fish) were measured every two weeks. The feed consumption of each treatment was recorded and readjusted according to the obtained biomass at the treatment for two week. The average body weight gain (g/fish) was estimated according to the following equations according to Uten (1978): -Weight increase (g/fish) = Final weight (g) – Initial weight (g) -Daily weight increase (g/fish/day) = Weight increase (g/fish) / Period (day) -Relative Growth Rate (RGR %) = Final weight (g) – Initial weight (g) / Initial weight (g) × 100 (Brown, 1957) -Specific Growth Rate (SGR) = (In W1 – In W0) / T In: Natural logarithm

29

Chapter Three

Materials and Methods

W1: Final weight; W0: Initial weight; T: time between W1 and W0 -Feed conversion Ratio (FCR) = Total feed intake (g) / Total weight Gain (g/fish) -Feed Efficiency Ratio (FEC) = Total weight gain (g/fish) / Total feed intake × 100 -Protein Efficiency Ratio (PER) = Total wet weight gain (g/fish) / protein Intake (g) -Survival rate (%) = Fish No. at the end period / Fish No. at the initial period ×100. 3-7 Biological parameters After the blood samples collected, all the fish samples were sacrificed and soon the abdominal cavity was opened immediately to remove, gonads and liver to be weighed at once. The gonad and liver index were calculated according to Lagler (1956), as follow: -Gonadosomatic Index (GSI) % = Gonads weight (g)/ Body weight (g) x100 -Liver index % = liver weight (g)/body weight (g) x 100 3-8 Haematological examination Blood collection was done via cardiac puncture technique, blood was withdrawn directly from tail at the end of experiment using sterile disposable syringe (needle gauge 23). The blood was transferred into a tube containing EDTA solution for hematological tests. 3-8-1 Red Blood Cells Counting (RBC) Modified Dices Fluid was followed to dilute blood to count RBCs by haemocytometer chamber, in the presence of cover slide, special pipette for dilution fluid, and microscope. Blood was filled up to level 0.5, while dilution

30

Chapter Three

Materials and Methods

fluid was filled up to the level 101of the pipette then mixed in 8 figure movements. First few drops were neglected. A drop was dripped on the covered haemocytometer then covered. Five squares were used to count the number of red blood cells Dacie and Lewis (1995). 3-8-2: Hemoglobin Test (Hb) • Principle:The

determination

of

hemoglobin

which

is

converted

into

cyanomethemoglobin under the influence of potassium ferricyanide and potassium cyanide Coles (1986). • Reagent:Drapkin's reagent Haemoglobin standard 15 g/dl. Procedure • 5.0 ml 0f Drapkin’s solution was mixed with 20 µl of blood sample • The mixture was allowed to stand for 5minuts. • Haemoglobin concentration was determined spectrophotometrically under wave length of visible light of 540 nm. Reagent Amount Sample Drapkin's Reagent

5 ml

20µl

3.8.3 Packed Cell Volume (PCV %) Packed cell volume was measured by using microhematocrit capillary tubes, after being filled with blood up to2/3 of their length, The other end of the tubes was plugged by clay and set in microhematocrit centrifuge for (5) minutes, speed

31

Chapter Three

Materials and Methods

1500 round/minute, then read by using micro-hematocrit reader according to the method mentioned by Archer et al., 1998). 3.8.4 White Blood Cells Counting (WBC) Modified Dices Fluid was used to dilute blood to count WBC's by haemocytometer method as mentioned before in RBC's counting, the only difference was the type of pipette that used to fill up dilution fluid. WBC's were counted in four squares (squares of 4 corners) (Dacie and Lewis, 1995). (Mean Corpuscular Hemoglobin; pg); (Mean Corpuscular Volume; fl); 9

(Granulocyte; %); (Lymphocyte; %); (Monocyte; %); (Platelet; 10 cells/l). 3.8.5 Measuring the concentration of cholesterol Method followed by Nilain (1974) was applied to measure the concentration of cholesterol using a special reagent Company Giesse Haly, Roma as follows: 1. Three samples were prepared; the first is the officer blank sample, the second and third samples are the standard and examined samples, respectively. 2. Ten micro liter of distilled water was kept in the control tube and 10 micro liter of standard material in the standard tube and 10 micro liter of serum in the Sample tube. 3. Total volume of 1.5 ml of reagent solution was add to each tube and mixed well , then placed in a water bath for 5 minutes at 37 ˚C 4. The absorbance result was recorded using a spectrophotometer at the optical wavelength of 500 nanometer after resetting the device using the control sample. Cholesterol concentration was calculated according to the following Equation: Sample - Reagent blank Cholesterol Conc. (mg / dI) = ‫ــــــــــــــــــــــــــــــــــــــــ‬ Standard - Reagent blank

32

× Stan. Conc.

Chapter Three

Materials and Methods

3-9 Chemical composition Chemical analysis of the fish muscles conducted les was c for crude protein %, ether extract % ash % and fiber % according to Association of Official Analytical chemists (AOAC, 2002), all fish samples of different treatments were utilized before and after experiment. 3-9-1 Crude protein Crude protein was calculated from sample nitrogen content was determined using Kjeldahl apparatus (Gerhardt Kjeldatherm method, N % x 6.25). 3-9-2 Ether extract Crude lipid using ether extraction in multi-unit extraction Soxtec apparatus (dicloromethane extraction by Soxlhet method), the sample was put in thimble to extract cellulose, diethyl ether was added for 8 hours with heating, then the solvent was evaporated in electric oven at a temperature of 105 ˚ C for half an hour, and then the sample was weighed to determine the percentage of fat before and after extraction. 3-9-3 Ash Ash Content was analysed using a muffle furnace (incineration at 550 ºC for 12 h).

33

Chapter Three

Materials and Methods

3-10 Statistical analysis Results were expressed as Mean± SE. Statistical analysis of data was performed on the basis of one- way analysis of variance (ANOVA).Treatments differences were determined using least significant difference (LSD). The model of analysis was as follows: Yij = µ + Ti + Eij µ = The overall mean. Ti = The effect of treatment. Eij= The random error.

34

CHAPTER FOUR Results and Discussions 4.1. Water quality: The water quality displayed in table (4) Dissolved oxygen mg/L and pH showed that there were no a significant difference (P>0.05) of other treatment, but in Temperature c˚ There were significant differences (P≤ 0.05) between Cont. and LN 2%, LN 2.5%, SFO 1.5% and SFO 2%. Table 4: Water parameter during the experimental trails. Treatment

Temperature C˚

Dissolved

oxygen

pH

mg/L Cont.

27±0. 02b

7.32 ± 0.20a

6.82 ± 0.31a

LN 1.5%

29±0.10ab

7.01±0.11a

6.03±0.20a

LN 2%

30±0.22a

7.41±0.21a

6.34±0.33a

LN 2.5%

33±0.21a

6.75±0.15a

7.01±0.25a

SFO 1.5%

30±0.22a

7.22±0.37a

6.01±0.22a

SFO 2%

32±0.24a

6.78±0.30a

7.22±0.23a

SFO 2.5%

28±0.04b

7.57±0.34a

6.65±0.21a

Data are presented as mean ± S.D. Data in the same row with different superscript are significantly different (P<0.05).

Dissolve oxygen values ranged between 6.75-7.57 mg/l and pH values ranged between 7.5-8.5. The water temperature 27-33°C was not suitable for the growth of common carp. The common carp prefer moderate temperatures and any increasing caused the reduction in their appetite to food intake, causing a negative impact on the growth (Lagler et al., 1977).

36

Chapter Four

Results and Discussions

Wu et al., (1993) found that a 7.0-8.0 pH range produced the best functioning of many physiological responses and enzyme activities in the carp. The differences in growth performance among treatments in the current study may be due to food availability and competition for food (Albaster and Lioyd, 1982). Common carp, euriterm species, tolerates a temperature range 32°C, which makes it the most widespread species in freshwaters. When water temperature drops below 10°C, the intensity of feeding common carp is reduced, and at temperatures below 7°C the feeding process stops. For optimal thermal comfort spread growth is faster, more efficient feed conversion and increased resistance to disease (Masseria et al., 1999). 4.2. Growth Performances 4.2.1. Weight Gain Growth performance and feed utilization displayed in (Table 5). The initial average body weights of common carp (Cyprinus carpio) of this experimental were ranged between 35-40g. After 12 weeks of feeding trail on safflower and linseed oil there were significant differences between fish treatments. The average primary weight in Table (5) was ranged between 35.61 in LN 2% to 40.45g in LN 1.5%, while these ranges were increased at the end of experiment between 59.38 in LN 2% to 77.69g in SFO 2%. The final weight of fish fed SFO 2% and SFO 2.5% showed that there were a significant difference (P≤ 0.05) compared to control (T1), and other treatment of LN 1.5%, LN 2%, LN 2.5%, and SFO 1.5%. Fish fed SFO diet had the highest weight gain which were significantly different (P ≤ 0.05) from those fish receiving the other diets, followed by fish on LN diet.

37

Chapter Four

Results and Discussions

The result of statistical analysis showed significant increase (P≤0.05) in T6 compared to other treatments; SFO 2.5% showed no significant variations compared to Control and LN 2.5%. The final biomass in table (5) was ranged between 1068.84 g in LN 2% to 1368.42 g in SFO 2%. Increasing total weight of experimental fish in table (5) was ranged between (427. 86g and 710.46g) in LN 2% and in SFO 2% respectively. The specific growth rates (SGR %) of C. carpio fed on SFO and LN diets were 0.68%, 0.64%, 0.29%, 0.27%, 0.24%, 0.23% and 0.22%, in SFO 2%,SFO 2.5%,LN 2.5%, LN 2%,SFO 1.5%, Control and LN 1.5% respectively (Table 5). There were significant differences (P≤ 0.05) between experimental treatments, SFO 2% and SFO 2.5% were showed significant increase of SGR% compared to fish fed on control diets and to fish fed on LN (LN 1.5%, LN 2% and LN 2.5%). The results of Relative Growth Rates (RGR) showed that there was lowest value in control reached (14%), while the highest value was found in SFO 2% (36%). The statistical analysis showed that there were no significant different (P>0.05) between control LN 1.5% and LN 2%. The survival rate showed that the control treatment reached 100%, while the survival

rates

for

other

dietary

38

treatments

were

88%.

