CHAPTER ll

MATERIALS AND METHODS

Experimental Site and Duration of the Study

The study was conducted at DNSC Freshwater fishpond, New Visayas, Panabo

City. The duration of the study was 60 days.

Experimental Set-up

The experimental Set-up consist of twelve hapa nets immerse in the concrete

tank and has the dimension of 1m x 1m x 1.5m with a distance 0.5 between hapa net.

The materials that will be used in this study are economically available and durable

enough to withstand weather fluctuation all throughout the culture period.

Experimental Treatments and Design

The study has four (4) treatments and each treatment will be replicated 3 times in

completely randomize designed (CRD). T1-Formulate diet control, T2-Formulated feed

with 1% of FFPH, T3-Formulated feed with 2% of FPH, T4-Formulated feed with 3% of

FPH.
T3R1 T2R3

T3R3 T4R2

T4R1 T3R2

T1R2 T1R1

T2R2 T4R3

T2R1 T1R3
Figure 2. The representation of the experimental design.

Experimental Species

The African catfish (Clarias gariepinus) fingerlings was used as experimental

species in this study. They have cylindrical body that reach an average length of 1.5

inches (4 cm). It has an elongated body and a smooth, scaleless skin. They also have

long barbels around their mouth. Additionally, they have a dorsal fin, coloration can vary

from gray to brownish-black. The 150 pcs of African Catfish will be purchased at Aqua

Penon Hatchery, Brgy. Ising, Carmen, Davao del Norte. In the selection of the

experimental species, the fish must be healthy, must be free from injury and no history

of disease within the facility which may lead to mortality of the fish.

Preparation of Fish Protein Hydrolysate

Acid hydrolysis was carried out using the method of Wisuthiphaet, Kongruang, &

Chamcheun, (2015) with minor modifications (Figure 1). The milkfish by-products were

thawed at 4 °C for 16 h. A combination of ground viscera (50g) and ground bones and

fins (50 g) were used in this study. Afterwards, the mixture (100 g) was added with 50

mL distilled water in a 250 mL Erlenmeyer flask. Three different hydrochloric acid

concentrations (4, 6, and 8M) were used in this study. Fifty milliliters of acid were added

to the fish slurry. The mixture was autoclaved at 121 °C under 15 psi for 90 min. The

reaction was then terminated by adjusting the pH of the solution to 5 using 6M NaOH.

The slurry was then filtered to remove any remaining debris. The filtered liquid was then

dried in an oven at 80 °C for 48 hrs. The dried sample was powdered and stored in a

desiccator until further analysis.

Homogenization

Acid hydrolysis

(6 M HCI, 1:1 w/v)

121 °C, 15 psi for 90

Reaction termination
(pH adjustment to 5)

Milkfish by-products
Filtration protein hydrolysate

Over drying at 80°C for

Table 1. Composition of the control and experimental diets

Ingredients/
Test diets (%)
Chemical
Composition T1 T2 T3 T4
f
(0%) (1%) (2%) (3%)

Fish meal 22.00% 18.00 18.00 18.00

FPH 0.00 1.00 2.00 3.00

Soybean meal 30.00 31.08 30.08 29.08

Rice bran 27.00 27.00 27.00 27.00

Corn 18.00 18.00 18.00 18.00

Fish oil 3.2 3.2 3.2 3.2

Vitamin mix 0.5 0.5 0.5 0.5

Mineral mix 0.5 0.5 0.5 0.5

Total 100 100 100 100

Feed Preparation Procedure

This will be sieve or sifted until the fine liquid will obtained. The ground commercial feed

will be weighed into a desired amount and placed in a basin for mixing within 5 minutes.

Determine the correct percentage of the amount of Fish Protein hydrolysate to be added

to the feed. The Fish Protein Hydrolysate would then be mixed with feed. Then once the

Fish Protein Hydrolysate has been incorporated, the feed would likely be tested.
Grinding Sieving Weighing Mixing

Pelleting Sun Drying Packing

Feeds and Feeding Scheme

One day after stocking, 10% feeding rate was adopted in the first 15 days of

culture while 8% feeding rate was used in the next 15 days and was adjusted to 6%

feeding rate in the last phase of the culture period. Feeding of the GET-Excel catfish

fingerlings was done (3) times a day at 8:00 A.M with 30% of the feeds, at 12:00 noon,

30% and 40% at 4:00 p.m.in the afternoon.

