Journal ofInternational Cooperation for
Agricultural Development

J Intl Cooper Agric Dev 2019; 17: 8–13

Original

Growth and Feed Utilization ofJuvenile Nile Tilapia Fed with
Boiled Moringa oleifera Meal Diets:
A Preliminary Report
Albert V. Manuel1), A. M. Shahabuddin1), Debasish Saha1), Miguel H. Chele1),
Kelvin O. A. da Conceicao1) Naoaki Tsutsui1) and Takao Yoshimatsu1)
1) Laboratory of Shallow Sea Aquaculture, Graduate School of Bioresources, Mie University, Mie 514-8507, Japan

Received September 21, 2018 Accepted: December 10, 2018
Abstract. Juveniles of Nile tilapia were stocked (4.47 ± 0.26 g, eight fish per tank) and fed with test diets containing different
inclusion levels of boiled Moringa oleifera leaf meal powder (0–30%: MLM 0, MLM 10, MLM 20, MLM 30), at 6% bodyweight for a total for six weeks. MLM 10 recorded the best average weight gain, specific growth rate, food conversion rate
and protein efficiency ratio but there were no significance differences in all parameters with the MLM 30 diet which had thehighest MLM inclusion. As there were no differences in fish growth and feed utilization between MLM 0 diet and MLM
10–30 diet, it would be possible to replace fish meal as a main dietary protein source by MLM up to 30% from 40% without
any negative influence on the growth of juvenile Nile tilapia and is applicable as a practical rearing diet.
Key words: Nile tilapia, Moringa oleifera, growth, feed ingredient, fish meal

Introduction
Aquaculture is believed to reduce pressure on the wildstock in fisheries and meet the needs of on-growing world
food supply1). Fish feed is the single largest expenditure insemi-intensive and intensive fish culture operations. One
major reason for its high cost is the animal protein content
used in feed formulations. Animal proteins, for example,
fish meal is usually the biggest proportion of commercial
fish feeds. Global fish meal and fish oil production averaged 6.5 and 1.3 million metric tonnes, respectively, overthe past 20 years, however, some years’ productions werehigher and in others lower2). Recently, researchers havebeen considering plant proteins as a substitute for animal
proteins in fish feed to lower cost. For example, a study inCorresponding author: T Yoshimatsu, e-mail: takaoyos@bio.
mie-u.ac.jp
8 J Intl Cooper Agric Dev 2019

Egypt was carried out to replace fish meal with a mixture(soyabean, cotton seed, sunflower and linseed meals) ofdifferent plant protein sources in juvenile Nile Tilapia
feed and concluded that a replacement of 100% fish meal
protein would have no adverse effects for up to 16 weeks
old juvenile3). Furthermore, another study showed that cottonseed meal can partially replace fish meal as a source ofprotein in compound feed at a limited amount of no morethan 50% for tilapia raised in recirculation systems4). Plant
sourced proteins to replace fishmeal have successfully
been carried out but there are some challenges. One beingthat these plant proteins need to be purchased at a certain
cost which may be lower than fish meal but still carries aprice to use as part of the fish feed ingredient.
Moringa oleifera, is a resilient tree that can survive ina variety of climates and substandard soils. Moringa plant
is native to the sub-Himalayan regions of Northwest India
and the plant thrives in many countries of Africa, Arabia,

South East Asia, the Pacific and Caribbean Islands as well
as South America5, 6). It is readily available at no cost in
many of these countries and is fast growing and hardy.
Moringa leaves have a high nutritional value and containvarious amino acids, which are rarely found in other plants.
From those practical merits, in agriculture, Moringa leaves
are used to feed cattle7), pigs8), chickens and poultry9). The
use of Moringa leaf meal (MLM) also might be a good
choice for the aquaculture field. There are several feeding
trials that have used Moringa leaf meal as a replacement
to animal protein (fish meal) for fish feed in many placesin the world. For example, a preliminary study by Richteret al. (2003) carried out feeding trials using Nile tilapia
and results indicated that raw Moringa leaf meal can beused to substitute up to 10% of dietary protein in Niletilapia without significant reduction in growth10). A study
by Afuang et al. (2003) showed a comparative nutritional
evaluation of raw, methanol extracted residues and methanol extracts of Moringa leaves on the growth performanceand feed utilization in Nile tilapia and results showed thatthe solvent extracted Moringa leaf meal could be included
up to 33% in tilapia diets without any negative effects on
growth11). The solvent extraction method resulted in theinactivation and reduction of antinutrients and improved
the palatability of Moringa leaf meal diets. Therefore, eventhough the antinutrient levels were not measured, the current experiment treated Moringa leaf by boiling and sun
drying, a simple practical method for developing countries

that has not been used before in treating Moringa leaves,hoping that this would also reduce negative influences byantinutrients in the test diets.
In Mozambique and most parts of Africa, tilapia farmers cannot afford expensive imported fish feed and are
in dire need for cheaper feed, so ingredients that wereused to prepare test diets were chosen to be practical andaffordable for farmers in Mozambique. The choice of thesenutrient levels, particularly protein, was intended to reflect
the practical diets used in developing countries. Therefore,
this preliminary experiment focuses on substituting fishmeal at different inclusion levels with boiled MLM to help
overcome problems like high feed cost and reduce wild
stock exploitation.

