Fisheries Journal, 15 .
, 1368-1379 .
http://doi.
org/10.
29303/jp.
PROTEIN LEVEL EFFICIENCY OF TILAPIA Oreochromis niloticus
FEED WITH SELENOMETHIONINE SUPPLEMENTATION
Efisiensi Tingkat Protein Pakan Ikan Nila Oreochromis niloticus dengan Suplementasi Selenometionin Christian Ernsz Pattipeilohy1*.
Bethsy Jane Pattiasina1.
Joice Welly Loupatty1.
Chrisoetanto Patrick Pattirane2 Department of Aquaculture.
Pattimura University, 2Aquaculture Study Program.
Polytechnic of Marine Affairs and Fisheries Karawang Mr.
Chr.
Soplanit Street.
Poka.
Ambon Bay.
Ambon City.
Maluku 97233 Coresponding author: christian.
pattipeilohy@lecturer.
(Received May 1st 2025.
Accepted June 20th 2.
ABSTRACT One of the nutrients that play an important role for fish growth is protein, one of which is in the formation of body tissues.
The amount and quality of protein in fish feed affects growth performance, feed utilization, water quality and of course the price of the feed itself.
Excessive feed protein or whose quality does not meet the needs will be excreted as nitrogen waste, especially in the form of ammonia.
This study aimed to reduce protein consumption in tilapia (Oreochromis niloticu.
diets through selenomethionine supplementation.
The test feed was designed using two different protein concentrations: 26% and 30%.
Selenomethionine supplementation levels were 0 mg/kg .
for 30% feed protein, and 3 mg/kg and 6 mg/kg Se for 26% feed protein.
This study used a completely randomized design (CRD) with three treatments and three replicates.
Tilapia with an average initial weight of 8.
05 A 0.
25 g were reared in an aquarium measuring 100y50y50 cmA with a stocking density of 15 fish per The fish were reared for 60 days and fed three times a day until full.
The results showed that supplementation of 3 mg/kg Selenomethionine to 26% feed protein was better than either the control or 6 mg/kg Se supplementation.
Supplementation of 3 mg/kg Se to 26% feed protein level significantly increased feed intake (FI), final biomass weight (B.
, feed conversion ratio (FCR), feed efficiency (FE), protein retention (PR), and daily growth rate (DGR) in A 100% survival rate was recorded in all treatment groups.
Keywords: Feed.
Protein Efficiency.
Selenomethionine.
Tilapia
ABSTRAK
Salah satu nutrien yang memainkan peranan penting bagi pertumbuhan ikan adalah protein, salah satu peranannya yakni dalam pembentukan jaringan tubuh.
Jumlah dan kualitas protein dalam pakan ikan berpengaruh terhadap kinerja pertumbuhan, pemanfaatan pakan, kualitas air dan tentunya harga pakan itu sendiri.
Protein pakan yang berlebih atau yang kualitasnya tidak sesuai dengan kebutuhan akan diekskresikan sebagai buangan nitrogen terutama dalam bentuk Penelitian ini bertujuan untuk mengurangi konsumsi protein pada makanan ikan nila e-ISSN : 2622-1934, p-ISSN : 2302-6049 Fisheries Journal, 15 .
, 1368-1379.
http://doi.
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Pattipeilohy et al.
, .
(Oreochromis niloticu.
melalui suplementasi selenometionin.
Pakan uji dirancang menggunakan dua konsentrasi protein berbeda: 26% dan 30%.
Tingkat suplementasi Selenomethionine yakni 0 mg/kg .
untuk protein pakan 30%, dan 3 mg/kg dan 6 mg/kg Se untuk protein pakan 26%.
Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) dengan tiga perlakuan dan tiga ulangan.
Ikan nila dengan bobot awal rata-rata 8,05 A 0,25 g dipelihara dalam akuarium berukuran 100y50y50 cmA dengan padat tebar 15 ekor per Ikan-ikan tersebut dipelihara selama 60 hari dan diberi pakan tiga kali sehari sampai Hasil penelitian menunjukkan bahwa suplementasi 3 mg/kg Selenomethionine ke dalam protein pakan 26% lebih baik dibandingkan perlakuan lainya baik itu kontrol ataupun suplementasi 6 mg/kg Se dalam pakan.
