6 Different Experimental Diets Were Formulated Biology Essay

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In this experiment, 6 different experimental diets were formulated labeled as Diet 1, Diet 2, Diet 3, Diet 4, Diet 5 and Diet 6 had been use throughout this experiment. All of the diets were prepared in the Lab 307, Fish Nutritional Lab and Fish House. The ingredient (Table 3.1) for all these diets were the same but the proportion of ingredient that used were different in each diets. The ingredient that use to prepare the diets were fish meal, soy bean meal, dextrin, enzyme premix, fish oil, vitamin premix, mineral premix, carboxylmethytl cellulose (CMC), chromic oxide and bentonite clay.

Fish meal and soybean meal that were used as the protein source in this experiment contain the same amounts in diet formulation. Dextrin was used as the carbohydrate source and carboxyl methyl cellulose (CMC) as a binder. CMC, vitamin premix, mineral premix and chromic oxide (Cr2O3) were added in diet formulation at 1.5% , 3%, 4% and 1% respectively. The addition of chromic oxide as an inert marker for the determination of nutrient digestibility and it can be determined by using wet-acid digestion method that was described by Fukurawa and Tsukahara (1966). The addition of bentonite clay was added as an inert filler to allow the adjustment of the energy content of the diets.

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All these diet were different in terms of their enzyme level and energy level (Plate 3.1). The pellets of Diet 1, Diet 2 and Diet 3 have same energy level of 350kcal within 0.00% enzyme in the Diet 1, 0.02% enzyme in Diet 2 and 0.04% enzyme in Diet 3.Meanwhile the pellets of Diet 4, Diet 5 and Diet 6 have same energy level of 250kcal with 0.00% enzyme in the Diet 4, 0.02% enzyme in Diet 5 and 0.04% enzyme in Diet 6. Decreasing the energy level from 350kcal to 250kcal in diet is due to the amount of dextrin that added into the Diet 1, Diet 2 and Diet 3 as compared to Diet 4, Diet 5 and Diet 6 (Table 3.1).

Table 3.1 The Composition of the experimental diets (g/100g dry diet)

Ingredient

Composition (%)

Diet 1

Diet 2

Diet 3

Diet 4

Diet 5

Diet 6

Fish Meal

7.94

7.94

7.94

7.94

7.94

7.94

Soybean Meal

51.13

51.13

51.13

51.13

51.13

51.13

Dextrin

26.45

26.45

26.45

1.45

1.45

1.45

Enzyme premix1

0.00

0.50

1.00

0.00

0.50

1.00

Fish oil

1.95

1.95

1.95

1.95

1.95

1.95

soybean oil

1.46

1.46

1.46

1.46

1.46

1.46

Vitamin pre-mix2

3.00

3.00

3.00

3.00

3.00

3.00

Mineral pre-mix3

4.00

4.00

4.00

4.00

4.00

4.00

CMC

1.50

1.50

1.50

1.50

1.50

1.50

Chromic oxide

1.00

1.00

1.00

1.00

1.00

1.00

Bentonite clay

1.58

1.00

0.58

26.58

26.08

25.58

1Contained 4% of allzyme VegproTM and 96% of α-cellulose

2Contained(as kg-1) ascorbic acid,45; inositol,5; choline chloride,75; niasin,4.5; riboflavin,1; pyridoxine HCL,1;thiamin HCL,1;d-calcium pantothenate,3; retinyl acetate,0.6; vitamin D3,0.083;menadione,1.67;DL-α-topopherol acetate (500IU/g),8;d-biotin,0.02;folic acid,0.09;vitamin B12,0.00135;cellulose,855.0357.

3Contained (as kg-1) calcium phosphate monobasic,270.98; calcium lactate,327; ferrous sulphate,25; magnesium sulphate,132;potassium chloride,50; sodium chloride,60; potassium iodide,0.15; copper sulphate,0.785; manganese oxide,0.8;colbalt carbonate,1;zinc oxide,3; sodium selenite,0.011; calcium carbonate,129.274.

According the diet formulations in Table 3.1, all the ingredients of each diet were weighed approximately and added into the plastic containers except for fish oil and soybean oils, all the ingredient were mixed in the commercial mixer (Hobart mixer) about 30 minutes. The fish oil is added into the Hobart mixer while the entire ingredient was still stirred as well as the soybean oil. After that, distilled water was slowly added into the mixer until the dough was formed. Then, all the dough was put into the meat machine to form a 3-mm of noodle-like long strands. Then, the strand was dried on the meshed-shelf cabinet with two fans at room temperature for overnight. Since the dried long strands diets are too big to be eaten by fish, the strands was cut into small pellets, sieved, sealed it in the polyethylene bags with labels and stored in the refrigerator. Some pelleted diets were taken for dietary proximate analysis. The pellets that use in this experiment are show in Plate 3.1.