Materials and Methods
Feed Formulation and Preparation

Moringa leaves were picked, boiled, dried and ground to
powder form in the Instituto Superior Politécnicode Gaza
(ISPG) in Chokwe, Mozambique by ISPS academic staff
and sent to Mie University in Japan where feed formulation and preparation were done. A control (0%: MLM 0)
and three test diets were prepared with different inclusionlevels (10%: MLM 10, 20%: MLM 20, 30%: MLM 30)
and analysis of their proximate compositions were carriedout and is shown in Table 1.

Table 1 Feed formulation and proximate composition of MLM diets fed to O. niloticus during the
growth trial
Ingredients (%)

Moringa Leaf

MLM 0

Fish Meal		
40
Processed Moringa		
0Cassava Powder		20
Rice Bran		
28
Fish Oil		
4
Binder (CMC)1		 4
Vitamin Premix2		 2
Trace Mineral Premix3		 2

MLM 10

MLM 20

MLM 30

37
10
20
21
4
4
2
2

33
20
20
15
4
4
2
2

30
30
20
8
4
4
2
2

Proximate Composition (%)					Moisture
Ash
Crude Protein
Crude Lipid
Carbohydrate

9.1
8.7
26.8
8.5
46.9

8.3
11.6
33.6
7.8
38.7

7.8
12.9
32.5
9.0
37.8

7.5
13.7
32.8
9.9
36.1

7.8
15.0
30.4
11.4
35.4

1 Carboxymethylcellulose.

2 Vitamin Premix, diluted in cellulose, provided the following in mg. kg–1 diet according to the requirementof O. niloticus12): vitamin A 1.8; vitamin D3 0.025; vitamin E 25; vitamin K 5.2; thiamine 2.5; riboflavin 6;
pyridoxine 16; pantothenic acid10; niacin 121; folic acid 0.82; vitamin B12 0.024; biotin 0.06; chlorine
chloride 1000.
3 Trace Mineral Mix provided by the following minerals (mg. kg–1 diet): zinc (as ZnSO .7H O) 150; iron (as4
2
FeSO4. 7H2O) 40; manganese (as MnSO4. 7H2O) 25; copper (as CuCl2) 3; iodine (as KI) 5; cobalt (as CoCl2.
6H2O) 0.05; selenium (as Na2SeO3) 0.09.

J Intl Cooper Agric Dev 2019 9

Fish Sampling and Experimental Setup

Juveniles of Nile tilapia Oreochromis niloticus were
sampled from the wild stock in Nikko river, Aichi Prefecture, Japan and then transferred to Mie University
where they were kept for a week to acclimatize. Following
acclimatization, fish weighing an average weight of 4.47
± 0.26 g were distributed (eight fish per tank) into twelve
10 L-volume tanks (three tanks per treatment) in a closed
recirculating system set indoor. Fish were fed with test
diets containing different levels of MLM at 6% (wet basis)
body weight for a total of six weeks at temperature, 25 ±2ºC. In a two-week interval, individual weights were measured until the end of the experiment where all fish weresacrificed and sent for whole body composition proximate
analysis (Table 3).

Fig. 1. Change in body weight of O. niloticus over a periodof six weeks after been fed with different levels of
MLM diets.

Growth Performance and Utilization

Growth and feed utilization were determined in terms of
average daily weight gain (ADWG), specific growth rate(SGR), feed conversion ratio (FCR), protein efficiency
ratio (PER) as follows:
ADWG (g fish–1 day–1) =final weight – initial weight/ number of days
SGR (% day–1) = 100 × (ln [final body weight] –ln [initial body weight])/ no. of days
FCR = feed intake / live weight gain
PER = live weight gain/ crude protein intake
Whole Body Composition

After the sixth week of rearing, fish were sacrificed andsent for a whole-body proximate analysis to determine theamount of crude protein, crude lipid, ash, moisture andcarbohydrates and results expressed as a percentage of live
weight.