Conditioning, Stocking and Sampling

Before stocking, the initial body weight of the fish was measured using electronic

weighing scale, respectively. The Get-Excel Catfish fingerlings upon arrival was

acclimatized by floating the bags for at least 30 minutes before stocking to equalize the

water temperature in the plastic bags with the surrounding water. The plastic bags was

opened and gradually filled with the surrounding water to acclimatize the fish in field

conditions.

Sampling for growth performance was done every 15th day starting at day 1 until

the end of the study. Sampling was done by randomly scooping the fish from the

aquarium using scoop net, after which the fish were placed in the basin with enough

water and aeration. After all, the fish were placed in the basin, it was placed in a beaker
and weighed, the body weight of the fish was measured using electronic weighing scale.

The outcome of sampling was the basis for the adjustment of feeds to be given for the

next 15 days of the culture.

Feeding Management

According to Gule and Geremew (2022), the implementation of effective feed

management practices in catfish (Clarias gariepinus) culture production has significant

economic implications, as it directly impacts production performance and long-term

aquaculture development. To maximize revenue and optimize production, it is crucial to

adopt efficient feed management strategies. The feeding requirements for cat fish

fingerlings, as stated by BFAR (2000), suggest that they should be fed within the range

of 30-15% of their biomass requirements. In this study, the catfish fingerlings was fed a

diet containing Fish Protein Hydrolysate. Feeding was conducted three times daily at

specific times: 0800H (30% of the daily feed intake), 1200H (30% of the daily feed

intake), and 1600H (40% of the daily feed intake), throughout the 60 days culture

period. The percentage of feeds consumed by the catfish was determined based on

their feeding requirement.

Composition (%) Earthen Pond Reservoir Tank Flow-Through Rivers

Moisture 71.7I±1.65a 74.20±1.39a 73.60±2.37a 72.29±0.68a

Crude Protein 19.77±2.66a 20.99±0.68a 19.80±0.53a 18.84±1.00a

Crude Fat 3.85±0.5c 5.68±0.81a 4.95±0.38b 1.35±0.29d

Ash 1.52±0.40b 1.49±0.40b 2.23±0.91a 1.47±0.60b

Maintenance and Monitoring of Set-Up

The hapa net will be maintained by washing it every 15 days to ensure that the

unwanted matter and unconsumed feeds will be removed to avoid suffocation of the

stock, and ensure also the culture units to be clean. It will be done through brushing the

nets for the circulation of water. The hapa net will be monitored daily, to avoid the

escape of the cultured species.

Growth and Feed Utilization Parameters

Growth performances and feed utilization of fish was determined as described by

Alhassan et al., 2012 and Chavez et al., 2014 as follows:

Survival (%) = (final number of fish/ (initial number of fish) x 100

Weight Gain (g) = (final body weight-initial body weight)

Feed Conversion Ratio (FCR) = [Total feed given (g)] / [Total weight gain (g)]
Daily Feed Allowance (DFA) = weight of sample x number of sample x survival rate x

feeding rate

Daily Feed Ration (DFR) = Biomass x Feeding Rate

Feed Conversion Ratio

The feed conversion ratio (FCR) of the three types of feed which were utilized in

this study was evaluated after 60 days of culture. The total amount of feed consumed by

the fish in each treatment were sum-up and compared with the total weight gain after

which the FCR was computed.

Economic Analysis

Cost and return analyses were conducted to determine the profitability of the

project. Cost-benefit analysis translated as return on investments (ROI) and cash

payback period was the determinant factors to package this technology. All expenses

incurred throughout the study were recorded as a basis in calculating the production

cost. ROI was determined to measure the economic profitability using the formula:

ROI Net Income/ Production Cost X 100%

Statistical Analysis

The mean values of the African catfish fingerlings will be statistically compared

using a Parametric and Non-parametric. Since I did not conduct the study yet, I don’t

have the statistical data so it is depending on if the assumptions are being met or

satisfied.