Suplementasi 3 mg/kg Se terhadap kadar protein pakan 26% secara signifikan meningkatkan asupan pakan (FI), berat biomassa akhir (B.
, rasio konversi pakan (FCR), efisiensi pakan (FE), retensi protein (PR), dan laju pertumbuhan harian (DGR) pada ikan nila.
Tingkat kelangsungan hidup 100% tercatat di semua kelompok Kata Kunci: Efisiensi Protein.
Ikan Nila.
Pakan.
Selenometionin
INTRODUCTION
Protein is an important ingredient crucial for tissue development and proliferation in organisms (Halver, 1.
The price of protein in feed substantially affects the total cost of fish The amount and quality of dietary protein directly influence fish growth rates, survival rates, and production expenses in aquaculture.
Inadequate dietary protein undermines the availability necessary for the production of new bodily tissues.
In contrast, high or inadequate protein levels result in ineffective use for tissue synthesis, leading to nitrogen excretion, predominantly as ammonia (Pattipeilohy et al.
, 2.
Selenium (S.
has recently attracted significant interest in animal nutrition as an essential micromineral (Lin, 2014.
Baidya & Murthy, 2.
Selenium and selenoproteins are essential for the antioxidant defense mechanisms of organisms.
Research indicates that elevated selenium levels mitigate oxidative stress induced by oxidized fish oil (Malandrakis et al.
, 2.
Furthermore, it has been observed for the first time that the mRNA expression of several selenoproteins, including gpx1, selenof, selenon, selenoi, selenoo, selenoe, msrb1, sephs2, selenot, selenot2, and selenop, is influenced by the interaction between selenium and oxidized fish oil (Liu et al.
, 2.
Selenium is a key component of the enzyme glutathione peroxidase (Rotruck et al.
, 1.
, crucial for cellular defense against oxidative damage by promoting the reduction of hydrogen peroxide and lipid peroxides (Watanabe et al.
, 1.
Selenium is an essential trace element that functions as a constituent of iodothyronine deiodinase enzymes.
Iodothyronine deiodinase (ID), a selenoprotein, facilitates the transformation of thyroxine (T.
into the biologically active thyroid hormone, 3,5,3'-triiodothyronine (T.
(Brown & Arthur, 2.
Thyroxine levels modulate insulin production, with increased insulin levels enhancing glucose uptake into cells, thus channeling protein-derived energy towards growth activities (Pattipeilohy et al.
, 2.
Fish serve as a superior source of selenium for humans, possessing greater amounts of selenium than alternative food sources.
Furthermore, selenium in fish is primarily found in an organic form, which is effectively absorbed and retained by humans (Wang et al.
, 2.
Administering suitable forms of selenium at optimal concentrations in fish feed not only promotes growth and health but also improves the efficacy of aquaculture products by yielding selenium-enriched, health-beneficial seafood (Cotter et al.
, 2.
Supplementation of organic selenium at 2 and 4 mg Se/kg of feed has shown remarkable growth performance in red tilapia (Pattipeiluhu et al.
, 2.
The inclusion of selenium in feed serves as a method to save protein, hence improving the utilization of non-protein energy when provided at appropriate dosages (Zhang et al.
, 2.
e-ISSN : 2622-1934, p-ISSN : 2302-6049 Fisheries Journal, 15 .
, 1368-1379.
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Pattipeilohy et al.
, .
METHODS
Time and Place of Research This research was conducted from November 2024 to January 2025 at the Cultivation Laboratory.
Aquaculture Study Program.
Faculty of Fisheries and Marine Science.
Pattimura University.
Ambon.
Tools and Materials The tools and materials used in this study were aquarium, aerator hose, aerator stone, aerator pump, digital scale, ruler, water pump, spoons, red tilapia, selenomethionine.
Fish meal.
MBM.
Soy flour.
Wheat flour.
Tapioca flour.
Pollard flour.
Fish oil.
Corn oil Premix .
ithout S.
, and Binder (PMC).
Experimental Diets Se supplementation in accordance with the optimal Se level in fish feed has an impact on the growth, health condition of fish and provides aquaculture products enriched with Selenium content (Wang et al.