Diet 6

Diet 5

Diet 4

Diet 3

Diet 2

Diet 1

Plate 3.1: The 6 experimental diet that used in these study

3.2 Experimental Fish and Feeding Trial

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At start this experiment, the red hybrid tilapia, Oreochromis sp. were obtained from the local hatchery were stocked firstly into 1000L fiberglass tanks for them to being able to acclimatize to the laboratory conditions and feed by a commercially diet that available twice a day for one week before they were used for actual experiment.

At the beginning of feeding, 12 the red hybrid tilapia (Oreochromis sp) fingerlings was selected and distributed randomly into 18 glass tanks within an early average weight for each fish is 5.70 ± 0.05 g (Plate 3.2). The entire cleaned glass tank was provided with dechlorinated water and air-stones for continuous aeration throughout the experiment. Every day the whole water mass in the glass tank change and the aeration of each tank was checked to maintain the water quality. The diet was given randomly into the tank within 3 replicates. The entire glass tanks were cleaned weekly during the sampling.

The feeding trial was carried out for 9 weeks. All the 6 different diet were weighted every week in each single day according to some satiation calculation and keep it in the polyethylene bags. Throughout of the experiment, the fishes were fed twice a day, once in the morning around 0700 to 0900 and 1500 to 1700 in the afternoon. The fishes were slowly fed with the diet and being observed every day.

Plate 3.2: The glass tank within the red hybrid tilapia used in this study

3.3 Sampling

Throughout 9 weeks of the experiment, all the fish from each tank were weighed on a weekly basis in order to obtain the apparent feeding satiation along the experiment. Before the experiment was carried out, some fishes were killed and dried in oven for the initial proximate composition of whole body of the fish.

At the end of the experiment after 9 weeks, 5-8 fish were randomly selected from each tank was randomly weighted under moderate anaesthesia (ethylene glycol monophenyl ether)

, killed and dissected to remove the tissues. Tissues such as liver, viscera and fat were dissected and weighted to determine the hepatosomatic index (HSI), viscerosomatic index (VSI) and intraperitoneal fat index (IPF). The liver and muscle from each tank were wrapped, keep it in the polyethylene bags and then stored in the freezer for further analysis.

The remaining fish of each tank that unsevered and undissected are stored in freezer for whole-body composition analysis. For whole-body composition, the fishes were cut into small size, dried in the oven, and blended it into fine powder before run the analysis of whole body proximate analysis by using the standard Association of Official Analytical Chemist (AOAC, 1997) method.

3.4 Proximate Composition

The proximate analysis was carried out for both the diets and the fishes. The proximate analysis including the moisture, crude protein, crude lipid, crude fiber and ash content were determined by standard Association of Official Analytical Chemist (AOAC, 1997) method with slight modifications (Refer to Appendix 1).

Briefly, moisture content was determined by drying the samples in an oven at 105°C until it showed the constant weight. The crude protein was determined by Kjedahl method (AOAC, 1997) by digesting the sample with concentrated H2SO4 follow by 40% alkali distillation and titration by 0.1N HCL. The crude lipid content was determined by the Solution A (Chloroform/methanol) extraction in separating tunnel then followed by drying the extraction of lipid in oven to evaporate the solvent. Ash content was determined by dry ashing in ash crucible in muffle furnace for 5 hours at 550°C until there is no black particle. Then, the crude fiber was determined by the dried lipid free sample will go through the acid extraction (0.225N), alkali extraction (0.313N NaOH) then the drying ashing process in the muffle furnace.

The moisture, crude protein, lipid and ash content were determined by both diets and the fishes. Meanwhile the crude fiber only required for the diets but not the fishes. All the experimental diets tested for their nutritional value before the start of feeding trial to ensure the nutrient levels of each pellets prepared were correct and meet the requirement or purpose of the experiment. Same with the whole body composition, some fish were collected, cut, dried in the oven and blended into finely powder for the subsequent initial analysis of the whole body proximate analysis.

3.5 Calculation

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The livers, viscera and interaperitorial fat separated and weighted to determine Hepatosomatic Index (HIS), Viscerasomatic Index (VSI) and Intraperitoneal Fat Index (IPF). The formulas for the calculation for these parameters are calculated as below;

Hepatosomatic Index (HSI) % = Liver weight (g) x 100

Fish weight (g)

Viscerosomatic Index (VSI) %= Viscera weight (g) x 100

Fish weight (g)

Intraperitoneal Fat Index Index (IPF) %= Intraperitoneal fat weight (g) x 100

Fish weight (g)

3.6 Statistical Analysis

The available data obtained from the experiment were subjected to one way analysis of variances (ANOVA) and two way analyses of variances (ANOVA) to test the effect of different dietary on proximate composition and body-organ indices. The multiple comparison of means were determined by using Duncan's Multiple Range Test (Duncan, 1955) with a statistical significant when P-values were <0.05. All analysis was performed using the SPSS program, 11.5 for Windows (SPSS Inc., Chicago, IL, USA).