Table 5: Effect of safflower oil and linseed oil on growth performance of common carp 12 weeks fed on experimental diets (mean ±SE). Treatme nts

Average primary weight (g)

Average final weight (g)

Primary biomass (g)

Final biomass (g)

Control

37.33±2.60 b

65.42±4.44 c

671.94

1177.38

Increasin g total weight (g) 505.44

Specific growth rate SGR (%) 0.23±0.08d

Relative growth rate RGR ( %) 14±0.32d

Survival rate (%)

100.00 LN 1.5%

40.45±2.22 a

71.11±5.78 b

782.10

1279.98

497.88

0.22±0.05d

15±0.09d

LN 2%

35.61±1.88 bc

59.38±5.99 e

640.98

1068.84

427.86

0.27±0.04c

15±0.04d

88.88

88.88 LN 2.5%

37.22±4.33 b

66.64±7.12 c

669.96

1199.52

529.56

0.29±0.04c

17±0.04c

SFO 1.5%

39.44±3.67 a

63.27±5.55 d

709.92

1138.86

428.94

0.24±0.06d

16±0.06cd

SFO 2%

38.44±3.76 ab

77.69±3.67 a

687.96

1398.42

710.46

0.68±0.03a

36±0.07a

88.88

SFO 2.5%

35.88±2.22 bc

68.92±3.90 c

645.84

1240.56

594.72

0.64±0.04b

30±0.09b

88.88

88.88

88.88

Data are presented as mean ± S.D. Data in the same row with different superscript are significantly different (P<0.05).

Growth rate of fish is a good quantitative parameter in measuring the amount of energy available to an organism for various functions, furthermore, measurement of the growth can often be used to provide an index of physiological status and growth performance, based on the growth performance, safflower oil was better utilized by C. carpio than linseed oil, among plant oils, oil rich in omega-6 fatty acid (safflower oil) promoted better growth of C. carpio than those containing high levels of omega-3 (linseed oil) which performed very poorly. A similar observation was found by Guary et al. (1976) for Penaeus japonicus. This

40

Chapter Four

Results and Discussions

suggests that omega-6 had better nutritional value for C. carpio than omega-3 as has been reported by Kanazawa et al. (1980) for Penaeus japonicus. Wilson et al. (1987) pointed out that channel catfish grew better with diets containing sunflower oil than those with linseed oils rich in either omega-6 or omega-3. It has previously been reported that omega-6 PUFA, including both omega-6 and HUFAs, was required for optimal growth and prevention of signs of EFA deficiency in rainbow trout (New, 1999). Several studies are confirming similar results found in some experiments with vegetable oils (canola oil, soybean oil, linseed oil; rapeseed oil and palm oil) in different fish species (brook charr, trout, atlantic salmon and European sea bass), it seems that the use of various safflower oil has improved the nutritional value of the experimental diets, in the present study, no adverse effect was observed on the growth performance of common carp, the replacement of fish oil by some vegetable oils in fish diets have been studied in turbot (Regost et al., 2003), gilthead sea bream (Caballero et al., 2002) and sea bass (Izquierdo et al., 2003) without negative effect on the growth performances of fish. which may be due to the containment of the omega-6 (Guillou et al., 1995; Regost et al. 2003 and Richard et al., 2006). The results are similar with the result obtained by Tidwell and Robinette (1990) who added a palm oil (1.5%) as a source for the omega-6 and fish oil (1%) as a source for omega-3 in diets of catfish (Ictalurus punctatus), omega-3 and omega-6 had made valuable and similar increases in total weight and in the daily weight. Thus, in the present study, no adverse effect was observed on the growth performance of common carp fed diet contained safflower oil.

41

Chapter Four

Results and Discussions

Bell et al., (1994) and Regost et al., (2003) were reported that there were no negative effect on the growth of fish fed diets contained safflower oil and linseed oil. Also investigated three diets containing fish oil, safflower oil and linseed oil for juvenile turbot, reported that there were no differences in final weight. Regost et al. (2003) pointed out three diets, containing fish oil, soybean oil and linseed oil, for marketable size turbot. They reported the high growth rate of turbot; however the incorporation of vegetable oils in the diets resulted in a slight decrease in growth as compared to those fed on fish oil-based diet, On the contrary, the weight gain and SGR of SFO treatment were higher than in LN treatment. Okoye and Eyo (2002) pointed out that values for the specific growth rate of the qualitative amounted to 0.012 and 0.008 and 0.007 and 0.005% g / day when used 1% of different oily sources. A Shea butter oil and pork lard as sources for arachidonic acid and palm oil as a source for linoleic acid fixed rate of 1% in the diets of fingerlings (Clarias anguillaris), noted that the criteria for growth rates obtained from this study is higher than the results of the research reviewed may be due to the difference of fish species, the components of the pellets and the different oils used, it is correspondence to the results of our study. Guary et al. (1976) the growth and survival of fingerlings of Clarias anguillaris (1.28 g/ fish) in the experiment lasted 56 days, the growth rates were (3.83) and (2.64) and (1.66) and 1.25 g/ fish respectively. The result showed clearly the importance of oil in the diets of fish. Rodolfo et al. (2006) also studied replacement of fish oil with linseed oil rich in linoliec acid in fish rainbow trout (Oncorhynchus myksis) in the experiment lasted 231 days. Four experimental diets containing increased levels of linseed oil ( 0 and 6, 12 and 13%) were used as a replacement for fish oil, , and the increased 42

Chapter Four

Results and Discussions

daily weight measure was more than 0.3 g/ day and the feed conversion ratio was less than 1.5. The results of present study are similar to study by Al-Ashab (2011) on the growth of common carp, different ratio of sunflower oil and corn oil as omega-6 sources in the diets.

43

Chapter Four

Results and Discussions

Takeuchi (1996) conducted that the grass carp (Ctenopharyngodon idella) fed on diets without essential fatty acids; He found symptoms of deficiencies, including curvature of the spine with low growth rates. Glencross et al. (2002) showed the balance effect fatty acids omega-3and omega-6 and from the growth of Penaeus monodon prawn they found that the optimal ratio of omega-3 and omega-6 was 2.5, the results of their study showed that overlap omega-3 and omega-6 are important factors to feed prawn to support growth. Karalazos et al. (2007) noted an improvement in the values of relative growth rate and qualitative adding of safflower oil as a source of linoliec acid levels 0, 3 and 6% in the Atlantic salmon Salmo salar L. done at low temperature in water using low and high protein diets. They scored on the sequence values amounted to (76, 79 and 82 %) for the relative growth rate and (0.49, 0.52 and 0.56) for the qualitative growth rate when used a high protein diet (38.7 -39.1 %). While the lowprotein diet (34.4 %) value ( 66, 69, and 73%) for the relative growth rate 0.50, 0.52 and 0.53 g/ day for qualitative growth rate, consecutively. Gumus and Fatime (2010) reported that using of fish oil as a source for omega-3 in the diets of young tilapia (Oreochromis niloticus) is one of the main ingredients in the diets of young fish. It refers to its vital role in supporting and promoting growth rates, as improved the values at level of (0.9% - 1.1%)of the fish oil with the use of fish meal as a source of protein in the diets. It has also been reported that partial replacement of fish oil by vegetable oils such as rapeseed, soybean, linseed or palm oils in fish feeds has no negative impacts on survival of Atlantic salmon (Salmo salar) (Rosenlund et al., 2001). Copeman (2002) demonstrated that eating fatty acids are not enough guarantees to build the immune cells solid but also work more effectively. The results reported

44

Chapter Four

Results and Discussions

above are consistent with the results of cur vent study in terms of the role of essential fatty acids from a family omega-6 in supporting the growth rates depends on species and source of the fish omega-6 as well as the components of experience pellets. Comparing the results of the current study with a study done by Nick et al. (1991) in replacing fish oil with palm and sunflower oils in the diets of running African catfish( Clarius gariepinus). They found that a complete replacement of fish oil had improved the growth of fish. In fact no differences between palm oil and safflower oil suggest that omega-6 has a limited impact of essential fatty acid on these species. Physiological functions that could be played by fatty acid in the processes of representative in the body depending on the length of the fatty acid chain, the number of double bonds and the location of their first link for a treatment representation. As such the recent very great importance for the nature and physiological characteristics and carried out by the body and that of which can result in fatty acid role in supporting and contemporizing relative qualitative growth rates. Through the fact that fatty acids are involved in the installation of lipid phosphorylation in cell membranes, which affects in controlling metabolic processes and thus supports the growth rates (Yildirim-Aksoy et al.,2007 ). Skalli and Robin (2004) noted in an experiment conducted for a period of 12 weeks on the larvae of European fish bass Dicentrar chuslabrax fed on a different level of omega-3 of the pellets. The lowest level of omega-3 (0.2%) showed a significant decrease in the growth, while the rate of growth rate improved when upgrading omega-3 to (0.7%), and become better at the level (1.9%). The results were compatible with what was referred by Lee et al. (1993) when they studied the effect of HPUFA omega-3 variably (0.5, 1, 1.5, 2, 2.5 and 3%) on growth and body composition in rocky fish (Sebastes schlegeli). The increase in 45

Chapter Four

Results and Discussions

the values of relative growth rates was above the increase ratios being the best value at the level of 2.5%, which is comparable to the results of current research and the difference that happened may be due to the difference in fish and the conditions of the experiment. The optimum growth and feed efficiency was obtained in juvenile common carp (Cyprinus carpio) when fed diets that supplied 1% of omega-3 and omega-6 fatty acids (Takeuchi and Watanabe, 1977). Growth performance for tilapia hybrid was considerably raised by feeding cod liver oil compared to corn oil, for optimum growth of tilapia are require omega-3 fatty acid (Chou and Shiau,1996). YU and Sinnhuber (1972) noticed the impact of linolenic acid and Docosahexaenoic acid on the growth and composition of fatty acids for fish rainbow trout (Salmo gairdneri). The highest growth rate was given when added 1% of former acids compared to non acid fed which caused symptoms trauma or stroke, poor appetite and low growth rates, the analysis also showed fast conversion of α-linolienic acid in rainbow trout (Salmo gairdneri). 4.2.2. Food Conversion Ratio Food conversion ratio was ranged between 1.33 - 1.90(g /fish) in cont. and SFO 2%, respectively. Food efficiency ratio was ranged from 20.46% - 53.42% in LN 1.5% and cont. respectively, while the values of protein efficiency ratio varied from 1.14% in SFO 2% to 1.39% in SFO 1.5%. The food efficiency ratio in fish fed on SFO 2% and control there were higher significantly different (P≤ 0.05) than other treatments. The FCR and PER in the experimental diets showed no significant differences between each other treatments.

46

Chapter Four

Results and Discussions

Table 6: Feed utilization; FCR, PER and FER of common carp after 12 weeks feeding Experimental diets (±SE) in C. carpio L. Treatments

Feed Conversion Feed Efficiency Ratio Protein Efficiency Ratio (FCR)(g/fish) (FER)% Ratio (PER)%

Cont.

1.33 ±0.40a

53.42±0.01a

1.26 ±0.11a

LN 1.5%

1.68 ±0.15a

20.46±0.12d

1.31±0.12a

LN 2%

1.69 ±0.75a

33.48±0.56c

1.38±0.25a

LN 2.5%

1.59± 0.23a

22.58±0.49d

1.20 ±0.25a

SFO 1.5%

1.71± 0.71a

49.60±0.36a

1.39±0.35a

SFO 2%

1.90± 0.12a

41.17±0.89b

1.14±0.42a

SFO 2.5%

1.82± 0.18a

43.33± 0.34b

1.16±0.60a

Data are presented as mean ± S.D. Data in the same row with different superscript are significantly different (P<0.05).