, 2.
The fish diet was formulated to provide 26% protein, supplemented with two levels of selenomethionine: 3 and 6 mg Se/kg feed.
A study was undertaken to evaluate the effectiveness of selenium in protein utilization compared to a 30% protein diet lacking selenomethionine supplementation.
(Pattipeilohy et al.
, 2.
showed that a feed protein level of 28% supplemented with selenium 3 mg/kg feed was able to increase tilapia growth compared to a feed protein level of 28% with selenium supplementation of 6 mg/kg feed.
The diets were formulated to provide an energy content of 3,700Ae4,000 kcal GE/kg and included powdered selenomethionine.
The selenomethionine was carefully combined with other feed ingredients using a mixer, homogenized, and subsequently pelletized.
The pellets were dehydrated in an oven at 35AC for 24 hours.
A proximate analysis was performed on the created meal to determine its nutritional composition, and a selenium assay was executed to quantify selenium levels.
Table 1 delineates the composition of raw materials, the proximate analysis of the experimental meals on a dry weight basis, and the selenium concentration of the The study's test subjects were Nile tilapia (Oreochromis niloticu.
, measuring 10Ae12 cm in length, with an initial average weight of 8.
05 A 0.
25 g.
Experimental Design The study employed a Completely Randomized Design (CRD) with three treatments and three replications.
The treatments consisted of P26S3 .
% protein 3 mg seleniu.
P26S6 .
% protein 6 mg seleniu.
, and P30S0 .
% protein 0 mg seleniu.
Table 1.
Composition of Test Feed Formulations and Proximate Results of Test Feeds Selenomethionine Addition .
g/k.
Raw Material P26S3
P26S6
P30S0
Raw Material Composition (%) Fish meal
MBM
Soy flour Wheat flour Tapioca flour Pollard flour Fish oil Corn oil Premix .
ithout S.
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Pattipeilohy et al.
, .
Raw Material Binder (PMC) Se .
g/kg fee.
Proximate analysis .
% Dry Weigh.
Protein
Lipid
Ash
Crude Fiber
BETN
GE .
kal/kg fee.
C/P Selenium feed .
g/k.
Selenomethionine Addition .
g/k.
P26S3
P26S6
P30S0
Description: 1.
MBM: Meat bone meal, 2.
PMC: Polymethylolcarbamide.
Se: selenomethionine, 4.
BETN: Extract material without nitrogen, 5.
GE: Gross energy (Watanabe 1.
Maintenance and Data Collection The test subjects in this study were red tilapia (Oreochromis niloticu.
, averaging 5A7 cm in size and an initial weight of 8.
05A0.
25 g.
Nine aquaria, each sized 100y50y50 cmA, were utilized to house the fish, with each aquarium containing 175 liters of water.
The stocking density was 15 fish per 175 liters of water, utilizing a total of 270 red tilapia fingerlings.
Feeding occurred ad libitum three times daily at 08:00, 12:00, and 16:00 WIB.
Fish biomass was measured at the commencement and conclusion of the maintenance period, alongside proximate analysis of fish body composition and selenium content assessment.
Biomass measurements were conducted following a 24-hour fasting interval.
Upon conclusion of the maintenance phase, blood samples were obtained from a subset of fish within each treatment group for biochemical analysis.
Total of Feed Consumption The calculation of feed consumption was determined by calculating the amount of feed given during the experiment minus the amount of uneaten and dried leftover feed.
Final Body Weight Final Boddy weight can be calculated based on the NRC .
formula as follows:
OIW = Wt Oe Wo Description:
OIW : Final Boddy Weight : Weight of test animals at the end of the study .
: Weight of test animals at the beginning of the study .
Feed Conversion Ratio Feed conversion is the ratio of the amount of feed in dry conditions given during the maintenance process minus the weight of dead fish and the initial weight of fish at the end of maintenance (NRC,1.
With the following equation:
FCR =
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, .
Description:
FCR : Feed Conversion Ratio : Weight of test animals at the end of the study .
: Weight of test animals at the beginning of the study .
: Weight of dead fish .
: Amount of feed consumed .