to be similar, for example, a study showed that Moringa
leaf had a crude protein content of 31–35%13), 25%10),
and 25.4%11). By adding MLM with different inclusionlevels, test diets including 30.4–33.6% crude protein were
formulated in the laboratory (Table 1).
The overall performance of our feeding trial is summarized in Table 2 together with the nutrient utilization.
Although, fish fed with the MLM 10 diet clearly showed
the best performance in average daily weight gain
(ADWG) of 1.78 ± 0.78 g fish–1 day–1, specific growth rate(SGR) of 0.48 ± 0.07% day–1, feed conversion rate (FCR)
of 2.79 ± 0.61, and protein efficiency ratio (PER) of 1.20
± 0.23, there were no significant difference in comparison
to other diets, mainly due to the varied weight gain values
and limitation of individual number used in each treatment.
Whole Body Composition

Statistical analysis (SPSS vers. 16. IBM®, New York,
USA) was carried out using Tukey’s test after one-way
analysis of variance (ANOVA) to determine differencesamong treatment groups which were deemed significant
at P<0.05.

As for the effect of MLM supplementation on proximatebody compositions of juvenile Tilapia, crude protein andlipid contents were measured 14.1% and 1.2%, respectively in initial fish body. After 6 weeks feeding, the valuesof 18.8% and 3.6% were noted in MLM 0 group and
16.7–19.2% and 3.0–3.3% were obtained in each MLM
supplemented groups.

Results

Discussion

Growth Performance and Feed Utilization

The rearing results obtained in the present experimentsuggested that an inclusion of MLM up till 30% would
be possible without any negative influence on feed intake
and fish growth. One factor that limits the high inclusionsof MLM in fish diets is the presence of antinutrients.
According to a previous study, 30% inclusion of Moringa leaf caused a depression in growth and this could

Statistical Analysis

Body weight gain during the six weeks of growth trial isshown in the Fig. 1. Fish fed actively on the prepared test
diets but not at a consistent rate and this resulted in a low
weight gain.
Moringa leaf has high levels of crude protein (26.8% in
the present study) and previous studies have proved this10 J Intl Cooper Agric Dev 2019

Table 2 Growth performance and nutrient utilization of O. niloticus fed with MLM diets
Initial Body Weight (g)Final Body Weight (g)ADWG (g fish-1 day–1)
SGR (% day–1)
FCR
PER

MLM 0

MLM10

MLM 20

MLM 304.53 ± 0.44a

4.36 ± 0.10a

4.44 ± 0.13a

4.54 ± 0.29a
10.07 ± 0.46b
0.72 ± 0.01c
0.41 ± 0.01d
3.26 ± 0.83e
1.01 ± 0.61f

10.88 ± 0.60b

11.52 ± 1.68b

0.79 ± 0.07c
0.45 ± 0.02d
2.94 ± 0.16e
1.01 ± 0.55f

1.78 ± 0.78c
0.48 ± 0.07d
2.79 ± 0.61e
1.20 ± 0.23f

11.13 ± 0.43b
0.96 ± 0.52c
0.47 ± 0.04d
2.84 ± 0.26e
1.09 ± 0.11f

Mean ± SE (n=3) in each column followed by different letters (P˂0.05) indicates a significant difference.
AWG: Average daily weight gain, SGR: specific growth rate, FCR: feed conversion rate, and PER:
protein efficiency ratio.

Table 3 Whole body proximate composition of O. niloticus fed test diets
before and after the experiment (% fresh weight)
Moisture
Crude Protein
Crude Lipid
Ash
Carbohydrate

Initial

MLM 0

MLM 10

MLM 20

MLM 30

81.2
14.1
1.2
3.2
0.3

74.3
18.8
3.6
3.2
0.1

77
16.7
3.1
2.4
0.8

74.9
19.2
3.0
2.8
0.1

74.9
18.9
3.3
2.8
0.1

likely be attributed to several factors, most importantly,
antinutrients7). However, this problem could be solved bydrying and/or grinding of moringa leaves before aqueous
extraction that would improve the removal of soluble
antinutrients13). The study further mentions that rejection
was prominent in (45%: MLM 45) and (60%: MLM 60)
fish diets, so therefore it is safe to say that an inclusionof (30%: MLM 30) would have no negative effect on fish
intake and growth.
The best FCR value of 2.79 ± 0.61, obtained from fish
fed with the MLM 10 diet in the current experiment wasconsidered to be quite high, however, FCR values reported
in a previous study showed that an inclusion of 30% aqueous extracted MLM in fish diets recorded a FCR of 4.97 ±0.3713) and in comparison, to the current experiment, a 30%
MLM inclusion in fish diet recorded a much lower FCR
value of 3.26 ± 0.83. The experiment was conducted overmonths of October to December and room temperatures
ranged from 18–25°C but heaters were placed in the recirculating tanks to control water temperature. The optimum
range of culturing O. niloticus is from 26°C to 29°C14)
and it appears that low temperatures may have affected
FCR. A study carried out in winter reported FCRs ranging
from 3.3–4.715) for different sizes of O. niloticus reared
during winter and compared to the current experiment,
FCR values ranged from 2.79–3.26. Therefore, to obtain amuch better FCR value with the same experimental design,
a study with more individuals per treatment, bigger culturetanks, an effective antinutrient removal technique, anda temperature of 28 ± 1°C, would provide an optimum