Mraz et al. (2010) pointed the importance of sesame oil as a source for omega6 in the diets of common carp as improved the values of conversion rates and food efficiency ratio food at the levels of (0%, 0.8%, 1.2 %, 1.8 %) gained (2.98, 2.54, 2.06 and 2.61 g/fish) for feed conversion rates, and (0.56%, 0.62%, 0.74% and 0.59%) for the feed efficiency ratio. Catacutan and Coloso (1995) and Steffens (1996) both linseed and safflower oils share in securing an important part of the power requirements

for the fish and

using the protein feed for construction and growth, the oils added are contributed in transferring the dissolved nutrients, such as soluble vitamins, as the essential fatty acids, as the necessary sterols.FCR was not negatively affected using SFO in fed.

47

Chapter Four

Results and Discussions

Izquierdo et al., (2003) showed that it is possible to replace fish oil by safflower oil or linseed oil in the diets for carp, the results denoted the higher ability of carp to accept vegetable oils in comparison with other marine fish species such as gilthead sea bream (Sparus aurata). Beef tallow also performed out fish fed diets supplemented with safflower oil (high in omega-6). Palm oil contains both omega-3 and omega- 6 and is also the richest natural source tocopherols and tocotrienols which function as antioxidants. These offer beneficial effect to growth and flesh quality when fish is fed high levels of palm oil in their diets (Lim et al., 2001). The levels of omega-3 and omega-6 fatty acids are higher in vegetable oil than pork lard (Maynard et al., 1979) and probably palm oil in this experiment. Caballero et al. (2002) and Bell and Dick (2005) reported a reduced percentage of eicosapentanoic acid and docosapentaenoic acid of fish oil in the muscle of trout that were compared to fed diets containing vegetable oils. This result suggests the possible metabolic competition between linoleic acid and alphalinolenic acid since both fatty acids are substrates for the same enzymes. Furthermore, it was concluded that a high content of linoleic acid in soybean oil dietary might have inhibited the conversion of alpha-linolenic acid in linseed oil into the longer chain essential fatty acid in fish oil. Although fish cannot synthesize linolenic and linoleic polyunsaturated fatty acids, their body has the requirement of them which should be provided from exogenous dietary sources (ICAR, 2006). These fatty acids along with stearic acid, palmitic acid and oleic acid series are probably important for better growth of fish, higher weight gains and the deposition of tissue protein have been associated with increased dietary fats and oil in fish fresh water, therefore lipid quality should be provided in the diet of fish. The better performance of Heterobranchus longifilis fingerlings fed on soybean oil diets could be due to their preference of omega-3 48

Chapter Four

Results and Discussions

and omega- 6 fatty acids which are of significant amounts and in required ratio in soybean oil to meet up metabolic needs (Sarkar, 2002). Since all animals, including marine fish do not have the ability to configure the linoliec acid due to a lack of necessary enzymes for the composition must be provided from an external source (NRC, 1993). Different levels of feeding had improved the nutritional value of the diets in an experiment. This improvement could be due to the containment of oil, linseed oil, and safflower oil on the linoleic acid, as distinguished by appropriate quantity of which amounted to 20.06% and 66.2% (Regost et al., 2003). Lee et al. (1997) indicated that fish have the ability to digest and metabolism fat more effectively than some carbohydrates as an energy source, and perhaps the reason is the lipase enzyme is more effective than the amylase enzyme and oils have better effect on providing the protein for growth than carbohydrates. Castell et al. (1972) pointed out that the lack of essential fatty acids in the diets of rainbow trout (Salmo gairdneri) cause the low growth weakness in the rate of food conversion, increases the respiration rates and reduces the bit rate of hemoglobin as well as increasing the water content in the muscle. Stickney (1983) explained the importance of essential fatty acids from the treatment of omega-3 for rate and conversion and efficiency rate of diet high in tilapia. They reported that when fish oil and contents of high nutritional value prevent a shortage of essential fatty acids promoting growth which improves the conversion rates of food and food efficiency ratio, being consistent with the findings current research study. Adding linseed oil in the diets of fish shares to increase palatability diets of fish. This is what has been seen in the increase and accepted quantities of feed intake with increasing the levels of linseed oil, which may have contributed to the

49

Chapter Four

Results and Discussions

increase in protein intake. As noted by Karalazos et al. (2007) an increase in the values of protein intake with the addition of oil, linseed oil and fish oil in the diets of Atlantic salmon are coincided with the amounts of feed intake. This reflected positively on the values of the efficiency ratio of protein and it was confirmed by Kim and Lee (2004) who described that quantities increase of feed intake has enhanced affordable protein and efficiency ratio of protein when meet the nutritional needs of HPUFA omega-3 Paralichthys olivaceus. The proportion of protein efficiency was improved among fish fed on container pellets of flax oil and a source of omega-6. The reason is might be due to a decline in the rate of passage of food through the gut, allowing for good absorption of most of the food in the pellets, and the matching results of research carried out by Rodriguez et al.(1998 ), it stated that the long-chain fatty acids in linseed oil improves the ability of intestinal absorption and improves the immune response and contributes to weight gain, increases feed consumption and improve the efficiency of feed conversion and protein efficiency ratio which is due to the increase protein intake. Lee et al. (2003) indicated in experiment lasted ten weeks that food additives from liver oil, liver oil containing a high level of omega-3 of EPA, DHA. They may increase the weight and the proportion of food efficiency and efficiency ratio of protein in platichthys stellatus (1.9 g/fish). It suggested that omega-3 dietary is important for the development of the natural growth that recorded in the larvae and platichthys stellatus. They concluded that the needs of the omega-3 for stellar fish flounder amounted to 1.5% and the ratio is better to increase the weight and food efficiency ratio and protein efficiency ratio which is similar to the results obtained omega-3 in this study.

50

Chapter Four

Results and Discussions

4.3. Biological parameters Results presented in the Table (7) showed the lowest value of liver somatic index found in SFO 2.5% (1.60), while the highest value found in LN 2.5% reached 2.94. The result of statistical analysis showed no significant variations (P>0.05) between all treatments. Gonado somatic index ranged between (2.12 – 2.62) in Control and LN 2.5%, respectively. No significant variations (P>0.05) were found between all dietary treatments. Table 7: Effect of feeding trail of linseed oil and safflower oil on liver somatic index and gonado somatic index(± SE) in C. carpio. Treatments Liver somatic index Gonado somatic index Cont.

2.60±0.53a

2.12±0.34a

LN 1.5%

2.40±0.18 a

2.26±0.67a

LN 2%

2.80±0.38a

2.60±0.23a

LN 2.5%

2.94±0.25a

2.62±0.00a

SFO 1.5%

2.47±0.35a

2.32±0.54a

SFO 2%

1.97±0.04a

2.48±0.26a

SFO 2.5%

1.60±0.92a

2.18±0.28a

Data are presented as mean ± S.D. Data in the same row with different superscript are significantly different (P<0.05).

51

Chapter Four

Results and Discussions

Fatty acids are predominantly formed in liver from two-carbon body acetyl-Co enzyme through an action of cytosolic multi enzyme complex called fatty acid synthesis. Caballero et al. (2004) reported that the reduction of the dietary fatty acids due to the inclusion of vegetable oils in the diets tends to promote fat accumulation in the livers of fish. Genc et al. (2005) studied the effects of dietary SFO and LN on hepatic lipid of hybrid tilapia. Differences observed between samples of wild and farmed fish were statistically no significant (p > 0.05) of the liver. A high omega-3 and omega-6 content in farmed fish has also been reported by Alasalvar et al. (2002); Saglik et al. (2003) for Sea bass.

4.4. Hematological parameters 4.4.1 Hematology Hematological parameters are presented in Table (8). A comparison between treatments showed that RBC varied from 1.40 in LN 2.5% to 2.59 cells x 106µl in SFO 1.5%, with no significant variations at (P>0.05) between Cont., LN 1.5%, LN 2% and LN 2.5%, also at (P>0.05) between the treatments of SFO 1.5%,SFO 2% and SFO 2.5%. Same result was found in WBC count which ranged between 11.23 to 17.93 cellx103 µl in LN 2.5% and SFO 2%, respectively. Statistical analysis of RBC showed no significant variations between LN treatments compared to control diet (P>0.05). Hemoglobin concentration varied from 9.66 in LN 2.5% to 11.89 g/dl in SFO 2%. Results showed no significant variations (P>0.05) between SFO treatments, while high ratio of LN diet (LN 2.5%) showed a significant decrease (P≤0.05) compared to cont., LN 1.5% and LN 2%. Table (8) displayed a variation in PCV within treatments ranged from 24.33 in control to 31.55% in SFO 2%. Results of statistical analysis showed no significant 52

Chapter Four

Results and Discussions

variations between LN 2%, LN 2.5% and SFO 1.5%; and between SFO 2% and SFO 2.5% (P>0.05). The table (8)

showed variations in MCH within treatments ranged from

22.23pg in SFO 1.5% to 54.83 pg in SFO 2.5%, SFO 2.5%, SFO 2% and control showed a significant increase at P≤0.05 compared to other treatment. MCV within treatments ranged from 107µm3 in LN 2% to 160.80µm3 in an SFO 1.5% and SFO 1.5% higher significant increase at P≤0.05 with other treatment. The results of statistical analysis showed a significant increase at P≤ 0.05 between SFO 2.5% and SFO 1.5%, while no significant differences at P>0.05 were found between LN treatments.

53

Chapter Four

Results and Discussions

Table 8: Haematological parameter of common carp fed on Experimental diets. (Cont. LN 1.5%, LN 2%, LN 2.5%, SFO 1.5%, SFO 2% and SFO 2.5%). Treatments

RBC WBC 6 (cells x 0 µl) (cells x 03µl)

Hb (g/dl)

Cont.

1.86±0.20b

12.40±0.34b

LN 1.5%

1.51±0.28b

LN 2%

PCV %

MCH (pg)

MCV (µm3)

11.33±1.500a 24.33±1.70c

35.56 a±1.35a

130.26± 1.45b

13.83±0.43ab

10.00±2.00a

24.65± 1.99c

24.30 ±3.50b

121.13±8.60b

1.69 ±0.02b

13.96±1.18ab

11.66±1.50a

27.76 ± 2.06b

25.36±5.90b

107.00 ±7.60c

LN 2.5%

1.40±0.07b

11.23±0.55b

9.66±2.50b

29.09± 0.78b

25.00 ±5.30b

129.53 ±7.45b

SFO 1.5%

2.59 ±0.39a

15.73±0.90a

10.00±2.50a

29.65±0.77b

22.23 ±2.30b

160.80±6.76a

SFO 2%

2.27±0.05a

17.23±1.50a

11.89±2.50a

31.55± 1.90a

36.43±2.25a

127.00 ±3.65b

SFO 2.5%

2.14±0.41a

17.93±1.40 a

10.00 ±1.00a

31.33± 2.88a

54.83±2.65a

119.73±6.87b

Data are presented as mean ± S.D. Data in the same row with different superscript are significantly different (P<0.05).