Feed Efficiency The feed utilization efficiency (FE) value is calculated using the formula (NRC,1.
as Wt Oe Wo
FE =
X 100%
Description:
: Feed utilization efficiency (%) : Weight of test animals at the end of the study .
: Weight of test animals at the beginning of the study .
: Amount of feed consumed during the study .
Protein Efficiency Ratio Protein efficiency ratio value is calculated using the formula (NRC,1.
PER =
ycOyc Oe ycOycu ycU 100% ycEycn Description:
PER : Protein efficiency ratio (%) : Weight of test animals at the end of the study .
: Weight of test animals at the beginning of the study .
: The consumed feed protein weight .
Growth Rate The daily relative growth rate of tilapia seeds observed in the study was calculated using the formula:
LnWt Oe LnWo
SGR =
X 100%
Description:
SGR : Daily weight growth rate (%/da.
: Weight of test animals at the end of the study .
: Weight of test animals at the beginning of the study .
: Maintenance time .
Survival Rate The percentage of survival rate was calculated using the formula from (NRC,1.
as SR = x 100% Description:
: Survival (%) : Number of fish alive at the end of maintenance .
: Number of fish at the beginning of maintenance e-ISSN : 2622-1934, p-ISSN : 2302-6049 Fisheries Journal, 15 .
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Pattipeilohy et al.
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Blood Glucose Analysis of blood glucose levels using a glucose test device.
Gluco Dr AGM-2100 with a kit in the form of Gluco Dr strip Code 8.
Selenium Retention Se retention can be calculated based on the formula of Rider et al.
as follows:
RSe = FOeI x 100% Description:
RSe : Se retention (%) : Total body Se at the end of the experiment .
: Total body Se at the beginning of the experiment .
: Total amount of Se consumed .
Blood Proteins This analysis was performed using the principle of the biuret test.
The absorbance reading value was analyzed based on the formula in Mushawwir et al.
as follows:
BP =
yayc Oe yayca ycu 6yci.
yccya 10 yayc Oe yayca Oe1 Description:
: Blood proteins : Absorbance of sample tested : Absorbance of blank : Absorbance of standard Total Red Blood Cells Total erythrocyte examination aims to determine the health condition of fish by counting total erythrocytes in the blood.
The method of taking blood samples is sucked with a pipette with a scale of up to 0.
5, then Hayem's solution is sucked up to a scale of 101, the pipette is shaken to form a figure eight for 3-5 minutes so that it mixes homogeneously.
The first drop was discarded.
the next drop was dripped into the hemocytometer and covered with a glass cover (Blaxhall & Daisley 1.
Counts were made on 5 small squares of the hemocytometer and the number was calculated using the following formula.
Total erythrocyte = Calculated cell x 1x dilution factor small box volume Total White Blood Cells The method of calculating total leukocytes is the same as calculating total erythrocytes, which distinguishes the solution used, namely Turk's solution.
The total leukocyte count is expressed as n x 105/mm3.
Total leukocytes = e-ISSN : 2622-1934, p-ISSN : 2302-6049 Calculated cell x 1x dilution factor small box volume Fisheries Journal, 15 .
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Hematocrit Levels Hematocrit measurement was performed using a hematocrit micro-tube in the form of a heparin-coated capillary tube.
Blood samples were drawn using a capillary tube up to 3AE4 of the capillary, then covered with a covering material .
The capillary tube containing the blood was centrifuged at 3500 rpm for 15 minutes.
The reading was done by comparing the part of blood that settled with the whole part of blood in the micro hematocrit tube, using a micro The scale and results are expressed in percent (%), (Anderson & Siwicki 1.
Chemical Analysis The chemical analysis encompassed the quantification of selenium levels and the proximate makeup.
The ICP-OES method was employed to assess selenium levels in the feed and fish tissues.
Proximate analysis was performed on the raw feed ingredients, test feed, and fish bodies at both the commencement and conclusion of the study, adhering to the technique outlined by Takeuchi .
The analysis entailed quantifying moisture content via ovendrying at 105-110AC, protein content employing the Kjeldahl method, fat content utilizing the Soxhlet method for feed and the Folch method for fish bodies, ash content through incineration in a furnace at 400-600AC, and crude fiber content by dissolution in strong acids and bases followed by heating.