culture condition which would maximize feed in-take andconversion, resulting in obtaining a reasonable FCR value.
If all these parameters are achieved and a low FCR value
is obtained, MLM substitution instead of fish meal could
be carried out in developing countries to reduce fish feed
cost and encourage more farmers to tap into the fish farming business. Encouraging farmers to begin fish farming
businesses would not only boost the local economy but bea favorable protein provider which could help tackle theissue of malnutrition faced in African countries.
PER was best recorded in fish fed with MLM 10 dietbut there were no significant differences to other MLM
diets. PER also might have been affected by the suboptimal
temperature during the rearing period. PER decreased withdecreasing weight of fish and protein level and this was
the same case in an experiment by Richter et al. (2003)10).
The level of whole-body moisture as shown in Table 3
was higher in fish fed with MLM 10, MLM 20 and MLM
30 diets compared to the control MLM 0 group. Thisagreed with results of a reported study which had similar
lose in whole body lipid content and energy of common
carp fed with plant-based protein, Mucuna16). Furthermore,another study that studied factors affecting the proximate
composition of cultured fishes observed that body moisture and body lipid were inversely related17), which was thecase in this experiment.
The results of the present study showed that MLM 10
diet recorded the best ADWG, SGR, FCR and PER but
the results were not that statistically different from theMLM 30 diet. Due to these experimental results, a 30%
J Intl Cooper Agric Dev 2019 11

inclusion of MLM in fish diets replacing fish meal would
have no interference in the growth and performance ofjuvenile Nile tilapia. The use of Moringa leaf as a dietary
source in fish diets is limited due to the presence of antinutritional factors, particularly saponins and to a lesser
extent tannin, phytic acid and hydrogen cyanide18). The
present study processed Moringa leaves by boiling andthis may have led to a reduction of anti-nutrients due to theleaching of soluble fractions in water. To clarify this point,
more detailed experiments and biochemical analyses fromvarious viewpoints would be necessary in future. Also in
the next step, larger scale rearing experiments would beindispensable to establish the usage of boiled Moringa
meal as a cheap and sustainable feed source for fish.
Acknowledgement
This experiment was carried out as one part of a Japan
International Cooperation Agency (JICA) project (2016,
BOP Business Survey Project in Mozambique) carried out
under an academic MOU between ISPG and Mie university. We would like to thank JICA for its financial supportand ISPG in Mozambique for preparing the key ingredientfor this laboratory-level feeding experiment, Moringa leaf
meal. We acknowledge that the country of origin of thematerial is Mozambique and that the country’s biodiversity
had been managed by various legislations, in particular
the Convention on Biological Diversity and the Nagoya
Protocol which need to be adhered to.
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モリンガ飼料を与えた際のナイルティラピアの成長と飼料利用効率（予報）Albert V. Manuel, A. M. Shahabuddin, Debasish Saha, Miguel H. Chele, Kelvin O. A. da Conceicao，
筒井

直昭，吉松

隆夫

三重大学大学院生物資源学研究科 浅海増殖学研究室
要旨
ナイルティラピア幼魚（4.47 ± 0.26 g）を各水槽に 8 尾ずつ収容し，煮沸による熱処理を施したモリンガ Moringa oleifera
葉体の乾燥粉砕物（MLM）を 0 － 30％添加した試験飼料を体重の 6％分 6 週間の試験期間中毎日給餌して，成長，生残，飼料成分の利用効率，体成分に与える影響を予備的に検討した。その結果，MLM を 10％添加した試験区で最高の平均増重，比増殖率および飼料成分の利用効率が得られたが，MLM 無添加，あるいは 20–30％添加した区との間で統計的有意差は認められず，また給餌試験終了後の魚体の一般分析値にも大きな差異はなかった。本実験により，ナイルティラピア幼魚に対して，MLM を 30％添加することにより飼料タンパク質源である魚粉を 10％分削減することが可能であることが本試
験で分かった。

キーワード : ティラピア、モリンガ、成長、飼料成分、魚粉

J Intl Cooper Agric Dev 2019 13