54

Chapter Four

Results and Discussions

The results of present study indicate that there is a slight increase in the values of the blood studied when added LN and SFO oils. These results are in agreement with many of other studies in this field (Montero et al.,2001; Bransden et al., 2003 ; Al-Ashaab, 2011). Calder (2001) noted that the use of vegetable oils as a source of essential fatty acids affects blood cell count (RBC, WBC and Hb content) and represents the size percentage of red blood cells to the blood volume. In addition to soluble vitamins in fats that has a role in supporting the efficacy or activity of the spleen to produce red pellets and white blood. The current results are consistent with what was found by Montero et al. (2001) which is an increase in the number of white blood cells in fish fed on a pellets contained on omega-3and omega-6 with different concentrations. It is attributed to the fact that these acids have an important role in maintaining the factor of chemical control for the number of physiological functions in the body, such as growth, and also to maintain pressure coagulation blood, immune function and inflammatory. Lee et al. (1993) noted that there are signs appeared in the blood of Sebastes schlegeli which are improved number of red blood cells and hemoglobin content at the level of 0.9% omega-3, and also improve the physiological response to stress on the growth rate in fish. In this study hemoglobin and total erythrocyte count were observed among the (pork lard, sun flower oil, fish oil) feeding the different levels of omega-3 and omega-6 fatty acid, which is in agreement with the result of Choudhury et al. (2005). Leukocytes play an important role in non-specific or innate immunity and their count can be considered as an indicator of the health status of fish. A significantly

55

Chapter Four

Results and Discussions

higher WBC count was recorded at supplementation of a high level of omega-3. In the present study the increase in respective leukocyte count of dietary omega-3 fatty acid levels represents a possible inflammatory increased response mediated by leucocytes against bacterial infection (Roberts, 1978). 4.4.2 White blood cell The content of white blood cells presented in the Table (9) results of granulocyte the LN 2% significant decreases at P≤0.05 all other treatment. Lymphocyte were ranged between 3.0 in Control to 16.86% in SFO 2.5% and SFO 2.5% higher significant different at P≤0.05 all other treatment. Monocyte revealed no significant different with all dietary treatments except LN 2.5%. The percentage of Platelet% were ranged from 15.33 in SFO 2.5% to 42.05% in LN 2%, however there were no significant differences between all other treatments (P>0.05). Blood cholesterol varied from 2.34 in LN 2.5% to 3.45 mg/dl in SFO 2%, the results of statistical analysis showed no significant variations between the other treatment (P>0.05).

56

Chapter Four

Results and Discussions

Table 9: Effect of linseed oil and safflower oil (± SE) White blood cell Blood Treatments

Granulocyte %

Lymphocyte % Monocyte %

Platelet %

Cholesterl mg/dl

Cont.

LN 1.5%

LN 2%

LN 2.5%

SFO 1.5%

SFO 2%

SFO 2.5%

68.00±8.00a

3.00±1.50c

29.00 ±5.90b

28.00±9.50a

2.92±0.25a

63.13±1.40a

5.40±0.40bc

34.63 ±1.05a

20.33±1.50a

2.38±0.30a

59.86±2.90b

6.96±1.05bc

33.16 ±2.55a

42.00±5.50a

3.34±0.20a

66.56±6.35a

3.60±2.50bc

29.83±4.35ab

18.00±5.50a

2.34±0.60a

60.83±0.25a

6.90±3.50bc

32.26 ±0.70a

37.66±2.50a

2.83±0.30a

68.56±1.25a

8.21±0.80b

36.20±0.90a

15.64±3.00a

3.45±0.10a

66.96±4.05a

16.86±1.70a

34.86±0.95a

15.33±1.00a

2.66±0.20a

Data are presented as mean ± S.D. Data in the same row with different superscript are significantly different (P<0.05).

The low rates of synthesis of cholesterol inside cells leads to increased rates of taking molecules of blood cholesterol by body cells, which turns into bile acids by the enzyme7α-hydroxylase, which are flowing on the front of the intestines and absorbed , which holds under natural conditions as well as a few cholesterol food (Larsson and Fange,1977; Kennish et al.,1992). 57

Chapter Four

Results and Discussions

Williams (2007) demonstrated that the supply of animal pellets or vegetable oils containing fatty acids of the familyomega-3 and omega-6 leads to control the level of blood lipid and cholesterol, as well as the quality of fatty acid component for the fatty tissue in animals fed on these oils . There is also a significant impact on the quality or sources of oil added to the diet of fish and some of the qualities of vessels to fish, especially these oils long chain unsaturated fatty acid as reduce the level of blood cholesterol Olsen et al., (1991). Richard et al. (2006) noted the low level of blood cholesterol when the level of omega-3 and omega-6 increased the study of fat effect and the lack of essential fatty acids on growth in Salmo gairdneri. Larsson and Fange (1977) concluded that the essential fatty acids of omega-3 and omega-6 constitute the important parts of health status in the organism because they contribute significantly to the reduction of cholesterol. 4.5 Body composition Body composition presented in table (10) pointed out that crude protein values varied from 52.04% to 53.66 % in SFO 1.5% and control, respectively. Values of other extracts were ranged between 33.89% in control to 37.82% in SFO 2.5%, and the values of ash and fiber ranged between 9.00% in SFO 1.5% to 9.78% in control. Results showed a SFO positive relation between lipids in fish oil and the ratio of safflower oil in pellets, while LN showed a negative relation between lipids in fish oil and moisture and ash. Table (10) indicated the relationship between crude protein and oil in treatments.

58

Chapter Four

Results and Discussions

Results including that crude protein showed no significant variations at P>0.05 between the treatments. The dietary lipid content showed a significant increase at P≤ 0.05 between LN 2%, SFO 2% and SFO 2.5% compared with other treatments. Results of ash values showed no significant variations between other treatments (P>0.05).

Table 10: Effect of linseed oil and safflower oil on whole body composition at the end of experiment on dry matter basis (±SE) mean. Treatments

Crude Protein %

Ether Extract %

Ash %

Cont.

55.34 a±1.35

33.89b±1.03

9.78 a±0.67

LN 1.5%

53.74 a±0.84

35.74 ab±0.4

9.78 a±0.54

LN 2%

54.18 a±0.54

37.69 a±1.95

9.70 a±0.45

LN 2.5%

54.22 a±0.48

35.06 b±0.81

11.69 a±1.04

SFO 1.5%

53.75 a±0.36

34.32 b±0.19

9.00 a±0.54

SFO 2%

53.91 a±0.53

37.12 a±1.95

9.54 a±0.69

SFO 2.5%

54.73 a±0.92

37.82 a±0.33

9.03 a±0.37

Data are presented as mean ± S.D. Data in the same row with different superscript are significantly different (P<0.05).

Nandi et al. (2007) stated that the components of the diet are reflected on the formulation of nutrients in the body of Catla catla when studied the effect of adding of polyunsaturated fatty acids of omega-3 and omega-6. They realized the increased fat content in the whole body, which was accompanied by low moisture content. This is consistent with results of the study, showing the increased proportion of fat in the whole body due to the decrease of moisture content. This increasing may be due to the addition of different levels of various oils in the

59

Chapter Four

Results and Discussions

experiment. Common carp tend by their nature to the deposition of fat when fed on diets containing carbohydrates and fats (Csengeri et al., 1978; Hepher, 1988), while Skalli and Robin (2004) pointed out that carnivores fish tend to the deposition of fat in their tissues. Ben-Shan and Shi-Yen (1999) confirmed the needs for the maximum growth in Oreochromis niloticus × Oncorhynchus aureus components and found that nutrients in fish are generally reflect the components of their feed ingredients. Atalah et al. (2007) explained that the nature of additive fatty acid in the starter feeds for fish (Sparus aurata) had an impact on body composition of other nutrients when marine algae Crypthecodinium cohnii and Phaeodactylum tricornutum used as a source of essential fatty acids. The results were also consistent with the findings of Hansen et al. (2007) who studied the effect of total fish oil substitution with vegetarian protein diets on growth and deposition of protein in Atlantic cod Gadus morhua. The result showed a decrease in the percentage of fat with the increased rates of protein content and moisture. Sevket and Nazmi (2007) noted the changes in the composition of nutrients in tissues and liver of rainbow trout with seasonal and monthly changes. This happened alteration due to in natural food, which obtains in the aquatic environment, confirming that the changes in natural food had their effects on installing fish body of DHA nutrients. The present study agrees with Al-Ashaab (2011) who added various ratios of sunflower oil and corn oil as omega-6 sources in the diets for common carp. The study also in accordance with what was suggested by Nusrat et al. (2010) on the addition of oils containing group omega-3 and omega-6 in experiments which had

60

Chapter Four

Results and Discussions

increased the proportions of fat with a slight change in the proportions of protein and reduced the rates of moisture and ash in the bodies of fish.

61

“CONCLUSIONS AND RECOMMENDATIONS” -Conclusions 1- The current study shows that vegetable linseed oil and safflower oil used in the Experimental diets had given positive results by increasing the growth rates of fish, especially safflower oil. 2- Using linseed oil and safflower oil have improved the feed efficiency ratio. 3- Non significant increase was found in the values of blood parameters when added linseed oil and safflower oil, although the health status of experimental fish were improved compared to control treatment. 4- Results indicated that in the values of Gonado somatic index and Liver somatic index gradually decrease with increasing the ratio of safflower oil, on the contrary of linseed oil compared to control treatmentN 5- The linseed oil and safflower oil used in the current study had no affected the proportion of protein; however the proportion of ether extract with Safflower oil was significantly increased with decreasing the proportions of ash.@

62

-Recommendations

1- Further studies will require implementing similar work to optimize different levels of vegetable oils to identify the ideal level of inclusion in the fish diets.

2- Possibility of using mixture of vegetable oils and/or animal oils in fish diets. 3- Different environmental condition should be applied in future studies such as; water quality, temperature, pH and photoperiod in Kurdistan Region, IRAQ. 4-Using LN and SFO as a source of omega-3 and omega-6 for farming fish; grass carp, silver carp and rainbow trout and other local fish species in Kurdistan Region.