Data Analysis This research utilized a Completely Randomized Design (CRD) with three treatments and three replications.
The parameters analyzed statistically encompassed growth performance measures and chemical composition data.
Data were organized using MS Office Excel 2013, and normality and homogeneity assessments were performed prior to ANOVA.
Statistical analysis was conducted with SPSS 16.
0 software.
Variations among treatments were assessed using analysis of variance (ANOVA) at a 95% confidence level.
Should the F-test reveal substantial differences, the study was subsequently augmented with Duncan's multiple range RESULTS The supplementation of selenomethionine significantly affected FCR.
FBW.
FE.
PER.
SGR, and SR .
<0.
Table 2 indicates that selenium supplementation at 3 mg Se/kg meal with 26% protein content generally improved hunger and yielded better growth performance than alternative treatments.
This was demonstrated by elevated values in feed intake, protein retention, feed efficiency, specific growth rate, and total biomass compared to the other Table 2.
Total of Feed Consumption (TFC).
Final Body Weight (FBW).
Feed Conversion Ratio (FCR).
Feed Efficiency (FE).
Protein Efficiency Ratio (PER).
Specific Growth Rate (SGR), and Survival Rate (SR) in Tilapia
Treatment
Parameters P26S3
P26S6
P30S0
TFC .
381A1b FWB .
269A1b
FCR
38A0.
09A0.
65A0.
FE (%)
37A0.
32A0.
63A0.
PER (%)
07A2.
78A3.
61A0.
SGR (%)
27A0.
64A0.
93A0.
SR (%)
100A0a e-ISSN : 2622-1934, p-ISSN : 2302-6049 Fisheries Journal, 15 .
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The blood glucose analysis results in tilapia demonstrated considerable variation in blood glucose levels across treatments .
<0.
The minimum blood glucose level was recorded in the P30S0 treatment at 55 mg/L, but the P26S3 and P26S6 treatments exhibited blood glucose values of 65 mg/L and 72 mg/L, respectively.
Figure 1.
Blood Glucose Levels in Tilapia Subjected to Meals with Varying Protein Concentrations and Selenomethionine Supplementation (Distinct Letters on the Bar Chart Denote Significant Differences Across Treatments .
<0.
The results of the selenium retention study demonstrated that selenomethionine supplementation in diets with differing protein levels significantly influenced body selenium retention .
<0.
The highest selenium retention was observed in the 26% dietary protein treatment supplemented with 3 mg Se/kg feed, followed by 6 mg Se/kg.
The therapy without selenomethionine supplementation had the lowest selenium retention compared to the other Figure 2.
Retention of Selenium in Nile Tilapia Subjected to Diets with Varying Protein Concentrations and Selenomethionine Supplementation Levels (Distinct Letters on the Bar Chart Denote Significant Differences Between Treatments .
<0.
Figure 3 illustrates the blood protein concentration across three treatments, revealing the lowest concentration in the 26% protein diet supplemented with 6 mg Se/kg feed, followed by the 30% protein diet without Se supplementation.
The 26% protein diet supplemented with 3 mg Se/kg meal demonstrated the highest blood protein content of 6.
4 mg/L among all e-ISSN : 2622-1934, p-ISSN : 2302-6049 Fisheries Journal, 15 .
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Figure 3.
Blood protein Concentrations in Nile Tilapia Subjected to Diets with Varying Protein Levels and Selenomethionine Supplementation (Distinct Letters on the Bar Chart Denote Significant Differences Between Treatments .
<0.
Table 3 demonstrates that the lowest red blood cell count was observed in the 26% protein diet supplemented with 6 mg Se/kg of feed.
The lowest white blood cell count occurred in the 26% protein diet supplemented with 3 mg Se/kg feed, while the highest white blood cell count was recorded in the 30% protein diet without selenomethionine supplementation.
The highest hematocrit value was seen in the 26% protein diet supplemented with 3 mg Se/kg of feed.
Table 3.
Red Blood Cell (HR).