63

63

References Al-Rudainy,A.J. (2007). Approach to support simple fishermen and producers in Arabian Nation. Arab.Org. Food and Agric.,19P. Ana, C.A.; Damila, R.A.; Leandra, P. S.; Flavia, B. S.; Eliete, L. V.; Nilson, E.S. and Jesui, V.V. (2007). Effect of flaxseedoil in diet on fatty acid in the liver of Nile tilapia (Oreochromis niloticus). Vol. 57:273-277. AOAC. (2002). In: Hortwitz W (Ed) Official Methods of Analysis of AOAC International, 17th ed. Gaithersburg. Atalah, E.; Cruz , C.M.H.; Izquierdo, M.S.; Rosenlund, G.; Caballero, M.J.; Valencia, A. and Robaina, L. (2007). Tow micro algae Crythecodinium cohnii and Phaeodactylum tricornutumas alternative source of essential fatty acids in starter feeds for sea bream (Sparus aurata). Aquaculture, 270:178-185. Bell, G.; Torstensen, B. and Sargent, J. (2005).Replacement of marine fish oils with vegetable oils in feeds for farmed salmon. Lipid Technology, 17:7-11. Bell, J.G.; McGhee, F.; Campbell, P.J.; and Sargent, J.R.(2003). Rapeseed oil as an alternative to marine fish oil in diets of post-smolt Atlantic salmon (Salmo salar): changes in fish fatty acid composition and effectiveness of subsequent fish oil Aquaculture, 218:515-528. Ben-shan, C. and Shi-Yen, S.(1999). Both ω-3 and ω-6 fatty acids are required for maximal growth of juvenile Hybrid Tilapia. North American journal. Aquaculture, 61:13-20. Beveridge, M.C.M. and McAndrew, B.J. (2000).Tilapias: Biology and Exploitation, Great Britain: Kluwer Academic Publishers, 327-375. Billard, R. (1995) Carp: Biology and Culture. IRNA, Paris1995. Bransden, M.P.; Carter, C.G. and Nichols, P.D. (2003). Replacement of fish oil with sunflower oil in feeds for Atlantic salmon (Salmo salar L.): effecton

growth performance, tissue fatty acid composition and disease resistance. Comparative Biochemistry and physiology-part, B 135:611-625. Bureau, D.P; Hua, K. and Harris, A.M. (2008). The effect of dietary lipid and Long- chain n-3 PUFA levels on growth, energy utilization, carcass quality, and immune function of rainbow trout Oncorhynchus mykiss.J.The World Aqua. Soc., 39:1-21. Castell, J.D.; Sinnhuber, R.O; Wales, J.H. and Lee, D.J. (1972). Essential Fatty acids in the diet of rainbow trout Salmo gairneri : Growth, Feed Conversion and some gross deficiency symptoms. Journal of Nutrition, 102:110-118. Catacutan, M.R. and Coloso, R.M. (1995). Effect of dietary protein to energy ratios on growth, survival, and body composition of juvenile Asian seabass, L atescalcarifer. Aquaculture, 131:125-133. Chou, B.S. and Shiau, S.Y. (1999). optimal dietary lipid level for growth of Juvenile hybrid tilapia Oreochromis niloticus ×Oreochromis aureus. Aquaculture, 143:185-192. Chua,

B.

and

Kiron,

V.

(2006).Archophthalmol.

124:981-986

(http://en.wikipedia.org/wiki/Omega-3_fatty_acid). Coles, E.H. (1986). Veterinary clinical pathology.4th edition.Saunders , W.B. Company, Philadelphia.110-115,122. Copeman, L.A.; parrish, C.C.; Brown, J.A. and Harel, M. (2002). Effects of docosahexenoic, eicosapentaenoic and arachidonic acids on the early growth, survival, lipid composition and pigmentation of yellowtail flounder (Limandafer ruginea): a live food

enrichment experiment. Aquaculture,

210:285-304. Csengeri, I. (1996). Dietary effects on fatty acid metabolism of common carp. Arch Tierernahr, 49:73-92.

Csengeri, I.; Larkas, T.; Majoros, F.; Olah, J. and Szalay, M. (1978). Effect of on the fatty acids composition of carp Cyprinus carpio L. Aquaculture. Hung.,1:24-34. Dacie, J.V. and Lewis, S.M.(1995). Practical haematology, 8th ed. Churchill Livingstone, Edinburgh , New York, 609p. El-Khalil, F.F.M. (2005) . Scientific and practical principles of Fish farms.Chapter 2, Rearing, production and management of fish farms, first print, College of Agriculture, University of Al-Mansura, Pp 482. (In Arabic) El-Sayed, A-F.M. (1994).Principles of fish culture.Kuwait Organisation for Scientific Development, Pp 224. Fan,E.(1996).Major trends in global aquaculture.Production and summery overview of the Gulf area (1984-1985) K.Rana, M. Perotti, M. Pedini, A. Tacon. FAO (1996).The sixth world food- survey- Rome: 2-4. FAO ( 2010).The State of world fisheries and Aquaculture, (SOFIA) FAO Fisheries and Aquaculture. Department Food and Agriculture Organization of the United Nations Rome. Farkas, T.;Csegri,I.; Majors, F. and Olah, J. (1980). Metabolism of fatty acids in fish.III.Combind effect of environmental temperature and diet on formation and deposition of fatty acids in the carp, Cyprinus carpio L., 1758. Aquaculture, 20:29-40. Gowdet, A.I. (2007) .Mittelzurentwicklung der fischerei in der Kurdustan RegionIrak, Berliner Verlag, 115p. (In German). Guary, J.C., Kayama, K., Murakami, Y. and Ceccaldi, H.J.(1976). The effect of a fat-free diet and compounded diets supplemented with various oils on molt, growth and fatty acid composition of prawn, Penaeus japonicus. Aquaculture, 7: 245-259.

Gumus, E. and Fatim, E. (2010). Effect of partial Substitution of fish meal with Tuna Lier Meal on the fatty Acid profilof Nile Tilapia Fry, Oreochromis niloticus. KafkasUniv . Vet Fak. Derg., 16(Suppl-B):S283-S290. Hepher, B. (1988).Nutrition of pond fishes. Cambridge University press, Cambridge. 27p. Henderson, R.J. and Tocher, D. R. (1987).The lipid composition and biochemistry of freshwater fish.Progr. Lipid, Res., 26:281-347. Henderson, R.J.(1996).Fatty acid metabolism in freshwater fish with patticular reference to polyunsaturated fatty acids. Archives Animal Nutrition, 49:5-22. Ibeas, C.; Izquierdo, and Lorenzo, F. (1994). Effect of different levels of ω3 highly unsaturated fatty acids on growth and fatty acid composition of juvenile gilthead seabream (Cyprinus carpio), Aquaculture, 127:177-188. Ismael, S.A., (2004) Water policy in countries of Euphrates and Tigris and their effects on Kurdish case.Kurdistan Centre for Strategic Studies, 574 P. Janucey,K.and Ross, B. (1982). A quide to Tilipia Feeds and feeding institute of Aquaculture. Sterling Univ., Scotland, Sterling, 111p. Jobling, M. (1996).Environmental biology of fishes. Fish and fisheries series 16, Chapman and Hall.Kanazawa, A., Tokiwa, S., Kayama, M. and Hirata, M., 1977.Essential fatty acids in the diet of prawn. I. Effect of linoleic and linolenic acids ongrowth. Bull. Jpn. Sot. Sci. Fish., 43: 111 l-l114. Kanazawa, A.; Teshima, S.; Sakamoto, M. and Awal, M.A. (1980).Requirement of Tilapia zilliifor essential fatty acids. Bulletin of the Japanese Society of Scientific Fisheries, 33;47-55. Karalazos, V. ; Bendiksen, E.A. ; Dick, J.R. and Bell, J.G.(2007). Effect of dietary proteinand fat level and rapeseed oil on growth and tissue fatty acid composition and metabolism in Atlantic salmon Salmo salar L. reared at low water temperatures.Aquaculture Nutrition, 13: 256 –265.

Karapanagiotidis, I.T.; Bell, M.V.;Little, D.C. and Yakupityage, A.(2007). Replacement of dietary fish oils by alpha- linolenic acid – rich oils lowers omega-3 content in tilapia flesh. Lipids, 42:547-597. Kayama, M.;Hirata, M. and Hisai, T. (1986). Effect of water temperature on the desaturation of fatty acids in carp. Bulletin of the Japanese Society of Scientific Fisheries, 52:853-857. Kennish, J.M.; Sharp-Dahl, J.L.; Chambers, KA.Thrower, F. and Rice S.D. (1992).The effect of a herring diet on lipid composition, fatty acid composition, and cholesterol level in the muscle tissue of penreared Chinook salmon (Oncorhynchusts hawytscha).Aquaculture, 108:309-322. Kiessling, K.H. and Kiessling, A.(1993).Selective utilization of fatty acids in rainbow trout (Oncorhynchus mykiss Walbaum) red muscle mitochondria. Canadian Journal of Zoology, 71:248-251. Kim, Kyoung-Duck, and Lee,Sang-Min. (2004). Requirement of dietary n-3 highly

unsaturated

fatty

acids

for

juvenile

flouder(Paralichthys

oliaceus).Aquaculture, 229:315-323. Kitto, M.R. and Tabish, M. (2004).Aquaculture and food security in Iraq.Aquaculture Asia, 1: 31-33. Kris-Etherton, P.M.; Harris, W.S. and Appel, L.J. (2002). Fish consumption, fish oil, Omega-3 fatty acids and cardiovascular disease. Circulation, 106: 2747 2757.Leopold, M. (1981). problems of fish culture economics in Easten Europe, EIFAC/Tech.P.40, 9p. Lagler, K.F.; Bardach, J.E.; Miller, R.E. and May, D.R. (1977). Ichthyology, 2nd ed. John W. and Sons. New York. Larsson, A. and Fange, R.(1977).Cholesterol and free fatty acids (FFA) in the blood of marine fish. Comparative Biochemistry and Physiology. 57B: 191196.

Leaver, M.J.; Ezaz M.T.; Fontagne S.; Tocher, D.R.; Boukouvvala, E. and Krey,M. (2007). Multiple peroxisome proliferator-activated receptor

beta

subtypes from Atlantic salmon (Salmo salar). Journal of Molecular Endocrinology, 38:391-400. Lee,S.M. (1997). Effects of dietary lipid source and water temperature on nutrient digestibility in juvenile and adult Korean rockfish (Sebastes schlegeli). Korean Journal of Animal Nutrition and Feedstuffs, 21:381-390. Lee,D.N. ; Roehm , J.N. ;Yu, T.C. and Sinnhuber , B. (1967). Effect of ω-3 fatty acids on the growth rate of rainbow trout (Salmon gairdnerii). Journal of Nutrition, 92:93-98. Lee, S. ; Jong, H. L. and Kyoung-Duck K.(2003). Effect of dietary essential fatty acids on growth, body composition and blood chemistry of juvenile starry flounder (Platichthys stellatus) Aquaculture, 25:269-281. Lee, S.M.; Lee J.Y.; Kang Y.J. and Hur S.B. (1993).Effects of n-3 highly unsaturated fatty acids on growth and biochemical changes in the Korean rockfish Sebastes schlegeli. Changes of blood chemistry and properties of liver cells. Journal of Aquaculture, 6: 107-123. Lehninger, L.A.( 1982). Principles of Biochemistry. Worth publishers Inc.,607p. Montero, D.; Robaina, L.E.; Socorro, J.; Vergara, J.M.; Tort, L. and Izquierdo, M.S.(2001).Alteration of liver and muscle fatty acid composition in gilthead sea bream (Sparus aurata) juveniles held at high stocking density and fed an essential fatty acid deficient diet. Fish Physiology and Biochemistry, 24: 6372. Moyle, P.B. and Cech, J.R. (1982).Fishes: An introduction to ichthyology. Prentice-Hall,inc., New Jersey.

Mraz, J.; Schlechtriem, C.; Fang Y.; Cossins, A.; Zlabek, V.;samuelssen, T. and Pickova, J. (2010). Sesame as a potential modulator of fatty acid composition in common carp Cyprinus carpioL. Aquaculture Ressearch, 41:851-861. Muona, M. and Soivio, A. (1992).Changes in plasma lysozyme and blood leukocyte levels of hatchery-reared Atlantic salmon (Salmo salar L.) and sea trout (Salmo trutta L.) during parr-smolt transformation.Aquacalture, 106:7587. Nakamura, M.T. and Nara, T.Y. (2004). Structure, function, and dietary regulation of delta-6, delta-5, and delta-9 desaturases. Annual

review

Nutrition, 24:345-376. Nandi, S.; Routray, P.; Gupta, S.D. and

Rath, S.C. (2007). Reproduction

performance of carp, catla catla(ham.), reared on a formulated diet with PUFA supplementation. Journal of Applied Icthyology, 23:684-69 Nelson, D.L. and Cox, M.M.(2000). Lehninger, principles of biochemistry “3rd Ed. Worth publishing :New York. Nick, K.; Maria N.A. and Henderson, R.J. (1991).Effect of dietary soybean and cod-liver oil level on growth and body composition of gilthead bream Sparus aurata.Aquaculture, 104:293-308. NRC, (National Research Council) (1993).Nutrient Requirements of fish.National Acad. Press, Washington DC.,114p. Nusrat, N. M.; Farah, N. T.; Bhanger, M.I. and Aamna, B. (2010). Changes in fatty acid composition in muscle of three farmed carp fish species (Labeo rohita, Cirrhinus mrigala, Catla catla) raised under the same conditions. Food Chemistry.126:405-410. Okoye, F. C. and Eyo, A. A. (2002). The Growth and survival of Clarias anguillaris fingerlings fed on various lipid sources. Journal of Applied Sciences and Environmental Management, 6: 27-31.

Olsen,R. E.; Henderson,R. J. and Ringo, E.(1991).Lipids in Arctic charr Salvenins alpines L. 1.Dietry induced changes in lipid class and fatty acid composition. Fish physiology and Biochemistry, 9:151-164 Owen, J.M.; Adron, J.W.; Middleton,C. and Cowey, C.B. (1975).Elongation and desaturation of dietary fatty acids in turbot Scophthalmus maximus L. and rainbow trout Salmo gairderii Rich. Lipids,10:528-531. Pillay T.V. R. and Kutty, M. N. (2005). Aquaculture principles and practices. Oxford, UK. Pillay, T.V.R. (1990). Aquaculture: Principle and Practices. Fishing News Books. Radunz-Neto, J.; Corraze, G.; Bergot, P. and Kaushik,S.(1996). Estimation of essential faaty acid requirements of common carp larvae using semi-purified artificial diets. Arch Tierernahr, 49:41-48. Rinchard,J.; Czesny, S. and Dabrowski, K. (2007). Influence of lipid class and fatty acid deficiency on survival, growth, and fatty acid composition in rainbow trout juveniles. Aquaculture, 264:363-371. Robert, A.M.; Sang, L.; Danielle, A.A.; Charlotte, A.; Kathleen, M.G.; Bruce, R.B. and Mark, P.(2008). Fish oil prevents essential fatty acid deficiency and enhances growth: clinical and biochemical implications. Metabolism Clinical and Experimental, 57:698-707. Rodoifo, B. ;Simona,R. and Marco, M. (2006). The replacement of fish oil with linseed oil in the diet of large rainbow trout Oncorhynchus mykiss.Italian Journal of Animal Science, 5:155-165. Rodriguez, C.; Perez, J.A.; Badia, P.; Izquierdo, M.S.;Fernandez-palacios; H. and Hernandez, A.L.(1998).The n-3 highly unsaturated fatty acids requirements of gilthead sea bream (Sparus Aurata L.) Larvae When Using an Appropriate DHA/EPA Ratio in the Diet”, Aquaculture, 169:9-23.).

Ruyter, B.;Einen O. and Thomassen, M. (2000). Essential fatty acids in atlantic salmon: time course of changes in fatty acids composition of liver, bllod carcass by a diet deficient in n-3 and n-6 fatty acids, Aquaculture Nutrition , 6:109-117. Sargent, J.R.; Bell, J.G.; Bell M.V.; Henderson, R.J. and Tocher, D.R. (1993).The metabolism of phospholipids and polyunsaturated fatty acids in fish. In: Callou B, Vittelo P (eds) Coastal and Estuarine Studies- Aquaculture: Fundamental and Applied Research, PP.103-124. American Geophysical. Sargent, J.R.; Bell J.G.; Henderson,R.J. and Tocher, D.R. (1995).Requirement criteria for essential fatty acids. Journal of Applied Ichthyology, 11:183-198. Scott, M.L.; Nesheim, M.C. and Young, R.J. (1982).Nutrition of the Influence of different fats with varying addition of alpha-tocopherol acetate on growth and body composition of carp (Cyprinus carpioL.).Aquaculture, 69:57-67. Sevket, K. and Nazmi, P. (2007). Seasonal variation of Total lipid and Total Fatty acid in Muscle and Liver of Rainbow Trout (Oncorhynchus mykiss.; 1792). Reared in derbentdam lake. Turkish journal of fisheries and Aquatic Sciences, 7:27-31. Simopoulos, A. P. (2002).The importance of the ratio ofomega-6/ omega-3 essential fatty acids, Biomedicine and Pharmacotheraphy,56: 365-379 Skalli, A. and Robin, H. (2004). Requirement of ω-3 long chain polyunsaturated fatty acids for European sea bass Dicentrar chuslabrax juveniles: growth and fatty acid composition. Aquaculture, 240:399-415. Steffens, W. (1996). Protein sparing effect and nutritive significance of lipid supplementation in carp (Cyprinus carpio L.), Arch Tierernahar, 47:93-8. Stephanie, F.; Anne-Marie, E. and pierre, B. (1998). Histological changes induced by dietary phospholipids in intestine and liver of common carp (Cyprinus carpio L.) larvae. Aquaculture, 161:213-223.

Stickney, R.R. and Hardy, R.W. (1989). Lipid requirements of some warm water species. Aquaculture, 79:149-156. Stickney, R.R. and McGeachin, R.B.(1983).Effects of dietary lipid quality on growth and food conversion of tilapia. Proceedings of Wildlife Agencies, 1983:352-357. Stillwell,

W.

and

Wassall,

S.R.(2003).

Docosahexaenoicacid:membrane

properties of a unique fatty acid. Chem. Phys. Lipids, 126:1-27. Swift, D.R. (1993).Aquaculture training manual, Hartnols Ltd, Cornwall, UK. MSc Thesis, 22:907-910. Takeuchi, T.; Satoh, S. and Kiron, V. (2002).Common carp,Cyprius carpio.In: Webster CD, Lim CE (eds) Nutrient Requirements and Feeding of finfish for Aquaculture, pp. 245-261.CABI publishing, Oxon. Takeuchi, T.; Arai, S.; Watanabe, T. and Shimma, Y.(1980).Requirement of eel Anguilla japonica, for essential fatty acids. Nippon Suisan Gakkaishi, 46:345353. Takeuchi, T. and Watanabe, T. (1977). Requirement of carp for essential fatty acids. Nippon Suisan Gakkaishi, 43:541-551 Takeuchi, T. (1996).Essential fatty acid requirement in carp. Arch Tierernahr, 49:23-32. Tidwell, J. H.M. and Robinette, H.R. (1990).Changes in proximate and fatty acid composition of fillets from channel catfish during a two-year growth period.Transactions of the American Fisheries Society, 119:31-40. Utne, F. (1978). Standard methods and terminology in fin-fish nutrition from: proc. World Symp. On finfish nutrition and fish feed Technology, Hamburg, 20-23.June,Vol.2. Villalta,M.; Estevez, A.;Bransden, M.P. and Bel, J.G.(2008). Effects of dietary eicosapentaenoic acid on growth, survival, pigmentation and fatty acid

composition in Senegal sole (Soleaseneg alensis) larvae during the Artemia feeding period. Aquaculture Nutrition, 14:232-241. Wedemeyer, G.A.(1996). Basic physiological function.In:physiology of fish in intensive culture systems, Chapman and Hall. Williams, K.C. (2007). Nutritional requirements and feed development for postlarval spiny lobster: a review. Aquaculture,263:1-14. Yildirim-Aksoy, M.; Lim C.; Davis,A. and Klesius, P.H.(2007).Influence of dietary lipid sources on the growth performance, immune response and resistance of Nile tilapia (Oreochromis niloticus)to Streptococcus iniae challenge. Journal Applied of Aquacalture, 19:29-49. Yu, T.C. and Sinnhuber, R.O. (1979). Effect of dietary ω-3 and ω-6 fatty acids on growth and feed conversion efficiency of Coho salmon Oncorhynchus kisutchy. Aquaculture, 16:31-38. Yu,T.C.and

Sinnhuber,R.O.(1972).Effect

of

dietary

linolenic

acid

and

docosahexaenoicacid on growth and fatty acid composition of rainbow trout. J. Lipids, 7:450-454. Zheng, X.; Torstensen, B.E.; Tocher, D.R.; Dick, J.R.; Henderson, R.J. and Bell, J.G.(2005). Environmental and dietary influences on highly unsaturase fatty acid biosynthesis and expression of fatty acid desaturase and elongase genes in liver of Atlantic salmon (Salmo salar).Biochimicaet Biophysica Acta, 1734:13-24.