White Blood Cell (WBC), and Hematocrit Counts of Tilapia
Treatments Parameters P26S3
P26S6
P30S0
HR .
el/mm x10 ) 39A0.
13A0.
42A0.
WBC .
el/mm x10 ) 98A3.
93A1.
95A0.
Hematocrit (%) 33A8.
3A1.
43A1.
DISCUSSION
The supplementation of selenium (S.
not only stimulates insulin secretion but also elevates blood glucose levels, so triggering glycogenesis, which leads to the storage of energy as glycogen in the body.
Similar findings have been reported in tilapia by Pattipeilohy et al.
and in red tilapia by Pattipeiluhu et al.
These data suggest that organic selenium supplementation in feed, at a given dosage, can influence feed palatability.
The survival rate for all therapies reached 100% and was not influenced by selenium supplementation.
Marked changes were noted, with elevated blood glucose levels in tilapia receiving 3 and 6 mg Se/kg feed at a 26% dietary protein level, in contrast to the 30% dietary protein treatment devoid of Se supplementation.
Selenium supplementation in the diet resulted in heightened blood glucose levels, along with increased insulin release in the bloodstream.
The augmented insulin secretion facilitated the accumulation of energy reserves as body glycogen, a process termed glycogenesis (Suprayudi et al.
The incorporation of 3 and 6 mg Se/kg into the meal with a 26% protein content elevated the body Se concentration relative to the 30% protein diet treatment.
The elevation in body selenium levels signifies that selenium has a biological function, especially as selenoproteins.
Se retention plays a crucial role in growth rates in relation to protein storage and the utilization of non-protein energy as the primary energy source (Pattipeilohy et al.
, 2.
Plasma proteins consist of 60% albumin, 35% globulin, and 4% fibrinogen.
Albumin aids in the transport of e-ISSN : 2622-1934, p-ISSN : 2302-6049 Fisheries Journal, 15 .
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ions, chemicals, minerals, hormones, and metabolic waste.
fibrinogen is crucial for blood clotting following damage.
and globulin is involved in immune system activities (Ainsworth.
These components signify the extent of the immune reaction or the health status of the test fish.
The red blood cell (RBC) count of Nile tilapia in this study varied from 1.
13 to 1.
42 y 10 cells/mmA.
The erythrocyte count in healthy fish generally varies from 1.
05 to 3.
00 y 106 cells/mmA (Robert, 1.
Erythrocytes, or red blood cells, are the most prevalent form of blood cells relative to other cellular types.
Under typical circumstances, erythrocytes comprise around fifty percent of the blood volume.
The typical erythrocyte count in Nile tilapia varies from 20,000 to 3,000,000 cells/mmA (Hartika et al.
, 2.
White blood cells have a crucial role in the immune system, particularly in the production of antibodies in response to foreign substances (Purwanti et al.
, 2.
The typical hematocrit percentage in healthy Nile tilapia ranges from 27.
3% to 37.
8% (Hardi et al.
, 2.
The hematocrit levels during the three treatments in this study remained within the normal range, varying from 11.
3% to 18.
presumably influenced by selenium's function as glutathione peroxidase (GP.
GPx is essential for cellular defense against oxidative damage in cytoplasmic structures by reducing hydrogen peroxide and lipid peroxides (Watanabe et al.
, 1.
CONCLUSION
Selenomethionine supplementation in fish feed markedly affects feed intake (FI), final biomass weight (FBW), feed conversion ratio (FCR), feed efficiency (FE), protein retention (PR), and daily growth rate (DGR) in Nile tilapia (Oreochromis niloticu.
This addition also affects the protein efficiency of Nile tilapia feed.
This study's results indicate that a 3 mg/kg selenomethionine supplementation in a 26% protein diet produces superior growth performance relative to a 6 mg/kg selenomethionine supplementation or a 30% protein diet lacking selenomethionine supplementation.
ACKNOWLEDGEMENT
The researcher would like to thank all parties who have contributed and provided input in the preparation stage for the progress of this research.
Thanks are conveyed to fellow Lecturers of the Department of Aquaculture.
Faculty of Fisheries and Marine Sciences.
Pattimura University who have jointly assisted in the process of data collection and data analysis carried out.
REFERENCES