  $ 3% $ # (Linseed oil)              9   $ 456  7' 8$ 3$. / 012 6% $ # (Safflower oil)'#  G .8 9H 3:8 I F E  CD E $ Cyprinus carpio < 28 = :;  K 7$ 3J    9 1H  . 1 3 . 1  C 3 2 9$1  1 3E.$  126 9F  I  IC  5 '. 9 5 .9$ 21 Q 1HD 7 I$ P$ 1  11 0     19   L MD P I$ .9$ I   UH$I$ 9C $  %3 I.I :8  56 3:8D T5  I#D .9 14% 12 I  1 L$[$ 40% 35EW I $  \K UH$I$ '#   $      P  '1 F  $   K   I F 1  E$ 3  % 0 $  ^ 1 I :8  5`_ 7[  1  E : E1 $ 1  P  I 1 $     % 2    1 $ 3    % 1.5^ 1 I :8  5`_  .   1 $ '#    %1.5 012 F 1  $  . I 3    % 2.5 .'#    %2.5  I 1  .$ 3'#    %2 7  $C$ :K  WL.   1  012 '#         aW . L/D aW . 7L/D cF8 K  U .^  I$  1   I (P≤ 0.05 )  $.     1  E  (P≤ 0.05 )  $.  7  $ C$ E2$ . $ . P  EW #8 P 1  P  E  1   aW . . Ig .E$  012 (%/ 9$ / 9` 0.68, 0.64)  I$ aW . 7 L/D ,cFD K  U   .^ (P<0.05)  $.  0 128 E (%36) EI . $. (49.60) F 1 $ (53.42) 0 $8D 1  EW56 P $  74' EW#8 P 1 

i

‫_______________________________________________الخالصة‬ aW . E   $.   H$ < L/ 9  ,c FD K  U   .^ (P<0.05)  $.  0 128 .EW56 P $  8 $  $I 74' 9 ^ EW#8 P 1   F  ^ 1  0 12 75 6 : j ( \ 1F 106)   9     2 9  H  L/D 9 ^  2 (P<0.05)  $.  H$' E$  012 2.14 $ 2.27$ 2.59  I$ $ l I 9     . I .cF8  I C  JWK  012 756 :8 9 E 1C  $.  E$  012 17.23$ 17.93  $  I 1  L  '1 F 0  ( \ 1F 103) 9  = F   E   $.   H$  L/ 9  ,c F8D J WK  0 12 75 6 : 8  2 ( \9`$ I )0[$    I$1`$    1C 9 ^ PD  .K   I ( \9`) [$  1C 9 ^ 012 . .(54.83)  I  ^ 1  012 :8  56 .2 (MCH) = E  $.  K F :. . .(160.80)F 1  E(MCV)   9C '1 F . I$ $ 0$8D 1  E (68.00% ,68.56% ) . 0  II  KF  KF  D 3 I 1  E ( 16.86 %) .$ K   I  $'1  KF   '# 9 H E L/ 9 .(36.20% ) .$  1  E  $.  p1 F 7$. 7 $ $C$ /$ 9 P$ $ $ P .$  I P   I .I$ 3 I C  K  E   $ . K  E  $.   ` K F E $  $I I. E  $.  H$ L/ 9 3IC  :D 91 EW   P 1  E 1  $ %37.82 I.I  I 1  E  I.  $.  '1 F$ٕ ,IC  K  .37.69 % I.I   

ii

‫_______________________________________________الخالصة‬

‫‪iii‬‬

‫    ‪       6 3‬‬ ‫ ‪(Cyprinus carpio L.) ./0 1 , -‬‬ ‫ ‬ ‫‪  4   5    34   2‬‬ ‫‪ > ? -  4 < =0 ; 89:  7 654‬‬

‫‪, - ?  -‬‬

‫ ‬ ‫‪7   @ 7  6A 7/‬‬ ‫      )‪  #  "    !      (2009‬‬

‫‪$%‬‬

‫ ‪   4 1: 9.‬‬ ‫(‪& #  '#‬‬

‫السنة الميالدية ‪2014‬‬

‫السنة الھجرية ‪1436‬‬

@@ón‚íq @òŒI@òì@H3@Mb ïàûaI@ôØóîòìbšŠó"@Ûòì@HçbmóØ@ônî@ òŒI@õŠó îŠbØ@ôåïäaŒ@üi@òìaŠ†@ãb−ó÷@óîòìóåîím@ãó÷ @Hcommon carpI@ õb"b÷@ ôiŠbØ@ ô"bà@ Šó"óÜ@ H6Mb ïàûaI@ ôØóîòìbšŠó"@ Ûòì@ (ŠóÑ–óÈ@ ônî @ôáŽîŠóè@ \@ôäbáŽïÝ"@õüÙäaŒ@ \@çbØóï-ÜbØín“Ø@ôn/äaŒ@ôn-ÜóØbÐ@ \@ô"bà@õó ïÔbm@óÜ@HCyprinus carpioI @11@bmóè@a‹ÙŽïq@o"ò†@ça‹î@òŒíy@19@óÜ@ìíi@ãaìò†Šói@Âäbà@Žô"@õòìbà@üi@óØòìó䉎îím@@L×a‹ŽïÈ@\@çbn"†ŠíØ @40@ @M35I@ô“ŽïØ@ói@a‹åŽïè@ŠbØói@ô"bà@ 126@@ L@ˆû<Š@ 21@óÜ@ìíi@ônî‹i@çbØóï"bà@ôåmbèa<Š@õòìbàòì@ ßíÝî@ó÷ @õó-Üóàbà@óÜ@ LŠ@ óÑ"óÈì@çbmóØ@ônî@òŒ@óÜ@o"b÷@ f"@õŠó îŠbØ@õòìóå-ÜüÙŽïÜ@üi@ Hâ"@ 14M12@Iñ‫يژ‬Š†@òì@Hâ  @ôØaŠü‚óÜ@ çbmóØ@ ônîòŒ@ õ‫ @بڕ‬a‡à@ òìì†@ õó-Üóàbà@ óÜ@ ~ìíi@ E0@ a‡ï"bà@ ôØaŠü‚óÜ@ oîòŒ@ õ‫ @بڕ‬a‡Ü‫ۆ‬Bä‫کۆ‬ @õó-Üóàbà@ óÜ@ òì@ ìíi@ E2@ a‡ï"bà@ ôØaŠü‚óÜ@ çbmóØ@ ônîòŒ@ õ‫ @بڕ‬a‡àóîf"@ õó-Üóàbà@ óÜ@ ì@ ìíi@ E1.5a‡ï"bà @ônîòŒ@ õ‫ @بڕ‬a‡à@ a‡àóvåïq@ õó-Üóàbà@ óÜ@ òì@ ìíiE2.5@ a‡ï"bà@ ôØaŠü‚óÜ@ çbmóØ@ ônîòŒ@ õ‫ @بڕ‬a‡à@ a‡àòŠaíš @E2@a‡ï"bà@ôØaŠü‚óÜ@ŠóÑ–óÈ@ônîòŒ@õ‫@بڕ‬a‡àó’ó’@õó-Üóàbà@óÜ@òì@ìí[email protected]@a‡ï"bà@ôØaŠü‚óÜ@ŠóÑ–óÈ @Žô"@ÛóîóÜ- óàbà@Šóè@üi@ìí[email protected]@a‡ï"bà@ôØaŠü‚óÜ@ŠóÑ–óÈ@ônîòŒ@õ‫@بڕ‬a‡àómìóy@ õó-Üóàbà@óÜ@ òì@ ìíi @ãòˆ@ìì†@óäaˆû<Š ÚŽîŒìóy@Šóè@òì@p‹ ò†@õü‚óÜ@ô"bà@ •ó’@ÛóîòŠbiìì†@Šóè@òì@ìíióè@HŠa‹ÙmI@õòŠbiìì† @@ @.çbï’óÜ@ô“ïØóÜ 3@%@õò‰Žî<Š@ói@aŠ†ò†@Žðq@õòìóåîím@ô䆊aí‚ @çbîŠó îŠbØ@E2.5@ õò‰Žî<Š@ ói@ ŠóÑ–óÈ@LîòŒ@ ì@ E2@õò‰Žî<Š@ ói@ ŠóÑ–óÈ@ LîòŒ@ óØ@ çó‚ò†Šò†@ çbØóàb−ó÷ @Na†‹m@ ðäbØó-Üóàbà@ ÿó óÜ@ †ŠìaŠóiói@ çbØóï"bà@ õbmüØ@ õìì‡åîŒ@ õón"Šbi@ üi@ Hp<0.05I@ ôn"b÷óÜ@ ò‹mbîŒ @E2.5@õò‰Žî<Š@ói@ ŠóÑ–óÈ@LîòŒ@ ì@ E2@õò‰Žî<Š@ói@ ŠóÑ–óÈ@LîòŒ@óØ@ o/²Šò†@ ímìóØ@o"ò†@ ðäbØóàb−ó÷ @H% Oˆû<ŠO@â IH0.64,@ 0.68I@õŠüu@õó’ó @õ‡äòìbä@üi@Hp<0.05I@ôn"b÷@óÜ@ò‹mbîŒ@õŠó îŠbØ@ õŒaìbïu @Hp<0.05I@ ôn"b÷@ óÜ@ ò‹mbîŒ@ õŠó îŠbØ@ õŒaìbïu@ E2@ õò‰Žî<Š@ ói@ ŠóÑ–óÈ@ LîòŒ@ òì@ Ûóî@ õaì†@ óÜ@ Ûóî @õŒaìbïu@ @ E1.5@ õò‰Žî<Š@ ói@ ŠóÑ–óÈ@ LîòŒ@ ì@ ÿûBäüØ@ òì@ H%36I@ õò‰Žî<Š@ói@õò‰Žî<Š@õó’ó @õ‡äòìbäóÜ @Sïè@òì@ (ˆû<Š@ Oâ IH49.60@~@ 53.42I@ÛaŠü‚@ôåî<Šü @ ôØýbš@ üi@Hp<0.05I@ôn"b÷@óÜ@ò‹mbîŒ@õŠó îŠbØ

A

@Na‡äbØóÜ-óàbà@çaíïä@óÜ@µmû‹q@ôØýbš@õ@ò‰Žî‫@ڕ‬òì@ ÛaŠü‚@ôåî<Šü @ôn"b÷@ ì@@óÜ@ ìíi@óä@Šó îŠbØ@ðØóîóîŒaìbïu @ô䆋Ø@Šbàˆóè@óÜ@óîóè@õŠó îŠbØ@ôØóîŒaìbïu@óØ@ pìóØŠò†@a†óØòìó䉎îím@ôäbØó"bà@ôåŽîí‚@ôäbØóäaíŽïq@óÜ @ói@ ŠóÑ–óÈ@LîòŒ@òìH2.59I@ói@ E1.5@õò‰Žî<Š@ói@ ŠóÑ–óÈ@LîòŒ@óÜ@HBïÜ\óäb‚@ 106I@a‡äbØòŠí"@óØû‹‚ @óØû‹‚@ô䆋Ø@Šbàˆóè@a‡mbØ@çbàóè@óÜ@H2.14I@ E2.5@õò‰Žî<Š@ói@ ŠóÑ–óÈ@LîòŒ@òì@ (2.27I@ E2@õò‰Žî<Š @ŠóÑ–óÈ@LîòŒ@òì@H17.93I@ói@a†@E2.5@õò‰Žî<Š@ói@ŠóÑ–óÈ@LîòŒ@óÜ@HBïÜ\óäb‚@103 I@æŽîí‚@ôäbØóïr" @ônî‹i@H@BïÜOâÌI@@æŽîí‚@ôåïiü-Ý üáïè@õ<‹i@òì@L@ìíióè@çbîŠó îŠbØ@õŒaìbï[email protected]@ói@ E2@õò‰Žî<Š@ói @ôäbØóØû‹‚@ õòŠbióÔ@ óÜ@ L@ ìíióè@ õŠó îŠbØ@ õŒaìbïu@ óØE2.5@ @ çbmóØ@ LîòŒ@ üi@ H9.66I@ óÜ@ ìíi I@ E2.5@ õò‰Žî<Š@ ói@ ŠóÑ–óÈ@ LîòŒ@ ‫( و‬31.55@ %I@ E2@ õò‰Žî<Š@ ói@ ŠóÑ–óÈ@ LîòŒ@ üi (PCVIæŽîí‚ @ói@ ŠóÑ–óÈ@LîòŒ@ üi@ HBÜ\@â @üÙïqIHMCH@I@µiü-Ý üáïè@ôäbØóØû‹‚@õa<‹ÙŽïm@õ<‹i@òì ìíi@H 31.33% @õò‰Žî<Š@ói@ŠóÑ–óÈ@LîòŒ@üi@@HMCV@I@æŽîí‚@ôäbØóØû‹‚@õòŠbióÔ@õa<‹ÙŽïm@òì@ìí[email protected]@E2.5@õò‰Žî<Š @LîòŒ@óÜ@ çbØòŠa†ó-ÜüÙäò†@óäb‚@ô䆋Ø@Šbàˆóè@óÜ@ìíióè@Šó îŠbØ@ôØóîŒaìbïu@òì N@ìí[email protected]@ E1.5 @óÜ@ã-ýói@ìíi@H 68.00 %I @ @ õò‰Žî<Š@ói@a†ÿûBäüØ@óÜ@òì@H 68.56@ %I@ói@a†@ @ E2@õò‰Žî<Š@ói@ ŠóÑ–óÈ @ói@ ìíióè@ a†@ E2@ õò‰Žî<Š@ ói@ ŠóÑ–óÈ@ LîòŒ@ óÜ@ õŠó îŠbØ@ õŒaìbïu@ a‡äbØóØìbä@ Ûbm@ óäb‚@ ô䆋Ø@ Šbàˆóè @òì@@~ìíiH 16.86 %I@ói@@E2.5@õò‰Žî<Š@ói@ŠóÑ–óÈ@LîòŒ üi@çbØóîìbÑáïÜ@óäb‚@óÜ@òìH36.20 %I@õò‰Žî<Š @¶ûB/ÜüØì@ çbØóåïŽïà@ óåîˆ‫ڕ‬ìŠó u@ñŠójŽî<Š@õòŠbiŠò†@òì@Nìíióä@ÛóîŒaìbïu@Sïè@ æŽîí‚@ôäbØò<Šóq@õ<‹i@óÜ @Sïè@óØòìóåîím@ô"bà@ôn’ü @üi@a‡îìbïáïØ@ôåïåÙ“q@óÜòì@ Nóïä@Šó îŠbØ@õŒaìbïu@Sïè@a‹Ø@ôåïjŽïm@ æŽîí‚ @LîòŒ@‫@و‬óÜ@ìíióè@õŠó îŠbØ@õŒaìbïu@õŠìóš@õò‰Žî<Š@òì@~†bàì<Š@µmû‹q@õò‰Žî<ŠóÜ@ìíi@óäŠó îŠbØ@ôØóîŒaìbïu [email protected]% I@õò‰Žî<Šói@E2@õò‰Žî<Š@ói@çbmóØ@LîòŒ@‫@و‬òì@H37.82 %I@õò‰Žî<Š@ó[email protected]@õò‰Žî<Š@ói@ŠóÑ–óÈ

B

@æŽîí‚@ôäbØòŠòíŽïqìó’ó ŠóóÜ@6M@bïàü÷ì@3M@bïàü÷@ðäbåŽïèŠbØói@õŠóîŠbØ (Cyprinus carpio L.) ìþi@ðiŠbØ@ôbà@üi@@ @@óØóîóàbä @óÜ@Úï’ói@Ûòì@ðäbáŽïÝ@ñüÙäaŒ@óÜ@çbØóïÜbØín“Ø@ón,äaŒ@-ÜóØbÐ@ðäóàí−ó÷@ói@òìa‹Ø@•óÙ“Žïq ðÜòˆb÷@ðàìíiìŠói@çbØóïÜbØín“Øón,äaŒóÜŠónbà@ñóàbäaì2‹i@ðäbåŽïénò†ói@ðäbØóïn,îìa‡Žïq@

ðbà@ðàóèŠói@ @@ @@çóîýóÜ @@æî‡ÜaŠ‡–@æî‡ÜaþÈ@çbÄb÷ @ @ðäbáïÝ@ñüÙäaŒ@LçbØóïÜbØín“Øón,äaŒ@ônÜóØbÐ@LH2009I@@ðÜòˆb÷@ðàìíiìŠói@óÜ@‘ŽíîŠíÜbÙi @@

@@ @@@-’ŠóqŠó@ói

@ @Üa@â b§a@kݾa‡jÈ@N† Šò†ò‡îŠbî@õŠü,ïÐû‹q@ @ @ @2714@ñ†ŠíØ@ðÜb@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@2014@ îaŒ@ðÜb @@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@

EFFECT OF USING OMEGA-3 AND OMEGA-6 ON GROWTH AND ...

EFFECT OF USING OMEGA-3 AND OMEGA-6 ON G ... ATOLOGICAL PARAMETERS OF COMMON CARP.pdf. EFFECT OF USING OMEGA-3 AND OMEGA-6 ...

4MB Sizes 7 Downloads 242 Views

Recommend Documents

Effect Of Ecological Factors On The Growth And Chlorophyll A ...
Effect Of Ecological Factors On The Growth And Chlor ... ed Kappaphycus alvarezii In Coral Reef Ecosystem.pdf. Effect Of Ecological Factors On The Growth And ...

Effect of plant growth regulators and chemicals on ...
In India, banana accounts for higher fruit production .... The data were subjected to statistical analysis as per the procedure of Panse and Sukhatme (1961).

Effect of saline water on growth, biochemical parameters and yield ...
to grow and yield under saline conditions show certain physiological attributes which make the crops adapted to salinity. Irrigation water with high chloride ...

Effect of shade and spacing on growth, yield and quality of black ...
Irrespective of the quantum of shade, the senescence was delayed by about a ... shade and spacing on growth, yield and quality of black musli.pdf. Open. Extract.

Effect of leaf retention on rooting of cuttings and growth of cuttings of ...
tree species for timber production in our country. Propagation by stem cuttings may be the most. promising technique of vegetative propagation for establishment of indigenous forest tree species,. chikrasi plantations on a large-scale. *Institute of

Effect of growth regulators on callus induction in Rice ... - CiteSeerX
CSR 10, W.Ponni, BPT 5204 and IR 29. The medium used for this callus induction was Murashige and Skoog (1962) commonly referred as MS medium with six ...

The effect of vascular endothelial growth factor and brain‐derived ...
from the site of injury [6]. ... adhered to charged slides (Superfrost Plus, ... *all comparisons P < 0.05, except 3 vs 4; †number of nerve fibres positive per ...

Effect of growth regulators on yield, nutrient uptake ...
Table 2. Effect of genotypes and growth regulators on N,P,K and protein content and yield of seed and ... management factors and renewable energy sources.

VELOCITY EFFECT ON INFLATIONARY GROWTH OF ...
Error Correction Mechanism (ECM) is employed to test for short run adjustment ... negative and statistically significant error correction coefficient is a necessary.

Omega3 and skeletal health.pdf
tomy potentiated PGE2 production in mouse calvarial os- teoblastic cell ... strated that LTB4 inhibited cell proliferation in cultured os- ...... Arend WP, Dayer JM.

Study and Investigate Effect of Input Parameters on Temperature and ...
Equipment Components AB SE–631 85 Eskilstuna, Sweden 2007. [12]. www.fuchs-europe.de. [13]. Industrial Gearbox Service Manual of LOCTITE. [14]. Industry Catalogue of HI-TECH DRIVES Pvt. Ltd. [15].Carl Byington, Ryan Brawrs, Sanket Amin, James Hopki

Effect of salt stress on germination and seedling growth ...
Abstract : Five rice genotypes viz. White Ponni, ADT 38, ADT 39, IR 20 and .... The data was analysed for its significance as per Panse and Sukhatme (1961).

Effect of media and growth regulators on multiple shoot ...
Indian Silk, 40: 13-14. ... (MS macro half strength and Heller's micronutrients) ... Cultivars. Number of multiple shoots. MS. G. ½ MSH. Mean. Grand. Mean. BA3.

Effect of jasmonic acid on in vitro explant growth and ...
Oct 17, 2005 - The shoot fresh mass, root length and root numbers of two potato (Solanum tuberosum L.) cultivars Favorita and. Helanwuhua were increased significantly by the application of 0.2 - 2 mg dm-3 jasmonic acid (JA) in the Murashige and. Skoo

Influence of different levels of spacing and manuring on growth ...
Page 1 of 8. 1. Influence of different levels of spacing and manuring on growth, yield and. quality of Alpinia calcarata (Linn.) Willd. Baby P Skaria, PP Joy, Samuel Mathew and J Thomas. 2006. Kerala Agricultural University, Aromatic and Medicinal Pl

EFFECT OF HIGH CALCIUM AND PHOSPHORUS ON THE ...
EFFECT OF HIGH CALCIUM AND PHOSPHORUS ON THE GROWTH.pdf. EFFECT OF HIGH CALCIUM AND PHOSPHORUS ON THE GROWTH.pdf. Open.

Effect of Porogen Residue on Chemical, Optical, and ...
crete changes are observed: the redshift in the Si–CH3 absorbance and a minimal H2O amplitude increase (around 3200 cm−1) pre- sumably due to an increase in the low-k pore radii after the He/H2. DSP plasma exposure. The observations are typical o

Effect of electron acceptor structure on stability and ...
The generic structure of an organic solar cell, a bulk heterojunction has two distinct and continuous layers. One consists of an electron donor, this layer is usually.

Effect of Melatonin on Sleep, Behavior, and Cognition ...
Article Plus (online only) materials for this article appear on the Journal's Web .... (degree of resemblance between the activity patterns on individual days) ..... Classification of Sleep Disorders, Revised: Diagnostic and Coding Manual. Chicago: .

Effect of storage containers and seed treatments on ...
renewable energy in general and biomass energy .... resources was carried out. ... Table 2. Effect of containers and seed treatments on bruchid damage (%) in ...

effect of nacl priming duration and concentration on ... - Core
coefficient of velocity of fenugreek seeds and the best result was obtained with (4 .... Statistical analysis ... variance, using SPSS 13.0 software and the difference.

Study and Investigate Effect of Input Parameters on ... - IJRIT
to apply DOE techniques to achieve desired design of gearbox for control the temperature and noise .... replication total 32 experiments will be performed as shown in table ΙΙ. ... will be carried out using dB meter or by using ultrasonic sensor.

Effect of Seed Pre-treatments on Germination and ...
(1998) stated graphing relationship between the relative yield of cotton ... Data were subjected to statistical analysis according to ..... Israel Program for Scientific.